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Gallo MC, Elias A, Reynolds J, Ball JR, Lieberman JR. Regional Gene Therapy for Bone Tissue Engineering: A Current Concepts Review. Bioengineering (Basel) 2025; 12:120. [PMID: 40001640 PMCID: PMC11852166 DOI: 10.3390/bioengineering12020120] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/24/2024] [Revised: 01/20/2025] [Accepted: 01/24/2025] [Indexed: 02/27/2025] Open
Abstract
The management of segmental bone defects presents a complex reconstruction challenge for orthopedic surgeons. Current treatment options are limited by efficacy across the spectrum of injury, morbidity, and cost. Regional gene therapy is a promising tissue engineering strategy for bone repair, as it allows for local implantation of nucleic acids or genetically modified cells to direct specific protein expression. In cell-based gene therapy approaches, a variety of different cell types have been described including mesenchymal stem cells (MSCs) derived from multiple sources-bone marrow, adipose, skeletal muscle, and umbilical cord tissue, among others. MSCs, in particular, have been well studied, as they serve as a source of osteoprogenitor cells in addition to providing a vehicle for transgene delivery. Furthermore, MSCs possess immunomodulatory properties, which may support the development of an allogeneic "off-the-shelf" gene therapy product. Identifying an optimal cell type is paramount to the successful clinical translation of cell-based gene therapy approaches. Here, we review current strategies for the management of segmental bone loss in orthopedic surgery, including bone grafting, bone graft substitutes, and operative techniques. We also highlight regional gene therapy as a tissue engineering strategy for bone repair, with a focus on cell types and cell sources suitable for this application.
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Affiliation(s)
- Matthew C. Gallo
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
| | - Aura Elias
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
| | - Julius Reynolds
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
| | - Jacob R. Ball
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
| | - Jay R. Lieberman
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
- Alfred E. Mann Department of Biomedical Engineering, Viterbi School of Engineering, University of Southern California, Los Angeles, CA 90089, USA
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Fred EJ, Minardi S, Goodwin AM, Nandurkar TS, Plantz MA, Lyons JG, Paul JT, Foley JP, Wintring AJ, Furman AA, Jeong S, Yun C, Stock SR, Hsu WK, Hsu EL. A Mechanistic and Preclinical Assessment of BioRestore Bioactive Glass as a Synthetic Bone Graft Extender and Substitute for Osteoinduction and Spine Fusion. Clin Spine Surg 2024; 37:315-321. [PMID: 38531819 DOI: 10.1097/bsd.0000000000001597] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/21/2023] [Accepted: 11/29/2023] [Indexed: 03/28/2024]
Abstract
STUDY DESIGN Preclinical animal study. OBJECTIVE Evaluate the osteoinductivity and bone regenerative capacity of BioRestore bioactive glass. SUMMARY OF BACKGROUND DATA BioRestore is a Food and Drug Administration (FDA)-approved bone void filler that has not yet been evaluated as a bone graft extender or substitute for spine fusion. METHODS In vitro and in vivo methods were used to compare BioRestore with other biomaterials for the capacity to promote osteodifferentiation and spinal fusion. The materials evaluated (1) absorbable collagen sponge (ACS), (2) allograft, (3) BioRestore, (4) Human Demineralized Bone Matrix (DBM), and (5) MasterGraft. For in vitro studies, rat bone marrow-derived stem cells (BMSC) were cultured on the materials in either standard or osteogenic media (SM, OM), followed by quantification of osteogenic marker genes ( Runx2, Osx, Alpl, Bglap, Spp1 ) and alkaline phosphatase (ALP) activity. Sixty female Fischer rats underwent L4-5 posterolateral fusion (PLF) with placement of 1 of 5 implants: (1) ICBG from syngeneic rats; (2) ICBG+BioRestore; (3) BioRestore alone; (4) ICBG+Allograft; or (5) ICBG+MasterGraft. Spines were harvested 8 weeks postoperatively and evaluated for bone formation and fusion via radiography, blinded manual palpation, microCT, and histology. RESULTS After culture for 1 week, BioRestore promoted similar expression levels of Runx2 and Osx to cells grown on DBM. At the 2-week timepoint, the relative ALP activity for BioRestore-OM was significantly higher ( P <0.001) than that of ACS-OM and DBM-OM ( P <0.01) and statistically equivalent to cells grown on allograft-OM. In vivo, radiographic and microCT evaluation showed some degree of bridging bone formation in all groups tested, with the exception of BioRestore alone, which did not produce successful fusions. CONCLUSIONS This study demonstrates the capacity of BioRestore to promote osteoinductivity in vitro. In vivo, BioRestore performed similarly to commercially available bone graft extender materials but was incapable of producing fusion as a bone graft substitute. LEVEL OF EVIDENCE Level V.
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Affiliation(s)
- Elianna J Fred
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Silvia Minardi
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Alyssa M Goodwin
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Tejas S Nandurkar
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Mark A Plantz
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Joseph G Lyons
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Jonathan T Paul
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - James P Foley
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Allison J Wintring
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Andrew A Furman
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Soyeon Jeong
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Chawon Yun
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Stuart R Stock
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Cell and Developmental Biology, Northwestern University, Chicago, IL
| | - Wellington K Hsu
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
| | - Erin L Hsu
- Simpson Querrey Institute (SQI), Northwestern University, Chicago, IL
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
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Lambrechts MJ, D'Antonio ND, Heard JC, Yalla G, Karamian BA, Markova DZ, Kepler CK. The Inhibitory Effect of NSAIDs and Opioids on Spinal Fusion: An Animal Model. Spine (Phila Pa 1976) 2024; 49:821-828. [PMID: 38348858 DOI: 10.1097/brs.0000000000004959] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/28/2023] [Accepted: 02/02/2024] [Indexed: 06/25/2024]
Abstract
STUDY DESIGN Translational research. OBJECTIVE To evaluate the relative effects of NSAIDs, opioids, and a combination of the two on spinal fusion inhibition in a rodent model. SUMMARY OF BACKGROUND DATA Nonsteroidal anti-inflammatory drugs (NSAIDs) and opioids are common postoperative analgesic agents. Since NSAIDs inhibit the cyclooxygenase (COX) pathway, they are seldom prescribed following spinal fusion. Opioids may be given instead, but recent evidence suggests opioids also adversely affect spinal fusion quality and success. METHODS Eighty male Sprague-Dawley rats underwent L4-5 posterior lumbar fusion and were given one of the following analgesia regimens: saline, morphine (6 mg/kg), ketorolac (4 mg/kg), or morphine (3 mg/kg) and ketorolac (2 mg/kg). Serum samples were drawn to evaluate systemic pro-osteoblastic cytokines and vascular endothelial growth factor-A (VEGF-A) levels, which were measured through enzyme-linked immunosorbent assays (ELISA). After six weeks, the rats were sacrificed, and the operated spinal segments underwent manual palpation, microCT, and histologic analysis. RESULTS Manual palpation scores were significantly diminished in the opioid, NSAID, and multimodal groups when compared with control ( P <0.001). MicroCT fusion scores ( P <0.001) and fusion rates (control: 75% vs . NSAID: 35% vs . opioid: 0% vs . combination: 15%, P <0.001) were significantly diminished in the treatment groups. The bone volume (BV) to tissue volume (TV) ratio (BV/TV) ( P <0.001) and bone mineral density (BMD) ( P <0.001) were all lower in the treatment groups, with the opioid and combined groups having the lowest BMD. Although statistically insignificant ( P <0.09), the concentration of VEGF-A was greater in the control group compared with opioids, NSAIDs, and the combined group. CONCLUSION Opioids and NSAIDs, both independently and combined, inhibited spinal fusion and caused inferior bony callus. Administration of opioids resulted in the lowest rate of spinal fusion. We propose this may be due to the inhibition of VEGF-A, which limits angiogenesis to the burgeoning fusion mass.
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Affiliation(s)
- Mark J Lambrechts
- Department of Orthopaedic Surgery, Rothman Orthopaedic Institute at Thomas Jefferson University, Philadelphia, PA
- Department of Orthopaedic Surgery, Washington University in St. Louis School of Medicine, St. Louis, MO
| | - Nicholas D D'Antonio
- Department of Orthopaedic Surgery, Rothman Orthopaedic Institute at Thomas Jefferson University, Philadelphia, PA
| | - Jeremy C Heard
- Department of Orthopaedic Surgery, Rothman Orthopaedic Institute at Thomas Jefferson University, Philadelphia, PA
| | - Goutham Yalla
- Department of Orthopaedic Surgery, Rothman Orthopaedic Institute at Thomas Jefferson University, Philadelphia, PA
| | - Brian A Karamian
- Department of Orthopaedic Surgery, University of Utah, Salt Lake City, UT
| | - Dessislava Z Markova
- Department of Orthopaedic Surgery, Rothman Orthopaedic Institute at Thomas Jefferson University, Philadelphia, PA
| | - Christopher K Kepler
- Department of Orthopaedic Surgery, Rothman Orthopaedic Institute at Thomas Jefferson University, Philadelphia, PA
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Cottrill E, Pennington Z, Sattah N, Jing C, Salven D, Johnson E, Downey M, Varghese S, Rocos B, Richardson W. Gene Therapy and Spinal Fusion: Systematic Review and Meta-Analysis of the Available Data. World Neurosurg 2024; 186:219-234.e4. [PMID: 38583566 DOI: 10.1016/j.wneu.2024.03.174] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/17/2024] [Accepted: 03/31/2024] [Indexed: 04/09/2024]
Abstract
OBJECTIVE To analyze the extant literature describing the application of gene therapy to spinal fusion. METHODS A systematic review of the English-language literature was performed. The search query was designed to include all published studies examining gene therapy approaches to promote spinal fusion. Approaches were classified as ex vivo (delivery of genetically modified cells) or in vivo (delivery of growth factors via vectors). The primary endpoint was fusion rate. Random effects meta-analyses were performed to calculate the overall odds ratio (OR) of fusion using a gene therapy approach and overall fusion rate. Subgroup analyses of fusion rate were also performed for each gene therapy approach. RESULTS Of 1179 results, 35 articles met criteria for inclusion (all preclinical), of which 26 utilized ex vivo approaches and 9 utilized in vivo approaches. Twenty-seven articles (431 animals) were included in the meta-analysis. Gene therapy use was associated with significantly higher fusion rates (OR 77; 95% confidence interval {CI}: [31, 192]; P < 0.001); ex vivo strategies had a greater effect (OR 136) relative to in vivo strategies (OR 18) (P = 0.017). The overall fusion rate using a gene therapy approach was 80% (95% CI: [62%, 93%]; P < 0.001); overall fusion rates were significantly higher in subjects treated with ex vivo compared to in vivo strategies (90% vs. 42%; P = 0.011). For both ex vivo and in vivo approaches, the effect of gene therapy on fusion was independent of animal model. CONCLUSIONS Gene therapy may augment spinal fusion; however, future investigation in clinical populations is necessary.
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Affiliation(s)
- Ethan Cottrill
- Department of Orthopaedic Surgery, Duke University Health System, Durham, NC, USA; Department of Biomedical Engineering, Duke University, Durham, NC, USA.
| | | | - Nathan Sattah
- Department of Orthopaedic Surgery, Duke University Health System, Durham, NC, USA
| | - Crystal Jing
- Department of Orthopaedic Surgery, Duke University Health System, Durham, NC, USA
| | - Dave Salven
- Department of Orthopaedic Surgery, Duke University Health System, Durham, NC, USA
| | - Eli Johnson
- Department of Neurosurgery, Duke University Health System, Durham, NC, USA
| | - Max Downey
- Department of Surgery, NYU Grossman School of Medicine, NY, USA
| | - Shyni Varghese
- Department of Orthopaedic Surgery, Duke University Health System, Durham, NC, USA; Department of Biomedical Engineering, Duke University, Durham, NC, USA
| | - Brett Rocos
- Department of Orthopaedic Surgery, Duke University Health System, Durham, NC, USA
| | - William Richardson
- Department of Orthopaedic Surgery, Duke University Health System, Durham, NC, USA
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Rolsma JL, Darch W, Higgins NC, Morgan JT. The tardigrade-derived mitochondrial abundant heat soluble protein improves adipose-derived stem cell survival against representative stressors. Sci Rep 2024; 14:11834. [PMID: 38783150 PMCID: PMC11116449 DOI: 10.1038/s41598-024-62693-w] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/14/2024] [Accepted: 05/20/2024] [Indexed: 05/25/2024] Open
Abstract
Human adipose-derived stem cell (ASC) grafts have emerged as a powerful tool in regenerative medicine. However, ASC therapeutic potential is hindered by stressors throughout their use. Here we demonstrate the transgenic expression of the tardigrade-derived mitochondrial abundant heat soluble (MAHS) protein for improved ASC resistance to metabolic, mitochondrial, and injection shear stress. In vitro, MAHS-expressing ASCs demonstrate up to 61% increased cell survival following 72 h of incubation in phosphate buffered saline containing 20% media. Following up to 3.5% DMSO exposure for up to 72 h, a 14-49% increase in MAHS-expressing ASC survival was observed. Further, MAHS expression in ASCs is associated with up to 39% improved cell viability following injection through clinically relevant 27-, 32-, and 34-gauge needles. Our results reveal that MAHS expression in ASCs supports survival in response to a variety of common stressors associated with regenerative therapies, thereby motivating further investigation into MAHS as an agent for stem cell stress resistance. However, differentiation capacity in MAHS-expressing ASCs appears to be skewed in favor of osteogenesis over adipogenesis. Specifically, activity of the early bone formation marker alkaline phosphatase is increased by 74% in MAHS-expressing ASCs following 14 days in osteogenic media. Conversely, positive area of the neutral lipid droplet marker BODIPY is decreased by up to 10% in MAHS-transgenic ASCs following 14 days in adipogenic media. Interestingly, media supplementation with up to 40 mM glucose is sufficient to restore adipogenic differentiation within 14 days, prompting further analysis of mechanisms underlying interference between MAHS and differentiation processes.
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Affiliation(s)
- Jordan L Rolsma
- Department of Bioengineering, University of California, 900 University Ave, Riverside, CA, 92521, USA
| | - William Darch
- Department of Bioengineering, University of California, 900 University Ave, Riverside, CA, 92521, USA
| | - Nicholas C Higgins
- Department of Bioengineering, University of California, 900 University Ave, Riverside, CA, 92521, USA
| | - Joshua T Morgan
- Department of Bioengineering, University of California, 900 University Ave, Riverside, CA, 92521, USA.
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Virk S, Vaishnav A, Kumagai H, Yao YC, Dowdell J, Sandhu H, Schwab F, Qureshi S. Pilot Study on Percutaneous Delivery of Recombinant Human Bone Morphongenetic Protein-2 Augments Fusion in a Nicotine-impaired Rabbit Fusion Model. Clin Spine Surg 2023; 36:E512-E518. [PMID: 37651560 DOI: 10.1097/bsd.0000000000001516] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/21/2022] [Accepted: 06/21/2023] [Indexed: 09/02/2023]
Abstract
STUDY DESIGN A nicotine-impaired spinal fusion rabbit model. OBJECTIVE To examine whether controlled delivery of morselized absorbable collagen sponge recombinant human bone morphogenetic protein-2 (rhBMP2) in a delayed manner postsurgery would allow for improved bone healing. SUMMARY OF BACKGROUND DATA The current delivery method of rhBMP-2 during surgery causes a burst of rhBMP-2, which is not sustained. Given that bone morphogenetic protein 2 (BMP-2) expression peaks later in the fusion process, there may be the benefit of delivery of rhBMP-2 later in the healing process. METHODS Sixteen male 1-year-old rabbits underwent a posterolateral spinal fusion with iliac crest bone graft at L5-L6 while being given nicotine to prevent spinal fusion as previously published. Eight were controls, whereas 8 had morselized rhBMP-2 (4.2 mg) injected at the fusion site at 4 weeks postoperatively. Histologic, radiologic, and palpation examinations were performed at 12 weeks to determine fusion status and the volume of bone formed. Hematoxylin and eosin stains were used for histology. A Student t test was used to compare the computed tomography scan measured volume of bone created between the control cohort (CC) and rhBMP-2 delayed delivery cohort (BMP-DDC). RESULTS Of the total, 7/8 rabbits in the BMP-DDC and 5/8 rabbits in the CC formed definitive fusion with a positive palpation examination, bridging bone between transverse processes on computed tomography scan, and an x-ray showing fusion. Histologic analysis revealed newly remodeled bone within the BMP-DDC. There was an increased average volume of bone formed within the BMP-DDC versus the CC (22.6 ± 13.1 vs 11.1 ± 3.6 cm 3 , P = 0.04). CONCLUSION Our study shows that injectable morselized absorbable collagen sponge/rhBMP-2 can create twice as much bone within a nicotine-impaired rabbit spine fusion model when delivered 4 weeks out from the time of surgery.
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Affiliation(s)
- Sohrab Virk
- Department of Orthopedic Surgery, Northwell Health, New Hyde Park, NY
| | - Avani Vaishnav
- Department of Orthopedic Surgery, Boston University Medical Center, Boston, MA
| | - Hiroshi Kumagai
- Department of Orthopedic Surgery, Hospital for Special Surgery, New York, NY
| | - Yu-Cheng Yao
- Department of Orthopedic Surgery, Hospital for Special Surgery, New York, NY
| | - James Dowdell
- Department of Orthopedic Surgery, Hospital for Special Surgery, New York, NY
| | - Harvinder Sandhu
- Department of Orthopedic Surgery, Hospital for Special Surgery, New York, NY
| | - Frank Schwab
- Department of Orthopedic Surgery, Northwell Health, New Hyde Park, NY
| | - Sheeraz Qureshi
- Department of Orthopedic Surgery, Hospital for Special Surgery, New York, NY
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Broussolle T, Roux JP, Chapurlat R, Barrey C. Murine models of posterolateral spinal fusion: A systematic review. Neurochirurgie 2023; 69:101428. [PMID: 36871885 DOI: 10.1016/j.neuchi.2023.101428] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/22/2022] [Revised: 02/05/2023] [Accepted: 02/07/2023] [Indexed: 03/07/2023]
Abstract
BACKGROUND Rodent models are commonly used experimentally to assess treatment effectiveness in spinal fusion. Certain factors are associated with better fusion rates. The objectives of the present study were to report the protocols most frequently used, to evaluate factors known to positively influence fusion rate, and to identify new factors. METHOD A systematic literature search of PubMed and Web of Science found 139 experimental studies of posterolateral lumbar spinal fusion in rodent models. Data for level and location of fusion, animal strain, sex, weight and age, graft, decortication, fusion assessment and fusion and mortality rates were collected and analyzed. RESULTS The standard murine model for spinal fusion was male Sprague Dawley rats of 295g weight and 13 weeks' age, using decortication, with L4-L5 as fusion level. The last two criteria were associated with significantly better fusion rates. On manual palpation, the overall mean fusion rate in rats was 58% and the autograft mean fusion rate was 61%. Most studies evaluated fusion as a binary on manual palpation, and only a few used CT and histology. Average mortality was 3.03% in rats and 1.56% in mice. CONCLUSIONS These results suggest using a rat model, younger than 10 weeks and weighing more than 300 grams on the day of surgery, to optimize fusion rates, with decortication before grafting and fusing the L4-L5 level.
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Affiliation(s)
- T Broussolle
- Department of Spine Surgery, P. Wertheimer University Hospital, GHE, hospices civils de Lyon, université Claude-Bernard Lyon 1, Lyon, France; Inserm UMR 1033, université Claude-Bernard Lyon 1, Lyon, France.
| | - Jean-Paul Roux
- Inserm UMR 1033, université Claude-Bernard Lyon 1, Lyon, France
| | - R Chapurlat
- Inserm UMR 1033, université Claude-Bernard Lyon 1, Lyon, France
| | - C Barrey
- Department of Spine Surgery, P. Wertheimer University Hospital, GHE, hospices civils de Lyon, université Claude-Bernard Lyon 1, Lyon, France; Arts et métiers ParisTech, ENSAM, 151, boulevard de l'Hôpital, 75013 Paris, France
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Holmes C, Ishida W, Perdomo-Pantoja A, Elder BD, Cottrill E, Locke J, Witham TF. Comparing the efficacy of adipose-derived and bone marrow-derived cells in a rat model of posterolateral lumbar fusion. J Orthop Res 2022; 40:909-916. [PMID: 34081344 DOI: 10.1002/jor.25111] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/25/2021] [Revised: 05/26/2021] [Accepted: 05/31/2021] [Indexed: 02/04/2023]
Abstract
Although bone marrow-derived mesenchymal stem cells (BMCs) have been widely used in spinal fusion procedures, adipose-derived stem cells (ASCs) offer a number of advantages as an alternative clinical cell source. This study directly compares the efficacy of ASCs and BMCs from the same donor animals to achieve successful fusion when combined with a clinical-grade bone graft substitute in a rat lumbar fusion model. ASCs and BMCs were isolated from the same Lewis donor rats and grown to passage 2 (P2). Single-level bilateral posterolateral intertransverse process lumbar fusion surgery was performed on syngeneic rats divided into three experimental groups: clinical-grade bone graft substitute alone (CBGS); CBGS+ rat ASCs (rASC); and, CBGS+ rat BMCs (rBMC). Eight weeks postoperatively, fusion was evaluated via micro-CT, manual palpation and histology. In vitro analysis of the osteogenic capacity of rBMCs and rASCs was also performed. Results indicated that the average fusion volume in the rASC group was the largest and was significantly larger than the CBGS group. Although the rASC group displayed the highest fusion rates via micro-CT and manual palpation, this difference was not statistically significant. Cell-seeded grafts showed more histological bone formation than cell-free grafts. P2 rASCs and rBMCs displayed similar in vitro osteogenic differentiation capacities. Overall, this study showed that, when combined with a clinical-grade bone graft substitute in a rat model, rASCs cells yielded the largest fusion masses and comparable fusion results to rBMCs. These results add to growing evidence that ASCs provide an attractive alternative to BMCs for spinal fusion procedures.
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Affiliation(s)
- Christina Holmes
- Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.,Department of Chemical and Biomedical Engineering, Florida A&M University-Florida State University College of Engineering, Tallahassee, Florida, USA
| | - Wataru Ishida
- Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
| | | | - Benjamin D Elder
- Department of Neurosurgery, Mayo Clinic, Rochester, Minnesota, USA
| | - Ethan Cottrill
- Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
| | - John Locke
- Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
| | - Timothy F Witham
- Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
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Characterization and functional analysis of the adipose tissue-derived stromal vascular fraction of pediatric patients with osteogenesis imperfecta. Sci Rep 2022; 12:2414. [PMID: 35165317 PMCID: PMC8844034 DOI: 10.1038/s41598-022-06063-4] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/07/2021] [Accepted: 01/12/2022] [Indexed: 11/08/2022] Open
Abstract
AbstractPediatric patients with Osteogenesis Imperfecta (OI), a heritable connective tissue disorder, frequently suffer from long bone deformations. Surgical correction often results in bone non-unions, necessitating revision surgery with autogenous bone grafting using bone-marrow-derived stem cells (BM-SC) to regenerate bone. BM-SC harvest is generally invasive and limited in supply; thus, adipose tissue's stromal vascular fraction (SVF) has been introduced as an alternative stem cell reservoir. To elucidate if OI patients' surgical site dissected adipose tissue could be used as autologous bone graft in future, we investigated whether the underlying genetic condition alters SVF's cell populations and in vitro differentiation capacity. After optimizing SVF isolation, we demonstrate successful isolation of SVF of pediatric OI patients and non-OI controls. The number of viable cells was comparable between OI and controls, with about 450,000 per gram tissue. Age, sex, type of OI, disease-causing collagen mutation, or anatomical site of harvest did not affect cell outcome. Further, SVF-containing cell populations were similar between OI and controls, and all isolated SVF's demonstrated chondrogenic, adipogenic, and osteogenic differentiation capacity in vitro. These results indicate that SVF from pediatric OI patients could be used as a source of stem cells for autologous stem cell therapy in OI.
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10
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Plantz M, Lyons J, Yamaguchi JT, Greene AC, Ellenbogen DJ, Hallman MJ, Shah V, Yun C, Jakus AE, Procissi D, Minardi S, Shah RN, Hsu WK, Hsu EL. Preclinical Safety of a 3D-Printed Hydroxyapatite-Demineralized Bone Matrix Scaffold for Spinal Fusion. Spine (Phila Pa 1976) 2022; 47:82-89. [PMID: 34115714 PMCID: PMC8765284 DOI: 10.1097/brs.0000000000004142] [Citation(s) in RCA: 12] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/01/2023]
Abstract
STUDY DESIGN Prospective, randomized, controlled preclinical study. OBJECTIVE The objective of this study was to compare the host inflammatory response of our previously described hyperelastic, 3D-printed (3DP) hydroxyapatite (HA)-demineralized bone matrix (DBM) composite scaffold to the response elicited with the use of recombinant human bone morphogenetic protein-2 (rhBMP-2) in a preclinical rat posterolateral lumbar fusion model. SUMMARY OF BACKGROUND DATA Our group previously found that this 3D-printed HA-DBM composite material shows promise as a bone graft substitute in a preclinical rodent model, but its safety profile had yet to be assessed. METHODS Sixty female Sprague-Dawley rats underwent bilateral posterolateral intertransverse lumbar spinal fusion using with the following implants: 1) type I absorbable collagen sponge (ACS) alone; 2) 10 μg rhBMP-2/ACS; or 3) the 3DP HA-DBM composite scaffold (n = 20). The host inflammatory response was assessed using magnetic resonance imaging, while the local and circulating cytokine expression levels were evaluated by enzyme-linked immunosorbent assays at subsequent postoperative time points (N = 5/time point). RESULTS At both 2 and 5 days postoperatively, treatment with the HA-DBM scaffold produced significantly less soft tissue edema at the fusion bed site relative to rhBMP-2-treated animals as quantified on magnetic resonance imaging. At every postoperative time point evaluated, the level of soft tissue edema in HA-DBM-treated animals was comparable to that of the ACS control group. At 2 days postoperatively, serum concentrations of tumor necrosis factor-α and macrophage chemoattractant protein-1 were significantly elevated in the rhBMP-2 treatment group relative to ACS controls, whereas these cytokines were not elevated in the HA-DBM-treated animals. CONCLUSION The 3D-printed HA-DBM composite induces a significantly reduced host inflammatory response in a preclinical spinal fusion model relative to rhBMP-2.Level of Evidence: N/A.
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Affiliation(s)
- Mark Plantz
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - Joseph Lyons
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - Jonathan T. Yamaguchi
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - Allison C. Greene
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - David J. Ellenbogen
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - Mitchell J. Hallman
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - Vivek Shah
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - Chawon Yun
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | | | | | - Silvia Minardi
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - Ramille N. Shah
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
- Dimension Inx Corp, Chicago, IL
| | - Wellington K. Hsu
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
| | - Erin L. Hsu
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL
- Center for Regenerative Nanomedicine, Simpson Querrey Institute, Chicago, IL
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11
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Kannan A, Minardi S, Ellenbogen DJ, Hallman MJ, Greene AC, Yamaguchi JT, Plantz MA, Jeong S, Sana KC, Shah V, Yun C, Hsu EL, Hsu WK. The effect of local steroid application on bony fusion in a rat posterolateral spinal arthrodesis model. JOR Spine 2021; 4:e1177. [PMID: 35005443 PMCID: PMC8717102 DOI: 10.1002/jsp2.1177] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/08/2021] [Revised: 09/26/2021] [Accepted: 10/25/2021] [Indexed: 11/23/2022] Open
Abstract
INTRODUCTION Local steroid administration during anterior cervical spine surgery has been shown to improve postoperative dysphagia. However, concerns over potential complications remain. This study aims to evaluate the effect of local steroid administration on bone regeneration and spine fusion in a preclinical model, as well as the impact on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) in a 3D culture system. MATERIALS AND METHODS Forty-five rats underwent bilateral L4-L5 posterolateral lumbar fusion (PLF) utilizing local delivery of low-dose recombinant human bone morphogenetic protein-2 (rhBMP-2; 0.5 μg/implant). Rats were divided into three groups: no steroid (control), low dose (0.5 mg/kg), and high dose (2.5 mg/kg) of triamcinolone. Bone growth and fusion were assessed using radiography, blinded manual palpation, and micro-CT analysis and were visualized by histology. The impact of triamcinolone exposure on osteogenic differentiation of hBM-MSCs was evaluated by gene expression analysis, alkaline phosphatase activity assay, and alizarin red staining. RESULTS No significant differences in fusion scores or rates were seen in the low- or high-dose steroid treatment groups relative to untreated controls. Quantification of new bone formation via micro-CT imaging revealed no significant between-group differences in the volume of newly regenerated bone. Triamcinolone also had no negative impact on pro-osteogenic gene transcript levels, and ALP activity was enhanced in the presence of triamcinolone. Mineral deposition appeared comparable in cultures grown with and without triamcinolone. CONCLUSIONS Local steroid application does not seem to inhibit rhBMP-2-mediated spine fusion in rats, though our study may not be adequately powered to detect differences in fusion as measured by manual palpation or bone volume as measured by micro-CT. These findings suggest that local triamcinolone may not increase pseudarthrosis in spine fusion procedures. Further large animal and clinical studies to verify its safety and efficacy are warranted.
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Affiliation(s)
- Abhishek Kannan
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
| | - Silvia Minardi
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - David J. Ellenbogen
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Mitchell J. Hallman
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Allison C. Greene
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Jonathan T. Yamaguchi
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Mark A. Plantz
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Soyoen Jeong
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Kennedy C. Sana
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Vivek Shah
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Chawon Yun
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Erin L. Hsu
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
| | - Wellington K. Hsu
- Department of Orthopaedic SurgeryNorthwestern UniversityChicagoIllinoisUSA
- Center for Regenerative NanomedicineSimpson Querrey InstituteChicagoIllinoisUSA
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12
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Golubovsky JL, Ejikeme T, Winkelman R, Steinmetz MP. Osteobiologics. Oper Neurosurg (Hagerstown) 2021; 21:S2-S9. [PMID: 34128067 DOI: 10.1093/ons/opaa383] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/11/2020] [Accepted: 09/06/2020] [Indexed: 12/17/2022] Open
Abstract
BACKGROUND Osteobiologics are engineered materials that facilitate bone healing and have been increasingly used in spine surgery. Autologous iliac crest bone grafts have been used historically, but morbidity associated with graft harvesting has led surgeons to seek alternative solutions. Allograft bone, biomaterial scaffolds, growth factors, and stem cells have been explored as bone graft substitutes and supplements. OBJECTIVE To review current and emerging osteobiologic technologies. METHODS A literature review of English-language studies was performed in PubMed. Search terms included combinations of "spine," "fusion," "osteobiologics," "autologous," "allogen(e)ic," "graft," "scaffold," "bone morphogenic protein," and "stem cells." RESULTS Evidence supports allograft bone as an autologous bone supplement or replacement in scenarios where minimal autologous bone is available. There are promising data on ceramics and P-15; however, comparative human trials remain scarce. Growth factors, including recombinant human bone morphogenic proteins (rhBMPs) 2 and 7, have been explored in humans after successful animal trials. Evidence continues to support the use of rhBMP-2 in lumbar fusion in patient populations with poor bone quality or revision surgery, while there is limited evidence for rhBMP-7. Stem cells have been incredibly promising in promoting fusion in animal models, but human trials to this point have only involved products with questionable stem cell content, thereby limiting possible conclusions. CONCLUSION Engineered stem cells that overexpress osteoinductive factors are likely the future of spine fusion, but issues with applying viral vector-transduced stem cells in humans have limited progress.
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Affiliation(s)
- Joshua L Golubovsky
- Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Education Institute, Cleveland Clinic, Cleveland, Ohio, USA
| | - Tiffany Ejikeme
- Case Western Reserve University School of Medicine, Cleveland, Ohio, USA
| | - Robert Winkelman
- Department of Neurosurgery, Neurological Institute, Cleveland Clinic, Cleveland, Ohio, USA
| | - Michael P Steinmetz
- Center for Spine Health, Department of Neurosurgery, Neurologic Institute, Cleveland Clinic Foundation, Cleveland, Ohio, USA
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13
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Yun C, Haleem MS, Jeong S, Oyer MA, Driscoll AJ, Chang KY, Yun J, Paul J, Lubbe RJ, Stock SR, Hsu WK, Hsu EL. Effect of Postoperative Analgesic Exposure to the Cannabinoid Receptor Agonist WIN55 on Osteogenic Differentiation and Spinal Fusion in Rats. J Bone Joint Surg Am 2021; 103:984-991. [PMID: 33759484 DOI: 10.2106/jbjs.20.00573] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/01/2023]
Abstract
BACKGROUND After spinal surgery and other orthopaedic procedures, most patients receive opioids for pain, leading to potential complications such as pseudarthrosis and opioid abuse associated with long-term use. As an alternative, the endocannabinoid system has been shown to have antinociceptive activity, while contributing to bone homeostasis via the CB1 and CB2 cannabinoid receptors. This study evaluates the impact of the cannabinoid receptor agonist WIN55,212-2 (WIN55) on osteogenic differentiation in vitro as well as bone regeneration and spinal fusion in a preclinical rat model. METHODS Primary rat bone marrow stromal cells were cultured in standard or osteogenic media and exposed to vehicle alone or WIN55. Runx2 and Alkaline phosphatase (Alpl) were quantified via qPCR (quantitative real-time polymerase chain reaction), followed by assessment of ALP activity and matrix mineralization. For in vivo evaluation, 45 female Sprague Dawley rats (n = 15 per group) underwent L4-L5 posterolateral spinal fusion with bilateral placement of collagen scaffolds preloaded with low-dose rhBMP-2 (recombinant human bone morphogenetic protein-2; 0.5 μg/implant). Postoperatively, rats received the vehicle alone or 0.5 or 2.5 mg/kg WIN55 via daily intraperitoneal injections for 5 days. Bone regeneration and spinal fusion were assessed using radiography, manual palpation-based fusion scoring, microcomputed tomography imaging, and histology. RESULTS mRNA expression levels of Runx2 and Alp were similar among cells treated with vehicle alone and WIN55. Likewise, exposure to WIN55 did not inhibit ALP activity or bone matrix mineralization. In this animal model, no significant differences were found among groups with regard to mean fusion score, fusion rate, or new bone volume. CONCLUSIONS WIN55 showed no adverse impact on osteogenic differentiation, bone regeneration, and spinal fusion. This supports that cannabinoid receptor agonists should be further investigated as a potential alternative approach for postoperative analgesia following spinal fusion and other orthopaedic procedures requiring bone-healing. CLINICAL RELEVANCE The identification of alternative treatments for postoperative pain following orthopaedic surgical procedures is crucial in combating the ongoing opioid abuse crisis. The endocannabinoid system may represent a viable alternative target for addressing orthopaedic postoperative pain.
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Affiliation(s)
- Chawon Yun
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Meraaj S Haleem
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Soyeon Jeong
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Mark A Oyer
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
| | - Adam J Driscoll
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Kevin Y Chang
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Jonghwa Yun
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Jonathan Paul
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Ryan J Lubbe
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Stuart R Stock
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Wellington K Hsu
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
| | - Erin L Hsu
- Departments of Orthopaedic Surgery (C.Y., M.S.H., S.J., M.A.O., A.J.D., K.Y.C., J.Y., J.P., R.J.L., W.K.H., and E.L.H.) and Cell and Molecular Biology (S.R.S.), Northwestern University Feinberg School of Medicine, Chicago, Illinois
- Simpson Querrey Institute, Northwestern University, Chicago, Illinois
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Plantz MA, Minardi S, Lyons JG, Greene AC, Ellenbogen DJ, Hallman M, Yamaguchi JT, Jeong S, Yun C, Jakus AE, Blank KR, Havey RM, Muriuki M, Patwardhan AG, Shah RN, Hsu WK, Stock SR, Hsu EL. Osteoinductivity and biomechanical assessment of a 3D printed demineralized bone matrix-ceramic composite in a rat spine fusion model. Acta Biomater 2021; 127:146-158. [PMID: 33831576 DOI: 10.1016/j.actbio.2021.03.060] [Citation(s) in RCA: 16] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/12/2021] [Revised: 03/09/2021] [Accepted: 03/25/2021] [Indexed: 12/11/2022]
Abstract
We recently developed a recombinant growth factor-free bone regenerative scaffold composed of stoichiometric hydroxyapatite (HA) ceramic particles and human demineralized bone matrix (DBM) particles (HA-DBM). Here, we performed the first pre-clinical comparative evaluation of HA-DBM relative to the industry standard and established positive control, recombinant human bone morphogenetic protein-2 (rhBMP-2), using a rat posterolateral spinal fusion model (PLF). Female Sprague-Dawley rats underwent bilateral L4-L5 PLF with implantation of the HA-DBM scaffold or rhBMP-2. Fusion was evaluated using radiography and blinded manual palpation, while biomechanical testing quantified the segmental flexion-extension range-of-motion (ROM) and stiffness of the fused segments at 8-weeks postoperatively. For mechanistic studies, pro-osteogenic gene and protein expression at 2-days and 1-, 2-, and 8-weeks postoperatively was assessed with another cohort. Unilateral fusion rates did not differ between the HA-DBM (93%) and rhBMP-2 (100%) groups; however, fusion scores were higher with rhBMP-2 (p = 0.008). Both treatments resulted in significantly reduced segmental ROM (p < 0.001) and greater stiffness (p = 0.009) when compared with non-operated controls; however, the degree of stabilization was significantly higher with rhBMP-2 treatment relative to the HA-DBM scaffold. In the mechanistic studies, PLGA and HA scaffolds were used as negative controls. Both rhBMP-2 and HA-DBM treatments resulted in significant elevations of several osteogenesis-associated genes, including Runx2, Osx, and Alp. The rhBMP-2 treatment led to significantly greater early, mid, and late osteogenic markers, which may be the mechanism in which early clinical complications are seen. The HA-DBM scaffold also induced osteogenic gene expression, but primarily at the 2-week postoperative timepoint. Overall, our findings show promise for this 3D-printed composite as a recombinant growth factor-free bone graft substitute for spinal fusion. STATEMENT OF SIGNIFICANCE: Despite current developments in bone graft technology, there remains a significant void in adequate materials for bone regeneration in clinical applications. Two of the most efficacious bone graft options are the gold-standard iliac crest bone graft and recombinant human-derived bone morphogenetic protein-2 (rhBMP-2), available commercially as Infuse™. Although efficacious, autologous graft is associated with donor-site morbidity, and Infuse™ has known side effects related to its substantial host inflammatory response, possibly associated with a immediate, robust osteoinductive response. Hence, there is a need for a bone graft substitute that provides adequate osteogenesis without associated adverse events. This study represents a significant step in the design of off-the-shelf growth factor-free devices for spine fusion.
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Comparison of Freshly Isolated Adipose Tissue-derived Stromal Vascular Fraction and Bone Marrow Cells in a Posterolateral Lumbar Spinal Fusion Model. Spine (Phila Pa 1976) 2021; 46:631-637. [PMID: 32991510 DOI: 10.1097/brs.0000000000003709] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/01/2023]
Abstract
STUDY DESIGN Rat posterolateral lumbar fusion model. OBJECTIVE The aim of this study was to compare the efficacy of freshly isolated adipose tissue-derived stromal vascular fraction (A-SVF) and bone marrow cells (BMCs) cells in achieving spinal fusion in a rat model. SUMMARY OF BACKGROUND DATA Adipose tissue-derived stromal cells (ASCs) offer advantages as a clinical cell source compared to bone marrow-derived stromal cells (BMSCs), including larger available tissue volumes and reduced donor site morbidity. While pre-clinical studies have shown that ex vivo expanded ASCs can be successfully used in spinal fusion, the use of A-SVF cells better allows for clinical translation. METHODS A-SVF cells were isolated from the inguinal fat pads, whereas BMCs were isolated from the long bones of syngeneic 6- to 8-week-old Lewis rats and combined with Vitoss (Stryker) bone graft substitute for subsequent transplantation. Posterolateral spinal fusion surgery at L4-L5 was performed on 36 female Lewis rats divided into three experimental groups: Vitoss bone graft substitute only (VO group); Vitoss + 2.5 × 106 A-SVF cells/side; and, Vitoss + 2.5 × 106 BMCs/side. Fusion was assessed 8 weeks post-surgery via manual palpation, micro-computed tomography (μCT) imaging, and histology. RESULTS μCT imaging analyses revealed that fusion volumes and μCT fusion scores in the A-SVF group were significantly higher than in the VO group; however, they were not significantly different between the A-SVF group and the BMC group. The average manual palpation score was highest in the A-SVF group compared with the BMC and VO groups. Fusion masses arising from cell-seeded implants yielded better bone quality than nonseeded bone graft substitute. CONCLUSION In a rat model, A-SVF cells yielded a comparable fusion mass volume and radiographic rate of fusion to BMCs when combined with a clinical-grade bone graft substitute. These results suggest the feasibility of using freshly isolated A-SVF cells in spinal fusion procedures.Level of Evidence: N/A.
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Lo WC, Tsai LW, Yang YS, Chan RWY. Understanding the Future Prospects of Synergizing Minimally Invasive Transforaminal Lumbar Interbody Fusion Surgery with Ceramics and Regenerative Cellular Therapies. Int J Mol Sci 2021; 22:3638. [PMID: 33807361 PMCID: PMC8037583 DOI: 10.3390/ijms22073638] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/16/2021] [Revised: 03/22/2021] [Accepted: 03/26/2021] [Indexed: 12/14/2022] Open
Abstract
Transforaminal lumber interbody fusion (TLIF) is the last resort to address the lumber degenerative disorders such as spondylolisthesis, causing lower back pain. The current surgical intervention for these abnormalities includes open TLIF. However, in recent years, minimally invasive TLIF (MIS-TLIF) has gained a high momentum, as it could minimize the risk of infection, blood loss, and post-operative complications pertaining to fusion surgery. Further advancement in visualizing and guiding techniques along with grafting cage and materials are continuously improving the safety and efficacy of MIS-TLIF. These assistive techniques are also playing a crucial role to increase and improve the learning curve of surgeons. However, achieving an appropriate output through TLIF still remains a challenge, which might be synergized through 3D-printing and tissue engineering-based regenerative therapy. Owing to their differentiation potential, biomaterials such as stem/progenitor cells may contribute to restructuring lost or damaged tissues during MIS-TLIF, and this therapeutic efficacy could be further supplemented by platelet-derived biomaterials, leading to improved clinical outcomes. Thus, based on the above-mentioned strategies, we have comprehensively summarized recent developments in MIS-TLIF and its possible combinatorial regenerative therapies for rapid and long-term relief.
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Affiliation(s)
- Wen-Cheng Lo
- Department of Surgery, Division of Neurosurgery, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan; (Y.-S.Y.); (R.W.Y.C.)
- Department of Neurosurgery, Taipei Medical University Hospital, Taipei 11031, Taiwan
- Taipei Neuroscience Institute, Taipei Medical University, Taipei 11031, Taiwan
| | - Lung-Wen Tsai
- Department of Medical Education and Research, Taipei Medical University Hospital, Taipei 11031, Taiwan;
| | - Yi-Shan Yang
- Department of Surgery, Division of Neurosurgery, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan; (Y.-S.Y.); (R.W.Y.C.)
- Department of Neurosurgery, Taipei Medical University Hospital, Taipei 11031, Taiwan
- Taipei Neuroscience Institute, Taipei Medical University, Taipei 11031, Taiwan
| | - Ryan Wing Yuk Chan
- Department of Surgery, Division of Neurosurgery, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan; (Y.-S.Y.); (R.W.Y.C.)
- Department of Neurosurgery, Taipei Medical University Hospital, Taipei 11031, Taiwan
- Taipei Neuroscience Institute, Taipei Medical University, Taipei 11031, Taiwan
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Hallman M, Driscoll JA, Lubbe R, Jeong S, Chang K, Haleem M, Jakus A, Pahapill R, Yun C, Shah R, Hsu WK, Stock SR, Hsu EL. Influence of Geometry and Architecture on the In Vivo Success of 3D-Printed Scaffolds for Spinal Fusion. Tissue Eng Part A 2020; 27:26-36. [PMID: 32098585 DOI: 10.1089/ten.tea.2020.0004] [Citation(s) in RCA: 17] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
We previously developed a recombinant growth factor-free, three-dimensional (3D)-printed material comprising hydroxyapatite (HA) and demineralized bone matrix (DBM) for bone regeneration. This material has demonstrated the capacity to promote re-mineralization of the DBM particles within the scaffold struts and shows potential to promote successful spine fusion. Here, we investigate the role of geometry and architecture in osteointegration, vascularization, and facilitation of spine fusion in a preclinical model. Inks containing HA and DBM particles in a poly(lactide-co-glycolide) elastomer were 3D-printed into scaffolds with varying relative strut angles (90° vs. 45° advancing angle), macropore size (0 μm vs. 500 μm vs. 1000 μm), and strut alignment (aligned vs. offset). The following configurations were compared with scaffolds containing no macropores: 90°/500 μm/aligned, 45°/500 μm/aligned, 90°/1000 μm/aligned, 45°/1000 μm/aligned, 90°/1000 μm/offset, and 45°/1000 μm/offset. Eighty-four female Sprague-Dawley rats underwent spine fusion with bilateral placement of the various scaffold configurations (n = 12/configuration). Osteointegration and vascularization were assessed by using microComputed Tomography and histology, and spine fusion was assessed via blinded manual palpation. The 45°/1000 μm scaffolds with aligned struts achieved the highest average fusion score (1.61/2) as well as the highest osteointegration score. Both the 45°/1000 μm/aligned and 90°/1000 μm/aligned scaffolds elicited fusion rates of 100%, which was significantly greater than the 45°/500 μm/aligned iteration (p < 0.05). All porous scaffolds were fully vascularized, with blood vessels present in every macropore. Vessels were also observed extending from the native transverse process bone, through the protrusions of new bone, and into the macropores of the scaffolds. When viewed independently, scaffolds printed with relative strut angles of 45° and 90° each allowed for osteointegration sufficient to stabilize the spine at L4-L5. Within those parameters, a pore size of 500 μm or greater was generally sufficient to achieve unilateral fusion. However, our results suggest that scaffolds printed with the larger pore size and with aligned struts at an advancing angle of 45° may represent the optimal configuration to maximize osteointegration and fusion capacity. Overall, this work suggests that the HA/DBM composite scaffolds provide a conducive environment for bone regeneration as well as vascular infiltration. This technology, therefore, represents a novel, growth-factor-free biomaterial with significant potential as a bone graft substitute for use in spinal surgery. Impact statement We previously developed a recombinant growth factor-free, three-dimensional (3D)-printed composite material comprising hydroxyapatite and demineralized bone matrix for bone regeneration. Here, we identify a range of 3D geometric and architectural parameters that support the preclinical success of the scaffold, including efficient vascularization, osteointegration, and, ultimately, spinal fusion. Our results suggest that this material holds great promise as a clinically translatable biomaterial for use as a bone graft substitute in orthopedic procedures requiring bone regeneration.
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Affiliation(s)
- Mitchell Hallman
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - J Adam Driscoll
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - Ryan Lubbe
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - Soyeon Jeong
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - Kevin Chang
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - Meraaj Haleem
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - Adam Jakus
- Simpson Querrey Institute, Chicago, Illinois, USA.,Northwestern University Department of Materials Science and Engineering, Evanston, Illinois, USA.,Transplant Division, Northwestern University Department of Surgery, Chicago, Illinois, USA
| | - Richard Pahapill
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - Chawon Yun
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - Ramille Shah
- Simpson Querrey Institute, Chicago, Illinois, USA.,Northwestern University Department of Materials Science and Engineering, Evanston, Illinois, USA.,Transplant Division, Northwestern University Department of Surgery, Chicago, Illinois, USA.,Orthopaedic Research Laboratory, Beaumont Health, Royal Oak, Michigan, USA.,Northwestern University Department of Biomedical Engineering, Evanston, Illinois, USA
| | - Wellington K Hsu
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
| | - Stuart R Stock
- Simpson Querrey Institute, Chicago, Illinois, USA.,Argonne National Laboratory, Argonne, Illinois, USA.,Northwestern University Department of Cell and Molecular Biology, Chicago, Illinois, USA
| | - Erin L Hsu
- Northwestern University Department of Orthopaedic Surgery, Chicago, Illinois, USA.,Simpson Querrey Institute, Chicago, Illinois, USA
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Driscoll JA, Lubbe R, Jakus AE, Chang K, Haleem M, Yun C, Singh G, Schneider AD, Katchko KM, Soriano C, Newton M, Maerz T, Li X, Baker K, Hsu WK, Shah RN, Stock SR, Hsu EL. 3D-Printed Ceramic-Demineralized Bone Matrix Hyperelastic Bone Composite Scaffolds for Spinal Fusion. Tissue Eng Part A 2020; 26:157-166. [PMID: 31469055 PMCID: PMC7044791 DOI: 10.1089/ten.tea.2019.0166] [Citation(s) in RCA: 34] [Impact Index Per Article: 6.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/24/2019] [Accepted: 08/23/2019] [Indexed: 12/31/2022] Open
Abstract
Although numerous spinal biologics are commercially available, a cost-effective and safe bone graft substitute material for spine fusion has yet to be proven. In this study, "3D-Paints" containing varying volumetric ratios of hydroxyapatite (HA) and human demineralized bone matrix (DBM) in a poly(lactide-co-glycolide) elastomer were three-dimensional (3D) printed into scaffolds to promote osteointegration in rats, with an end goal of spine fusion without the need for recombinant growth factor. Spine fusion was evaluated by manual palpation, and osteointegration and de novo bone formation within scaffold struts were evaluated by laboratory and synchrotron microcomputed tomography and histology. The 3:1 HA:DBM composite achieved the highest mean fusion score and fusion rate (92%), which was significantly greater than the 3D printed DBM-only scaffold (42%). New bone was identified extending from the host transverse processes into the scaffold macropores, and osteointegration scores correlated with successful fusion. Strikingly, the combination of HA and DBM resulted in the growth of bone-like spicules within the DBM particles inside scaffold struts. These spicules were not observed in DBM-only scaffolds, suggesting that de novo spicule formation requires both HA and DBM. Collectively, our work suggests that this recombinant growth factor-free composite shows promise to overcome the limitations of currently used bone graft substitutes for spine fusion. Impact Statement Currently, there exists a no safe, yet highly effective, bone graft substitute that is well accepted for use in spine fusion procedures. With this work, we show that a three-dimensional printed scaffold containing osteoconductive hydroxyapatite and osteoinductive demineralized bone matrix that promotes new bone spicule formation, osteointegration, and successful fusion (stabilization) when implemented in a preclinical model of spine fusion. Our study suggests that this material shows promise as a recombinant growth factor-free bone graft substitute that could safely promote high rates of successful fusion and improve patient care.
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Affiliation(s)
- J. Adam Driscoll
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | - Ryan Lubbe
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | - Adam E. Jakus
- Simpson Querrey Institute, Chicago, Illinois
- Department of Materials Science and Engineering, Northwestern University Evanston, Illinois
- Transplant Division, Department of Surgery, Northwestern University, Chicago, Illinois
| | - Kevin Chang
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | - Meraaj Haleem
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | - Chawon Yun
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | - Gurmit Singh
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | - Andrew D. Schneider
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | - Karina M. Katchko
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | | | - Michael Newton
- Orthopaedic Research Laboratory, Beaumont Health, Royal Oak, Michigan
| | - Tristan Maerz
- Orthopaedic Research Laboratory, Beaumont Health, Royal Oak, Michigan
- Department of Orthopaedic Surgery, University of Michigan, Ann Arbor, Michigan
| | - Xin Li
- Simpson Querrey Institute, Chicago, Illinois
| | - Kevin Baker
- Orthopaedic Research Laboratory, Beaumont Health, Royal Oak, Michigan
- Department of Orthopaedic Surgery, Oakland University—William Beaumont School of Medicine, Rochester, Michigan
| | - Wellington K. Hsu
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
| | - Ramille N. Shah
- Simpson Querrey Institute, Chicago, Illinois
- Department of Materials Science and Engineering, Northwestern University Evanston, Illinois
- Transplant Division, Department of Surgery, Northwestern University, Chicago, Illinois
- Orthopaedic Research Laboratory, Beaumont Health, Royal Oak, Michigan
- Department of Biomedical Engineering, Northwestern University, Evanston, Illinois
| | - Stuart R. Stock
- Simpson Querrey Institute, Chicago, Illinois
- Department of Cell and Molecular Biology, Northwestern University, Evanston, Illinois
| | - Erin L. Hsu
- Department of Orthopaedic Surgery, Northwestern University, Chicago, Illinois
- Simpson Querrey Institute, Chicago, Illinois
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Medhat D, Rodríguez CI, Infante A. Immunomodulatory Effects of MSCs in Bone Healing. Int J Mol Sci 2019; 20:ijms20215467. [PMID: 31684035 PMCID: PMC6862454 DOI: 10.3390/ijms20215467] [Citation(s) in RCA: 63] [Impact Index Per Article: 10.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/07/2019] [Revised: 10/29/2019] [Accepted: 10/30/2019] [Indexed: 12/29/2022] Open
Abstract
Mesenchymal stem cells (MSCs) are capable of differentiating into multilineage cells, thus making them a significant prospect as a cell source for regenerative therapy; however, the differentiation capacity of MSCs into osteoblasts seems to not be the main mechanism responsible for the benefits associated with human mesenchymal stem cells hMSCs when used in cell therapy approaches. The process of bone fracture restoration starts with an instant inflammatory reaction, as the innate immune system responds with cytokines that enhance and activate many cell types, including MSCs, at the site of the injury. In this review, we address the influence of MSCs on the immune system in fracture repair and osteogenesis. This paradigm offers a means of distinguishing target bone diseases to be treated with MSC therapy to enhance bone repair by targeting the crosstalk between MSCs and the immune system.
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Affiliation(s)
- Dalia Medhat
- Medical Biochemistry Department, National Research Centre, Dokki, Giza 12622, Egypt.
| | - Clara I Rodríguez
- Stem Cells and Cell Therapy Laboratory, Biocruces Bizkaia Health Research Institute, Cruces University Hospital, Plaza de Cruces S/N, 48903 Barakaldo, Bizkaia, Spain.
| | - Arantza Infante
- Stem Cells and Cell Therapy Laboratory, Biocruces Bizkaia Health Research Institute, Cruces University Hospital, Plaza de Cruces S/N, 48903 Barakaldo, Bizkaia, Spain.
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20
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Bhamb N, Kanim LEA, Drapeau S, Mohan S, Vasquez E, Shimko D, McKAY W, Bae HW. Comparative Efficacy of Commonly Available Human Bone Graft Substitutes as Tested for Posterolateral Fusion in an Athymic Rat Model. Int J Spine Surg 2019; 13:437-458. [PMID: 31745449 DOI: 10.14444/6059] [Citation(s) in RCA: 20] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/08/2023] Open
Abstract
Background Insufficient data exist on bone graft substitute materials efficacy; two thirds lack any clinical data.1,2 This prospective animal study identified efficacy differences among commercially available materials of several classes. Methods Historically validated muscle pouch osteoinduction study (OIS) and posterolateral fusion (PLF) were performed in an athymic rat model. Grafting material products implanted were demineralized bone matrix (DBM)-based allografts (Accell EVO3, DBX Mix, DBX Strip, Grafton Crunch, Grafton Flex, Grafton Matrix, Grafton Putty, Magnifuse, and Progenix Plus), allografts (OsteoSponge, MinerOss), cellular allograft (Osteocel Plus), ceramics (Mozaik Strip), or activated ceramics (Actifuse ABX Putty, Vitoss BA). After 4 weeks, OIS specimens were evaluated ex vivo by histologic osteoinductivity. After 8 weeks, PLF ex vivo specimens were evaluated for fusion by manual palpation (FMP), radiography (FXR), and histology (FHISTO). Results OIS: No materials exhibited a rejection reaction on histology. All DBM-based materials exhibited osteoinductive potential as new bone formation at > 88% of implanted sites. One plain allograft (OsteoSponge) formed bone at 25% of sites. No bone formed for one ceramic (Mozaik Strip), three activated ceramics (Actifuse ABX Putty), or one cellular allograft, regardless of human bone marrow aspirate (hBMA) when added. PLF: Among the 10 DBMs, 6 had FMP of 100% (Accell EVO3, DBX Mix, DBX Strip, Grafton Flex, Grafton Putty, Magnifuse), 2 had FMP of 94% (Grafton Crunch, Grafton Matrix), and 2 conditions had FMP of 0% (Progenix Plus, Progenix Plus + athymic rat iliac crest bone graft [arICBG]). Ceramics (Mozaik Strip), activated ceramics (Actifuse ABX Putty, Vitoss BA), plain allograft (OsteoSponge, MinerOss (PLF study), and cellular allograft (Osteocel Plus) demonstrated 0% FMP. ArICBG demonstrated 13% FMP. Conclusions Eight DBM-based materials (Accell EVO3, DBX Mix, DBX Strip, Grafton Crunch, Grafton Flex, Grafton Matrix, Grafton Putty, Magnifuse) demonstrated excellent (> 90% FMP) efficacy in promoting fusion via bone healing. Two DBM conditions (Progenix Plus, Progenix Plus + arICBG) showed no manual palpation fusion (FMP). Systematically, over the 2 studies (OIS and PLF), cellular (Osteocel Plus), plain allografts (OsteoSponge, MinerOss; PLF study), ceramic (Mozaik Strip), and activated ceramics (Actifuse ABX Putty, Vitoss BA) demonstrated poor FMP efficacy (< 10%). Clinical Relevance When selecting DBMs, clinicians must be cognizant of variability in DBM efficacy by product and lot. While theoretically osteoinductive, cellular allograft and activated ceramics yielded poor in vivo efficacy. Whole allograft and ceramics may provide osteoconductive scaffolding for mixed-material grafting; however, surgeons should be cautious in using them alone. Direct clinical data are needed to establish efficacy for any bone graft substitute.
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Affiliation(s)
- Neil Bhamb
- Cedars-Sinai Medical Center, Los Angeles, California
| | - Linda E A Kanim
- Translational and Clinical Research, Spine Center, Cedars-Sinai Medical Center, Los Angeles, California
| | | | | | | | | | | | - Hyun W Bae
- Cedars-Sinai Medical Center, Los Angeles, California
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21
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Loozen LD, Kruyt MC, Kragten AHM, Schoenfeldt T, Croes M, Oner CF, Dhert WJA, Alblas J. BMP-2 gene delivery in cell-loaded and cell-free constructs for bone regeneration. PLoS One 2019; 14:e0220028. [PMID: 31365542 PMCID: PMC6668905 DOI: 10.1371/journal.pone.0220028] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2018] [Accepted: 07/08/2019] [Indexed: 11/18/2022] Open
Abstract
To induce osteogenicity in bone graft substitutes, plasmid-based expression of BMP-2 (pBMP-2) has been successfully applied in gene activated matrices based on alginate polymer constructs. Here, we investigated whether cell seeding is necessary for non-viral BMP-2 gene expression in vivo. Furthermore, to gain insight in the role of BMP-producing cells, we compared inclusion of bone progenitor cells with non-osteogenic target cells in gene delivery constructs. Plasmid DNA encoding GFP (pGFP) was used to trace transfection of host tissue cells and seeded cells in a rat model. Transgene expression was followed in both cell-free alginate-ceramic constructs as well as constructs seeded with syngeneic fibroblasts or multipotent mesenchymal stromal cells (MSCs). Titration of pGFP revealed that the highest pGFP dose resulted in frequent presence of positive host cells in the constructs. Both cell-loaded groups were associated with transgene expression, most effectively in the MSC-loaded constructs. Subsequently, we investigated effectiveness of cell-free and cell-loaded alginate-ceramic constructs with pBMP-2 to induce bone formation. Local BMP-2 production was found in all groups containing BMP-2 plasmid DNA, and was most pronounced in the groups with MSCs transfected with high concentration pBMP-2. Bone formation was only apparent in the recombinant protein BMP-2 group. In conclusion, we show that non-viral gene delivery of BMP-2 is a potentially effective way to induce transgene expression in vivo, both in cell-seeded as well as cell-free conditions. However, alginate-based gene delivery of BMP-2 to host cells or seeded cells did not result in protein levels adequate for bone formation in this setting, calling for more reliable scaffold compatible transfection methods.
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Affiliation(s)
- Loek D. Loozen
- Dept. Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands
| | - Moyo C. Kruyt
- Dept. Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands
| | | | - Ted Schoenfeldt
- Dept. Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands
| | - Michiel Croes
- Dept. Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands
| | - Cumhur F. Oner
- Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
| | - Wouter J. A. Dhert
- Dept. Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands
- Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
| | - Jacqueline Alblas
- Dept. Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands
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22
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Abstract
STUDY DESIGN Rat posterolateral arthrodesis model. OBJECTIVE Quantify the impact of administration of a proton pump inhibitor on spine fusion. SUMMARY OF BACKGROUND DATA Proton pump inhibitors (PPIs) are widely used for gastrointestinal disorders and for ulcer prophylaxis in patients taking non-steroidal anti-inflammatory drugs. PPIs cause chronic acid suppression which has been found to result in decreased bone mineral density, increased fracture risk, and impaired fracture healing. Despite advances in surgical techniques, pseudarthrosis still occurs in up to 24% of patients requiring revision surgery following spinal fusion procedures. Thus, there are likely many unidentified risk factors. While PPIs have been hypothesized to impact fracture healing, no study has evaluated their effect on spine arthrodesis rates. METHODS Thirty-eight female rats underwent posterolateral lumbar spinal fusion. Rats were divided into two groups: normal saline control and pantroprazole, which was administered by daily intraperitoneal injections. At 8 weeks postoperative spines were evaluated with manual palpation, microCT, histologic analysis, and biomechanical testing. RESULTS Fusion rates of the control group and PPI group were not significantly different (100% vs. 94%). Average fusion scores were significantly lower in the pantoprazole group. New bone formation identified on microCT imaging of bilaterally fused specimens demonstrated a lower average volume of newly generated bone in the PPI group, but this difference was not significant. Biomechanical testing demonstrated no significant difference in strength or stiffness of the fusion mass between the groups. CONCLUSION This study demonstrates that administration of PPIs does not inhibit fusion rates, bone formation, or affect biomechanical integrity of fusion. However, lower fusion scores in the PPI group suggest that a negative impact may still exist. Future studies will explore growth factor and protein expression in the fusion masses as well as utilize higher doses of PPI to fully discern their effect on spine fusion. LEVEL OF EVIDENCE N/A.
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23
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24
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Dang ABC, Hong H, Lee K, Luan T, Reddy S, Kuo AC. Repurposing Human Osteoarthritic Cartilage as a Bone Graft Substitute in an Athymic Rat Posterolateral Spinal Fusion Model. Int J Spine Surg 2018; 12:735-742. [PMID: 30619678 DOI: 10.14444/5092] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/29/2023] Open
Abstract
Background Spinal fusion involves both endochondral and intramembranous bone formation. We previously demonstrated that endochondral cartilage grafts that were derived from human osteoarthritic (OA) articular cartilage can be used as a bone graft in mouse models. We hypothesized that OA cartilage could also be recycled and repurposed as a bone graft substitute in a posterolateral lumbar spinal fusion model in athymic rats. Methods OA articular cartilage was obtained from the femoral resection of a healthy 60-year-old man undergoing elective total knee arthroplasty. The chondrocytes recovered from this tissue were dedifferentiated in monolayer tissue culture and then transitioned to culture conditions that promote chondrocyte hypertrophy. The resultant cell pellets were then used as bone graft substitute for single-level posterolateral spinal fusion in 5 athymic rats. Decortication alone was used as the control group. Spinal fusion was assessed with manual palpation, micro-computed tomography, and histologic analysis. Results In the experimental group, micro-computed tomography at 4 and 8 weeks demonstrated bilateral fusion in 4 of 5 animals and unilateral fusion in 1 animal. At 8 weeks, manual palpation and histologic analysis showed direct correlation with the radiographic findings. Animals undergoing decortication alone failed to generate any spinal fusion. The difference in the fusion rate between groups was statistically significant with respect to both unilateral fusion (P = .047) and bilateral fusion (P = .007). Conclusions In the absence of additional surgically implanted bone graft, hypertrophic chondrocyte grafts are sufficient for generating single-level posterolateral lumbar fusion in athymic rats. Clinical Relevance This animal study demonstrates that cartilage harvested from OA knees can be used as a bone graft substitute. Commercially available cell-based bone grafts have previously only used mesenchymal stem cells or osteoblast precursor cells.
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Affiliation(s)
- Alan B C Dang
- Orthopaedic Section, Surgical Service, San Francisco VA Medical Center, San Francisco, California.,Department of Orthopaedic Surgery, University of California, San Francisco, California
| | - Helena Hong
- Washington University School of Medicine, St Louis, Missouri
| | - Katie Lee
- Department of Orthopaedic Surgery, University of California, San Francisco, California
| | - Tammy Luan
- Orthopaedic Section, Surgical Service, San Francisco VA Medical Center, San Francisco, California
| | - Sanjay Reddy
- Orthopaedic Section, Surgical Service, San Francisco VA Medical Center, San Francisco, California
| | - Alfred C Kuo
- Orthopaedic Section, Surgical Service, San Francisco VA Medical Center, San Francisco, California.,Department of Orthopaedic Surgery, University of California, San Francisco, California
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25
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Jin YZ, Lee JH. Mesenchymal Stem Cell Therapy for Bone Regeneration. Clin Orthop Surg 2018; 10:271-278. [PMID: 30174801 PMCID: PMC6107811 DOI: 10.4055/cios.2018.10.3.271] [Citation(s) in RCA: 100] [Impact Index Per Article: 14.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/08/2018] [Accepted: 06/27/2018] [Indexed: 12/16/2022] Open
Abstract
Mesenchymal stem cells (MSCs) have been used in clinic for approximately 20 years. During this period, various new populations of MSCs have been found or manipulated. However, their characters and relative strength for bone regeneration have not been well known. For a comprehensive understanding of MSCs, we reviewed the literature on the multipotent cells ranging from the definition to the current research progress for bone regeneration. Based on our literature review, bone marrow MSCs have been most widely studied and utilized in clinical settings. Among other populations of MSCs, adipose-derived MSCs and perivascular MSCs might be potential candidates for bone regeneration, whose efficacy and safety still require further investigation.
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Affiliation(s)
- Yuan-Zhe Jin
- Department of Orthopedic Surgery, Seoul National University College of Medicine, Seoul, Korea
| | - Jae Hyup Lee
- Department of Orthopedic Surgery, Seoul National University College of Medicine, Seoul, Korea.,Department of Orthopedic Surgery, SMG-SNU Boramae Medical Center, Seoul, Korea.,Institute of Medical and Biological Engineering, Seoul National University Medical Research Center, Seoul, Korea
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26
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Koerner JD, Markova DZ, Schroeder GD, Calio BP, Shah A, Brooks CW, Vaccaro AR, Anderson DG, Kepler CK. The local cytokine and growth factor response to recombinant human bone morphogenetic protein-2 (rhBMP-2) after spinal fusion. Spine J 2018; 18:1424-1433. [PMID: 29550606 DOI: 10.1016/j.spinee.2018.03.006] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/09/2017] [Revised: 02/12/2018] [Accepted: 03/09/2018] [Indexed: 02/03/2023]
Abstract
BACKGROUND CONTEXT The systemic response regarding cytokine expression after the application of recombinant human bone morphogenetic protein-2 (rhBMP-2) in a rat spinal fusion model has recently been defined, but the local response has not been defined. Defining the local cytokine and growth factor response at the fusion site will help explain the roles of these molecules in the fusion process, as well as that of rhBMP-2. Our hypothesis is that the application of rhBMP-2 to the fusion site will alter the local levels of cytokines and growth factors throughout the fusion process, in a manner that is different from the systemic response, given the tissue-specific effects of rhBMP-2. PURPOSE The purpose of this study was to evaluate the local cytokine and growth factor response after the application of rhBMP-2 in a rat spinal fusion model. STUDY DESIGN/SETTING This was a basic science animal model study. METHODS This study was partially funded by a physician-sponsored grant from Medtronic. A total of 135 Wistar rats (age 8 weeks, weighing approximately 300-400 g) underwent L4-L5 posterolateral intertransverse fusion with demineralized bone graft (approximately 0.4-cm3 rat demineralized bone matrix [DBM] per side). In the first group, 10 µg of rhBMP-2 on an allograft collagen sponge (ACS) was added to the fusion site with approximately 0.4-cm3 rat DBM per side. In the second group, 100 µg of rhBMP-2 on an ACS was added to the fusion site with approximately 0.4-cm3 rat DBM per side, and the third experiment was the control group, which consisted of only an ACS plus 0.4-cm3 DBM per side. There were nine groups of five animals each per experiment. Each group was sacrificed at time points up to 4 weeks (1, 6, 24, and 48 hours, and 4, 7, 14, 21, and 28 days after surgery). At sacrifice, the DBM, transverse processes, and any new bone formed were harvested, immediately frozen in liquid nitrogen, and prepared for protein extraction. ELISA was performed to compare the levels of various cytokines (interleukin [IL]-1β, tumor necrosis factor alpha, IL-6, IL-1RA [IL-1 receptor antagonist], IL-4, and IL-10) and growth factors (vascular endothelial growth factor [VEGF], endothelia growth factor [EGF], insulin-like growth factor-1 [IGF-1], platelet derived growth factor [PDGF], transforming growth factor beta [TGF-β]) that are known to be involved in the fusion-fracture healing process. Fusion was evaluated on the rats sacrificed at 28 days by manual palpation and microcomputed tomography (microCT) by two independent observers. RESULTS The expression of cytokines and growth factors varied throughout the fusion process at each time point. In the groups treated with rh-BMP-2, IL-6 and IL-1RA had higher expression in the early time points (1 and 6 hours). Tumor necrosis factor alpha demonstrated significantly lower expression in the groups treated with rhBMP-2 at Days 1, 2, and 4. At the early time points (1 and 6 hours), in the groups treated with rhBMP-2, all of the growth factors IGF-1, VEGF, platelet derived growth factor AB (PDGF-AB), TGF-β had equal or lower expression compared with controls. At 24 hours, there was a peak in IGF-1, VEGF, and PDGF-AB. These growth factors then declined, with IGF-1 and PDGF-AB having a second peak at Day 7. At 4 weeks, all of the rhBMP-2-treated animals fused based on manual palpation and microCT. The control group had four of five rats fused based on manual palpation and two of five rats based on microCT. CONCLUSIONS There is significant variability in the expression of cytokines throughout the fusion process after treatment with rhBMP-2. The inflammatory response appears to peak early (1 and 6 hours), followed by a significant decrease with rhBMP-2 treatment. However, the growth factor expression appears to be suppressed early (1 and 6 hours), followed by a peak at 24 hours, and a second peak at Day 7.
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Affiliation(s)
- John D Koerner
- Seton Hall-Hackensack Meridian School of Medicine, Hackensack, NJ, USA.
| | - Dessislava Z Markova
- Thomas Jefferson University Department of Orthopaedic Surgery, Philadelphia, PA, USA
| | - Greg D Schroeder
- Rothman Institute, Thomas Jefferson University, 925 Chestnut St, Philadelphia, PA 19, USA
| | - Brian P Calio
- Sidney Kimmel Medical College, Philadelphia, PA, USA
| | - Anuj Shah
- Sidney Kimmel Medical College, Philadelphia, PA, USA
| | | | - Alexander R Vaccaro
- Rothman Institute, Thomas Jefferson University, 925 Chestnut St, Philadelphia, PA 19, USA
| | - D Greg Anderson
- Rothman Institute, Thomas Jefferson University, 925 Chestnut St, Philadelphia, PA 19, USA
| | - Christopher K Kepler
- Rothman Institute, Thomas Jefferson University, 925 Chestnut St, Philadelphia, PA 19, USA
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Tissue Engineered Bone Differentiated From Human Adipose Derived Stem Cells Inhibit Posterolateral Fusion in an Athymic Rat Model. Spine (Phila Pa 1976) 2018; 43:533-541. [PMID: 28816826 PMCID: PMC5812848 DOI: 10.1097/brs.0000000000002384] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/01/2023]
Abstract
STUDY DESIGN Biological augmentation spinal arthrodesis trial in athymic rats. OBJECTIVE To assess the efficacy of tissue-engineered bone to promote L4-L5 intertransverse process fusion in an athymic rat model. SUMMARY OF BACKGROUND DATA Each year in the United States, over 400,000 spinal fusion surgeries are performed requiring bone graft. The current gold standard for posterolateral lumbar fusion is autogenous iliac crest bone graft (ICBG), but the harvesting of ICBG is associated with increased operative time and significant complications. This being the case, an alternative cost-effective bone graft source is needed. METHODS Bovine bone cores were sterilized and decellularized for scaffold production. Human adipose derived mesenchymal stem cells (ADSC) were obtained and verified by tridifferentiation testing and seeded onto dried scaffolds. The seeded cores were cultured for 5 weeks in culture medium designed to mimic endochondral ossification and produce hypertrophic chondrocytes. Single-level intertransverse process fusions were performed at the L4-L5 level of 31 athymic rats. Fifteen rats were implanted with the hypertrophic chondrocyte seeded scaffold and 16 had scaffold alone. Half of the study rats were sacrificed at 3 weeks and the other half at 6 weeks. Spinal fusion was assessed using 2D and 3D micro computed tomography (μCT) analysis and tissue histology. RESULTS At 3 weeks, none of the tissue engineered rats had partial or complete fusion, whereas 62.5% of the decellularized rats fused and another 12.5% had partial fusions (P = 0.013). At 6 weeks, none of the tissue engineered rats fused and 50% had partial fusions, whereas 87.5% of the decellularized rats fused (P = 0.002). CONCLUSION Tissue engineered bone composed of hypertrophic chondrocytes inhibits posterolateral fusion in an athymic rat model and therefore does not represent a promising cost-effective bone graft substitute. LEVEL OF EVIDENCE N/A.
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Bougioukli S, Sugiyama O, Pannell W, Ortega B, Tan MH, Tang AH, Yoho R, Oakes DA, Lieberman JR. Gene Therapy for Bone Repair Using Human Cells: Superior Osteogenic Potential of Bone Morphogenetic Protein 2-Transduced Mesenchymal Stem Cells Derived from Adipose Tissue Compared to Bone Marrow. Hum Gene Ther 2018; 29:507-519. [PMID: 29212377 DOI: 10.1089/hum.2017.097] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022] Open
Abstract
Ex vivo regional gene therapy strategies using animal mesenchymal stem cells genetically modified to overexpress osteoinductive growth factors have been successfully used in a variety of animal models to induce both heterotopic and orthotopic bone formation. However, in order to adapt regional gene therapy for clinical applications, it is essential to assess the osteogenic capacity of transduced human cells and choose the cell type that demonstrates the best clinical potential. Bone-marrow stem cells (BMSC) and adipose-derived stem cells (ASC) were selected in this study for in vitro evaluation, before and after transduction with a lentiviral two-step transcriptional amplification system (TSTA) overexpressing bone morphogenetic protein 2 (BMP-2; LV-TSTA-BMP-2) or green fluorescent protein (GFP; LV-TSTA-GFP). Cell growth, transduction efficiency, BMP-2 production, and osteogenic capacity were assessed. The study demonstrated that BMSC were characterized by a slower cell growth compared to ASC. Fluorescence-activated cell sorting analysis of GFP-transduced cells confirmed successful transduction with the vector and revealed an overall higher but not statistically significant transduction efficiency in ASC versus BMSC (90.2 ± 4.06% vs. 80.4 ± 8.51%, respectively; p = 0.146). Enzyme-linked immunosorbent assay confirmed abundant BMP-2 production by both cell types transduced with LV-TSTA-BMP-2, with BMP-2 production being significantly higher in ASC versus BMSC (239.5 ± 116.55 ng vs. 70.86 ± 24.7 ng; p = 0.001). Quantitative analysis of extracellular deposition of calcium (Alizarin red) and alkaline phosphatase activity showed that BMP-2-transduced cells had a higher osteogenic differentiation capacity compared to non-transduced cells. When comparing the two cell types, ASC/LV-TSTA-BMP-2 demonstrated a significantly higher mineralization potential compared to BMSC/LV-TSTA-BMP-2 7 days post transduction (p = 0.014). In conclusion, this study demonstrates that transduction with LV-TSTA-BMP-2 can significantly enhance the osteogenic potential of both human BMSC and ASC. BMP-2-treated ASC exhibited higher BMP-2 production and greater osteogenic differentiation capacity compared to BMP-2-treated BMSC. These results, along with the fact that liposuction is an easy procedure with lower donor-site morbidity compared to BM aspiration, indicate that adipose tissue might be a preferable source of MSCs to develop a regional gene therapy approach to treat difficult bone-repair scenarios.
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Affiliation(s)
- Sofia Bougioukli
- 1 Department of Orthopedic Surgery, Keck School of Medicine, University of Southern California , Los Angeles, California
| | - Osamu Sugiyama
- 1 Department of Orthopedic Surgery, Keck School of Medicine, University of Southern California , Los Angeles, California
| | - William Pannell
- 1 Department of Orthopedic Surgery, Keck School of Medicine, University of Southern California , Los Angeles, California
| | - Brandon Ortega
- 1 Department of Orthopedic Surgery, Keck School of Medicine, University of Southern California , Los Angeles, California
| | - Matthew H Tan
- 1 Department of Orthopedic Surgery, Keck School of Medicine, University of Southern California , Los Angeles, California
| | - Amy H Tang
- 1 Department of Orthopedic Surgery, Keck School of Medicine, University of Southern California , Los Angeles, California
| | - Robert Yoho
- 2 Cosmetic Surgery Practice , Pasadena, California
| | - Daniel A Oakes
- 1 Department of Orthopedic Surgery, Keck School of Medicine, University of Southern California , Los Angeles, California
| | - Jay R Lieberman
- 1 Department of Orthopedic Surgery, Keck School of Medicine, University of Southern California , Los Angeles, California
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Gene Therapy Strategies in Bone Tissue Engineering and Current Clinical Applications. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2018; 1119:85-101. [DOI: 10.1007/5584_2018_253] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
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Yuan W, Zheng J, Qian J, Zhou X, Wang M, Wang X. [Sustained release of recombinant human bone morphogenetic protein-2 combined with stromal vascular fraction cells in promoting posterolateral spinal fusion in rat model]. ZHONGGUO XIU FU CHONG JIAN WAI KE ZA ZHI = ZHONGGUO XIUFU CHONGJIAN WAIKE ZAZHI = CHINESE JOURNAL OF REPARATIVE AND RECONSTRUCTIVE SURGERY 2017; 31:862-869. [PMID: 29798533 DOI: 10.7507/1002-1892.201703043] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Subscribe] [Scholar Register] [Indexed: 11/03/2022]
Abstract
Objective To observe the effect of stromal vascular fraction cells (SVFs) from rat fat tissue combined with sustained release of recombinant human bone morphogenetic protein-2 (rhBMP-2) in promoting the lumbar fusion in rat model. Methods SVFs were harvested from subcutaneous fat of bilateral inguinal region of 4-month-old rat through the collagenase I digestion. The sustained release carrier was prepared via covalent bond of the rhBMP-2 and β-tricalcium phosphate (β-TCP) by the biominetic apatite coating process. The sustained release effect was measured by BCA method. Thirty-two rats were selected to establish the posterolateral lumbar fusion model and were divided into 4 groups, 8 rats each group. The decalcified bone matrix (DBX) scaffold+PBS, DBX scaffold+rhBMP-2/β-TCP sustained release carrier, DBX scaffold+SVFs, and DBX scaffold+rhBMP-2/β-TCP sustained release carrier+SVFs were implanted in groups A, B, C, and D respectively. X-ray films, manual spine palpation, and high-resolution micro-CT were used to evaluate spinal fusion at 8 weeks after operation; bone mineral density (BMD) and bone volume fraction were analyzed; the new bone formation was evaluated by HE staining and Masson's trichrome staining, osteocalcin (OCN) was detected by immunohistochemical staining. Results The cumulative release amount of rhBMP-2 was about 40% at 2 weeks, indicating sustained release effect of rhBMP-2; while the control group was almost released within 2 weeks. At 8 weeks, the combination of manual spine palpation, X-ray, and micro-CT evaluation showed that group D had the strongest bone formation (100%, 8/8), followed by group B (75%, 6/8), group C (37.5%, 3/8), and group A (12.5%, 1/8). Micro-CT analysis showed BMD and bone volume fraction were significantly higher in group D than groups A, B, and C ( P<0.05), and in group B than groups A and C ( P<0.05). HE staining, Masson's trichrome staining, and immunohistochemistry staining for OCN staining exhibited a large number of cartilage cells with bone matrix deposition, and an active osteogenic process similar to the mineralization of long bones in group D. The bone formation of group B was weaker than that of group D, and there was no effective new bone formation in groups A and C. Conclusion The combination of sustained release of rhBMP-2 and freshly SVFs can significantly promote spinal fusion in rat model, providing a theoretical basis for further clinical applications.
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Affiliation(s)
- Wei Yuan
- Department of Orthopedic Surgery, Affiliated Zhoupu Hospital, Shanghai University of Medicine & Health Sciences, Shanghai, 201318, P.R.China
| | - Jun Zheng
- Department of Orthopedics and Traumatology, Yueyang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200437,
| | - Jinyu Qian
- Department of Orthopedics and Traumatology, Yueyang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200437, P.R.China
| | - Xiaoxiao Zhou
- Department of Orthopedic Surgery, Affiliated Zhoupu Hospital, Shanghai University of Medicine & Health Sciences, Shanghai, 201318, P.R.China
| | - Minghui Wang
- Department of Orthopedic Surgery, Affiliated Zhoupu Hospital, Shanghai University of Medicine & Health Sciences, Shanghai, 201318, P.R.China
| | - Xiuhui Wang
- Department of Orthopedic Surgery, Affiliated Zhoupu Hospital, Shanghai University of Medicine & Health Sciences, Shanghai, 201318,
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Mesenchymal Stem Cells for the Treatment of Spinal Arthrodesis: From Preclinical Research to Clinical Scenario. Stem Cells Int 2017; 2017:3537094. [PMID: 28286524 PMCID: PMC5327761 DOI: 10.1155/2017/3537094] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/23/2016] [Accepted: 01/05/2017] [Indexed: 02/07/2023] Open
Abstract
The use of spinal fusion procedures has rapidly augmented over the last decades and although autogenous bone graft is the “gold standard” for these procedures, alternatives to its use have been investigated over many years. A number of emerging strategies as well as tissue engineering with mesenchymal stem cells (MSCs) have been planned to enhance spinal fusion rate. This descriptive systematic literature review summarizes the in vivo studies, dealing with the use of MSCs in spinal arthrodesis surgery and the state of the art in clinical applications. The review has yielded promising evidence supporting the use of MSCs as a cell-based therapy in spinal fusion procedures, thus representing a suitable biological approach able to reduce the high cost of osteoinductive factors as well as the high dose needed to induce bone formation. Nevertheless, despite the fact that MSCs therapy is an interesting and important opportunity of research, in this review it was detected that there are still doubts about the optimal cell concentration and delivery method as well as the ideal implantation techniques and the type of scaffolds for cell delivery. Thus, further inquiry is necessary to carefully evaluate the clinical safety and efficacy of MSCs use in spine fusion.
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Virk SS, Coble D, Bertone AL, Hussein HH, Khan SN. Experimental Design and Surgical Approach to Create a Spinal Fusion Model in a New Zealand White Rabbit (Oryctolagus cuniculus). J INVEST SURG 2016; 30:226-234. [PMID: 27739917 DOI: 10.1080/08941939.2016.1235748] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022]
Abstract
There are several animal models routinely used for study of the spinal fusion process and animal selection largely depends on the scientific question to be answered. This review outlines the advantages and disadvantages of various animal models used to study spinal fusion and describes the New Zealand White (NSW) rabbit which is the most popular preclinical model to study spinal fusion. We outline critical steps required in planning and performing spinal fusion surgery in this model. This includes determination of the required animal number to obtain statistical significance, an outline of appropriate technique for posterolateral fusion and other components of completing a study. As advances in drug delivery move forward and our understanding of the cascade of gene expression occurring during the fusion process grows, performing and interpreting preclinical animal models will be vital to validating new therapies to enhance spinal fusion.
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Affiliation(s)
- Sohrab S Virk
- a Department of Orthopaedics , Ohio State University Wexner Medical Center , Columbus , Ohio , USA
| | - Dondrae Coble
- b Office of Research, College of Veterinary Medicine, The Ohio State University , Columbus , Ohio , USA
| | - Alicia L Bertone
- a Department of Orthopaedics , Ohio State University Wexner Medical Center , Columbus , Ohio , USA.,c Comparative Orthopedic Research Laboratory, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University , Columbus , Ohio , USA
| | - Hayam Hamaz Hussein
- c Comparative Orthopedic Research Laboratory, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University , Columbus , Ohio , USA
| | - Safdar N Khan
- a Department of Orthopaedics , Ohio State University Wexner Medical Center , Columbus , Ohio , USA
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Betz VM, Betz OB, Rosin T, Keller A, Thirion C, Salomon M, Manthey S, Augat P, Jansson V, Müller PE, Rammelt S, Zwipp H. An expedited approach for sustained delivery of bone morphogenetic protein-7 to bone defects using gene activated fragments of subcutaneous fat. J Gene Med 2016; 18:199-207. [DOI: 10.1002/jgm.2892] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/29/2016] [Revised: 06/28/2016] [Accepted: 06/29/2016] [Indexed: 12/26/2022] Open
Affiliation(s)
- Volker M. Betz
- Department of Trauma and Reconstructive Surgery and Center for Translational Bone, Joint and Soft Tissue Research; University Hospital Carl Gustav Carus Dresden, TU Dresden; Dresden Germany
| | - Oliver B. Betz
- Department of Orthopedic Surgery, Physical Medicine and Rehabilitation; University Hospital Grosshadern, Ludwig-Maximilians-University Munich; Munich Germany
| | - Tom Rosin
- Department of Trauma and Reconstructive Surgery and Center for Translational Bone, Joint and Soft Tissue Research; University Hospital Carl Gustav Carus Dresden, TU Dresden; Dresden Germany
| | - Alexander Keller
- Department of Orthopedic Surgery, Physical Medicine and Rehabilitation; University Hospital Grosshadern, Ludwig-Maximilians-University Munich; Munich Germany
| | | | | | - Suzanne Manthey
- Department of Trauma and Reconstructive Surgery and Center for Translational Bone, Joint and Soft Tissue Research; University Hospital Carl Gustav Carus Dresden, TU Dresden; Dresden Germany
| | - Peter Augat
- Institute of Biomechanics; Trauma Center Murnau; Murnau Germany
- Paracelsus Medical University; Salzburg Austria
| | - Volkmar Jansson
- Department of Orthopedic Surgery, Physical Medicine and Rehabilitation; University Hospital Grosshadern, Ludwig-Maximilians-University Munich; Munich Germany
| | - Peter E. Müller
- Department of Orthopedic Surgery, Physical Medicine and Rehabilitation; University Hospital Grosshadern, Ludwig-Maximilians-University Munich; Munich Germany
| | - Stefan Rammelt
- Department of Trauma and Reconstructive Surgery and Center for Translational Bone, Joint and Soft Tissue Research; University Hospital Carl Gustav Carus Dresden, TU Dresden; Dresden Germany
| | - Hans Zwipp
- Department of Trauma and Reconstructive Surgery and Center for Translational Bone, Joint and Soft Tissue Research; University Hospital Carl Gustav Carus Dresden, TU Dresden; Dresden Germany
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Mendoza MC, Sonn KA, Kannan AS, Bellary SS, Mitchell SM, Singh G, Park C, Yun C, Stock SR, Hsu EL, Hsu WK. The effect of vancomycin powder on bone healing in a rat spinal rhBMP-2 model. J Neurosurg Spine 2016; 25:147-53. [DOI: 10.3171/2015.11.spine15536] [Citation(s) in RCA: 29] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Abstract
OBJECTIVE
This study aims to quantify the impact of vancomycin powder application on new bone formation and spine fusion rates in a rat posterolateral arthrodesis model.
METHODS
Thirty-six female Sprague-Dawley rats underwent a posterolateral lumbar spinal fusion (PLF) at the L-4 and L-5 vertebrae. Fusion was elicited via implantation of an absorbable collagen sponge containing 3 µg rhBMP-2. Rats were divided into 3 groups: no vancomycin (control), standard-dose vancomycin, and high-dose vancomycin, based on what was applied to the fusion bed. Clinical studies typically describe the application of 1 g vancomycin into the surgical wound. Presuming an average individual patient weight of 70 kg, a weight-based equivalent dose of vancomycin powder was applied subfascially in the PLF model constituting a “standard-dose” treatment group (14.3 mg/kg, n = 12). To determine whether there is a critical threshold beyond which vancomycin increases the risk of pseudarthrosis, a 10-fold higher dose was administered to a “high-dose” treatment group (143 mg/kg, n = 12). No vancomycin powder was applied to the surgical site in the control group (n = 12). Fusion was evaluated with plain radiographs at 4 and 8 weeks after surgery. The spines were harvested after the 8-week radiographs were obtained and evaluated using manual palpation, microCT analysis, and histological analysis.
RESULTS
Radiographs demonstrated equivalent bridging bone formation in all groups. No significant differences in fusion scores were seen in the standard-dose (mean 2.25) or high-dose (2.13) treatment groups relative to untreated control animals (1.78). Similarly, fusion rates did not differ significantly different between vancomycin-treated animals (100% for both groups) and control animals (92%). Quantification of new bone formation via microCT imaging revealed no significant between-groups differences in the volume of newly regenerated bone (control vs standard-dose vancomycin, p = 0.57; control vs high-dose vancomycin, p = 0.53).
CONCLUSIONS
This is the first in vivo study to specifically address the development of pseudarthrosis after intrawound application of vancomycin during fusion surgery. Our results demonstrate that vancomycin powder does not inhibit fusion rates at a dose that is the weight-percentage equivalent of what is routinely used by surgeons. Moreover, bone formation and fusion rates were not reduced even after administration of a vancomycin dose that is 10-fold higher than that which is typically administered clinically. Our findings suggest that if there is a critical threshold above which vancomycin inhibits bone healing, such a dose is out of the range which might be considered reasonable for clinical use.
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Affiliation(s)
| | | | | | | | | | | | | | | | | | - Erin L. Hsu
- 3Department of Orthopaedic Surgery, Simpson Querrey Institute for Bionanotechnology and Regenerative Nanomedicine, Northwestern University, Chicago, Illinois
| | - Wellington K. Hsu
- 3Department of Orthopaedic Surgery, Simpson Querrey Institute for Bionanotechnology and Regenerative Nanomedicine, Northwestern University, Chicago, Illinois
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Ishida W, Elder BD, Holmes C, Lo SFL, Witham TF. Variables Affecting Fusion Rates in the Rat Posterolateral Spinal Fusion Model with Autogenic/Allogenic Bone Grafts: A Meta-analysis. Ann Biomed Eng 2016; 44:3186-3201. [PMID: 27473706 DOI: 10.1007/s10439-016-1701-8] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2016] [Accepted: 07/21/2016] [Indexed: 01/14/2023]
Abstract
The rat posterolateral spinal fusion model with autogenic/allogenic bone graft (rat PFABG) has been increasingly utilized as an experimental model to assess the efficacy of novel fusion treatments. The objective of this study was to investigate the reliability of the rat PFABG model and examine the effects of different variables on spinal fusion. A web-based literature search from January, 1970 to September, 2015, yielded 26 studies, which included 40 rat PFABG control groups and 449 rats. Data regarding age, weight, sex, and strain of rats, graft volume, graft type, decorticated levels, surgical approach, institution, the number of control rats, fusion rate, methods of fusion assessment, and timing of fusion assessment were collected and analyzed. The primary outcome variable of interest was fusion rate, as evaluated by manual palpation. Fusion rates varied widely, from 0 to 96%. The calculated overall fusion rate was 46.1% with an I 2 value of 62.4, which indicated moderate heterogeneity. Weight >300 g, age >14 weeks, male rat, Sprague-Dawley strain, and autogenic coccyx grafts increased fusion rates with statistical significance. Additionally, an assessment time-point ≥8 weeks had a trend towards statistical significance (p = 0.070). Multi-regression analysis demonstrated that timing of assessment and age as continuous variables, as well as sex as a categorical variable, can predict the fusion rate with R 2 = 0.82. In an inter-institution reliability analysis, the pooled overall fusion rate was 50.0% [44.8, 55.3%], with statistically significant differences among fusion outcomes at different institutions (p < 0.001 and I 2 of 72.2). Due to the heterogeneity of fusion outcomes, the reliability of the rat PFABG model was relatively limited. However, selection of adequate variables can optimize its use as a control group in studies evaluating the efficacy of novel fusion therapies.
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Affiliation(s)
- Wataru Ishida
- Department of Neurosurgery, The Johns Hopkins University School of Medicine, 1800 Orleans St., Room 6007, Baltimore, MD, 21287, USA
| | - Benjamin D Elder
- Department of Neurosurgery, The Johns Hopkins University School of Medicine, 1800 Orleans St., Room 6007, Baltimore, MD, 21287, USA.
| | - Christina Holmes
- Department of Neurosurgery, The Johns Hopkins University School of Medicine, 1800 Orleans St., Room 6007, Baltimore, MD, 21287, USA
| | - Sheng-Fu L Lo
- Department of Neurosurgery, The Johns Hopkins University School of Medicine, 1800 Orleans St., Room 6007, Baltimore, MD, 21287, USA
| | - Timothy F Witham
- Department of Neurosurgery, The Johns Hopkins University School of Medicine, 1800 Orleans St., Room 6007, Baltimore, MD, 21287, USA
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Buser Z, Acosta FL. Stem cells and spinal fusion--are we there yet? Spine J 2016; 16:400-1. [PMID: 27063500 DOI: 10.1016/j.spinee.2015.11.013] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/02/2015] [Revised: 10/23/2015] [Accepted: 11/10/2015] [Indexed: 02/03/2023]
Abstract
Wheeler DL, Fredericks DC, Dryer RF, Bae HW. Allogeneic mesenchymal precursor cells (MPCs) combined with an osteoconductive scaffold to promote lumbar interbody spine fusion in an ovine model. Spine J 2016:16:389-99 (in this issue).
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Affiliation(s)
- Zorica Buser
- Department of Orthopaedic Surgery, University of Southern California, 1450 San Pablo St, Suite 5400, Los Angeles, CA, 90033, USA
| | - Frank L Acosta
- Department of Orthopaedic Surgery, University of Southern California, 1450 San Pablo St, Suite 5400, Los Angeles, CA, 90033, USA; Department of Neurological Surgery, University of Southern California, 1450 San Pablo St, Suite 5400, Los Angeles, CA, 90033, USA.
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Ghodasra JH, Nickoli MS, Hashmi SZ, Nelson JT, Mendoza M, Nicolas JD, Bellary SS, Sonn K, Ashtekar A, Park CJ, Babu J, Yun C, Ghosh A, Kannan A, Stock SR, Hsu WK, Hsu EL. Ovariectomy-Induced Osteoporosis Does Not Impact Fusion Rates in a Recombinant Human Bone Morphogenetic Protein-2-Dependent Rat Posterolateral Arthrodesis Model. Global Spine J 2016; 6:60-8. [PMID: 26835203 PMCID: PMC4733376 DOI: 10.1055/s-0035-1556582] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/22/2015] [Accepted: 04/24/2015] [Indexed: 11/16/2022] Open
Abstract
Study Design Randomized, controlled animal study. Objective Recombinant human bone morphogenetic protein-2 (rhBMP-2) is frequently utilized as a bone graft substitute in spinal fusions to overcome the difficult healing environment in patients with osteoporosis. However, the effects of estrogen deficiency and poor bone quality on rhBMP-2 efficacy are unknown. This study sought to determine whether rhBMP-2-induced healing is affected by estrogen deficiency and poor bone quality in a stringent osteoporotic posterolateral spinal fusion model. Methods Aged female Sprague-Dawley rats underwent an ovariectomy (OVX group) or a sham procedure, and the OVX animals were fed a low-calcium, low-phytoestrogen diet. After 12 weeks, the animals underwent a posterolateral spinal fusion with 1 μg rhBMP-2 on an absorbable collagen sponge. Representative animals were sacrificed at 1 week postoperative for alkaline phosphatase (ALP) and osteocalcin serum analyses. The remaining animals underwent radiographs 2 and 4 weeks after surgery and were subsequently euthanized for fusion analysis by manual palpation, micro-computed tomography (CT) imaging, and histologic analysis. Results The ALP and osteocalcin levels were similar between the control and OVX groups. Manual palpation revealed no significant differences in the fusion scores between the control (1.42 ± 0.50) and OVX groups (1.83 ± 0.36; p = 0.07). Fusion rates were 100% in both groups. Micro-CT imaging revealed no significant difference in the quantity of new bone formation, and histologic analysis demonstrated bridging bone across the transverse processes in fused animals from both groups. Conclusions This study demonstrates that estrogen deficiency and compromised bone quality do not negatively influence spinal fusion when utilizing rhBMP-2, and the osteoinductive capacity of the growth factor is not functionally reduced under osteoporotic conditions in the rat. Although osteoporosis is a risk factor for pseudarthrosis/nonunion, rhBMP-2-induced healing was not inhibited in osteoporotic rats.
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Affiliation(s)
- Jason H. Ghodasra
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Michael S. Nickoli
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Sohaib Z. Hashmi
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - John T. Nelson
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Marco Mendoza
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Joseph D. Nicolas
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Sharath S. Bellary
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Kevin Sonn
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Amruta Ashtekar
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Christian J. Park
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Jacob Babu
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Chawon Yun
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Anjan Ghosh
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Abhishek Kannan
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Stuart R. Stock
- Department of Materials Science and Engineering, Northwestern University Feinberg School of Medicine, Evanston, Illinois, United States
| | - Wellington K. Hsu
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States,Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
| | - Erin L. Hsu
- Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States,Address for correspondence Erin L. Hsu, PhD Department of Orthopaedic Surgery, Northwestern University Feinberg School of Medicine676 N. St. Clair Street, Suite 1350, Chicago, IL 60611United States
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LIPUS promotes spinal fusion coupling proliferation of type H microvessels in bone. Sci Rep 2016; 6:20116. [PMID: 26830666 PMCID: PMC4735589 DOI: 10.1038/srep20116] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/27/2015] [Accepted: 12/29/2015] [Indexed: 01/17/2023] Open
Abstract
Low-intensity pulsed ultrasound (LIPUS) has been found to accelerate spinal fusion. Type H microvessels are found in close relation with bone development. We analyzed the role of type H vessels in rat spinal fusion model intervened by LIPUS. It was found LIPUS could significantly accelerate bone fusion rate and enlarge bone callus. Osteoblasts were specifically located on the bone meshwork of the allograft, and were surrounded by type H microvessels. LIPUS could significantly increase the quantity of osteoblasts during spine fusion, which process was coupled with elevated angiogenesis of type H microvessels. Our results suggest that LIPUS may be a noninvasive adjuvant treatment modality in spinal fusion for clinical use. The treatment is recommended for usage for at least one month.
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Liao JC. Cell Therapy Using Bone Marrow-Derived Stem Cell Overexpressing BMP-7 for Degenerative Discs in a Rat Tail Disc Model. Int J Mol Sci 2016; 17:ijms17020147. [PMID: 26805824 PMCID: PMC4783881 DOI: 10.3390/ijms17020147] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/14/2015] [Revised: 01/13/2016] [Accepted: 01/14/2016] [Indexed: 01/08/2023] Open
Abstract
Degenerative discs can cause low back pain. Cell-based transplantation or growth factors therapy have been suggested as a strategy to stimulate disc regeneration. Bone marrow-derived mesenchymal stem cells (BMDMSC) containing bone morphogenetic protein-7 (BMP-7) gene were constructed. We evaluated the effectiveness of these BMP-7 overexpressing cells on degenerative discs in rat tails. In vitro and in vivo studies were designed. In the first stage, the rats were divided into two group according to discs punctured by different needle gauges (18 gauge and 22 gauge). In the second stage, the ideal size of needle was used to induce rat tail disc degeneration. These animals are divided into three groups according to timing of treatment (zero-week, two-week, four-week). Each group was divided into three treating subgroups: control group, BMDMSC group, and Baculo-BMP-7-BMDMSC group. Each rat undergoes radiography examination every two weeks. After eight weeks, the discs were histologically examined with hematoxylin and eosin stain and Alcian blue stain. The 18-gauge group exhibited significant decrease in disc height index (%) than 22-gauge group at eight weeks at both Co6-7 (58.1% ± 2.8% vs. 63.7% ± 1.0%, p = 0.020) and Co8-9 discs (62.7% ± 2.8% vs. 62.8% ± 1.5%, p = 0.010). Baculo-BMP-7-BMDMSCs group showed significant difference in disc height index compared to the BMDMSCs group at both Co6-7 (93.7% ± 1.5% vs. 84.8% ± 1.0%, p = 0.011) and Co8-9 (86.0% ± 2.1% vs. 81.8% ± 1.7%, p = 0.012). In Baculo-BMP-7-BMDMSCs group, the zero-week treatment subgroup showed significant better in disc height index compared to two-week treatment group (p = 0.044), and four-week treatment group (p = 0.011). The zero-week treatment subgroup in Baculo-BMP-7-BMDMSCs group also had significant lower histology score than two-week treatment (4.3 vs. 5.7, p = 0.045) and four-week treatment (4.3 vs. 6.0, p = 0.031). In conclusion, Baculo-BMP-7-BMDMSC can slow down the progression of disc degeneration, but could not provide evidence of regeneration. Early treatment might obtain more distinct results.
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Affiliation(s)
- Jen-Chung Liao
- Department of Orthopedics Surgery, Bone and Joint Research Center, Chang Gung Memorial Hospital, Chang Gung University, No._5, Fu-Shin Street; Kweishian, Taoyuan 333, Taiwan.
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Avril P, Le Nail LR, Brennan MÁ, Rosset P, De Pinieux G, Layrolle P, Heymann D, Perrot P, Trichet V. Mesenchymal stem cells increase proliferation but do not change quiescent state of osteosarcoma cells: Potential implications according to the tumor resection status. J Bone Oncol 2015; 5:5-14. [PMID: 26998421 PMCID: PMC4782020 DOI: 10.1016/j.jbo.2015.11.002] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/07/2015] [Revised: 11/13/2015] [Accepted: 11/30/2015] [Indexed: 12/11/2022] Open
Abstract
Conventional therapy of primary bone tumors includes surgical excision with wide resection, which leads to physical and aesthetic defects. For reconstruction of bone and joints, allografts can be supplemented with mesenchymal stem cells (MSCs). Similarly, adipose tissue transfer (ATT) is supplemented with adipose-derived stem cells (ADSCs) to improve the efficient grafting in the correction of soft tissue defects. MSC-like cells may also be used in tumor-targeted cell therapy. However, MSC may have adverse effects on sarcoma development. In the present study, human ADSCs, MSCs and pre-osteoclasts were co-injected with human MNNG-HOS osteosarcoma cells in immunodeficient mice. ADSCs and MSCs, but not the osteoclast precursors, accelerated the local proliferation of MNNG-HOS osteosarcoma cells. However, the osteolysis and the metastasis process were not exacerbated by ADSCs, MSCs, or pre-osteoclasts. In vitro proliferation of MNNG-HOS and Saos-2 osteosarcoma cells was increased up to 2-fold in the presence of ADSC-conditioned medium. In contrast, ADSC-conditioned medium did not change the dormant, quiescent state of osteosarcoma cells cultured in oncospheres. Due to the enhancing effect of ADSCs/MSCs on in vivo/in vitro proliferation of osteosarcoma cells, MSCs may not be good candidates for osteosarcoma-targeted cell therapy. Although conditioned medium of ADSCs accelerated the cell cycle of proliferating osteosarcoma cells, it did not change the quiescent state of dormant osteosarcoma cells, indicating that ADSC-secreted factors may not be involved in the risk of local recurrence.
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Affiliation(s)
- Pierre Avril
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France
| | - Louis-Romée Le Nail
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France; University Hospital, Service de Chirurgie Orthopédique et Traumatologique, Tours F-37044, France; Faculté de Médecine, Université François Rabelais, Tours F-37044, France
| | - Meadhbh Á Brennan
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France
| | - Philippe Rosset
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France; University Hospital, Service de Chirurgie Orthopédique et Traumatologique, Tours F-37044, France; Faculté de Médecine, Université François Rabelais, Tours F-37044, France
| | - Gonzague De Pinieux
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France; Faculté de Médecine, Université François Rabelais, Tours F-37044, France; University Hospital, Service d'Anatomie Pathologique, Tours F-37044, France
| | - Pierre Layrolle
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France
| | - Dominique Heymann
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France
| | - Pierre Perrot
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France; University Hospital, Service de Chirurgie Plastique et des Brûlés, Nantes F-44093, France
| | - Valérie Trichet
- INSERM, UMR 957, Equipe Labellisée LIGUE 2012, Nantes F-44035, France; Université de Nantes, Nantes Atlantique Universités, Laboratoire de Physiopathologie de la Résorption Osseuse et Thérapie des Tumeurs Osseuses Primitives, Faculté de Médecine, 1 rue Gaston Veil, Nantes F-44035, France
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Kömürcü E, Özyalvaçlı G, Kaymaz B, Gölge UH, Göksel F, Cevizci S, Adam G, Ozden R. Effects of Local Administration of Boric Acid on Posterolateral Spinal Fusion with Autogenous Bone Grafting in a Rodent Model. Biol Trace Elem Res 2015; 167:77-83. [PMID: 25728510 DOI: 10.1007/s12011-015-0274-y] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/22/2015] [Accepted: 02/13/2015] [Indexed: 11/29/2022]
Abstract
Spinal fusion is among the most frequently applied spinal surgical procedures. The goal of the present study was to evaluate whether the local administration of boric acid (BA) improves spinal fusion in an experimental spinal fusion model in rats. Currently, there is no published data that evaluates the possible positive effects if the local administration of BA on posterolateral spinal fusion. Thirty-two rats were randomly divided into four independent groups: no material was added at the fusion area for group 1; an autogenous morselized corticocancellous bone graft was used for group 2; an autogenous morselized corticocancellous bone graft with boric acid (8.7 mg/kg) for group 3; and only boric acid was placed into the fusion area for group 4. The L4-L6 spinal segments were collected at week 6, and the assessments included radiography, manual palpation, and histomorphometry. A statistically significant difference was determined between the groups with regard to the mean histopathological scores (p = 0.002), and a paired comparison was made with the Mann-Whitney U test to detect the group/groups from which the difference originated. It was determined that only the graft + BA practice increased the histopathological score significantly with regard to the control group (p = 0.002). Whereas, there was no statistically significant difference between the groups in terms of the manual assessment of fusion and radiographic analysis (respectively p = 0.328 and p = 0.196). This preliminary study suggests that BA may clearly be useful as a therapeutic agent in spinal fusion. However, further research is required to show the most effective dosage of BA on spinal fusion, and should indicate whether BA effects spinal fusion in the human body.
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Affiliation(s)
- Erkam Kömürcü
- Faculty of Medicine, Department of Orthopaedics and Traumatology, Çanakkale Onsekiz Mart University, Çanakkale, Turkey,
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Hsu EL, Sonn K, Kannan A, Bellary S, Yun C, Hashmi S, Nelson J, Mendoza M, Nickoli M, Ghodasra J, Park C, Mitchell S, Ashtekar A, Ghosh A, Jain A, Stock SR, Hsu WK. Dioxin Exposure Impairs BMP-2-Mediated Spinal Fusion in a Rat Arthrodesis Model. J Bone Joint Surg Am 2015; 97:1003-10. [PMID: 26085534 DOI: 10.2106/jbjs.n.01311] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/01/2023]
Abstract
BACKGROUND Cigarette smoking inhibits bone-healing and leads to increased rates of pseudarthrosis. However, the mechanisms behind these effects are controversial. Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin)--a cigarette smoke constituent and potent activator of the aryl hydrocarbon receptor (Ahr)--negatively impacts bone quality and osteoblast differentiation. We hypothesized that activation of the Ahr by dioxin would inhibit bone morphogenetic protein (BMP)-2-mediated spinal fusion in a rat arthrodesis model. METHODS Female Long-Evans rats were pretreated with dioxin or vehicle in six weekly doses, followed by bilateral posterior lumbar spinal fusion across the L4-L5 transverse processes using recombinant human BMP (rhBMP)-2. Treatments continued until sacrifice at four weeks postoperatively. A third group was treated with dioxin for six weeks, followed by a recovery period of four elimination half-lives to assess the reversible effects of dioxin exposure on spinal fusion capacity. Bone formation and fusion capacity were evaluated using fusion scoring, radiography, micro-computed tomography, and histologic analysis. RESULTS Fusion scores for dioxin-treated and dioxin-recovery rats were significantly lower than those for controls. Although fusion rates were also significantly reduced in dioxin-treated animals relative to controls (50% versus 100%, respectively), rates were not significantly reduced in dioxin-recovery animals (80%). CONCLUSIONS Dioxin treatment significantly inhibited spinal fusion in a rat arthrodesis model, and a prolonged cessation of dioxin exposure facilitated only a partial recovery of bone-healing capacity. This finding indicates that, although the effects of dioxin are persistent, an extended recovery from exposure could potentially restore bone regeneration in vivo. CLINICAL RELEVANCE Development of a pharmacologic agent that reduces the adverse effects of cigarette smoke on bone-healing could prove useful to orthopaedic surgeons. Since dioxin and other similar cigarette smoke toxins exert their effects through Ahr pathway activation, the receptor represents a potential therapeutic target to improve spinal fusion rates in patients who smoke.
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Affiliation(s)
- Erin L Hsu
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Kevin Sonn
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Abhishek Kannan
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Sharath Bellary
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Chawon Yun
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Sohaib Hashmi
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - John Nelson
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Marco Mendoza
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Michael Nickoli
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Jason Ghodasra
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Christian Park
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Sean Mitchell
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Amruta Ashtekar
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Anjan Ghosh
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Akshay Jain
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Stuart R Stock
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
| | - Wellington K Hsu
- Department of Orthopaedic Surgery, Northwestern University, 676 North Saint Claire Street, Suite #1350, Chicago, IL 60611. E-mail address for E.L. Hsu:
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Gao Y, Li C, Wang H, Fan G. Acceleration of bone-defect repair by using A-W MGC loaded with BMP2 and triple point-mutant HIF1α-expressing BMSCs. J Orthop Surg Res 2015; 10:83. [PMID: 26018771 PMCID: PMC4450843 DOI: 10.1186/s13018-015-0219-3] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/16/2014] [Accepted: 05/09/2015] [Indexed: 02/03/2023] Open
Abstract
Background The goal of this study is to explore the effects of A-W MGC (apatite-wollastonite magnetic bioactive glass-ceramic) loaded with BMP2 (bone morphogenetic protein 2)- and HIF1αmu (hypoxia-inducible factor 1 mutation)-expressing BMSCs (bone marrow mesenchymal stem cells) on the bone defect repair. Methods (1) BMSCs were infected with viral solution containing BMP2 and HIF1αmu with the best MOI (multiplicity of infection). The efficiency was observed via hrGFP (human renilla reniformis green fluorescent protein). (2) The cells were divided into five groups (A–E), and ALP (alkaline phosphatase) activity was measured. (3) BMP2 and HIF1α (hypoxia-inducible factor 1α) protein were measured. (4) A-W MGC was loaded with BMSCs that contain the genes and implanted into the bone defect model. The animals were sacrificed 8 and 12 weeks later. (5) The healing was measured with X-ray, histology, and biomechanics. Results (1) BMSCs in A–D showed high transfection efficiency. (2) ALP in A and B was higher than the others (p = 0.041 or 0.038); A was higher than B (p = 0.038); (3) BMP2 in A and B was higher than the others (p = 0.014). HIF1α in A and C was higher than the others (p = 0.020). (4) 8 and 12 weeks after, an X-ray indicated that bone defect was nearly fully repaired in A and C. (5) 12 weeks after, the bone remodeling was complete in A and C. (6) The flexural strength in A and C was stronger than the others (p = 0.043). Conclusion Engineered A-W MGC with BMP2 and HIF1αmu-expressing BMSCs exhibits comparable therapeutic effects of bone-defect repair as an autologous bone graft.
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Affiliation(s)
- Yuzhong Gao
- Department of Orthopedics, The Affiliated First Hospital of China Medical University, No. 155, Nanjing North Street, Heping District, 110001, Shenyang, Liaoning province, China. .,2nd Ward of Bone and Joint, The First Affiliated Hospital of Liaoning Medical University, No.2, Wuduan, Renmin Street, 121001, Jinzhou, China.
| | - Chen Li
- Biobank, The First Affiliated Hospital of Liaoning Medical University, No.2, Wuduan, Renmin Street, 121001, Jinzhou, China.
| | - Hao Wang
- 2nd Ward of Bone and Joint, The First Affiliated Hospital of Liaoning Medical University, No.2, Wuduan, Renmin Street, 121001, Jinzhou, China.
| | - Guangyu Fan
- Department of Orthopedics, The Affiliated First Hospital of China Medical University, No. 155, Nanjing North Street, Heping District, 110001, Shenyang, Liaoning province, China.
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Huang C, Dai J, Zhang XA. Environmental physical cues determine the lineage specification of mesenchymal stem cells. Biochim Biophys Acta Gen Subj 2015; 1850:1261-6. [PMID: 25727396 DOI: 10.1016/j.bbagen.2015.02.011] [Citation(s) in RCA: 67] [Impact Index Per Article: 6.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/12/2014] [Revised: 02/05/2015] [Accepted: 02/20/2015] [Indexed: 12/13/2022]
Abstract
BACKGROUND Physical cues of cellular environment affect cell fate and differentiation. For example, an environment with high stiffness drives mesenchymal stem cells (MSCs) to undergo osteogenic differentiation, while low stiffness leads to lipogenic differentiation. Such effects could be independent of chemical/biochemical inducers. SCOPE OF REVIEW Stiffness and/or topography of cellular environment can control MSC differentiation and fate determination. In addition, physical factors such as tension, which resulted from profound cytoskeleton reorganization during MSC differentiation, affect the gene expression essential for the differentiation. Although physical cues control MSC lineage specification probably by reorganizing and tuning cytoskeleton, the full mechanism is largely unclear. It also remains elusive how physical signals are sensed by cells and transformed into biochemical and biological signals. More importantly, it becomes pivotal to define explicitly the physical cue(s) essential for cell differentiation and fate decision. With a focus on MSC, we present herein current understanding of the interplay between i) physical cue and factors and ii) MSC differentiation and fate determination. MAJOR CONCLUSIONS Biophysical cues can initiate or strengthen the biochemical signaling for MSC fate determination and differentiation. Physical properties of cellular environment direct the structural adaptation and functional coupling of the cells to their environment. GENERAL SIGNIFICANCE These observations not only open a simple avenue to engineer cell fate in vitro, but also start to reveal the physical elements that regulate and determine cell fate.
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Affiliation(s)
- Chao Huang
- Stephenson Cancer Center and Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA
| | - Jingxing Dai
- Stephenson Cancer Center and Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; Department of Anatomy, Southern Medical University, Guangzhou, China
| | - Xin A Zhang
- Stephenson Cancer Center and Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
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Asatrian G, Pham D, Hardy WR, James AW, Peault B. Stem cell technology for bone regeneration: current status and potential applications. STEM CELLS AND CLONING-ADVANCES AND APPLICATIONS 2015; 8:39-48. [PMID: 25709479 PMCID: PMC4334288 DOI: 10.2147/sccaa.s48423] [Citation(s) in RCA: 39] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
Continued improvements in the understanding and application of mesenchymal stem cells (MSC) have revolutionized tissue engineering. This is particularly true within the field of skeletal regenerative medicine. However, much remains unknown regarding the native origins of MSC, the relative advantages of different MSC populations for bone regeneration, and even the biologic safety of such unpurified, grossly characterized cells. This review will first summarize the initial discovery of MSC, as well as the current and future applications of MSC in bone tissue engineering. Next, the relative advantages and disadvantages of MSC isolated from distinct tissue origins are debated, including the MSC from adipose, bone marrow, and dental pulp, among others. The perivascular origin of MSC is next discussed. Finally, we briefly comment on pluripotent stem cell populations and their possible application in bone tissue engineering. While continually expanding, the field of MSC-based bone tissue engineering and regeneration shows potential to become a clinical reality in the not-so-distant future.
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Affiliation(s)
- Greg Asatrian
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, Los Angeles, CA, USA
| | - Dalton Pham
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, Los Angeles, CA, USA ; Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, Los Angeles, CA, USA
| | - Winters R Hardy
- UCLA/Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Los Angeles, CA, USA
| | - Aaron W James
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, Los Angeles, CA, USA ; Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, Los Angeles, CA, USA ; UCLA/Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Los Angeles, CA, USA
| | - Bruno Peault
- UCLA/Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Los Angeles, CA, USA ; Medical Research Council Centre for Regenerative Medicine, Edinburgh, Scotland, UK
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Schroeder J, Kueper J, Leon K, Liebergall M. Stem cells for spine surgery. World J Stem Cells 2015; 7:186-194. [PMID: 25621119 PMCID: PMC4300930 DOI: 10.4252/wjsc.v7.i1.186] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/28/2014] [Revised: 10/08/2014] [Accepted: 10/29/2014] [Indexed: 02/06/2023] Open
Abstract
In the past few years, stem cells have become the focus of research by regenerative medicine professionals and tissue engineers. Embryonic stem cells, although capable of differentiating into cell lineages of all three germ layers, are limited in their utilization due to ethical issues. In contrast, the autologous harvest and subsequent transplantation of adult stem cells from bone marrow, adipose tissue or blood have been experimentally utilized in the treatment of a wide variety of diseases ranging from myocardial infarction to Alzheimer’s disease. The physiologic consequences of stem cell transplantation and its impact on functional recovery have been studied in countless animal models and select clinical trials. Unfortunately, the bench to bedside translation of this research has been slow. Nonetheless, stem cell therapy has received the attention of spinal surgeons due to its potential benefits in the treatment of neural damage, muscle trauma, disk degeneration and its potential contribution to bone fusion.
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Lee SS, Hsu EL, Mendoza M, Ghodasra J, Nickoli MS, Ashtekar A, Polavarapu M, Babu J, Riaz RM, Nicolas JD, Nelson D, Hashmi SZ, Kaltz SR, Earhart JS, Merk BR, McKee JS, Bairstow SF, Shah RN, Hsu WK, Stupp SI. Gel scaffolds of BMP-2-binding peptide amphiphile nanofibers for spinal arthrodesis. Adv Healthc Mater 2015; 4:131-141. [PMID: 24753455 PMCID: PMC4206675 DOI: 10.1002/adhm.201400129] [Citation(s) in RCA: 141] [Impact Index Per Article: 14.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/28/2014] [Revised: 03/17/2014] [Indexed: 11/12/2022]
Abstract
Peptide amphiphile (PA) nanofibers formed by self-assembly can be customized for specific applications in regenerative medicine through the use of molecules that display bioactive signals on their surfaces. Here, the use of PA nanofibers with binding affinity for the bone promoting growth factor BMP-2 to create a gel scaffold for osteogenesis is reported. With the objective of reducing the amount of BMP-2 used clinically for successful arthrodesis in the spine, amounts of growth factor incorporated in the scaffolds that are 10 to 100 times lower than that those used clinically in collagen scaffolds are used. The efficacy of the bioactive PA system to promote BMP-2-induced osteogenesis in vivo is investigated in a rat posterolateral lumbar intertransverse spinal fusion model. PA nanofiber gels displaying BMP-2-binding segments exhibit superior spinal fusion rates relative to controls, effectively decreasing the required therapeutic dose of BMP-2 by 10-fold. Interestingly, a 42% fusion rate is observed for gels containing the bioactive nanofibers without the use of exogenous BMP-2, suggesting the ability of the nanofiber to recruit endogenous growth factor. Results obtained here demonstrate that bioactive biomaterials with capacity to bind specific growth factors by design are great targets for regenerative medicine.
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Affiliation(s)
- Sungsoo S Lee
- Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208, USA
| | - Erin L Hsu
- Department of Orthopaedic Surgery, Institute for BioNanotechnology in Medicine, Northwestern University, Chicago, IL 60611, USA
| | - Marco Mendoza
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Jason Ghodasra
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Michael S Nickoli
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Amruta Ashtekar
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Mahesh Polavarapu
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Jacob Babu
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Rehan M Riaz
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Joseph D Nicolas
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - David Nelson
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Sohaib Z Hashmi
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Start R Kaltz
- Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208, USA
| | - Jeffrey S Earhart
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Bradley R Merk
- Department of Orthopaedic Surgery, Northwestern University, Chicago, IL 60611, USA
| | - Jeff S McKee
- Baxter international Inc., Deerfield, IL 60016, USA
| | | | - Ramille N Shah
- Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208, USA
| | - Wellington K Hsu
- Department of Orthopaedic Surgery, Department of Neurological Surgery, Institute for BioNanotechnology in Medicine, Northwestern University, Chicago, IL 60611, USA
| | - Samuel I Stupp
- Department of Materials Science and Engineering, Department of Chemistry, Northwestern University, Evanston, IL 60208, USA
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Skovrlj B, Guzman JZ, Al Maaieh M, Cho SK, Iatridis JC, Qureshi SA. Cellular bone matrices: viable stem cell-containing bone graft substitutes. Spine J 2014; 14:2763-72. [PMID: 24929059 PMCID: PMC4402977 DOI: 10.1016/j.spinee.2014.05.024] [Citation(s) in RCA: 54] [Impact Index Per Article: 4.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/30/2013] [Revised: 04/03/2014] [Accepted: 05/20/2014] [Indexed: 02/03/2023]
Abstract
BACKGROUND CONTEXT Advances in the field of stem cell technology have stimulated the development and increased use of allogenic bone grafts containing live mesenchymal stem cells (MSCs), also known as cellular bone matrices (CBMs). It is estimated that CBMs comprise greater than 17% of all bone grafts and bone graft substitutes used. PURPOSE To critically evaluate CBMs, specifically their technical specifications, existing published data supporting their use, US Food and Drug Administration (FDA) regulation, cost, potential pitfalls, and other aspects pertaining to their use. STUDY DESIGN Areview of literature. METHODS A series of Ovid, Medline, and Pubmed-National Library of Medicine/National Institutes of Health (www.ncbi.nlm.nih.gov) searches were performed. Only articles in English journals or published with English language translations were included. Level of evidence of the selected articles was assessed. Specific technical information on each CBM was obtained by direct communication from the companies marketing the individual products. RESULTS Five different CBMs are currently available for use in spinal fusion surgery. There is a wide variation between the products with regard to the average donor age at harvest, total cellular concentration, percentage of MSCs, shelf life, and cell viability after defrosting. Three retrospective studies evaluating CBMs and fusion have shown fusion rates ranging from 90.2% to 92.3%, and multiple industry-sponsored trials are underway. No independent studies evaluating spinal fusion rates with the use of CBMs exist. All the commercially available CBMs claim to meet the FDA criteria under Section 361, 21 CFR Part 1271, and are not undergoing FDA premarket review. The CBMs claim to provide viable MSCs and are offered at a premium cost. Numerous challenges exist in regard to MSCs' survival, function, osteoblastic potential, and cytokine production once implanted into the intended host. CONCLUSIONS Cellular bone matrices may be a promising bone augmentation technology in spinal fusion surgery. Although CBMs appear to be safe for use as bone graft substitutes, their efficacy in spinal fusion surgery remains highly inconclusive. Large, nonindustry sponsored studies evaluating the efficacy of CBMs are required. Without results from such studies, surgeons must be made aware of the potential pitfalls of CBMs in spinal fusion surgery. With the currently available data, there is insufficient evidence to support the use of CBMs as bone graft substitutes in spinal fusion surgery.
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Affiliation(s)
- Branko Skovrlj
- Department of Neurosurgery, Icahn School of Medicine at Mount Sinai, 1 Gustave L Levy Place, Box 1136, New York, NY 10029, USA
| | - Javier Z. Guzman
- Department of Orthopaedics, Icahn School of Medicine at Mount Sinai, 5 East 98th St, 9th Floor, Box 1188, New York, NY 10029, USA
| | - Motasem Al Maaieh
- Department of Orthopaedics, Icahn School of Medicine at Mount Sinai, 5 East 98th St, 9th Floor, Box 1188, New York, NY 10029, USA
| | - Samuel K. Cho
- Department of Orthopaedics, Icahn School of Medicine at Mount Sinai, 5 East 98th St, 9th Floor, Box 1188, New York, NY 10029, USA
| | - James C. Iatridis
- Department of Orthopaedics, Icahn School of Medicine at Mount Sinai, 5 East 98th St, 9th Floor, Box 1188, New York, NY 10029, USA
| | - Sheeraz A. Qureshi
- Department of Orthopaedics, Icahn School of Medicine at Mount Sinai, 5 East 98th St, 9th Floor, Box 1188, New York, NY 10029, USA,Corresponding author. Department of Orthopaedic Surgery, Ichan School of Medicine at Mount Sinai, 5 E. 98th St, Box 1188, New York, NY 10029, USA. Tel.: (212) 241-3909; fax: (212) 534-6202.
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Ahn J, Park S, Cha BH, Kim JH, Park H, Joung YK, Han I, Lee SH. Delivery of growth factor-associated genes to mesenchymal stem cells for cartilage and bone tissue regeneration. BIOMATERIALS AND BIOMECHANICS IN BIOENGINEERING 2014. [DOI: 10.12989/bme.2014.1.3.151] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
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Chung CG, James AW, Asatrian G, Chang L, Nguyen A, Le K, Bayani G, Lee R, Stoker D, Zhang X, Ting K, Péault B, Soo C. Human perivascular stem cell-based bone graft substitute induces rat spinal fusion. Stem Cells Transl Med 2014; 3:1231-41. [PMID: 25154782 DOI: 10.5966/sctm.2014-0027] [Citation(s) in RCA: 47] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022] Open
Abstract
Adipose tissue is an attractive source of mesenchymal stem cells (MSCs) because of its abundance and accessibility. We have previously defined a population of native MSCs termed perivascular stem cells (PSCs), purified from diverse human tissues, including adipose tissue. Human PSCs (hPSCs) are a bipartite cell population composed of pericytes (CD146+CD34-CD45-) and adventitial cells (CD146-CD34+CD45-), isolated by fluorescence-activated cell sorting and with properties identical to those of culture identified MSCs. Our previous studies showed that hPSCs exhibit improved bone formation compared with a sample-matched unpurified population (termed stromal vascular fraction); however, it is not known whether hPSCs would be efficacious in a spinal fusion model. To investigate, we evaluated the osteogenic potential of freshly sorted hPSCs without culture expansion and differentiation in a rat model of posterolateral lumbar spinal fusion. We compared increasing dosages of implanted hPSCs to assess for dose-dependent efficacy. All hPSC treatment groups induced successful spinal fusion, assessed by manual palpation and microcomputed tomography. Computerized biomechanical simulation (finite element analysis) further demonstrated bone fusion with hPSC treatment. Histological analyses showed robust endochondral ossification in hPSC-treated samples. Finally, we confirmed that implanted hPSCs indeed differentiated into osteoblasts and osteocytes; however, the majority of the new bone formation was of host origin. These results suggest that implanted hPSCs positively regulate bone formation via direct and paracrine mechanisms. In summary, hPSCs are a readily available MSC population that effectively forms bone without requirements for culture or predifferentiation. Thus, hPSC-based products show promise for future efforts in clinical bone regeneration and repair.
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Affiliation(s)
- Choon G Chung
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Aaron W James
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Greg Asatrian
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Le Chang
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Alan Nguyen
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Khoi Le
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Georgina Bayani
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Robert Lee
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - David Stoker
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Xinli Zhang
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Kang Ting
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Bruno Péault
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
| | - Chia Soo
- Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, Department of Pathology and Laboratory Medicine, UCLA Operation Mend, and Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles, Los Angeles, California, USA; Marina Plastic Surgery Associates, Marina del Rey, California, USA; Center for Cardiovascular Science and MRC Center for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom
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