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Liu X, Chen R, Cui G, Feng R, Liu K. Exosomes derived from platelet-rich plasma present a novel potential in repairing knee articular cartilage defect combined with cyclic peptide-modified β-TCP scaffold. J Orthop Surg Res 2024; 19:718. [PMID: 39497084 PMCID: PMC11533314 DOI: 10.1186/s13018-024-05202-z] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/17/2024] [Accepted: 10/23/2024] [Indexed: 11/06/2024] Open
Abstract
BACKGROUND The aim of this study was to investigate the therapeutic effects and mechanisms of PRP-exos combined with cyclic peptide-modified β-TCP scaffold in the treatment of rabbit knee cartilage defect. METHODS PRP-exos were extracted and characterized by TEM, NTA and WB. The therapeutic effects were evaluated by ICRS score, HE staining, Immunohistochemistry, qRT-PCR and ELISA. The repair mechanism of PRP-exos was estimated and predicted by miRNA sequencing analysis and protein-protein interaction network analysis. RESULTS The results showed that PRP-exos had a reasonable size distribution and exhibited typical exosome morphology. The combination of PRP-exos and cyclic peptide-modified β-TCP scaffold improved ICRS score and the expression level of COL-2, RUNX2, and SOX9. Moreover, this combination therapy reduced the level of MMP-3, TNF-α, IL-1β, and IL-6, while increasing the level of TIMP-1. In PRP-exos miRNA sequencing analysis, the total number of known miRNAs aligned across all samples was 252, and a total of 91 differentially expressed miRNAs were detected. The results of KEGG enrichment analysis and the protein-protein interaction network analysis indicated that the PI3K/AKT signaling pathway could impact the function of chondrocytes by regulating key transcription factors to repair cartilage defect. CONCLUSION PRP-exos combined with cyclic peptide-modified β-TCP scaffold effectively promoted cartilage repair and improved chondrocyte function in rabbit knee cartilage defect. Based on the analysis and prediction of PRP-exos miRNAs sequencing, PI3K/AKT signaling pathway may contribute to the therapeutic effect. These findings provide experimental evidence for the application of PRP-exos in the treatment of cartilage defect.
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Affiliation(s)
- Xuchang Liu
- Department of Orthopedic Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwuweiqi Road, Jinan, 250021, Shandong, China
- Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), 28789 East Jingshi Road, Jinan, 250103, Shandong, China
- Key Laboratory for Drug Screening Technology of Shandong Academy of Sciences, 28789 East Jingshi Road, Jinan, 250103, Shandong, China
| | - Rudong Chen
- Department of Orthopedic Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwuweiqi Road, Jinan, 250021, Shandong, China
| | - Guanzheng Cui
- Department of Orthopedic Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwuweiqi Road, Jinan, 250021, Shandong, China
| | - Rongjie Feng
- Department of Orthopedic Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwuweiqi Road, Jinan, 250021, Shandong, China.
| | - Kechun Liu
- Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), 28789 East Jingshi Road, Jinan, 250103, Shandong, China.
- Key Laboratory for Drug Screening Technology of Shandong Academy of Sciences, 28789 East Jingshi Road, Jinan, 250103, Shandong, China.
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Campbell K, Naire S, Kuiper JH. A mathematical model of signalling molecule-mediated processes during regeneration of osteochondral defects after chondrocyte implantation. J Theor Biol 2024; 592:111874. [PMID: 38908475 DOI: 10.1016/j.jtbi.2024.111874] [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: 10/12/2023] [Revised: 04/12/2024] [Accepted: 06/06/2024] [Indexed: 06/24/2024]
Abstract
Treating bone-cartilage defects is a fundamental clinical problem. The ability of damaged cartilage to self-repair is limited due to its avascularity. Left untreated, these defects can lead to osteoarthritis. Details of osteochondral defect repair are elusive, but animal models indicate healing occurs via an endochondral ossification-like process, similar to that in the growth plate. In the growth plate, the signalling molecules parathyroid hormone-related protein (PTHrP) and Indian Hedgehog (Ihh) form a feedback loop regulating chondrocyte hypertrophy, with Ihh inducing and PTHrP suppressing hypertrophy. To better understand this repair process and to explore the regulatory role of signalling molecules on the regeneration process, we formulate a reaction-diffusion mathematical model of osteochondral defect regeneration after chondrocyte implantation. The drivers of healing are assumed to be chondrocytes and osteoblasts, and their interaction via signalling molecules. We model cell proliferation, migration and chondrocyte hypertrophy, and matrix production and conversion, spatially and temporally. We further model nutrient and signalling molecule diffusion and their interaction with the cells. We consider the PTHrP-Ihh feedback loop as the backbone mechanisms but the model is flexible to incorporate extra signalling mechanisms if needed. Our mathematical model is able to represent repair of osteochondral defects, starting with cartilage formation throughout the defect. This is followed by chondrocyte hypertrophy, matrix calcification and bone formation deep inside the defect, while cartilage at the surface is maintained and eventually separated from the deeper bone by a thin layer of calcified cartilage. The complete process requires around 48 months. A key highlight of the model demonstrates that the PTHrP-Ihh loop alone is insufficient and an extra mechanism is required to initiate chondrocyte hypertrophy, represented by a critical cartilage density. A parameter sensitivity study reveals that the timing of the repair process crucially depends on parameters, such as the critical cartilage density, and those describing the actions of PTHrP to suppress hypertrophy, such as its diffusion coefficient, threshold concentration and degradation rate.
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Affiliation(s)
- Kelly Campbell
- School of Computing and Mathematics, Keele University, Keele, ST5 5BG, UK
| | - Shailesh Naire
- School of Computing and Mathematics, Keele University, Keele, ST5 5BG, UK
| | - Jan Herman Kuiper
- School of Pharmacy and Bioengineering, Keele University, Keele, ST5 5BG, UK; Robert Jones and Agnes Hunt Orthopaedic & District Hospital NHS Trust, Oswestry, SY10 7AG, UK.
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Yu L, Cavelier S, Hannon B, Wei M. Recent development in multizonal scaffolds for osteochondral regeneration. Bioact Mater 2023; 25:122-159. [PMID: 36817819 PMCID: PMC9931622 DOI: 10.1016/j.bioactmat.2023.01.012] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2022] [Revised: 12/30/2022] [Accepted: 01/14/2023] [Indexed: 02/05/2023] Open
Abstract
Osteochondral (OC) repair is an extremely challenging topic due to the complex biphasic structure and poor intrinsic regenerative capability of natural osteochondral tissue. In contrast to the current surgical approaches which yield only short-term relief of symptoms, tissue engineering strategy has been shown more promising outcomes in treating OC defects since its emergence in the 1990s. In particular, the use of multizonal scaffolds (MZSs) that mimic the gradient transitions, from cartilage surface to the subchondral bone with either continuous or discontinuous compositions, structures, and properties of natural OC tissue, has been gaining momentum in recent years. Scrutinizing the latest developments in the field, this review offers a comprehensive summary of recent advances, current hurdles, and future perspectives of OC repair, particularly the use of MZSs including bilayered, trilayered, multilayered, and gradient scaffolds, by bringing together onerous demands of architecture designs, material selections, manufacturing techniques as well as the choices of growth factors and cells, each of which possesses its unique challenges and opportunities.
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Affiliation(s)
- Le Yu
- Department of Chemical and Biomolecular Engineering, Ohio University, Athens, OH, 45701, USA
| | - Sacha Cavelier
- Department of Chemical and Biomolecular Engineering, Ohio University, Athens, OH, 45701, USA
| | - Brett Hannon
- Biomedical Engineering Program, Ohio University, Athens, OH, 45701, USA
| | - Mei Wei
- Biomedical Engineering Program, Ohio University, Athens, OH, 45701, USA
- Department of Mechanical Engineering, Ohio University, Athens, OH, 45701, USA
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Zhou Z, Cui J, Wu S, Geng Z, Su J. Silk fibroin-based biomaterials for cartilage/osteochondral repair. Am J Cancer Res 2022; 12:5103-5124. [PMID: 35836802 PMCID: PMC9274741 DOI: 10.7150/thno.74548] [Citation(s) in RCA: 88] [Impact Index Per Article: 29.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/29/2022] [Accepted: 06/18/2022] [Indexed: 02/07/2023] Open
Abstract
Osteoarthritis (OA) is a common joint disease with a high disability rate. In addition, OA not only causes great physiological and psychological harm to patients, but also puts great pressure on the social healthcare system. Pathologically, the disintegration of cartilage and the lesions of subchondral bone are related to OA. Currently, tissue engineering, which is expected to overcome the defects of existing treatment methods, had a lot of research in the field of cartilage/osteochondral repair. Silk fibroin (SF), as a natural macromolecular material with good biocompatibility, unique mechanical properties, excellent processability and degradability, holds great potential in the field of tissue engineering. Nowadays, SF had been prepared into various materials to adapt to the demands of cartilage/osteochondral repair. SF-based biomaterials can also be functionally modified to enhance repair performance further. In this review, the preparation methods, types, structures, mechanical properties, and functional modifications of SF-based biomaterials used for cartilage/osteochondral repair are summarized and discussed. We hope that this review will provide a reference for the design and development of SF-based biomaterials in cartilage/osteochondral repair field.
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Affiliation(s)
- Ziyang Zhou
- Institute of Translational Medicine, Shanghai University, Shanghai, 200444, China,Musculoskeletal Organoid Research Center, Shanghai University, Shanghai, 200444, China,School of Medicine, Shanghai University, Shanghai 200444, China,School of Life Sciences, Shanghai University, Shanghai 200444, China
| | - Jin Cui
- Institute of Translational Medicine, Shanghai University, Shanghai, 200444, China,Musculoskeletal Organoid Research Center, Shanghai University, Shanghai, 200444, China,Department of Orthopedics Trauma, Changhai Hospital, Second Military Medical University, Shanghai, 200433, China
| | - Shunli Wu
- Institute of Translational Medicine, Shanghai University, Shanghai, 200444, China,Musculoskeletal Organoid Research Center, Shanghai University, Shanghai, 200444, China,School of Medicine, Shanghai University, Shanghai 200444, China,School of Environmental and Chemical Engineering, Shanghai University, Shanghai 200444, China
| | - Zhen Geng
- Institute of Translational Medicine, Shanghai University, Shanghai, 200444, China,Musculoskeletal Organoid Research Center, Shanghai University, Shanghai, 200444, China,✉ Corresponding authors: Zhen Geng, ; Jiacan Su,
| | - Jiacan Su
- Institute of Translational Medicine, Shanghai University, Shanghai, 200444, China,Musculoskeletal Organoid Research Center, Shanghai University, Shanghai, 200444, China,✉ Corresponding authors: Zhen Geng, ; Jiacan Su,
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Small Ruminant Models for Articular Cartilage Regeneration by Scaffold-Based Tissue Engineering. Stem Cells Int 2021; 2021:5590479. [PMID: 34912460 PMCID: PMC8668357 DOI: 10.1155/2021/5590479] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/03/2021] [Revised: 10/10/2021] [Accepted: 11/08/2021] [Indexed: 11/29/2022] Open
Abstract
Animal models play an important role in preclinical studies, especially in tissue engineering scaffolds for cartilage repair, which require large animal models to verify the safety and effectiveness for clinical use. The small ruminant models are most widely used in this field than other large animals because they are cost-effective, easy to raise, not to mention the fact that the aforementioned animal presents similar anatomical features to that of humans. This review discusses the experimental study of tissue engineering scaffolds for knee articular cartilage regeneration in small ruminant models. Firstly, the selection of these scaffold materials and the preparation process in vitro that have been already used in vivo are briefly reviewed. Moreover, the major factors influencing the rational design and the implementation as well as advantages and limitations of small ruminants are also demonstrated. As regards methodology, this paper applies principles and methods followed by most researchers in the process of experimental design and operation of this kind. By summarizing and comparing different therapeutic concepts, this paper offers suggestions aiming to increase the effectiveness of preclinical research using small ruminant models and improve the process of developing corresponding therapies.
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Wu S, Kai Z, Wang D, Tao L, Zhang P, Wang D, Liu D, Sun S, Zhong J. Allogenic chondrocyte/osteoblast-loaded β-tricalcium phosphate bioceramic scaffolds for articular cartilage defect treatment. ARTIFICIAL CELLS, NANOMEDICINE, AND BIOTECHNOLOGY 2019; 47:1570-1576. [PMID: 31007085 DOI: 10.1080/21691401.2019.1604534] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/20/2018] [Revised: 03/29/2019] [Accepted: 04/01/2019] [Indexed: 10/27/2022]
Abstract
The medical community has expressed significant interest in the treatment of cartilage defect. Successful repair of articular cartilage defects remains a challenge in clinics. Due to the huge advantages of 3D micro/nanomaterials, 3D artificial micro/nano scaffolds have been widely developed and explored in the tissue repair of articular joints. In this study, chondrocyte/osteoblast-loaded β-tricalcium phosphate (β-TCP) bioceramic scaffold and chondrocyte-loaded β-TCP bioceramic scaffold were prepared by micromass stem cell culture and bioreactor-based cells-loaded scaffold culture for articular cartilage defect treatment. The results demonstrate chondrocyte and osteoblast can be successfully induced from allogeneic bone marrow stromal cells using micromass stem cell culture. Further, chondrocyte-loaded β-TCP scaffold and osteoblast-loaded β-TCP scaffold can be successfully prepared by bioreactor-based cells-loaded scaffold culture. Finally, the scaffolds are applied for Beagle articular cartilage defect treatment. The relative cartilage regeneration abilities on Beagle femoral trochleae were as follows: Chondrocyte/osteoblast-loaded β-TCP bioceramic scaffold group > chondrocyte-loaded β-TCP bioceramic scaffold group > β-TCP bioceramic scaffold. Therefore, micromass stem cell culture and bioreactor-based cells-loaded scaffold culture can be applied to prepare chondrocyte/osteoblast-loaded β-TCP bioceramic scaffold for articular cartilage defect treatment. It suggests allogenic chondrocyte/osteoblast-loaded β-TCP bioceramic scaffold could be potentially used in the treatment of patients with cartilage defects.
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Affiliation(s)
- Shuai Wu
- a Shandong Provincial Hospital , Shandong University , Jinan , People's Republic of China
| | - Zhiguo Kai
- b The No.4 hospital of Jinan , Jinan , People's Republic of China
| | - Dong Wang
- a Shandong Provincial Hospital , Shandong University , Jinan , People's Republic of China
| | - Lina Tao
- c Integrated Scientific Research Base on Comprehensive Utilization Technology for By-Products of Aquatic Product Processing , Ministry of Agriculture and Rural Affairs of the People's Republic of China , Shanghai , People's Republic of China
- d Laboratory of Quality & Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai) , Ministry of Agriculture and Rural Affairs of the People's Republic of China , Shanghai , People's Republic of China
- e Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, College of Food Science & Technology , Shanghai Ocean University , Shanghai , People's Republic of China
| | - Peng Zhang
- a Shandong Provincial Hospital , Shandong University , Jinan , People's Republic of China
| | - Dawei Wang
- a Shandong Provincial Hospital , Shandong University , Jinan , People's Republic of China
| | - Dongxing Liu
- a Shandong Provincial Hospital , Shandong University , Jinan , People's Republic of China
| | - Shui Sun
- a Shandong Provincial Hospital , Shandong University , Jinan , People's Republic of China
| | - Jian Zhong
- c Integrated Scientific Research Base on Comprehensive Utilization Technology for By-Products of Aquatic Product Processing , Ministry of Agriculture and Rural Affairs of the People's Republic of China , Shanghai , People's Republic of China
- d Laboratory of Quality & Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai) , Ministry of Agriculture and Rural Affairs of the People's Republic of China , Shanghai , People's Republic of China
- e Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, College of Food Science & Technology , Shanghai Ocean University , Shanghai , People's Republic of China
- f State Key Laboratory of Molecular Engineering of Polymers , Fudan University , Shanghai , People's Republic of China
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Tothova C, Novotny J, Nagy O, Hornakova P, Zert Z, Varga M, Medvecky L, Vdoviakova K, Danko J, Petrovova E. Changes in the Acute-Phase Protein Concentrations and Activities of Some Enzymes in Pigs Following the Repair of Experimentally Induced Articular Cartilage Defects Using Two Types of Biocement Powder. Animals (Basel) 2019; 9:ani9110931. [PMID: 31703315 PMCID: PMC6912659 DOI: 10.3390/ani9110931] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/22/2019] [Accepted: 11/05/2019] [Indexed: 12/19/2022] Open
Abstract
Simple Summary Articular cartilage reconstruction is aimed at the restoration of damaged joint cartilage. The use of bone cement is one type of method applicable for this reconstruction. The potential use of repair techniques must be evaluated by pre-clinical and clinical studies in animal models, including the assessment of some biochemical parameters. Acute-phase proteins are a class of proteins whose concentrations increase in response to injury or inflammation. They may serve as useful biomarkers for the evaluation of post-operative complications, as well as to reflect the extent of surgical trauma. Information regarding their usefulness after cartilage reconstruction are still limited. Similarly, little is known about the response of the organism to various reconstruction techniques and various biomaterials used for the repair of defects. This paper provides important information about the changes in the concentrations of acute-phase proteins and the activity of serum enzymes in pigs within the first 30 days following the repair of experimentally induced articular cartilage defects using tetracalcium phosphate/nanomonetite cement powder (C cement) and cement powder containing aminoacids (CAK cement). Marked inflammatory responses with increased acute-phase proteins concentrations were observed following the reconstruction of articular cartilage defects using both types of biocement powder. The results suggest, that the tetracalcium phosphate/nanomonetite cement powder without amino acids would be more suitable for possible cartilage repair in the human population. Abstract The objective of the study was to assess the usefulness of acute-phase proteins (APPs) and serum enzymes in the evaluation of post-operative state after cartilage reconstruction in an animal model (Sus scrofa domesticus). Fifteen clinically healthy female pigs were evaluated during the first 30 days after the repair of experimentally induced articular cartilage defects using two types of biocement powders. Animals were divided into groups according to the type of biocement powder used: CAK—with amino acids (n = 6), C—without amino acids (n = 6) and the control group (Ctr) was without biocement (n = 3). The concentrations of selected APPs—serum amyloid A (SAA), haptoglobin (Hp) and C-reactive protein (CRP), and the activities of some serum enzymes—creatine kinase (CK), alkaline phosphatase (AP), and lactate dehydrogenase (LD) were measured one day before the surgery and on days 7, 14, and 30 after the surgical intervention. The most significant changes during the evaluated period were observed in the concentrations of SAA (p < 0.001) and Hp (p < 0.001), with marked increase of values 7 days after surgery. There was a numerical, but not statistically significant, difference between CAK, C and Ctr groups (p > 0.05). Marked variations were observed also in the activities of the evaluated enzymes, with the most significant changes in the activity of AP in the CAK group (p < 0.001). Presented results suggest possible usefulness of some APPs and serum enzymes in the evaluation of post-operative inflammatory state after the reconstruction of articular cartilage defects.
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Affiliation(s)
- Csilla Tothova
- Clinic of Ruminants, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic;
- Correspondence: ; Tel.: +421-915-493-082
| | - Jaroslav Novotny
- Clinic of Swine, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic;
| | - Oskar Nagy
- Clinic of Ruminants, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic;
| | - Petra Hornakova
- Clinic of Horses, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic; (P.H.); (Z.Z.)
| | - Zdenek Zert
- Clinic of Horses, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic; (P.H.); (Z.Z.)
| | - Maros Varga
- Sport-arthro Centre, Privat Hospital Saca-Kosice, 040 15 Saca-Kosice, Slovak Republic;
| | - Lubomir Medvecky
- Institute of Materials Research SAS in Kosice, Watsonova 47, 040 01 Kosice, Slovak Republic;
| | - Katarina Vdoviakova
- Institute of Anatomy, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic; (K.V.); (J.D.); (E.P.)
| | - Jan Danko
- Institute of Anatomy, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic; (K.V.); (J.D.); (E.P.)
| | - Eva Petrovova
- Institute of Anatomy, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovak Republic; (K.V.); (J.D.); (E.P.)
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Lepage SIM, Robson N, Gilmore H, Davis O, Hooper A, St John S, Kamesan V, Gelis P, Carvajal D, Hurtig M, Koch TG. Beyond Cartilage Repair: The Role of the Osteochondral Unit in Joint Health and Disease. TISSUE ENGINEERING PART B-REVIEWS 2019; 25:114-125. [PMID: 30638141 PMCID: PMC6486663 DOI: 10.1089/ten.teb.2018.0122] [Citation(s) in RCA: 51] [Impact Index Per Article: 8.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 12/28/2022]
Abstract
Once believed to be limited to articular cartilage, osteoarthritis is now considered to be an organ disease of the “whole joint.” Damage to the articular surface can lead to, be caused by, or occur in parallel with, damage to other tissues in the joint. The relationship between cartilage and the underlying subchondral bone has particular importance when assessing joint health and determining treatment strategies. The articular cartilage is anchored to the subchondral bone through an interface of calcified cartilage, which as a whole makes up the osteochondral unit. This unit functions primarily by transferring load-bearing weight over the joint to allow for normal joint articulation and movement. Unfortunately, irreversible damage and degeneration of the osteochondral unit can severely limit joint function. Our understanding of joint pain, the primary complaint of patients, is poorly understood and past efforts toward structural cartilage restoration have often not been associated with a reduction in pain. Continued research focusing on the contribution of subchondral bone and restoration of the entire osteochondral unit are therefore needed, with the hope that this will lead to curative, and not merely palliative, treatment options. The purpose of this narrative review is to investigate the role of the osteochondral unit in joint health and disease. Topics of discussion include the crosstalk between cartilage and bone, the efficacy of diagnostic procedures, the origins of joint pain, current and emerging treatment paradigms, and suitable preclinical animal models for safety and efficacy assessment of novel osteochondral therapies. The goal of the review is to facilitate an appreciation of the important role played by the subchondral bone in joint pain and why the osteochondral unit as a whole should be considered in many cases of joint restoration strategies.
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Affiliation(s)
- Sarah I M Lepage
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Naomi Robson
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Hillary Gilmore
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Ola Davis
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Allyssa Hooper
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Stephanie St John
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Vashine Kamesan
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Paul Gelis
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Diana Carvajal
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
| | - Mark Hurtig
- 2 Department of Clinical Studies, University of Guelph, Guelph, Canada
| | - Thomas G Koch
- 1 Department of Biomedical Sciences, University of Guelph, Guelph, Canada
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Xiao H, Huang W, Xiong K, Ruan S, Yuan C, Mo G, Tian R, Zhou S, She R, Ye P, Liu B, Deng J. Osteochondral repair using scaffolds with gradient pore sizes constructed with silk fibroin, chitosan, and nano-hydroxyapatite. Int J Nanomedicine 2019; 14:2011-2027. [PMID: 30962685 PMCID: PMC6435123 DOI: 10.2147/ijn.s191627] [Citation(s) in RCA: 48] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022] Open
Abstract
Background One of the main problems associated with the development of osteochondral reparative materials is that the accurate imitation of the structure of the natural osteochondral tissue and fabrication of a suitable scaffold material for osteochondral repair are difficult. The long-term outcomes of single- or bilayered scaffolds are often unsatisfactory because of the absence of a progressive osteochondral structure. Therefore, only scaffolds with gradient pore sizes are suitable for osteochondral repair to achieve better proliferation and differentiation of the stem cells into osteochondral tissues to complete the repair of defects. Methods A silk fibroin (SF) solution, chitosan (CS) solution, and nano-hydroxyapatite (nHA) suspension were mixed at the same weight fraction to obtain osteochondral scaffolds with gradient pore diameters by centrifugation, freeze-drying, and chemical cross-linking. Results The scaffolds prepared in this study are confirmed to have a progressive structure starting from the cartilage layer to bone layer, similar to that of the normal osteochondral tissues. The prepared scaffolds are cylindrical in shape and have high internal porosity. The structure consists of regular and highly interconnected pores with a progressively increasing pore distribution as well as a progressively changing pore diameter. The scaffold strongly absorbs water, and has a suitable degradation rate, sufficient space for cell growth and proliferation, and good resistance to compression. Thus, the scaffold can provide sufficient nutrients and space for cell growth, proliferation, and migration. Further, bone marrow mesenchymal stem cells seeded onto the scaffold closely attach to the scaffold and stably grow and proliferate, indicating that the scaffold has good biocompatibility with no cytotoxicity. Conclusion In brief, the physical properties and biocompatibility of our scaffolds fully comply with the requirements of scaffold materials required for osteochondral tissue engineering, and they are expected to become a new type of scaffolds with gradient pore sizes for osteochondral repair.
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Affiliation(s)
- Hongli Xiao
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
| | - Wenliang Huang
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
| | - Kun Xiong
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
| | - Shiqiang Ruan
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
| | - Cheng Yuan
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
| | - Gang Mo
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
| | - Renyuan Tian
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
| | - Sirui Zhou
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
| | - Rongfeng She
- Department of Orthopedics, Guizhou Province People's Hospital, Guiyang 550002, Guizhou Province, People's Republic of China
| | - Peng Ye
- Emergency and Trauma Ward, Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China
| | - Bin Liu
- Surgical Laboratory, Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China
| | - Jiang Deng
- Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, People's Republic of China,
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Pelagalli A, Nardelli A, Lucarelli E, Zannetti A, Brunetti A. Autocrine signals increase ovine mesenchymal stem cells migration through Aquaporin-1 and CXCR4 overexpression. J Cell Physiol 2018; 233:6241-6249. [PMID: 29345324 DOI: 10.1002/jcp.26493] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/10/2017] [Accepted: 01/17/2018] [Indexed: 12/13/2022]
Abstract
Sheep is a relevant large animal model that is frequently used to test innovative tissue engineering (TE) approaches especially for bone reconstruction. Mesenchymal stem cells (MSCs) are used in TE applications because they represent key component of adult tissue repair. Importantly, MSCs from different species show similar characteristics, which facilitated their application in translational studies using animal models. Nowadays, many researches are focusing on the use of ovine mesenchymal stem cells (oMSCs) in orthopedic preclinical settings for regenerative medicine purposes. Therefore, there is a need to amplify our knowledge on the mechanisms underlying the behaviour of these cells. Recently, several studies have shown that MSC function is largely dependent on factors that MSCs release in the environment, as well as, in conditioned medium (CM). It has been demonstrated that MSCs through autocrine and paracrine signals are able to stimulate proliferation, migration, and differentiation of different type of cells including themselves. In this study, we investigated the effects of the CM produced by oMSCs on oMSCs themselves and we explored the signal pathways involved. We observed that CM caused an enhancement of oMSC migration. Furthermore, we found that CM increased levels of two membrane proteins involved in cell migration, Aquaporin 1 (AQP1), and C-X-C chemokine receptor type 4 (CXCR4), and activated Akt and Erk intracellular signal pathways. In conclusion, taken together our results suggest the high potential of autologous CM as a promising tool to modulate behaviour of MSCs thus improving their use in therapeutically approaches.
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Affiliation(s)
- Alessandra Pelagalli
- Department of Advanced Biomedical Sciences, University of Naples "Federico II", Naples, Italy
- Institute of Biostructure and Bioimaging, National Research Council, Naples, Italy
| | - Anna Nardelli
- Institute of Biostructure and Bioimaging, National Research Council, Naples, Italy
| | - Enrico Lucarelli
- Osteoarticolar Regeneration Laboratory, Rizzoli Orthopedic Institute, Bologna, Italy
| | - Antonella Zannetti
- Institute of Biostructure and Bioimaging, National Research Council, Naples, Italy
| | - Arturo Brunetti
- Department of Advanced Biomedical Sciences, University of Naples "Federico II", Naples, Italy
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11
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Brown WE, Huey DJ, Hu JC, Athanasiou KA. Functional self-assembled neocartilage as part of a biphasic osteochondral construct. PLoS One 2018; 13:e0195261. [PMID: 29634740 PMCID: PMC5892872 DOI: 10.1371/journal.pone.0195261] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/14/2017] [Accepted: 03/19/2018] [Indexed: 11/29/2022] Open
Abstract
Bone-to-bone integration can be obtained by osteoconductive ceramics such as hydroxyapatite (HAp) and beta-tricalcium phosphate (β-TCP), but cartilage-to-cartilage integration is notoriously difficult. Many cartilage repair therapies, including microfracture and mosaicplasty, capitalize on the reparative aspects of subchondral bone due to its resident population of stem cells and vascularity. A strategy of incorporating tissue engineered neocartilage into a ceramic to form an osteochondral construct may serve as a suitable alternative to achieve cartilage graft fixation. The use of a tissue engineered osteochondral construct to repair cartilage defects may also benefit from the ceramic’s proximity to underlying bone and abundant supply of progenitor cells and nutrients. The objective of the first study was to compare HAp and β-TCP ceramics, two widely used ceramics in bone regeneration, in terms of their ability to influence neocartilage interdigitation at an engineered osteochondral interface. Additional assays quantified ceramic pore size, porosity, and compressive strength. The compressive strength of HAp was six times higher than that of β-TCP due to differences in porosity and pore size, and HAp was thus carried forward in the second study as the composition with which to engineer an osteochondral construct. Importantly, it was shown that incorporation of the HAp ceramic in conjunction with the self-assembling process resulted in functionally viable neocartilage. For example, only collagen/dry weight and ultimate tensile strength of the chondral control constructs remained significantly greater than the neocartilage cut off the osteochondral constructs. By demonstrating that the functional properties of engineered neocartilage are not negatively affected by the inclusion of an HAp ceramic in culture, neocartilage engineering strategies may be directly applied to the formation of an osteochondral construct.
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Affiliation(s)
- Wendy E Brown
- Department of Biomedical Engineering, University of California Irvine, Irvine, California, United States of America
| | - Daniel J Huey
- Department of Biomedical Engineering, University of California Davis, Davis, California, United States of America
| | - Jerry C Hu
- Department of Biomedical Engineering, University of California Irvine, Irvine, California, United States of America
| | - Kyriacos A Athanasiou
- Department of Biomedical Engineering, University of California Irvine, Irvine, California, United States of America
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12
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Bioceramics for Osteochondral Tissue Engineering and Regeneration. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2018; 1058:53-75. [DOI: 10.1007/978-3-319-76711-6_3] [Citation(s) in RCA: 24] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/04/2023]
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13
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Dias IR, Viegas CA, Carvalho PP. Large Animal Models for Osteochondral Regeneration. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2018; 1059:441-501. [PMID: 29736586 DOI: 10.1007/978-3-319-76735-2_20] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
Namely, in the last two decades, large animal models - small ruminants (sheep and goats), pigs, dogs and horses - have been used to study the physiopathology and to develop new therapeutic procedures to treat human clinical osteoarthritis. For that purpose, cartilage and/or osteochondral defects are generally performed in the stifle joint of selected large animal models at the condylar and trochlear femoral areas where spontaneous regeneration should be excluded. Experimental animal care and protection legislation and guideline documents of the US Food and Drug Administration, the American Society for Testing and Materials and the International Cartilage Repair Society should be followed, and also the specificities of the animal species used for these studies must be taken into account, such as the cartilage thickness of the selected defect localization, the defined cartilage critical size defect and the joint anatomy in view of the post-operative techniques to be performed to evaluate the chondral/osteochondral repair. In particular, in the articular cartilage regeneration and repair studies with animal models, the subchondral bone plate should always be taken into consideration. Pilot studies for chondral and osteochondral bone tissue engineering could apply short observational periods for evaluation of the cartilage regeneration up to 12 weeks post-operatively, but generally a 6- to 12-month follow-up period is used for these types of studies.
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Affiliation(s)
- Isabel R Dias
- Department of Veterinary Sciences, Agricultural and Veterinary Sciences School, University of Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal. .,3B's Research Group - Biomaterials, Biodegradables and Biomimetics, Department of Polymer Engineering, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark - Parque da Ciência e Tecnologia, Zona Industrial da Gandra, Barco - Guimarães, 4805-017, Portugal. .,Department of Veterinary Medicine, ICVS/3B's - PT Government Associate Laboratory, Braga/Guimarães, Portugal.
| | - Carlos A Viegas
- Department of Veterinary Sciences, Agricultural and Veterinary Sciences School, University of Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal.,3B's Research Group - Biomaterials, Biodegradables and Biomimetics, Department of Polymer Engineering, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark - Parque da Ciência e Tecnologia, Zona Industrial da Gandra, Barco - Guimarães, 4805-017, Portugal.,Department of Veterinary Medicine, ICVS/3B's - PT Government Associate Laboratory, Braga/Guimarães, Portugal
| | - Pedro P Carvalho
- Department of Veterinary Medicine, University School Vasco da Gama, Av. José R. Sousa Fernandes 197, Lordemão, Coimbra, 3020-210, Portugal.,CIVG - Vasco da Gama Research Center, University School Vasco da Gama, Coimbra, Portugal
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14
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Okeagu CN, Baker EA, Barreras NA, Vaupel ZM, Fortin PT, Baker KC. Review of Mechanical, Processing, and Immunologic Factors Associated With Outcomes of Fresh Osteochondral Allograft Transplantation of the Talus. Foot Ankle Int 2017; 38:808-819. [PMID: 28385038 DOI: 10.1177/1071100717697649] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/01/2023]
Abstract
UNLABELLED Osteochondral lesions of the talus (OLTs) are an increasingly implicated cause of ankle pain and instability. Several treatment methods exist with varying clinical outcomes. Due in part to successful osteochondral allografting (OCA) in other joints, such as the knee and shoulder, OCA has gained popularity as a treatment option, especially in the setting of large lesions. The clinical outcomes of talar OCA have been inconsistent relative to the positive results observed in other joints. Current literature regarding OCA failure focuses mainly on 3 factors: the effect of graft storage conditions on chondrocyte viability, graft/lesion size, and operative technique. Several preclinical studies have demonstrated the ability for bone and cartilage tissue to invoke an immune response, and a limited number of clinical studies have suggested that this response may have the potential to influence outcomes after transplantation. Further research is warranted to investigate the role of immunological mechanisms as an etiology of OCA failure. LEVEL OF EVIDENCE Level V, expert opinion.
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Affiliation(s)
- Chikezie N Okeagu
- 1 Beaumont Health System, Department of Orthopaedic Research, Royal Oak, MI, USA
| | - Erin A Baker
- 1 Beaumont Health System, Department of Orthopaedic Research, Royal Oak, MI, USA
| | - Nicholas A Barreras
- 1 Beaumont Health System, Department of Orthopaedic Research, Royal Oak, MI, USA
| | - Zachary M Vaupel
- 2 Beaumont Health System, Department of Orthopaedic Surgery, Royal Oak, MI, USA
| | - Paul T Fortin
- 2 Beaumont Health System, Department of Orthopaedic Surgery, Royal Oak, MI, USA
| | - Kevin C Baker
- 1 Beaumont Health System, Department of Orthopaedic Research, Royal Oak, MI, USA
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15
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Vas WJ, Shah M, Al Hosni R, Owen HC, Roberts SJ. Biomimetic strategies for fracture repair: Engineering the cell microenvironment for directed tissue formation. J Tissue Eng 2017; 8:2041731417704791. [PMID: 28491274 PMCID: PMC5406151 DOI: 10.1177/2041731417704791] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/31/2017] [Accepted: 03/21/2017] [Indexed: 12/20/2022] Open
Abstract
Complications resulting from impaired fracture healing have major clinical implications on fracture management strategies. Novel concepts taken from developmental biology have driven research strategies towards the elaboration of regenerative approaches that can truly harness the complex cellular events involved in tissue formation and repair. Advances in polymer technology and a better understanding of naturally derived scaffolds have given rise to novel biomaterials with an increasing ability to recapitulate native tissue environments. This coupled with advances in the understanding of stem cell biology and technology has opened new avenues for regenerative strategies with true clinical translatability. These advances have provided the impetus to develop alternative approaches to enhance the fracture repair process. We provide an update on these advances, with a focus on the development of novel biomimetic approaches for bone regeneration and their translational potential.
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Affiliation(s)
- Wollis J Vas
- Department of Materials & Tissue, Institute of Orthopaedics & Musculoskeletal Science, University College London, Stanmore, UK
| | - Mittal Shah
- Department of Materials & Tissue, Institute of Orthopaedics & Musculoskeletal Science, University College London, Stanmore, UK
| | - Rawiya Al Hosni
- Department of Materials & Tissue, Institute of Orthopaedics & Musculoskeletal Science, University College London, Stanmore, UK
| | - Helen C Owen
- Department of Natural Sciences, School of Science & Technology, Middlesex University, London, UK
| | - Scott J Roberts
- Department of Materials & Tissue, Institute of Orthopaedics & Musculoskeletal Science, University College London, Stanmore, UK
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16
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Dias RP, Teixeira MF, Costa EC, Farias AC, Azevedo DA, Aguiar TD, Pinheiro MA. Potential for in vitro mesoderm differentiation of Wharton's jelly cells from ovine umbilical cord isolated in different culture media. PESQUISA VETERINARIA BRASILEIRA 2016. [DOI: 10.1590/s0100-736x2016001300012] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
Abstract: The mammalian Wharton's jelly of umbilical cord (WJUC) is a promising source of multipotent cells, providing advantages due to ethical implications, ease of collection and the absence of teratomas in pre-clinical trials. Ovine multipotent cells have already been isolated from various tissues, however there are no reports using umbilical cords in this species. This study aimed to investigate the best medium to transport the umbilical cord, to isolate and maintain ovine WJUC cells and to compare in vitro growth and mesodermal differentiation potential. Eight ovine umbilical cords were obtained during parturition, sectioned and transported in six different media: MEM, low glucose DMEM, M199, RPMI 1640, PBS and saline. For each transportation medium, four culture media were used and the tissue was explanted in 24-well plates and cultured in MEM, low glucose DMEM, M199 and RPMI 1640, all with 10% FBS. Every experiment was conducted with low-passage (P2), investigating MTT viability during four days and adipogenic, chondrogenic and osteogenesis differentiation was induced in vitro. The most effective transport medium (p<0.1) was low glucose DMEM. There was no bacterial or fungal contamination from collection. Cells from Wharton's jelly of ovine umbilical cords collected at natural birth possess fibroblastic morphology and the capacity for in vitro differentiation into adipogenic, chondrogenic and osteogenic cell lines. MTT tests and in vitro differentiation experiments revealed that cell culture medium modulates the behavior of cells and is an important factor for proliferation and maintenance of multipotency. Low glucose DMEM was the most suitable medium for the isolation of cells from Wharton's jelly of ovine umbilical cord.
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Affiliation(s)
| | | | - Edmara C. Costa
- Universidade da Integração Internacional da Lusofonia Afro-Brasileira, Brazil
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17
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Hwang IS, Bae HK, Cheong HT. Comparison of the characteristics and multipotential and in vivo cartilage formation capabilities between porcine adipose-derived stem cells and porcine skin-derived stem cell-like cells. Am J Vet Res 2016; 76:814-21. [PMID: 26309110 DOI: 10.2460/ajvr.76.9.814] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
OBJECTIVE To compare the characteristics and multipotential and in vivo cartilage formation capabilities of porcine adipose-derived stem cells (pASCs) with those of porcine skin-derived stem cell-like cells (pSSCs). ANIMALS Three 6-month-old female pigs and four 6-week-old female athymic mice. PROCEDURES Adipose and skin tissue specimens were obtained from each pig following slaughter and digested to obtain pASCs and pSSCs. For each cell type, flow cytometry and reverse transcription PCR assays were performed to characterize the expression of cell surface and mesenchymal stem cell markers, and in vitro cell cultures were performed to determine the adipogenic, osteogenic, and chondrogenic capabilities. Each cell type was then implanted into athymic mice to determine the extent of in vivo cartilage formation after 6 weeks. RESULTS The cell surface and mesenchymal stem cell marker expression patterns, multipotential capability, and extent of in vivo cartilage formation did not differ significantly between pASCs and pSSCs. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that pSSCs may be a viable alternative to pASCs as a source of progenitor cells for tissue engineering in regenerative medicine.
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18
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Tian Y, Tao L, Zhao S, Tai D, Liu D, Liu P. Isolation and morphological characterization of ovine amniotic fluid mesenchymal stem cells. Exp Anim 2015; 65:125-34. [PMID: 26616638 PMCID: PMC4873481 DOI: 10.1538/expanim.15-0031] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/25/2022] Open
Abstract
Mesenchymal stem cells (MSCs) are one of the most promising cell populations for tissue engineering and regenerative medicine. Of utmost importance to MSC research is identification of MSC sources that are easily obtainable and stable. Several studies have shown that MSCs can be isolated from amniotic fluid. The sheep is one of the main types of farm animal, and it has many biophysical and biochemical similarities to humans. Here, we obtained MSCs from ovine amniotic fluid and determined the expansion capacity, surface and intracellular marker expression, karyotype, and multilineage differentiation ability of these ovine amniotic fluid mesenchymal stem cells (oAF-MSCs). Moreover, expression levels of differentiation markers were measured using reverse transcription-qPCR (RT-qPCR). Our phenotypic analysis shows that the isolated oAF-MSCs are indeed MSCs.
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Affiliation(s)
- Yunyun Tian
- College of Life Sciences, Inner Mongolia University, Inner Mongolia, Hohhot 010021, P.R.China
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19
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Li JJ, Kim K, Roohani-Esfahani SI, Guo J, Kaplan DL, Zreiqat H. A biphasic scaffold based on silk and bioactive ceramic with stratified properties for osteochondral tissue regeneration. J Mater Chem B 2015; 3:5361-5376. [PMID: 26167284 PMCID: PMC4494762 DOI: 10.1039/c5tb00353a] [Citation(s) in RCA: 35] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 08/22/2023]
Abstract
Significant clinical challenges encountered in the effective long-term treatment of osteochondral defects have inspired advancements in scaffold-based tissue engineering techniques to aid repair and regeneration. This study reports the development of a biphasic scaffold produced via a rational combination of silk fibroin and bioactive ceramic with stratified properties to satisfy the complex and diverse regenerative requirements of osteochondral tissue. Structural examination showed that the biphasic scaffold contained two phases with different pore morphologies to match the cartilage and bone segments of osteochondral tissue, which were joined at a continuous interface. Mechanical assessment showed that the two phases of the biphasic scaffold imitated the load-bearing behaviour of native osteochondral tissue and matched its compressive properties. In vitro testing showed that different compositions in the two phases of the biphasic scaffold could direct the preferential differentiation of human mesenchymal stem cells towards the chondrogenic or osteogenic lineage. By featuring simple and reproducible fabrication and a well-integrated interface, the biphasic scaffold strategy established in this study circumvented the common problems experienced with integrated scaffold designs and could provide an effective approach for the regeneration of osteochondral tissue.
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Affiliation(s)
- Jiao Jiao Li
- Biomaterials and Tissue Engineering Research Unit, School of AMME, University of Sydney, Sydney, NSW 2006, Australia
| | - Kyungsook Kim
- Department of Biomedical Engineering, Tufts University, Medford, MA 02155, USA
| | - Seyed-Iman Roohani-Esfahani
- Biomaterials and Tissue Engineering Research Unit, School of AMME, University of Sydney, Sydney, NSW 2006, Australia
| | - Jin Guo
- Department of Biomedical Engineering, Tufts University, Medford, MA 02155, USA
| | - David L. Kaplan
- Department of Biomedical Engineering, Tufts University, Medford, MA 02155, USA
| | - Hala Zreiqat
- Biomaterials and Tissue Engineering Research Unit, School of AMME, University of Sydney, Sydney, NSW 2006, Australia
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20
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Otani Y, Komura M, Komura H, Ishimaru T, Konishi K, Komuro H, Hoshi K, Takato T, Tabata Y, Iwanaka T. Optimal amount of basic fibroblast growth factor in gelatin sponges incorporating β-tricalcium phosphate with chondrocytes. Tissue Eng Part A 2015; 21:627-36. [PMID: 25287675 DOI: 10.1089/ten.tea.2013.0655] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
BACKGROUND A gelatin sponge with slowly releasing basic fibroblast growth factor (b-FGF) enhances chondrogenesis. This study investigated the optimal amount of b-FGF in gelatin sponges to fabricate engineered cartilage. MATERIALS AND METHODS b-FGF (0, 10, 100, 500, 1000, and 2000 μg/cm(3))-impregnated gelatin sponges incorporating β-tricalcium phosphate (β-TCP) were produced. Chondrocytes were isolated from the auricular cartilage of C57B6J mice and expanded. The expanded auricular chondrocytes (10×10(6) cells/cm(3)) were seeded onto the gelatin sponges, which served as scaffolds. The construct assembly was implanted in the subcutaneous space of mice through a syngeneic fashion. Thereafter, constructs were retrieved at 2, 4, or 6 weeks. RESULTS (1) Morphology: The size of implanted constructs was larger than the size of the scaffold with 500, 1000, and 2000 μg/cm(3) b-FGF-impregnated gelatin sponges incorporating β-TCP at 4 and 6 weeks after implantation. (2) The weight of the constructs increased roughly proportional to the increase in volume of the b-FGF-impregnated scaffold at 2, 4, and 6 weeks after implantation, except in the 2000 μg/cm(3) b-FGF-impregnated constructs group. (3) Histological examination: Extracellular matrix in the center of the constructs was observed in gelatin sponges impregnated with more than 100 μg/cm(3) b-FGF at 4 weeks after implantation. The areas of cells with an abundant extracellular matrix were positive for cartilage-specific marker type 2 collagen in the constructs. (4) Protein assay: Glycosaminoglycan and collagen type 2 expression were significantly increased at 4 and 6 weeks on implantation of gelatin sponges impregnated with more than 100 μg/cm(3) b-FGF. At 6 weeks after implantation, the ratio of type 2 collagen to type 1 collagen in constructs impregnated with 100 μg/cm(3) or more b-FGF was higher than that in mice auricular cartilage. CONCLUSION Gelatin sponges impregnated with more than 100 μg/cm(3) b-FGF incorporating β-TCP with chondrocytes (10×10(6) cells/cm(3)) can fabricate engineered cartilage at 4 weeks after implantation.
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Affiliation(s)
- Yushi Otani
- 1 Department of Pediatric Surgery, Graduate School of Medicine, University of Tokyo , Tokyo, Japan
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21
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Li JJ, Kaplan DL, Zreiqat H. Scaffold-based regeneration of skeletal tissues to meet clinical challenges. J Mater Chem B 2014; 2:7272-7306. [PMID: 32261954 DOI: 10.1039/c4tb01073f] [Citation(s) in RCA: 60] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022]
Abstract
The management and reconstruction of damaged or diseased skeletal tissues have remained a significant global healthcare challenge. The limited efficacy of conventional treatment strategies for large bone, cartilage and osteochondral defects has inspired the development of scaffold-based tissue engineering solutions, with the aim of achieving complete biological and functional restoration of the affected tissue in the presence of a supporting matrix. Nevertheless, significant regulatory hurdles have rendered the clinical translation of novel scaffold designs to be an inefficient process, mainly due to the difficulties of arriving at a simple, reproducible and effective solution that does not rely on the incorporation of cells and/or bioactive molecules. In the context of the current clinical situation and recent research advances, this review will discuss scaffold-based strategies for the regeneration of skeletal tissues, with focus on the contribution of bioactive ceramic scaffolds and silk fibroin, and combinations thereof, towards the development of clinically viable solutions.
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Affiliation(s)
- Jiao Jiao Li
- Biomaterials and Tissue Engineering Research Unit, School of AMME, University of Sydney, Sydney, NSW 2006, Australia.
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22
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Zhang W, Lian Q, Li D, Wang K, Hao D, Bian W, He J, Jin Z. Cartilage repair and subchondral bone migration using 3D printing osteochondral composites: a one-year-period study in rabbit trochlea. BIOMED RESEARCH INTERNATIONAL 2014; 2014:746138. [PMID: 25177697 PMCID: PMC4142181 DOI: 10.1155/2014/746138] [Citation(s) in RCA: 57] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 02/19/2014] [Revised: 06/30/2014] [Accepted: 07/14/2014] [Indexed: 11/29/2022]
Abstract
Increasing evidences show that subchondral bone may play a significant role in the repair or progression of cartilage damage in situ. However, the exact change of subchondral bone during osteochondral repair is still poorly understood. In this paper, biphasic osteochondral composite scaffolds were fabricated by 3D printing technology using PEG hydrogel and β-TCP ceramic and then implanted in rabbit trochlea within a critical size defect model. Animals were euthanized at 1, 2, 4, 8, 16, 24, and 52 weeks after implantation. Histological results showed that hyaline-like cartilage formed along with white smooth surface and invisible margin at 24 weeks postoperatively, typical tidemark formation at 52 weeks. The repaired subchondral bone formed from 16 to 52 weeks in a "flow like" manner from surrounding bone to the defect center gradually. Statistical analysis illustrated that both subchondral bone volume and migration area percentage were highly correlated with the gross appearance Wayne score of repaired cartilage. Therefore, subchondral bone migration is related to cartilage repair for critical size osteochondral defects. Furthermore, the subchondral bone remodeling proceeds in a "flow like" manner and repaired cartilage with tidemark implies that the biphasic PEG/β-TCP composites fabricated by 3D printing provides a feasible strategy for osteochondral tissue engineering application.
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Affiliation(s)
- Weijie Zhang
- State Key Laboratory for Manufacturing Systems Engineering, Xi'an Jiaotong University, Xi'an 710054, China
- The First Department of Orthopaedics, The Second Affiliated Hospital, Health Science Center, Xi'an Jiaotong University, Xi'an 710004, China
- Department of Joint Surgery, Hong Hui Hospital, Health Science Center, Xi'an Jiaotong University, Xi'an 710054, China
| | - Qin Lian
- State Key Laboratory for Manufacturing Systems Engineering, Xi'an Jiaotong University, Xi'an 710054, China
| | - Dichen Li
- State Key Laboratory for Manufacturing Systems Engineering, Xi'an Jiaotong University, Xi'an 710054, China
| | - Kunzheng Wang
- The First Department of Orthopaedics, The Second Affiliated Hospital, Health Science Center, Xi'an Jiaotong University, Xi'an 710004, China
| | - Dingjun Hao
- Department of Spine Surgery, Hong Hui Hospital, Health Science Center, Xi'an Jiaotong University, Xi'an 710054, China
| | - Weiguo Bian
- State Key Laboratory for Manufacturing Systems Engineering, Xi'an Jiaotong University, Xi'an 710054, China
- Department of Orthopaedics, The First Affiliated Hospital, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, China
| | - Jiankang He
- State Key Laboratory for Manufacturing Systems Engineering, Xi'an Jiaotong University, Xi'an 710054, China
| | - Zhongmin Jin
- State Key Laboratory for Manufacturing Systems Engineering, Xi'an Jiaotong University, Xi'an 710054, China
- Institute of Medical and Biological Engineering, School of Mechanical Engineering, University of Leeds, Leeds LS2 9JT, UK
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Baghaban Eslaminejad M, Malakooty Poor E. Mesenchymal stem cells as a potent cell source for articular cartilage regeneration. World J Stem Cells 2014; 6:344-354. [PMID: 25126383 PMCID: PMC4131275 DOI: 10.4252/wjsc.v6.i3.344] [Citation(s) in RCA: 60] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/22/2013] [Revised: 12/07/2013] [Accepted: 04/29/2014] [Indexed: 02/06/2023] Open
Abstract
Since articular cartilage possesses only a weak capacity for repair, its regeneration potential is considered one of the most important challenges for orthopedic surgeons. The treatment options, such as marrow stimulation techniques, fail to induce a repair tissue with the same functional and mechanical properties of native hyaline cartilage. Osteochondral transplantation is considered an effective treatment option but is associated with some disadvantages, including donor-site morbidity, tissue supply limitation, unsuitable mechanical properties and thickness of the obtained tissue. Although autologous chondrocyte implantation results in reasonable repair, it requires a two-step surgical procedure. Moreover, chondrocytes expanded in culture gradually undergo dedifferentiation, so lose morphological features and specialized functions. In the search for alternative cells, scientists have found mesenchymal stem cells (MSCs) to be an appropriate cellular material for articular cartilage repair. These cells were originally isolated from bone marrow samples and further investigations have revealed the presence of the cells in many other tissues. Furthermore, chondrogenic differentiation is an inherent property of MSCs noticed at the time of the cell discovery. MSCs are known to exhibit homing potential to the damaged site at which they differentiate into the tissue cells or secrete a wide spectrum of bioactive factors with regenerative properties. Moreover, these cells possess a considerable immunomodulatory potential that make them the general donor for therapeutic applications. All of these topics will be discussed in this review.
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Repairing the osteochondral defect in goat with the tissue-engineered osteochondral graft preconstructed in a double-chamber stirring bioreactor. BIOMED RESEARCH INTERNATIONAL 2014; 2014:219203. [PMID: 25061604 PMCID: PMC4100384 DOI: 10.1155/2014/219203] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 03/31/2014] [Accepted: 04/28/2014] [Indexed: 11/17/2022]
Abstract
To investigate the reparative efficacy of tissue-engineered osteochondral (TEO) graft for repairing the osteochondral defect in goat, we designed a double-chamber stirring bioreactor to construct the bone and cartilage composites simultaneously in one β-TCP scaffold and observed the reparative effect in vivo. The osteochondral defects were created in goats and all the animals were divided into 3 groups randomly. In groups A, the defect was treated with the TEO which was cultured with mechanical stimulation of stir; in group B, the defect was treated with TEO which was cultured without mechanical stimulation of stir; in groups C, the defect was treated without TEO. At 12 weeks and 24 weeks after operation, the reparative effects in different groups were assessed and compared. The results indicated that the reparative effect of the TEO cultured in the bioreactor was better than the control group, and mechanical stimulation of stir could further improve the reparative effect. We provided a feasible and effective method to construct the TEO for treatment of osteochondral defect using autologous BMSCs and the double-chamber bioreactor.
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Stimulation of osteogenic and angiogenic ability of cells on polymers by pulsed laser deposition of uniform akermanite-glass nanolayer. Acta Biomater 2014; 10:3295-306. [PMID: 24726444 DOI: 10.1016/j.actbio.2014.03.035] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/09/2013] [Revised: 03/27/2014] [Accepted: 03/31/2014] [Indexed: 12/27/2022]
Abstract
Polymer biomaterials have been widely used for bone replacement/regeneration because of their unique mechanical properties and workability. Their inherent low bioactivity makes them lack osseointegration with host bone tissue. For this reason, bioactive inorganic particles have been always incorporated into the matrix of polymers to improve their bioactivity. However, mixing inorganic particles with polymers always results in inhomogeneity of particle distribution in polymer matrix with limited bioactivity. This study sets out to apply the pulsed laser deposition (PLD) technique to prepare uniform akermanite (Ca2MgSi2O7, AKT) glass nanocoatings on the surface of two polymers (non-degradable polysulfone (PSU) and degradable polylactic acid (PDLLA)) in order to improve their surface osteogenic and angiogenic activity. The results show that a uniform nanolayer composed of amorphous AKT particles (∼30 nm) of thickness 130 nm forms on the surface of both PSU and PDLLA films with the PLD technique. The prepared AKT-PSU and AKT-PDLLA films significantly improved the surface roughness, hydrophilicity, hardness and apatite mineralization, compared with pure PSU and PDLLA, respectively. The prepared AKT nanocoatings distinctively enhance the alkaline phosphate (ALP) activity and bone-related gene expression (ALP, OCN, OPN and Col I) of bone-forming cells on both PSU and PDLLA films. Furthermore, AKT nanocoatings on two polymers improve the attachment, proliferation, VEGF secretion and expression of proangiogenic factors and their receptors of human umbilical vein endothelial cells (HUVEC). The results suggest that PLD-prepared bioceramic nanocoatings are very useful for enhancing the physicochemical, osteogenic and angiogenic properties of both degradable and non-degradable polymers for application in bone replacement/regeneration.
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Orth P, Rey-Rico A, Venkatesan JK, Madry H, Cucchiarini M. Current perspectives in stem cell research for knee cartilage repair. STEM CELLS AND CLONING-ADVANCES AND APPLICATIONS 2014; 7:1-17. [PMID: 24520197 PMCID: PMC3897321 DOI: 10.2147/sccaa.s42880] [Citation(s) in RCA: 52] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Protocols based on the delivery of stem cells are currently applied in patients, showing encouraging results for the treatment of articular cartilage lesions (focal defects, osteoarthritis). Yet, restoration of a fully functional cartilage surface (native structural organization and mechanical functions) especially in the knee joint has not been reported to date, showing the need for improved designs of clinical trials. Various sources of progenitor cells are now available, originating from adult tissues but also from embryonic or reprogrammed tissues, most of which have already been evaluated for their chondrogenic potential in culture and for their reparative properties in vivo upon implantation in relevant animal models of cartilage lesions. Nevertheless, particular attention will be needed regarding their safe clinical use and their potential to form a cartilaginous repair tissue of proper quality and functionality in the patient. Possible improvements may reside in the use of biological supplements in accordance with regulations, while some challenges remain in establishing standardized, effective procedures in the clinics.
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Affiliation(s)
- Patrick Orth
- Department of Orthopaedic Surgery, Saarland University Medical Center, Homburg, Germany
| | - Ana Rey-Rico
- Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg, Germany
| | - Jagadeesh K Venkatesan
- Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg, Germany
| | - Henning Madry
- Department of Orthopaedic Surgery, Saarland University Medical Center, Homburg, Germany ; Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg, Germany
| | - Magali Cucchiarini
- Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg, Germany
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Schneider-Wald B, von Thaden AK, Schwarz MLR. [Defect models for the regeneration of articular cartilage in large animals]. DER ORTHOPADE 2013; 42:242-53. [PMID: 23575559 DOI: 10.1007/s00132-012-2044-2] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Abstract
BACKGROUND Several animal models are available for the analysis of regeneration of articular cartilage in large animals, such as sheep, pigs, goats, dogs and horses. The subchondral bone lamella must be considered when ACT and MACT techniques are examined in order to protect the implant against migration of cells from the bone marrow, although recruitment of cells is often desirable in the regeneration of human cartilage. MATERIAL AND METHODS The defects are mainly positioned at the condyles and the trochlea often bilaterally and spontaneous healing should be excluded. The follow-up period for assessment of the effectiveness of cartilage regeneration is 6-12 months. Shorter observation times up to 12 weeks can be used for pilot studies. Scores based on histological, immunohistological and biochemical staining are mostly used for assessing the regenerated tissue. Biomechanical tests with destructive features need isolated specimens from the animal but modern slice imaging techniques can reflect the progression of the healing processes over the time span of the study in vivo. CONCLUSION Approaches to standardize the evaluation of the regeneration of articular cartilage have been sporadically described whereas they are required from the point of view of the approval of new concepts for therapy and the protection of animals.
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Affiliation(s)
- B Schneider-Wald
- Sektion experimentelle Orthopädie und Unfallchirurgie, Orthopädisch-Unfallchirurgisches Zentrum, Universitätsmedizin Mannheim, Medizinische Fakultät Mannheim, Universität Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Deutschland.
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Lehmann M, Martin F, Mannigel K, Kaltschmidt K, Sack U, Anderer U. Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes. Eur J Histochem 2013; 57:e31. [PMID: 24441184 PMCID: PMC3896033 DOI: 10.4081/ejh.2013.e31] [Citation(s) in RCA: 40] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/08/2013] [Revised: 09/13/2013] [Accepted: 09/18/2013] [Indexed: 11/23/2022] Open
Abstract
Cartilage regeneration based on isolated and culture-expanded chondrocytes has been studied in various in vitro models, but the quality varies with respect to the morphology and the physiology of the synthesized tissues. The aim of our study was to promote in vitro chondrogenesis of human articular chondrocytes using a novel three-dimensional (3-D) cultivation system in combination with the chondrogenic differentiation factors transforming growth factor beta 2 (TGF-b2) and L-ascorbic acid. Articular chondrocytes isolated from six elderly patients were expanded in monolayer culture. A single-cell suspension of the dedifferentiated chondrocytes was then added to agar-coated dishes without using any scaffold material, in the presence, or absence of TGF-b2 and/or L-ascorbic acid. Three-dimensional cartilage-like constructs, called single spheroids, and microtissues consisting of several spheroids fused together, named as fusions, were formed. Generated tissues were mainly characterized using histological and immunohistochemical techniques. The morphology of the in vitro tissues shared some similarities to native hyaline cartilage in regard to differentiated S100-positive chondrocytes within a cartilaginous matrix, with strong collagen type II expression and increased synthesis of proteoglycans. Finally, our innovative scaffold-free fusion culture technique supported enhanced chondrogenesis of human articular chondrocytes in vitro. These 3-D hyaline cartilage-like microtissues will be useful for in vitro studies of cartilage differentiation and regeneration, enabling optimization of functional tissue engineering and possibly contributing to the development of new approaches to treat traumatic cartilage defects or osteoarthritis.
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Affiliation(s)
- M Lehmann
- Brandenburg University of Technology Cottbus - Senftenberg.
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Yang P, Huang X, Wang C, Dang X, Wang K. Repair of bone defects using a new biomimetic construction fabricated by adipose-derived stem cells, collagen I, and porous beta-tricalcium phosphate scaffolds. Exp Biol Med (Maywood) 2013; 238:1331-43. [PMID: 24157587 DOI: 10.1177/1535370213505827] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/22/2023] Open
Abstract
Adipose derived stem cells (ASCs) with multilineage differentiation capacities have been demonstrated as an alternative cell candidate for in vitro and in vivo bone regeneration. This suggests that they may be a potential candidate to repair the bone defects. We attempted to demonstrate the use of new biomimetic constructions of undifferentiated rabbit adipose-derived stem cells (rASCs) with fully interconnected porous beta-tricalcium phosphate (β-TCP) scaffolds encapsulated by collagen I hydrogel in the regeneration of a critical-sized defect of rabbit radii. Critical-sized defects in the left radii of rabbits were prepared and inserted with rASCs/collagen I/β-TCP scaffold composites or collagen I/β-TCP scaffold composites. The results were evaluated by histology, radiographs, micro-CT, Emission Computed Tomography (ECT), fluorochrome labeling, western blot, and mechanical testing at 4, 8, and 12 weeks postsurgery. Twelve weeks after implantation, the defects were almost completely repaired as confirmed by the presence of the cortical bone and medullary cavity, which was evaluated through radiologic, histologic, and biomechanical examination. Biodegradation of the biomaterials may be attributed to extracellular liquid dissolution together with cell-mediated phagocytosis. Our study shows that a greater number of rASCs in the porous β-TCP scaffold encapsulated by collagen I gel enhanced osteogenesis in critical-sized defects. We hope to garner new insight into the engineering of rASCs-based bone tissue for clinical application.
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Affiliation(s)
- Pei Yang
- Department of Orthopaedics, the Second Affiliated Hospital of Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi 710004, China
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30
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Lee EJ, Kasper FK, Mikos AG. Biomaterials for tissue engineering. Ann Biomed Eng 2013; 42:323-37. [PMID: 23820768 DOI: 10.1007/s10439-013-0859-6] [Citation(s) in RCA: 139] [Impact Index Per Article: 11.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/30/2013] [Accepted: 06/24/2013] [Indexed: 01/24/2023]
Abstract
Biomaterials serve as an integral component of tissue engineering. They are designed to provide architectural framework reminiscent of native extracellular matrix in order to encourage cell growth and eventual tissue regeneration. Bone and cartilage represent two distinct tissues with varying compositional and mechanical properties. Despite these differences, both meet at the osteochondral interface. This article presents an overview of current biomaterials employed in bone and cartilage applications, discusses some design considerations, and alludes to future prospects within this field of research.
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Affiliation(s)
- Esther J Lee
- Department of Bioengineering, Rice University, MS 142, P.O. Box 1892, Houston, TX, 77251-1892, USA
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31
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Nooeaid P, Salih V, Beier JP, Boccaccini AR. Osteochondral tissue engineering: scaffolds, stem cells and applications. J Cell Mol Med 2012; 16:2247-70. [PMID: 22452848 PMCID: PMC3823419 DOI: 10.1111/j.1582-4934.2012.01571.x] [Citation(s) in RCA: 201] [Impact Index Per Article: 15.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/04/2011] [Accepted: 03/21/2012] [Indexed: 12/17/2022] Open
Abstract
Osteochondral tissue engineering has shown an increasing development to provide suitable strategies for the regeneration of damaged cartilage and underlying subchondral bone tissue. For reasons of the limitation in the capacity of articular cartilage to self-repair, it is essential to develop approaches based on suitable scaffolds made of appropriate engineered biomaterials. The combination of biodegradable polymers and bioactive ceramics in a variety of composite structures is promising in this area, whereby the fabrication methods, associated cells and signalling factors determine the success of the strategies. The objective of this review is to present and discuss approaches being proposed in osteochondral tissue engineering, which are focused on the application of various materials forming bilayered composite scaffolds, including polymers and ceramics, discussing the variety of scaffold designs and fabrication methods being developed. Additionally, cell sources and biological protein incorporation methods are discussed, addressing their interaction with scaffolds and highlighting the potential for creating a new generation of bilayered composite scaffolds that can mimic the native interfacial tissue properties, and are able to adapt to the biological environment.
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Affiliation(s)
- Patcharakamon Nooeaid
- Department of Materials Science and Engineering Institute of Biomaterials, Friedrich-Alexander-University of Erlangen-NürnbergErlangen, Germany
| | - Vehid Salih
- Eastman Dental Institute, UCLLondon, United Kingdom
| | - Justus P Beier
- Department of Plastic and Hand Surgery, University Hospital of Erlangen Friedrich-Alexander-University of Erlangen-NürnbergErlangen, Germany
| | - Aldo R Boccaccini
- Department of Materials Science and Engineering Institute of Biomaterials, Friedrich-Alexander-University of Erlangen-NürnbergErlangen, Germany
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Lyahyai J, Mediano DR, Ranera B, Sanz A, Remacha AR, Bolea R, Zaragoza P, Rodellar C, Martín-Burriel I. Isolation and characterization of ovine mesenchymal stem cells derived from peripheral blood. BMC Vet Res 2012; 8:169. [PMID: 22999337 PMCID: PMC3514285 DOI: 10.1186/1746-6148-8-169] [Citation(s) in RCA: 59] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2012] [Accepted: 09/17/2012] [Indexed: 12/23/2022] Open
Abstract
BACKGROUND Mesenchymal stem cells (MSCs) are multipotent stem cells with capacity to differentiate into several mesenchymal lineages. This quality makes MSCs good candidates for use in cell therapy. MSCs can be isolated from a variety of tissues including bone marrow and adipose tissue, which are the most common sources of these cells. However, MSCs can also be isolated from peripheral blood. Sheep has been proposed as an ideal model for biomedical studies including those of orthopaedics and transmissible spongiform encephalopathies (TSEs). The aim of this work was to advance these studies by investigating the possibility of MSC isolation from ovine peripheral blood (oPB-MSCs) and by subsequently characterizing there in vitro properties. RESULTS Plastic-adherent fibroblast-like cells were obtained from the mononuclear fraction of blood samples. These cells were analysed for their proliferative and differentiation potential into adipocytes, osteoblasts and chondrocytes, as well as for the gene expression of cell surface markers. The isolated cells expressed transcripts for markers CD29, CD73 and CD90, but failed to express the haematopoietic marker CD45 and expressed only low levels of CD105. The expression of CD34 was variable. The differentiation potential of this cell population was evaluated using specific differentiation media. Although the ability of the cultures derived from different animals to differentiate into adipocytes, osteoblasts and chondrocytes was heterogeneous, we confirmed this feature using specific staining and analysing the gene expression of differentiation markers. Finally, we tested the ability of oPB-MSCs to transdifferentiate into neuronal-like cells. Morphological changes were observed after 24-hour culture in neurogenic media, and the transcript levels of the neurogenic markers increased during the prolonged induction period. Moreover, oPB-MSCs expressed the cellular prion protein gene (PRNP), which was up-regulated during neurogenesis. CONCLUSIONS This study describes for the first time the isolation and characterization of oPB-MSCs. Albeit some variability was observed between animals, these cells retained their capacity to differentiate into mesenchymal lineages and to transdifferentiate into neuron-like cells in vitro. Therefore, oPB-MSCs could serve as a valuable tool for biomedical research in fields including orthopaedics or prion diseases.
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Affiliation(s)
- Jaber Lyahyai
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, Miguel Servet 177, Zaragoza, 50013, Spain
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Abstract
It is a great honour that
Interface Focus
will publish a special issue to introduce the progress and achievements of biomaterials research in China. We feel it is a good opportunity to present the progress in Chinese biomaterials research to the international biomaterials community, and to promote the collaboration between international and Chinese biomaterials scientists.
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Affiliation(s)
- Xingdong Zhang
- National Engineering Research Center for Biomaterials, Sichuan University, Chengdu 610064, China
| | - Fuzhai Cui
- Institute of Regenerative Medical Materials, Tsinghua University, Beijing 100084, China
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Yunos DM, Ahmad Z, Salih V, Boccaccini AR. Stratified scaffolds for osteochondral tissue engineering applications: Electrospun PDLLA nanofibre coated Bioglass®-derived foams. J Biomater Appl 2011; 27:537-51. [DOI: 10.1177/0885328211414941] [Citation(s) in RCA: 49] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Abstract
This work focuses on designing bilayered constructs by combining electrospun poly-DL-Lactide (PDLLA) fibers and Bioglass®-derived scaffolds for development of osteochondral tissue replacement materials. Electrospinning was carried out using a solution of 5 wt/v% PDLLA in dimethyl carbonate. The PDLLA layer thickness increased from 2 to 150 µm with varying electrospinning time. Bioactivity studies in simulated body fluid showed that HA mineralization decreased as the thickness of the PDLLA layer increased. A preliminary in vitro study using chondrocyte cells (ATDC5) showed that cells attached, proliferated and migrated into the fibrous network, confirming the potential applicability of the bilayered scaffolds in osteochondral defect regeneration.
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Affiliation(s)
- DM Yunos
- Department of Materials, Imperial College London, London SW7 2BP, UK
- Present address: SIRIM Berhad, Malaysia
| | - Z Ahmad
- School of Pharmacy and Biomedical Science, University of Portsmouth, Portsmouth, PO1 2DT, UK
| | - V Salih
- UCL Eastman Dental Institute, London WC1X 8LD, UK
| | - AR Boccaccini
- Department of Materials, Imperial College London, London SW7 2BP, UK
- Institute of Biomaterials, Department of Materials Science and Engineering, University of Erlangen-Nuremberg, 91058 Erlangen, Germany
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Khanarian NT, Jiang J, Wan LQ, Mow VC, Lu HH. A hydrogel-mineral composite scaffold for osteochondral interface tissue engineering. Tissue Eng Part A 2011; 18:533-45. [PMID: 21919797 DOI: 10.1089/ten.tea.2011.0279] [Citation(s) in RCA: 79] [Impact Index Per Article: 5.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/12/2022] Open
Abstract
Osteoarthritis is the leading cause of physical disability among Americans, and tissue engineered cartilage grafts have emerged as a promising treatment option for this debilitating condition. Currently, the formation of a stable interface between the cartilage graft and subchondral bone remains a significant challenge. This study evaluates the potential of a hybrid scaffold of hydroxyapatite (HA) and alginate hydrogel for the regeneration of the osteochondral interface. Specifically, the effects of HA on the response of chondrocytes were determined, focusing on changes in matrix production and mineralization, as well as scaffold mechanical properties over time. Additionally, the optimal chondrocyte population for interface tissue engineering was evaluated. It was observed that the HA phase of the composite scaffold promoted the formation of a proteoglycan- and type II collagen-rich matrix when seeded with deep zone chondrocytes. More importantly, the elevated biosynthesis translated into significant increases in both compressive and shear moduli relative to the mineral-free control. Presence of HA also promoted chondrocyte hypertrophy and type X collagen deposition. These results demonstrate that the hydrogel-calcium phosphate composite supported the formation of a calcified cartilage-like matrix and is a promising scaffold design for osteochondral interface tissue engineering.
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Affiliation(s)
- Nora T Khanarian
- Biomaterials and Interface Tissue Engineering Laboratory, Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA
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Niyama K, Ide N, Onoue K, Okabe T, Wakitani S, Takagi M. Construction of osteochondral-like tissue graft combining β-tricalcium phosphate block and scaffold-free centrifuged chondrocyte cell sheet. J Orthop Sci 2011; 16:613-21. [PMID: 21713422 DOI: 10.1007/s00776-011-0120-9] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/18/2010] [Accepted: 06/09/2011] [Indexed: 02/09/2023]
Abstract
BACKGROUND The combination of a β-tricalcium phosphate (βTCP) block with a scaffold-free chondrocyte sheet formed by the centrifugation of chondrocytes in a well was investigated with the aim of constructing an osteochondral-like structure. METHODS Human and porcine articular cartilage chondrocytes were respectively centrifuged in a 96-well plate or cell culture insert (0.32 cm(2)) that was set in a 24-well plate, cultivated in the respective vessel for 3 weeks, and the cell sheets were harvested. In some cases, a cylindrical βTCP block (diameter 5 mm, height 3 mm) was placed on the sheet on days 1-7. The sheet size, cell number, and sulfated glycosaminoglycan accumulation were determined. RESULTS The use of a 96-well plate for not suspension but adhesion culture and the initial centrifugation of a well containing cells were crucial to obtaining a uniformly thick cell sheet. The glycosaminoglycan density of the harvested cell sheet was comparable to that of the pellet culture. An inoculum cell number of more than 31 × 10(5) cells tended to result in a curved cell sheet. Culture involving 18.6 × 10(5) cells and the 96-well plate for adhesion culture showed no curving of the cell sheet (thickness of 0.85 mm), and these were found to be the best of the culture conditions tested. The timing of the addition of a βTCP block to the cell sheet (1-7 days) markedly affected the balance between the thickness of cell sheet parts on and in the βTCP block. CONCLUSION Centrifugation and subsequent cultivation of chondrocytes (18.6 × 10(5) cells) in a 96-well plate for adhesion culture led to the production of a scaffold-free cartilage-like cell sheet with a thickness of 0.85 mm. A combined osteochondral-like structure was produced by putting a βTCP block on the cell sheet. The thickness of the cell sheet on the βTCP block and the binding strength between the cell sheet and the βTCP block could be optimized by adjusting the inoculum cell number and timing of βTCP block addition.
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Affiliation(s)
- Kouhei Niyama
- Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, Kita-ku N13W8, Sapporo, 060-8628, Japan
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Zhu M, Wang H, Liu J, He H, Hua X, He Q, Zhang L, Ye X, Shi J. A mesoporous silica nanoparticulate/β-TCP/BG composite drug delivery system for osteoarticular tuberculosis therapy. Biomaterials 2011; 32:1986-1995. [PMID: 21163519 DOI: 10.1016/j.biomaterials.2010.11.025] [Citation(s) in RCA: 61] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/23/2010] [Accepted: 11/14/2010] [Indexed: 10/18/2022]
Abstract
A composite scaffold drug delivery system (CS-DDS) for osteoarticular tuberculosis therapy has been prepared by loading bi-component drugs into a mesoporous silica nanoparticles (MSNs)-coated porous β-TCP scaffold, which was followed by an additional bioactive glass coating. Such a CS-DDS showed high performances in the local and extremely sustained delivery of the bi-component antitubercular drugs and excellent biocompatibility. N(2) sorption isotherms indicated greatly increased surface area of the composites compared to pure β-TCP scaffold, and the mesopores were around 2.6 nm which were large enough to encapsulate drugs such as isoniazide and rifampicin. The in vitro and in vivo release tests demonstrated extra sustained co-release profiles of rifampicin and isoniazide from such a CS-DDS, and both drug concentrations kept higher than their effective values to kill mycobacterium tuberculosis for as long as 42 days. The hepatic and renal function tests indicated that the CS-DDS had neglectable long-term lesions to liver and kidney.
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Affiliation(s)
- Min Zhu
- State Key Laboratory of High Performance Ceramics and Superfine Microstructure, Shanghai Institute of Ceramics, Chinese Academy of Sciences, 1295 Ding-Xi Road, Shanghai, 200050, PR China
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Xue D, Zheng Q, Zong C, Li Q, Li H, Qian S, Zhang B, Yu L, Pan Z. Osteochondral repair using porous poly(lactide-co-glycolide)/nano-hydroxyapatite hybrid scaffolds with undifferentiated mesenchymal stem cells in a rat model. J Biomed Mater Res A 2010; 94:259-70. [PMID: 20166224 DOI: 10.1002/jbm.a.32691] [Citation(s) in RCA: 70] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/24/2023]
Abstract
In this study, a novel three-dimensional poly (lactide-co-glycolide) (PLGA)/nano-hydroxyapatite (NHA) scaffold was fabricated by a thermally induced phase separation technique and its potential application in cartilage tissue-engineering was investigated. The PLGA scaffold was used as a control and mesenchymal stem cells (MSCs) were seeded in both scaffolds. After 12-days culture, SEM images and confocal laser scanning microscopy illustrated that MSCs attached more moderately and more cells distributed in PLGA/NHA scaffolds. MTT test and DNA assay showed that the viability and proliferation of MSCs in PLGA/NHA scaffolds were significantly superior to PLGA scaffolds during in vitro culture. Through in vivo study, the efficacy of this scaffold combining with MSCs for repairing articular osteochondral defects was evaluated in a rat model. Osteochondral defects in rats knees were left untreated, or treated with PLGA/NHA-MSCs composites or PLGA-MSCs composites. Twelve weeks after operation, histological examination revealed that the defects in the PLGA/NHA-MSCs treated group were filled with smooth and hyaline-like cartilage with abundant glycosaminoglycan and collagen type II deposition, but deficient in collagen type I at 12 weeks after operation. To investigate the final fate of MSCs transplanted into the defect areas, the fluorescent dye CM-DiI was used to prelabel cells. At 12 weeks after transplantation, we still observed the red fluorescence in the repair area. These findings suggest that the PLGA/NHA-MSCs composite may be potentially used for cartilage repair in clinical application. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.
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Affiliation(s)
- Deting Xue
- Department of Orthopaedics, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China
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Bal BS, Rahaman MN, Jayabalan P, Kuroki K, Cockrell MK, Yao JQ, Cook JL. In vivo outcomes of tissue-engineered osteochondral grafts. J Biomed Mater Res B Appl Biomater 2010; 93:164-74. [PMID: 20091911 DOI: 10.1002/jbm.b.31571] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Tissue-engineered osteochondral grafts have been synthesized from a variety of materials, with some success at repairing chondral defects in animal models. We hypothesized that in tissue-engineered osteochondral grafts synthesized by bonding mesenchymal stem cell-loaded hydrogels to a porous material, the choice of the porous scaffold would affect graft healing to host bone, and the quality of cell restoration at the hyaline cartilage surface. Bone marrow-derived allogeneic mesenchymal stem cells were suspended in hydrogels that were attached to cylinders of porous tantalum metal, allograft bone, or a bioactive glass. The tissue-engineered osteochondral grafts, thus created were implanted into experimental defects in rabbit knees. Subchondral bone restoration, defect fill, bone ingrowth-implant integration, and articular tissue quality were compared between the three subchondral materials at 6 and 12 weeks. Bioactive glass and porous tantalum were superior to bone allograft in integrating to adjacent host bone, regenerating hyaline-like tissue at the graft surface, and expressing type II collagen in the articular cartilage.
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Affiliation(s)
- B Sonny Bal
- Department of Orthopaedic Surgery, University of Missouri, Columbia, Missouri, USA.
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40
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Rentsch C, Hess R, Rentsch B, Hofmann A, Manthey S, Scharnweber D, Biewener A, Zwipp H. Ovine bone marrow mesenchymal stem cells: isolation and characterization of the cells and their osteogenic differentiation potential on embroidered and surface-modified polycaprolactone-co-lactide scaffolds. In Vitro Cell Dev Biol Anim 2010; 46:624-34. [PMID: 20490706 DOI: 10.1007/s11626-010-9316-0] [Citation(s) in RCA: 42] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2009] [Accepted: 03/31/2010] [Indexed: 12/13/2022]
Abstract
The current study was undertaken with the goal being isolation, cultivation, and characterization of ovine mesenchymal stem cells (oMSC). Furthermore, the objective was to determine whether biological active polycaprolactone-co-lactide (trade name PCL) scaffolds support the growth and differentiation of oMSC in vitro. The oMSC were isolated from the iliac crest of six merino sheep. Three factors were used to demonstrate the MSC properties of the isolated cells in detail. (1) Their ability to proliferate in culture with a spindle-shaped morphology, (2) presence of specific surface marker proteins, and (3) their capacity to differentiate into the three classical mesenchymal pathways, osteoblastic, adipogenic, and chondrogenic lineages. Furthermore, embroidered PCL scaffolds were coated with collagen I (coll I) and chondroitin sulfate (CS). The porous structure of the scaffolds and the coating with coll I/CS allowed the oMSC to adhere, proliferate, and to migrate into the scaffolds. The coll I/CS coating on the PCL scaffolds induced osteogenic differentiation of hMSC, without differentiation supplements, indicating that the scaffold also has an osteoinductive character. In conclusion, the isolated cells from the ovine bone marrow have similar morphologic, immunophenotypic, and functional characteristics as their human counterparts. These cells were also found to differentiate into multiple mesenchymal cell types. This study demonstrates that embroidered PCL scaffolds can act as a temporary matrix for cell migration, proliferation, and differentiation of oMSC. The data presented will provide a reliable model system to assess the translation of MSC-based therapy into a variety of valuable ovine experimental models under autologous settings.
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Affiliation(s)
- C Rentsch
- Department of Trauma and Reconstructive Surgery, University Hospital Carl Gustav Carus Dresden, Fetscher Strasse 74, 01307 Dresden, Germany.
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41
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Miyazaki T, Miyauchi S, Matsuzaka S, Yamagishi C, Kobayashi K. Formation of Proteoglycan and Collagen-Rich Scaffold-Free Stiff Cartilaginous Tissue Using Two-Step Culture Methods with Combinations of Growth Factors. Tissue Eng Part A 2010; 16:1575-84. [DOI: 10.1089/ten.tea.2009.0443] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Affiliation(s)
- Tatsuya Miyazaki
- Division of Research Institute, PG Research Co. Ltd., Tokyo, Japan
| | - Satoshi Miyauchi
- Division of Research Institute, PG Research Co. Ltd., Tokyo, Japan
| | | | - Chie Yamagishi
- Division of Research Institute, PG Research Co. Ltd., Tokyo, Japan
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42
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Yang P, Wang C, Shi Z, Huang X, Dang X, Li X, Lin SF, Wang K. rhVEGF 165 delivered in a porous beta-tricalcium phosphate scaffold accelerates bridging of critical-sized defects in rabbit radii. J Biomed Mater Res A 2010; 92:626-40. [PMID: 19235222 DOI: 10.1002/jbm.a.32403] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Segmental bone defects are a common obstacle in major orthopedic procedures, and the treatment of these defects remains a challenging clinical problem. Bone tissue engineering has been attracting much attention in recent years. We evaluated the ability of the specific combination of 3 microg rhVEGF(165) with a novel porous beta-tricalcium phosphate (beta-TCP) scaffold coated with fibrin sealant (FS) to facilitate bone regeneration. Unilateral 15-mm long critical-sized defects were prepared in the radial diaphysis of rabbits and treated with rhVEGF(165)/FS/scaffold or FS/scaffold. Healing of the defects was assessed at 4, 8, and 12 weeks, radiologically, histologically, and biomechanically. The results of the study demonstrated that the critical-sized defects in the midshaft of the rabbit radius, treated with rhVEGF(165) incorporated in porous beta-TCP scaffold by FS, can be completely bridged by cortical bone in 12 weeks. The bone marrow space was also reformed histologically and radiologically at 12 weeks postsurgery in the rhVEGF(165)-treated group. Furthermore, biomechanical examination demonstrated that the segmental bone defects were not only radiologically and histologically repaired but were also mechanically repaired. Interestingly, none of the defects was completely repaired at 12 weeks following treatment with FS/scaffold without rhVEGF(165). A solution-driven process is likely the predominant mechanism of accelerating biodegradation of the beta-TCP scaffold in the presence of rhVEGF(165); furthermore, cell-mediated phagocytosis also contributes to biodegradation of the biomaterials.
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Affiliation(s)
- Pei Yang
- Department of Orthopaedics, Second Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, Shaanxi, People's Republic of China
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43
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Hao T, Wen N, Cao JK, Wang HB, Lü SH, Liu T, Lin QX, Duan CM, Wang CY. The support of matrix accumulation and the promotion of sheep articular cartilage defects repair in vivo by chitosan hydrogels. Osteoarthritis Cartilage 2010; 18:257-65. [PMID: 19744589 DOI: 10.1016/j.joca.2009.08.007] [Citation(s) in RCA: 96] [Impact Index Per Article: 6.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/16/2009] [Revised: 08/25/2009] [Accepted: 08/26/2009] [Indexed: 02/02/2023]
Abstract
OBJECTIVE Chitosan has been widely used as an injectable scaffold in cartilage tissue engineering due to its characteristic biocompatibility and biodegradability. In this study, chitosan was used in its hydrogel form as a scaffold for chondrocytes that act to reconstruct tissue-engineered cartilage and repair articular cartilage defects in the sheep model. This study aims to find a novel way to apply chitosan in cartilage tissue engineering. METHODS Temperature-responsive chitosan hydrogels were prepared by combining chitosan, beta-sodium glycerophosphate (GP) and hydroxyethyl cellulose (HEC). Tissue-engineered cartilage reconstructions were made in vitro by mixing sheep chondrocytes with a chitosan hydrogel. Cell survival and matrix accumulation were analyzed after 3 weeks in culture. To collect data for in vivo repair, reconstructions cultured for 1 day were transplanted to the freshly prepared defects of the articular cartilage of sheep. Then at both 12 and 24 weeks after transplantation, the grafts were extracted and analyzed histologically and immunohistochemically. RESULTS The results showed that the chondrocytes in the reconstructed cartilage survived and retained their ability to secrete matrix when cultured in vitro. Transplanted in vivo, the reconstructions repaired cartilage defects completely within 24 weeks. The implantation of chitosan hydrogels without chondrocytes also helps to repair cartilage defects. CONCLUSIONS The chitosan-based hydrogel could support matrix accumulation of chondrocytes and could repair sheep cartilage defects in 24 weeks. This study showcased the success of a new technique in its ability to repair articular cartilage defects.
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Affiliation(s)
- T Hao
- Department of Tissue Engineering, Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, Beijing, PR China
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44
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Richardson SM, Hoyland JA, Mobasheri R, Csaki C, Shakibaei M, Mobasheri A. Mesenchymal stem cells in regenerative medicine: Opportunities and challenges for articular cartilage and intervertebral disc tissue engineering. J Cell Physiol 2010; 222:23-32. [DOI: 10.1002/jcp.21915] [Citation(s) in RCA: 150] [Impact Index Per Article: 10.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/08/2023]
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45
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O'Shea TM, Miao X. Bilayered scaffolds for osteochondral tissue engineering. TISSUE ENGINEERING PART B-REVIEWS 2009; 14:447-64. [PMID: 18844605 DOI: 10.1089/ten.teb.2008.0327] [Citation(s) in RCA: 87] [Impact Index Per Article: 5.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/12/2022]
Abstract
Osteoarthritis (OA) is a prevalent degenerative joint disease that places a significant burden on the socioeconomic efficacy of communities around the world. Tissue engineering repair of articular cartilage in synovial joints represents a potential OA treatment strategy superior to current surgical techniques. In particular, osteochondral tissue engineering, which promotes the simultaneous regeneration of articular cartilage and underlining subchondral bone, may be a clinically relevant approach toward impeding OA progression. The unique and complex functional demands of the two contrasting tissues that comprise osteochondral tissue require the use of bilayered scaffolds to promote individual growth of both on a single integrated implant. This paper reviews the three current bilayered scaffold strategies applied to solve this challenging problem, with a focus on the need for an innovative approach to design and fabrication of new optimized scaffold combinations to reinforce materials science as an important element of osteochondral tissue engineering.
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Affiliation(s)
- Timothy M O'Shea
- School of Engineering Systems, Queensland University of Technology, Brisbane, Australia
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46
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Bilayered chitosan-based scaffolds for osteochondral tissue engineering: influence of hydroxyapatite on in vitro cytotoxicity and dynamic bioactivity studies in a specific double-chamber bioreactor. Acta Biomater 2009; 5:644-60. [PMID: 18951857 DOI: 10.1016/j.actbio.2008.09.017] [Citation(s) in RCA: 107] [Impact Index Per Article: 6.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/27/2008] [Revised: 09/23/2008] [Accepted: 09/23/2008] [Indexed: 01/08/2023]
Abstract
Osteochondral tissue engineering presents a current research challenge due to the necessity of combining both bone and cartilage tissue engineering principles. In the present study, bilayered chitosan-based scaffolds are developed based on the optimization of both polymeric and composite scaffolds. A particle aggregation methodology is proposed in order to achieve an improved integrative bone-cartilage interface needed for this application, since any discontinuity is likely to cause long-term device failure. Cytotoxicity was evaluated by the MTS assay with the L929 fibroblast cell line for different conditions. Surprisingly, in composite scaffolds using unsintered hydroxyapatite, cytotoxicity was observed in vitro. This work reports the investigation that was conducted to overcome and explain this behaviour. It is suggest that the uptake of divalent cations may induce the cytotoxic behaviour. Sintered hydroxyapatite was consequently used and showed no cytotoxicity when compared to the controls. Microcomputed tomography (micro-CT) was carried out to accurately quantify porosity, interconnectivity, ceramic content, particle and pore sizes. The results showed that the developed scaffolds are highly interconnected and present the ideal pore size range to be morphometrically suitable for the proposed applications. Dynamical mechanical analysis (DMA) demonstrated that the scaffolds are mechanically stable in the wet state even under dynamic compression. The obtained elastic modulus was, respectively, 4.21+/-1.04, 7.98+/-1.77 and 6.26+/-1.04 MPa at 1 Hz frequency for polymeric, composite and bilayered scaffolds. Bioactivity studies using both a simulated body fluid (SBF) and a simulated synovial fluid (SSF) were conducted in order to assure that the polymeric component for chondrogenic part would not mineralize, as confirmed by scanning electron microscopy (SEM), inductively coupled plasma-optical emission spectroscopy (ICP) and energy-dispersive spectroscopy (EDS) for different immersion periods. The assays were carried out also under dynamic conditions using, for this purpose, a specifically designed double-chamber bioreactor, aiming at a future osteochondral application. It was concluded that chitosan-based bilayered scaffolds produced by particle aggregation overcome any risk of delamination of both polymeric and composite parts designed, respectively, for chondrogenic and osteogenic components that are mechanically stable. Moreover, the proposed bilayered scaffolds could serve as alternative, biocompatible and safe biodegradable scaffolds for osteochondral tissue engineering applications.
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47
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Surface modification of beta-tricalcium phosphate scaffolds with topological nanoapatite coatings. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2008. [DOI: 10.1016/j.msec.2008.02.004] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/19/2022]
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48
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Li Z, Gunn J, Chen M, Cooper A, Zhang M. On‐site alginate gelation for enhanced cell proliferation and uniform distribution in porous scaffolds. J Biomed Mater Res A 2008; 86:552-9. [DOI: 10.1002/jbm.a.31596] [Citation(s) in RCA: 34] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
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49
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Petersen JP, Ueblacker P, Goepfert C, Adamietz P, Baumbach K, Stork A, Rueger JM, Poertner R, Amling M, Meenen NM. Long term results after implantation of tissue engineered cartilage for the treatment of osteochondral lesions in a minipig model. JOURNAL OF MATERIALS SCIENCE. MATERIALS IN MEDICINE 2008; 19:2029-2038. [PMID: 17957447 DOI: 10.1007/s10856-007-3291-3] [Citation(s) in RCA: 39] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/28/2007] [Accepted: 09/25/2007] [Indexed: 05/25/2023]
Abstract
In present study we determined the long term in vivo integration and histological modeling of an in vitro engineered cartilage construct. Tissue engineered autologous cartilagenous tissue was cultured on calcium phosphate cylinders and implanted into osteochondral defects into the femoral condyles in minipigs. Radiological follow-up was performed at 2, 8, 26 and 52 weeks, condyles were harvested 26 and 52 weeks post-implantation. Thickness of cultivated tissue (1.10 +/- 0.55 mm) was comparable to in situ cartilage and cells produced in vitro cartilage specific proteins. In vivo, 26 and 52 weeks post-implantation defects were resurfaced with hyaline-like tissue, the implants were well integrated with no gap at the interface between the engineered neocartilage and the adjacent articular cartilage. Synthesis of type II collagen was detected 26 and 52 weeks after implantation. The modified ICRS score increased from 26 to 52 weeks. Histomorphometric evaluation revealed a decrease in cellularity in tissue engineered cartilage from 2.2-fold of native cartilage after 26 weeks to 1.5-fold after 52 weeks. In conclusion, these findings demonstrate the integration and maturation of tissue engineered cartilage pellets attached on a bone substitute carrier implanted in osteochondral defects over a long time.
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Affiliation(s)
- J P Petersen
- Department of Trauma, Hand and Reconstructive Surgery, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany
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Abstract
The application of autologous chondrocytes in cartilage repair procedures is associated with several disadvantages, including injury of healthy cartilage in a preceding surgery frequently resulting in formation of inferior fibrocartilage at defect sites. In order to improve the quality of regeneration, adult mesenchymal stem cells (MSC) are regarded as a promising alternative. The great challenge, when considering MSC for articular cartilage repair, is to generate cells with features of stable chondrocytes which are resistant to hypertrophy and terminal differentiation, as found in hyaline articular cartilage. Common in vitro protocols for chondrogenic differentiation of MSC successfully induce expression of multiple cartilage-specific molecules, including collagen type II and aggrecan, and result in a chondrocyte-like phenotype. However, in vitro chondrogenesis of MSC additionally promotes induction of fibrocartilage-like features such as expression of collagen type I, and hypertrophy, as demonstrated by up-regulation of collagen type X, MMP13 and ALP-activity. As a consequence, differentiated MSC pellets undergo mineralisation and vascularisation after ectopic transplantation in a process similar to endochondral ossification. This review discusses the complexity and entailed challenges when considering MSC from various sources for clinical application and the necessity to optimise chondrogenesis by repressing hypertrophy to obtain functional and suitable cells for cartilage repair.
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Affiliation(s)
- Karoliina Pelttari
- Division of Experimental Orthopaedics, Orthopaedic University Hospital Heidelberg, Schlierbacher Landstrasse 2002, Heidelberg, Germany
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