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Garcia‐Aponte OF, Kahlenberg S, Kouroupis D, Egger D, Kasper C. Effects of Hydrogels on Mesenchymal Stem/Stromal Cells Paracrine Activity and Extracellular Vesicles Production. J Extracell Vesicles 2025; 14:e70057. [PMID: 40091440 PMCID: PMC11911545 DOI: 10.1002/jev2.70057] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/07/2024] [Revised: 12/10/2024] [Accepted: 02/11/2025] [Indexed: 03/19/2025] Open
Abstract
Mesenchymal stem/stromal cells (MSCs) are a valuable source of paracrine factors, as they have a remarkable secretory capacity, and there is a sizeable knowledge base to develop industrial and clinical production protocols. Promising cell-free approaches for tissue regeneration and immunomodulation are driving research towards secretome applications, among which extracellular vesicles (EVs) are steadily gaining attention. However, the manufacturing and application of EVs is limited by insufficient yields, knowledge gaps, and low standardization. Facing these limitations, hydrogels represent a versatile three-dimensional (3D) culture platform that can incorporate extracellular matrix (ECM) components to mimic the natural stem cell environment in vitro; via these niche-mimicking properties, hydrogels can regulate MSCs' morphology, adhesion, proliferation, differentiation and secretion capacities. However, the impact of the hydrogel's architectural, biochemical and biomechanical properties on the production of EVs remains poorly understood, as the field is still in its infancy and the interdependency of culture parameters compromises the comparability of the studies. Therefore, this review summarizes and discusses the reported effects of hydrogel encapsulation and culture on the secretion of MSC-EVs. Considering the effects of cell-material interactions on the overall paracrine activity of MSCs, we identify persistent challenges from low standardization and process control, and outline future paths of research, such as the synergic use of hydrogels and bioreactors to enhance MSC-EV generation.
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Affiliation(s)
- Oscar Fabian Garcia‐Aponte
- Department of Biotechnology and Food Science, Institute of Cell and Tissue Culture TechnologiesUniversity of Natural Resources and Life SciencesViennaAustria
| | - Simon Kahlenberg
- Department of Biotechnology and Food Science, Institute of Cell and Tissue Culture TechnologiesUniversity of Natural Resources and Life SciencesViennaAustria
| | - Dimitrios Kouroupis
- Department of Orthopedics, UHealth Sports Medicine Institute, Miller School of MedicineUniversity of MiamiMiamiFloridaUSA
- Diabetes Research Institute & Cell Transplant Center, Miller School of MedicineUniversity of MiamiMiamiFloridaUSA
| | - Dominik Egger
- Institute of Cell Biology and BiophysicsLeibniz University HannoverHannoverGermany
| | - Cornelia Kasper
- Department of Biotechnology and Food Science, Institute of Cell and Tissue Culture TechnologiesUniversity of Natural Resources and Life SciencesViennaAustria
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Maji S, Aliabouzar M, Quesada C, Chiravuri A, Macpherson A, Pinch A, Kazyak K, Emara Z, Abeid BA, Kent RN, Midekssa FS, Zhang M, Baker BM, Franceschi RT, Fabiilli ML. Ultrasound-generated bubbles enhance osteogenic differentiation of mesenchymal stromal cells in composite collagen hydrogels. Bioact Mater 2025; 43:82-97. [PMID: 39345992 PMCID: PMC11439547 DOI: 10.1016/j.bioactmat.2024.09.018] [Citation(s) in RCA: 1] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/23/2024] [Revised: 08/30/2024] [Accepted: 09/13/2024] [Indexed: 10/01/2024] Open
Abstract
Hydrogels can improve the delivery of mesenchymal stromal cells (MSCs) by providing crucial biophysical cues that mimic the extracellular matrix. The differentiation of MSCs is dependent on biophysical cues like stiffness and viscoelasticity, yet conventional hydrogels cannot be dynamically altered after fabrication and implantation to actively direct differentiation. We developed a composite hydrogel, consisting of type I collagen and phase-shift emulsion, where osteogenic differentiation of MSCs can be non-invasively modulated using ultrasound. When exposed to ultrasound, the emulsion within the hydrogel was non-thermally vaporized into bubbles, which locally compacted and stiffened the collagen matrix surrounding each bubble. Bubble growth and matrix compaction were correlated, with collagen regions proximal (i.e., ≤ ∼60 μm) to the bubble displaying a 2.5-fold increase in Young's modulus compared to distal regions (i.e., > ∼60 μm). The viability and proliferation of MSCs, which were encapsulated within the composite hydrogel, were not impacted by bubble formation. In vitro and in vivo studies revealed encapsulated MSCs exhibited significantly elevated levels of RUNX2 and osteocalcin, markers of osteogenic differentiation, in collagen regions proximal to the bubble compared to distal regions. Additionally, alkaline phosphatase activity and calcium deposition were enhanced adjacent to the bubble. An opposite trend was observed for CD90, a marker of MSC stemness. Following subcutaneous implantation, bubbles persisted in the hydrogels for two weeks, which led to localized collagen alignment and increases in nuclear asymmetry. These results are a significant step toward controlling the 3D differentiation of MSCs in a non-invasive and on-demand manner.
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Affiliation(s)
- Somnath Maji
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
| | - Mitra Aliabouzar
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
- Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, USA
| | - Carole Quesada
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
| | - Anjali Chiravuri
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
- Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA
| | - Aidan Macpherson
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
- Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA
| | - Abigail Pinch
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
- Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA
| | - Karsyn Kazyak
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
| | - Ziyad Emara
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
| | - Bachir A Abeid
- Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, USA
| | - Robert N Kent
- Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA
| | - Firaol S Midekssa
- Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA
| | - Man Zhang
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
| | - Brendon M Baker
- Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA
- Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, USA
| | - Renny T Franceschi
- Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA
- Department of Periodontics and Oral Medicine, University of Michigan, Ann Arbor, MI, USA
| | - Mario L Fabiilli
- Department of Radiology, University of Michigan, Ann Arbor, MI, USA
- Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA
- Applied Physics Program, University of Michigan, Ann Arbor, MI, USA
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Kim C, Kang N, Min S, Thangam R, Lee S, Hong H, Kim K, Kim SY, Kim D, Rha H, Tag KR, Lee HJ, Singh N, Jeong D, Hwang J, Kim Y, Park S, Lee H, Kim T, Son SW, Park S, Karamikamkar S, Zhu Y, Hassani Najafabadi A, Chu Z, Sun W, Zhao P, Zhang K, Bian L, Song HC, Park SG, Kim JS, Lee SY, Ahn JP, Kim HK, Zhang YS, Kang H. Modularity-based mathematical modeling of ligand inter-nanocluster connectivity for unraveling reversible stem cell regulation. Nat Commun 2024; 15:10665. [PMID: 39715783 PMCID: PMC11666790 DOI: 10.1038/s41467-024-54557-8] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/01/2023] [Accepted: 11/14/2024] [Indexed: 12/25/2024] Open
Abstract
The native extracellular matrix is continuously remodeled to form complex interconnected network structures that reversibly regulate stem cell behaviors. Both regulation and understanding of its intricate dynamicity can help to modulate numerous cell behaviors. However, neither of these has yet been achieved due to the lack of designing and modeling such complex structures with dynamic controllability. Here we report modularity-based mathematical modeling of extracellular matrix-emulating ligand inter-cluster connectivity using the graph theory. Increasing anisotropy of magnetic nano-blockers proportionately disconnects arginine-glycine-aspartic acid ligand-to-ligand interconnections and decreases the number of ligand inter-cluster edges. This phenomenon deactivates stem cells, which can be partly activated by linearizing the nano-blockers. Remote cyclic elevation of high-anisotropy nano-blockers flexibly generates nano-gaps under the nano-blockers and augments the number of ligand inter-cluster edges. Subsequently, integrin-presenting stem cell infiltration is stimulated, which reversibly intensifies focal adhesion and mechanotransduction-driven differentiation both in vitro and in vivo. Designing and systemically modeling extracellular matrix-mimetic geometries opens avenues for unraveling dynamic cell-material interactions for tissue regeneration.
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Affiliation(s)
- Chowon Kim
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Nayeon Kang
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Sunhong Min
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Ramar Thangam
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Sungkyu Lee
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Hyunsik Hong
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Kanghyeon Kim
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Seong Yeol Kim
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Dahee Kim
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Hyunji Rha
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Kyong-Ryol Tag
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
- Advanced Analysis Center, Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
| | - Hyun-Jeong Lee
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
- Advanced Analysis Center, Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
| | - Nem Singh
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
- Department of Chemistry, Korea University, Seoul, Republic of Korea
| | - Daun Jeong
- Department of Orthopedic Surgery, Korea University Anam Hospital, Seoul, Republic of Korea
| | - Jangsun Hwang
- Department of Orthopedic Surgery, Korea University Anam Hospital, Seoul, Republic of Korea
| | - Yuri Kim
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Sangwoo Park
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea
| | - Hyesung Lee
- Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul, Republic of Korea
| | - Taeeon Kim
- Nano-Bio Convergence Department, Korea Institute of Materials Science (KIMS), Changwon, Gyeongnam, Republic of Korea
- Department of Future Convergence Materials, Korea University, Seoul, Republic of Korea
| | - Sang Wook Son
- Department of Dermatology, Korea University Ansan Hospital, Korea University College of Medicine, Ansan, Republic of Korea
| | - Steve Park
- Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea
| | | | - Yangzhi Zhu
- Terasaki Institute for Biomedical Innovation, Los Angeles, CA, USA
| | | | - Zhiqin Chu
- Department of Electrical and Electronic Engineering, Joint Appointment with School of Biomedical Sciences, The University of Hong Kong, Hong Kong, PR China
| | - Wujin Sun
- Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA, USA
| | - Pengchao Zhao
- School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou, PR China
| | - Kunyu Zhang
- School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou, PR China
| | - Liming Bian
- School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou, PR China
| | - Hyun-Cheol Song
- Electronic Materials Research Center, Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
- KIST-SKKU Carbon-Neutral Research Center, Sungkyunkwan University (SKKU), Suwon, Republic of Korea
| | - Sung-Gyu Park
- Nano-Bio Convergence Department, Korea Institute of Materials Science (KIMS), Changwon, Gyeongnam, Republic of Korea
- Department of Future Convergence Materials, Korea University, Seoul, Republic of Korea
| | - Jong Seung Kim
- Department of Chemistry, Korea University, Seoul, Republic of Korea
| | - Sang-Yup Lee
- Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul, Republic of Korea
| | - Jae-Pyoung Ahn
- Advanced Analysis Center, Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
| | - Hong-Kyu Kim
- Advanced Analysis Center, Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
| | - Yu Shrike Zhang
- Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital Harvard Medical School, Cambridge, MA, USA.
| | - Heemin Kang
- Department of Materials Science and Engineering, Korea University, Seoul, Republic of Korea.
- Department of Future Convergence Materials, Korea University, Seoul, Republic of Korea.
- College of Medicine, Korea University, Seoul, Republic of Korea.
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Barcelona-Estaje E, Oliva MAG, Cunniffe F, Rodrigo-Navarro A, Genever P, Dalby MJ, Roca-Cusachs P, Cantini M, Salmeron-Sanchez M. N-cadherin crosstalk with integrin weakens the molecular clutch in response to surface viscosity. Nat Commun 2024; 15:8824. [PMID: 39394209 PMCID: PMC11479646 DOI: 10.1038/s41467-024-53107-6] [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: 10/10/2023] [Accepted: 09/30/2024] [Indexed: 10/13/2024] Open
Abstract
Mesenchymal stem cells (MSCs) interact with their surroundings via integrins, which link to the actin cytoskeleton and translate physical cues into biochemical signals through mechanotransduction. N-cadherins enable cell-cell communication and are also linked to the cytoskeleton. This crosstalk between integrins and cadherins modulates MSC mechanotransduction and fate. Here we show the role of this crosstalk in the mechanosensing of viscosity using supported lipid bilayers as substrates of varying viscosity. We functionalize these lipid bilayers with adhesion peptides for integrins (RGD) and N-cadherins (HAVDI), to demonstrate that integrins and cadherins compete for the actin cytoskeleton, leading to an altered MSC mechanosensing response. This response is characterised by a weaker integrin adhesion to the environment when cadherin ligation occurs. We model this competition via a modified molecular clutch model, which drives the integrin/cadherin crosstalk in response to surface viscosity, ultimately controlling MSC lineage commitment.
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Affiliation(s)
- Eva Barcelona-Estaje
- Centre for the Cellular Microenvironment, Advanced Research Centre, University of Glasgow, Glasgow, UK
| | - Mariana A G Oliva
- Centre for the Cellular Microenvironment, Advanced Research Centre, University of Glasgow, Glasgow, UK
| | - Finlay Cunniffe
- Centre for the Cellular Microenvironment, Advanced Research Centre, University of Glasgow, Glasgow, UK
| | | | - Paul Genever
- Department of Biology, University of York, York, UK
| | - Matthew J Dalby
- Centre for the Cellular Microenvironment, Advanced Research Centre, University of Glasgow, Glasgow, UK
| | - Pere Roca-Cusachs
- Institute for Bioengineering of Catalonia (IBEC), the Barcelona Institute of Technology (BIST), Barcelona, Spain.
- University of Barcelona, Barcelona, Spain.
| | - Marco Cantini
- Centre for the Cellular Microenvironment, Advanced Research Centre, University of Glasgow, Glasgow, UK.
| | - Manuel Salmeron-Sanchez
- Centre for the Cellular Microenvironment, Advanced Research Centre, University of Glasgow, Glasgow, UK.
- Institute for Bioengineering of Catalonia (IBEC), the Barcelona Institute of Technology (BIST), Barcelona, Spain.
- Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain.
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5
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Arndt T, Chatterjee U, Shilkova O, Francis J, Lundkvist J, Johansson D, Schmuck B, Greco G, Nordberg ÅE, Li Y, Wahlberg LU, Langton M, Johansson J, Götherström C, Rising A. Tuneable Recombinant Spider Silk Protein Hydrogels for Drug Release and 3D Cell Culture. ADVANCED FUNCTIONAL MATERIALS 2024; 34:2303622. [PMID: 39355087 PMCID: PMC11440629 DOI: 10.1002/adfm.202303622] [Citation(s) in RCA: 8] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 04/01/2023] [Revised: 05/10/2023] [Indexed: 10/03/2024]
Abstract
Hydrogels are useful drug release systems and tissue engineering scaffolds. However, synthetic hydrogels often require harsh gelation conditions and can contain toxic by-products while naturally derived hydrogels can transmit pathogens and in general have poor mechanical properties. Thus, there is a need for a hydrogel that forms under ambient conditions, is non-toxic, xeno-free, and has good mechanical properties. A recombinant spider silk protein-derived hydrogel that rapidly forms at 37 °C is recently developed. The temperature and gelation times are well-suited for an injectable in situ polymerising hydrogel, as well as a 3D cell culture scaffold. Here, it is shown that the diffusion rate and the mechanical properties can be tuned by changing the protein concentration and that human fetal mesenchymal stem cells encapsulated in the hydrogels show high survival and viability. Furthermore, mixtures of recombinant spider silk proteins and green fluorescent protein (GFP) form gels from which functional GFP is gradually released, indicating that bioactive molecules are easily included in the gels, maintain activity and can diffuse through the gel. Interestingly, encapsulated ARPE-19 cells are viable and continuously produce the growth factor progranulin, which is detected in the cell culture medium over the study period of 31 days.
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Affiliation(s)
- Tina Arndt
- Department of Biosciences and NutritionKarolinska InstitutetNeoHuddinge14152Sweden
| | - Urmimala Chatterjee
- Department of Biosciences and NutritionKarolinska InstitutetNeoHuddinge14152Sweden
| | - Olga Shilkova
- Department of Biosciences and NutritionKarolinska InstitutetNeoHuddinge14152Sweden
| | - Juanita Francis
- Department of Biosciences and NutritionKarolinska InstitutetNeoHuddinge14152Sweden
| | | | - Daniel Johansson
- Department of Molecular SciencesSwedish University of Agricultural SciencesUppsala75007Sweden
| | - Benjamin Schmuck
- Department of Biosciences and NutritionKarolinska InstitutetNeoHuddinge14152Sweden
- Department of AnatomyPhysiology and BiochemistrySwedish University of Agricultural SciencesUppsala75007Sweden
| | - Gabriele Greco
- Department of AnatomyPhysiology and BiochemistrySwedish University of Agricultural SciencesUppsala75007Sweden
| | - Åsa Ekblad Nordberg
- Department of Clinical ScienceIntervention and TechnologyDivision of Obstetrics and GynecologyKarolinska InstitutetHuddinge14152Sweden
| | - Yan Li
- Department of Clinical ScienceIntervention and TechnologyDivision of Orthopedics and BiotechnologyKarolinska UniversitetssjukhusetHuddinge141 86Sweden
| | | | - Maud Langton
- Department of Molecular SciencesSwedish University of Agricultural SciencesUppsala75007Sweden
| | - Jan Johansson
- Department of Biosciences and NutritionKarolinska InstitutetNeoHuddinge14152Sweden
| | - Cecilia Götherström
- Department of Clinical ScienceIntervention and TechnologyDivision of Obstetrics and GynecologyKarolinska InstitutetHuddinge14152Sweden
| | - Anna Rising
- Department of Biosciences and NutritionKarolinska InstitutetNeoHuddinge14152Sweden
- Department of AnatomyPhysiology and BiochemistrySwedish University of Agricultural SciencesUppsala75007Sweden
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6
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Ferjaoui Z, López-Muñoz R, Akbari S, Chandad F, Mantovani D, Rouabhia M, D. Fanganiello R. Design of Alginate/Gelatin Hydrogels for Biomedical Applications: Fine-Tuning Osteogenesis in Dental Pulp Stem Cells While Preserving Other Cell Behaviors. Biomedicines 2024; 12:1510. [PMID: 39062083 PMCID: PMC11274465 DOI: 10.3390/biomedicines12071510] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/02/2024] [Revised: 07/03/2024] [Accepted: 07/05/2024] [Indexed: 07/28/2024] Open
Abstract
Alginate/gelatin (Alg-Gel) hydrogels have been used experimentally, associated with mesenchymal stromal/stem cells (MSCs), to guide bone tissue formation. One of the main challenges for clinical application is optimizing Alg-Gel stiffness to guide osteogenesis. In this study, we investigated how Alg-Gel stiffness could modulate the dental pulp stem cell (DPSC) attachment, morphology, proliferation, and osteogenic differentiation, identifying the optimal conditions to uncouple osteogenesis from the other cell behaviors. An array of Alg-Gel hydrogels was prepared by casting different percentages of alginate and gelatin cross-linked with 2% CaCl2. We have selected two hydrogels: one with a stiffness of 11 ± 1 kPa, referred to as "low-stiffness hydrogel", formed by 2% alginate and 8% gelatin, and the other with a stiffness of 55 ± 3 kPa, referred to as "high-stiffness hydrogel", formed by 8% alginate and 12% gelatin. Hydrogel analyses showed that the average swelling rates were 20 ± 3% for the low-stiffness hydrogels and 35 ± 2% for the high-stiffness hydrogels. The degradation percentage was 47 ± 5% and 18 ± 2% for the low- and high-stiffness hydrogels, respectively. Both hydrogel types showed homogeneous surface shape and protein (Alg-Gel) interaction with CaCl2 as assessed by physicochemical characterization. Cell culture showed good adhesion of the DPSCs to the hydrogels and proliferation. Furthermore, better osteogenic activity, determined by ALP activity and ARS staining, was obtained with high-stiffness hydrogels (8% alginate and 12% gelatin). In summary, this study confirms the possibility of characterizing and optimizing the stiffness of Alg-Gel gel to guide osteogenesis in vitro without altering the other cellular properties of DPSCs.
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Affiliation(s)
- Zied Ferjaoui
- Oral Ecology Research Group (GREB), Faculté de Médecine Dentaire, Université Laval, Québec City, QC G1V 0A6, Canada; (F.C.); (M.R.); (R.D.F.)
| | - Roberto López-Muñoz
- Laboratory for Biomaterials and Bioengineering, (CRC-Tier I), Department of Min-Met-Materials Eng and Regenerative Medicine, CHU de Quebec, Laval University, Quebec City, QC G1V 0A6, Canada; (R.L.-M.); (D.M.)
| | - Soheil Akbari
- Département de Génie Chimique, Université Laval, Québec City, QC G1V 0A6, Canada;
| | - Fatiha Chandad
- Oral Ecology Research Group (GREB), Faculté de Médecine Dentaire, Université Laval, Québec City, QC G1V 0A6, Canada; (F.C.); (M.R.); (R.D.F.)
| | - Diego Mantovani
- Laboratory for Biomaterials and Bioengineering, (CRC-Tier I), Department of Min-Met-Materials Eng and Regenerative Medicine, CHU de Quebec, Laval University, Quebec City, QC G1V 0A6, Canada; (R.L.-M.); (D.M.)
| | - Mahmoud Rouabhia
- Oral Ecology Research Group (GREB), Faculté de Médecine Dentaire, Université Laval, Québec City, QC G1V 0A6, Canada; (F.C.); (M.R.); (R.D.F.)
| | - Roberto D. Fanganiello
- Oral Ecology Research Group (GREB), Faculté de Médecine Dentaire, Université Laval, Québec City, QC G1V 0A6, Canada; (F.C.); (M.R.); (R.D.F.)
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7
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Lu P, Ruan D, Huang M, Tian M, Zhu K, Gan Z, Xiao Z. Harnessing the potential of hydrogels for advanced therapeutic applications: current achievements and future directions. Signal Transduct Target Ther 2024; 9:166. [PMID: 38945949 PMCID: PMC11214942 DOI: 10.1038/s41392-024-01852-x] [Citation(s) in RCA: 74] [Impact Index Per Article: 74.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/19/2023] [Revised: 04/02/2024] [Accepted: 04/28/2024] [Indexed: 07/02/2024] Open
Abstract
The applications of hydrogels have expanded significantly due to their versatile, highly tunable properties and breakthroughs in biomaterial technologies. In this review, we cover the major achievements and the potential of hydrogels in therapeutic applications, focusing primarily on two areas: emerging cell-based therapies and promising non-cell therapeutic modalities. Within the context of cell therapy, we discuss the capacity of hydrogels to overcome the existing translational challenges faced by mainstream cell therapy paradigms, provide a detailed discussion on the advantages and principal design considerations of hydrogels for boosting the efficacy of cell therapy, as well as list specific examples of their applications in different disease scenarios. We then explore the potential of hydrogels in drug delivery, physical intervention therapies, and other non-cell therapeutic areas (e.g., bioadhesives, artificial tissues, and biosensors), emphasizing their utility beyond mere delivery vehicles. Additionally, we complement our discussion on the latest progress and challenges in the clinical application of hydrogels and outline future research directions, particularly in terms of integration with advanced biomanufacturing technologies. This review aims to present a comprehensive view and critical insights into the design and selection of hydrogels for both cell therapy and non-cell therapies, tailored to meet the therapeutic requirements of diverse diseases and situations.
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Affiliation(s)
- Peilin Lu
- Nanomedicine Research Center, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, PR China
- Department of Minimally Invasive Interventional Radiology, and Laboratory of Interventional Radiology, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510260, PR China
| | - Dongxue Ruan
- State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, National Center for Respiratory Medicine, Department of Respiratory and Critical Care Medicine, Guangzhou Institute for Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510120, PR China
| | - Meiqi Huang
- Department of Minimally Invasive Interventional Radiology, and Laboratory of Interventional Radiology, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510260, PR China
| | - Mi Tian
- Department of Stomatology, Chengdu Second People's Hospital, Chengdu, 610021, PR China
| | - Kangshun Zhu
- Department of Minimally Invasive Interventional Radiology, and Laboratory of Interventional Radiology, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510260, PR China.
| | - Ziqi Gan
- Hospital of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, 510055, PR China.
| | - Zecong Xiao
- Nanomedicine Research Center, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, PR China.
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8
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Ghagre A, Delarue A, Srivastava LK, Koushki N, Ehrlicher A. Nuclear curvature determines Yes-associated protein localization and differentiation of mesenchymal stem cells. Biophys J 2024; 123:1222-1239. [PMID: 38605521 PMCID: PMC11140468 DOI: 10.1016/j.bpj.2024.04.008] [Citation(s) in RCA: 2] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/19/2023] [Revised: 01/17/2024] [Accepted: 04/08/2024] [Indexed: 04/13/2024] Open
Abstract
Controlling mesenchymal stem cell (MSC) differentiation remains a critical challenge in MSCs' therapeutic application. Numerous biophysical and mechanical stimuli influence stem cell fate; however, their relative efficacy and specificity in mechanically directed differentiation remain unclear. Yes-associated protein (YAP) is one key mechanosensitive protein that controls MSC differentiation. Previous studies have related nuclear mechanics with YAP activity, but we still lack an understanding of what nuclear deformation specifically regulates YAP and its relationship with mechanical stimuli. Here, we report that maximum nuclear curvature is the most precise biophysical determinant for YAP mechanotransduction-mediated MSC differentiation and is a relevant parameter for stem cell-based therapies. We employed traction force microscopy and confocal microscopy to characterize the causal relationships between contractility and nuclear deformation in regulating YAP activity in MSCs. We observed that an increase in contractility compresses nuclei anisotropically, whereby the degree of asymmetric compression increased the bending curvature of the nuclear membrane. We then examined membrane curvature and tension using thin micropatterned adhesive substrate lines and an FRET-based tension sensor, revealing the direct role of curvature in YAP activity driven by both active and passive nuclear import. Finally, we employed micropatterned lines to control nuclear curvature and precisely direct MSC differentiation. This work illustrates that nuclear curvature subsumes other biophysical aspects to control YAP-mediated differentiation in MSCs and may provide a deterministic solution to some of the challenges in mesenchymal stem cell therapies.
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Affiliation(s)
- Ajinkya Ghagre
- Department of Bioengineering, McGill University, Montreal, Canada
| | - Alice Delarue
- Department of Bioengineering, McGill University, Montreal, Canada
| | | | - Newsha Koushki
- Department of Bioengineering, McGill University, Montreal, Canada
| | - Allen Ehrlicher
- Department of Bioengineering, McGill University, Montreal, Canada; Department of Anatomy and Cell Biology, McGill University, Montreal, Canada; Department of Biomedical Engineering, McGill University, Montreal, Canada; Department of Mechanical Engineering, McGill University, Montreal, Canada; Rosalind and Morris Goodman Cancer Research Institute, McGill University, Montreal, Canada; Centre for Structural Biology, McGill University, Montreal, Canada.
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9
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Quan Y, Li J, Cai J, Liao Y, Zhang Y, Lu F. Transplantation of beige adipose organoids fabricated using adipose acellular matrix hydrogel improves metabolic dysfunction in high-fat diet-induced obesity and type 2 diabetes mice. J Cell Physiol 2024; 239:e31191. [PMID: 38219044 DOI: 10.1002/jcp.31191] [Citation(s) in RCA: 2] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/25/2023] [Revised: 12/22/2023] [Accepted: 12/28/2023] [Indexed: 01/15/2024]
Abstract
Transplantation of brown adipose tissue (BAT) is a promising approach for treating obesity and metabolic disorders. However, obtaining sufficient amounts of functional BAT or brown adipocytes for transplantation remains a major challenge. In this study, we developed a hydrogel that combining adipose acellular matrix (AAM) and GelMA and HAMA that can be adjusted for stiffness by modulating the duration of light-crosslinking. We used human white adipose tissue-derived microvascular fragments to create beige adipose organoids (BAO) that were encapsulated in either a soft or stiff AAM hydrogel. We found that BAOs cultivated in AAM hydrogels with high stiffness demonstrated increased metabolic activity and upregulation of thermogenesis-related genes. When transplanted into obese and type 2 diabetes mice, the HFD + BAO group showed sustained improvements in metabolic rate, resulting in significant weight loss and decreased blood glucose levels. Furthermore, the mice showed a marked reduction in nonalcoholic liver steatosis, indicating improved liver function. In contrast, transplantation of 2D-cultured beige adipocytes failed to produce these beneficial effects. Our findings demonstrate the feasibility of fabricating beige adipose organoids in vitro and administering them by injection, which may represent a promising therapeutic approach for obesity and diabetes.
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Affiliation(s)
- Yuping Quan
- Department of Plastic and Cosmetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, P. R. China
- Department of Plastic Surgery and Regenerative Medicine, Fujian Medical University Union Hospital, Fuzhou, China
| | - Jian Li
- Department of Plastic and Cosmetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, P. R. China
| | - Junrong Cai
- Department of Plastic and Cosmetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, P. R. China
| | - Yunjun Liao
- Department of Plastic and Cosmetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, P. R. China
| | - Yuteng Zhang
- Department of Plastic and Cosmetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, P. R. China
| | - Feng Lu
- Department of Plastic and Cosmetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, P. R. China
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10
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Ding L, Oh S, Shrestha J, Lam A, Wang Y, Radfar P, Warkiani ME. Scaling up stem cell production: harnessing the potential of microfluidic devices. Biotechnol Adv 2023; 69:108271. [PMID: 37844769 DOI: 10.1016/j.biotechadv.2023.108271] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/29/2023] [Revised: 10/08/2023] [Accepted: 10/13/2023] [Indexed: 10/18/2023]
Abstract
Stem cells are specialised cells characterised by their unique ability to both self-renew and transform into a wide array of specialised cell types. The widespread interest in stem cells for regenerative medicine and cultivated meat has led to a significant demand for these cells in both research and practical applications. Despite the growing need for stem cell manufacturing, the industry faces significant obstacles, including high costs for equipment and maintenance, complicated operation, and low product quality and yield. Microfluidic technology presents a promising solution to the abovementioned challenges. As an innovative approach for manipulating liquids and cells within microchannels, microfluidics offers a plethora of advantages at an industrial scale. These benefits encompass low setup costs, ease of operation and multiplexing, minimal energy consumption, and the added advantage of being labour-free. This review presents a thorough examination of the prominent microfluidic technologies employed in stem cell research and explores their promising applications in the burgeoning stem cell industry. It thoroughly examines how microfluidics can enhance cell harvesting from tissue samples, facilitate mixing and cryopreservation, streamline microcarrier production, and efficiently conduct cell separation, purification, washing, and final cell formulation post-culture.
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Affiliation(s)
- Lin Ding
- Smart MCs Pty Ltd, Ultimo, Sydney, 2007, Australia.
| | - Steve Oh
- Stem Cell Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore, 138668, Singapore
| | - Jesus Shrestha
- School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW 2007, Australia
| | - Alan Lam
- Stem Cell Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), Singapore, 138668, Singapore
| | - Yaqing Wang
- School of Biomedical Engineering, University of Science and Technology of China, Hefei 230026, China; Suzhou Institute for Advanced Research, University of Science and Technology of China, Suzhou 215123, China
| | - Payar Radfar
- Smart MCs Pty Ltd, Ultimo, Sydney, 2007, Australia
| | - Majid Ebrahimi Warkiani
- School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW 2007, Australia..
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11
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Lu C, Zheng J, Yoshitomi T, Kawazoe N, Yang Y, Chen G. How Hydrogel Stiffness Affects Adipogenic Differentiation of Mesenchymal Stem Cells under Controlled Morphology. ACS APPLIED BIO MATERIALS 2023; 6:3441-3450. [PMID: 37061939 DOI: 10.1021/acsabm.3c00159] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 04/17/2023]
Abstract
Matrix stiffness has been disclosed as an essential regulator of cell fate. However, it is barely studied how the matrix stiffness affects stem cell functions when cell morphology changes. Thus, in this study, the effect of hydrogel stiffness on adipogenic differentiation of human bone-marrow-derived mesenchymal stem cells (hMSCs) with controlled morphology was investigated. Micropatterns of different size and elongation were prepared by a photolithographical micropatterning technique. The hMSCs were cultured on the micropatterns and showed a different spreading area and elongation following the geometry of the underlying micropatterns. The cells with controlled morphology were embedded in agarose hydrogels of different stiffnesses. The cells showed a different level of adipogenic differentiation that was dependent on both hydrogel stiffness and cell morphology. Adipogenic differentiation became strong when the cell spreading area decreased and hydrogel stiffness increased. Adipogenic differentiation did not change with cell elongation. Therefore, cell spreading area and hydrogel stiffness could synergistically affect adipogenic differentiation of hMSCs, while cell elongation did not affect adipogenic differentiation. A change of cell morphology and hydrogel stiffness was accompanied by actin filament alignment that was strongly related to adipogenic differentiation. The results indicated that cell morphology could affect cellular sensitivity to hydrogel stiffness. The results will provide useful information for the elucidation of the interaction of stem cells and their microenvironmental biomechanical cues.
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Affiliation(s)
- Chengyu Lu
- Research Center for Functional Materials, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan
- Department of Materials Science and Engineering, Graduate School of Pure and Applied Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8577, Japan
| | - Jing Zheng
- Research Center for Functional Materials, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan
| | - Toru Yoshitomi
- Research Center for Functional Materials, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan
| | - Naoki Kawazoe
- Research Center for Functional Materials, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan
| | - Yingnan Yang
- Graduate School of Life and Environment Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan
| | - Guoping Chen
- Research Center for Functional Materials, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan
- Department of Materials Science and Engineering, Graduate School of Pure and Applied Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8577, Japan
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12
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Carballo-Pedrares N, Ponti F, Lopez-Seijas J, Miranda-Balbuena D, Bono N, Candiani G, Rey-Rico A. Non-viral gene delivery to human mesenchymal stem cells: a practical guide towards cell engineering. J Biol Eng 2023; 17:49. [PMID: 37491322 PMCID: PMC10369726 DOI: 10.1186/s13036-023-00363-7] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/13/2023] [Accepted: 06/27/2023] [Indexed: 07/27/2023] Open
Abstract
In recent decades, human mesenchymal stem cells (hMSCs) have gained momentum in the field of cell therapy for treating cartilage and bone injuries. Despite the tri-lineage multipotency, proliferative properties, and potent immunomodulatory effects of hMSCs, their clinical potential is hindered by donor variations, limiting their use in medical settings. To address this challenge, gene delivery technologies have emerged as a promising approach to modulate the phenotype and commitment of hMSCs towards specific cell lineages, thereby enhancing osteochondral repair strategies. This review provides a comprehensive overview of current non-viral gene delivery approaches used to engineer MSCs, highlighting key factors such as the choice of nucleic acid or delivery vector, transfection strategies, and experimental parameters. Additionally, it outlines various protocols and methods for qualitative and quantitative evaluation of their therapeutic potential as a delivery system in osteochondral regenerative applications. In summary, this technical review offers a practical guide for optimizing non-viral systems in osteochondral regenerative approaches. hMSCs constitute a key target population for gene therapy techniques. Nevertheless, there is a long way to go for their translation into clinical treatments. In this review, we remind the most relevant transfection conditions to be optimized, such as the type of nucleic acid or delivery vector, the transfection strategy, and the experimental parameters to accurately evaluate a delivery system. This survey provides a practical guide to optimizing non-viral systems for osteochondral regenerative approaches.
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Affiliation(s)
- Natalia Carballo-Pedrares
- Gene & Cell Therapy Research Group (G-CEL). Centro Interdisciplinar de Química y Biología - CICA, Universidade da Coruña, As Carballeiras, S/N. Campus de Elviña, 15071 A, Coruña, Spain
| | - Federica Ponti
- genT_LΛB, Department of Chemistry, Materials and Chemical Engineering "G. Natta", Politecnico Di Milano, 20131, Milan, Italy
- Laboratory for Biomaterials and Bioengineering, Canada Research Chair I in Biomaterials and Bioengineering for the Innovation in Surgery, Department of Min-Met-Materials Engineering & Research Center of CHU de Quebec, Division of Regenerative Medicine, Laval University, Quebec City, QC, Canada
| | - Junquera Lopez-Seijas
- Gene & Cell Therapy Research Group (G-CEL). Centro Interdisciplinar de Química y Biología - CICA, Universidade da Coruña, As Carballeiras, S/N. Campus de Elviña, 15071 A, Coruña, Spain
| | - Diego Miranda-Balbuena
- Gene & Cell Therapy Research Group (G-CEL). Centro Interdisciplinar de Química y Biología - CICA, Universidade da Coruña, As Carballeiras, S/N. Campus de Elviña, 15071 A, Coruña, Spain
| | - Nina Bono
- genT_LΛB, Department of Chemistry, Materials and Chemical Engineering "G. Natta", Politecnico Di Milano, 20131, Milan, Italy
| | - Gabriele Candiani
- genT_LΛB, Department of Chemistry, Materials and Chemical Engineering "G. Natta", Politecnico Di Milano, 20131, Milan, Italy.
| | - Ana Rey-Rico
- Gene & Cell Therapy Research Group (G-CEL). Centro Interdisciplinar de Química y Biología - CICA, Universidade da Coruña, As Carballeiras, S/N. Campus de Elviña, 15071 A, Coruña, Spain.
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13
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Zhang Y, Wang Z, Sun Q, Li Q, Li S, Li X. Dynamic Hydrogels with Viscoelasticity and Tunable Stiffness for the Regulation of Cell Behavior and Fate. MATERIALS (BASEL, SWITZERLAND) 2023; 16:5161. [PMID: 37512435 PMCID: PMC10386333 DOI: 10.3390/ma16145161] [Citation(s) in RCA: 15] [Impact Index Per Article: 7.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/22/2023] [Revised: 07/16/2023] [Accepted: 07/19/2023] [Indexed: 07/30/2023]
Abstract
The extracellular matrix (ECM) of natural cells typically exhibits dynamic mechanical properties (viscoelasticity and dynamic stiffness). The viscoelasticity and dynamic stiffness of the ECM play a crucial role in biological processes, such as tissue growth, development, physiology, and disease. Hydrogels with viscoelasticity and dynamic stiffness have recently been used to investigate the regulation of cell behavior and fate. This article first emphasizes the importance of tissue viscoelasticity and dynamic stiffness and provides an overview of characterization techniques at both macro- and microscale. Then, the viscoelastic hydrogels (crosslinked via ion bonding, hydrogen bonding, hydrophobic interactions, and supramolecular interactions) and dynamic stiffness hydrogels (softening, stiffening, and reversible stiffness) with different crosslinking strategies are summarized, along with the significant impact of viscoelasticity and dynamic stiffness on cell spreading, proliferation, migration, and differentiation in two-dimensional (2D) and three-dimensional (3D) cell cultures. Finally, the emerging trends in the development of dynamic mechanical hydrogels are discussed.
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Affiliation(s)
- Yuhang Zhang
- School of Mechanics and Safety Engineering, Zhengzhou University, Zhengzhou 450001, China (Q.L.)
- National Center for International Joint Research of Micro-Nano Moulding Technology, Zhengzhou University, Zhengzhou 450001, China
| | - Zhuofan Wang
- School of Mechanics and Safety Engineering, Zhengzhou University, Zhengzhou 450001, China (Q.L.)
- National Center for International Joint Research of Micro-Nano Moulding Technology, Zhengzhou University, Zhengzhou 450001, China
| | - Qingqing Sun
- School of Materials Science and Engineering, Zhengzhou University, Zhengzhou 450001, China
| | - Qian Li
- School of Mechanics and Safety Engineering, Zhengzhou University, Zhengzhou 450001, China (Q.L.)
- National Center for International Joint Research of Micro-Nano Moulding Technology, Zhengzhou University, Zhengzhou 450001, China
| | - Shaohui Li
- School of Materials Science and Engineering, Zhengzhou University, Zhengzhou 450001, China
| | - Xiaomeng Li
- School of Mechanics and Safety Engineering, Zhengzhou University, Zhengzhou 450001, China (Q.L.)
- National Center for International Joint Research of Micro-Nano Moulding Technology, Zhengzhou University, Zhengzhou 450001, China
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14
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Zhang J, Zeng Z, Chen Y, Deng L, Zhang Y, Que Y, Jiao Y, Chang J, Dong Z, Yang C. 3D-printed GelMA/CaSiO 3 composite hydrogel scaffold for vascularized adipose tissue restoration. Regen Biomater 2023; 10:rbad049. [PMID: 37274616 PMCID: PMC10234763 DOI: 10.1093/rb/rbad049] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/24/2023] [Revised: 03/24/2023] [Accepted: 04/23/2023] [Indexed: 06/06/2023] Open
Abstract
The increased number of mastectomies, combined with rising patient expectations for cosmetic and psychosocial outcomes, has necessitated the use of adipose tissue restoration techniques. However, the therapeutic effect of current clinical strategies is not satisfying due to the high demand of personalized customization and the timely vascularization in the process of adipose regeneration. Here, a composite hydrogel scaffold was prepared by three-dimensional (3D) printing technology, applying gelatin methacrylate anhydride (GelMA) as printing ink and calcium silicate (CS) bioceramic as an active ingredient for breast adipose tissue regeneration. The in vitro experiments showed that the composite hydrogel scaffolds could not only be customized with controllable architectures, but also significantly stimulated both 3T3-L1 preadipocytes and human umbilical vein endothelial cells in multiple cell behaviors, including cell adhesion, proliferation, migration and differentiation. Moreover, the composite scaffold promoted vascularized adipose tissue restoration under the skin of nude mice in vivo. These findings suggest that 3D-printed GelMA/CS composite scaffolds might be a good candidate for adipose tissue engineering.
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Affiliation(s)
| | | | - Yanxin Chen
- Joint Centre of Translational Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
- Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou 325000, China
| | - Li Deng
- Joint Centre of Translational Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
- Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou 325000, China
| | - Yanxin Zhang
- Joint Centre of Translational Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
- Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou 325000, China
| | - Yumei Que
- Joint Centre of Translational Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
- Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou 325000, China
| | - Yiren Jiao
- Joint Centre of Translational Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
- Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou 325000, China
| | - Jiang Chang
- Correspondence address. E-mail: (J.C.); (Z.D.); (C.Y.)
| | - Zhihong Dong
- Correspondence address. E-mail: (J.C.); (Z.D.); (C.Y.)
| | - Chen Yang
- Correspondence address. E-mail: (J.C.); (Z.D.); (C.Y.)
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15
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Elango J, Lijnev A, Zamora-Ledezma C, Alexis F, Wu W, Marín JMG, Sanchez de Val JEM. The Relationship of Rheological Properties and the Performance of Silk Fibroin Hydrogels in Tissue Engineering Application. Process Biochem 2022. [DOI: 10.1016/j.procbio.2022.12.012] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/24/2022]
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16
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Ze Y, Wang R, Deng H, Zhou Z, Chen X, Huang L, Yao Y. Three-dimensional bioprinting: A cutting-edge tool for designing and fabricating engineered living materials. BIOMATERIALS ADVANCES 2022; 140:213053. [PMID: 35964390 DOI: 10.1016/j.bioadv.2022.213053] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/28/2022] [Revised: 07/12/2022] [Accepted: 07/26/2022] [Indexed: 06/15/2023]
Abstract
The design of engineered living materials (ELMs) is an emerging field developed from synthetic biology and materials science principles. ELMs are multi-scale bulk materials that combine the properties of self-healing and organism adaptability with the designed physicochemical or mechanical properties for functional applications in various fields, including therapy, electronics, and architecture. Among the many ELM design and manufacturing methods, three-dimensional (3D) bioprinting stands out for its precise control over the structure of the fabricated constructs and the spatial distribution of cells. In this review, we summarize the progress in the field, cell type and material selection, and the latest applications of 3D bioprinting to manufacture ELMs, as well as their advantages and limitations, hoping to deepen our understanding and provide new insights into ELM design. We believe that 3D bioprinting will become an important development direction and provide more contributions to this field.
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Affiliation(s)
- Yiting Ze
- State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Ruixin Wang
- State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Hanzhi Deng
- State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Zheqing Zhou
- State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Xiaoju Chen
- State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Linyang Huang
- State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Yang Yao
- State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China.
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17
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Fuentes P, Torres MJ, Arancibia R, Aulestia F, Vergara M, Carrión F, Osses N, Altamirano C. Dynamic Culture of Mesenchymal Stromal/Stem Cell Spheroids and Secretion of Paracrine Factors. Front Bioeng Biotechnol 2022; 10:916229. [PMID: 36046670 PMCID: PMC9421039 DOI: 10.3389/fbioe.2022.916229] [Citation(s) in RCA: 23] [Impact Index Per Article: 7.7] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/08/2022] [Accepted: 06/01/2022] [Indexed: 11/13/2022] Open
Abstract
In recent years, conditioned medium (CM) obtained from the culture of mesenchymal stromal/stem cells (MSCs) has been shown to effectively promote tissue repair and modulate the immune response in vitro and in different animal models, with potential for application in regenerative medicine. Using CM offers multiple advantages over the implantation of MSCs themselves: 1) simpler storage, transport, and preservation requirements, 2) avoidance of the inherent risks of cell transplantation, and 3) potential application as a ready-to-go biologic product. For these reasons, a large amount of MSCs research has focused on the characterization of the obtained CM, including soluble trophic factors and vesicles, preconditioning strategies for enhancing paracrine secretion, such as hypoxia, a three-dimensional (3D) environment, and biochemical stimuli, and potential clinical applications. In vitro preconditioning strategies can increase the viability, proliferation, and paracrine properties of MSCs and therefore improve the therapeutic potential of the cells and their derived products. Specifically, dynamic cultivation conditions, such as fluid flow and 3D aggregate culture, substantially impact cellular behaviour. Increased levels of growth factors and cytokines were observed in 3D cultures of MSC grown on orbital or rotatory shaking platforms, in stirred systems, such as spinner flasks or stirred tank reactors, and in microgravity bioreactors. However, only a few studies have established dynamic culture conditions and protocols for 3D aggregate cultivation of MSCs as a scalable and reproducible strategy for CM production. This review summarizes significant advances into the upstream processing, mainly the dynamic generation and cultivation of MSC aggregates, for de CM manufacture and focuses on the standardization of the soluble factor production.
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Affiliation(s)
- Paloma Fuentes
- Escuela de Ingeniería Bioquímica, Facultad de Ingeniería, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile
| | - María José Torres
- Escuela de Ingeniería Bioquímica, Facultad de Ingeniería, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile
| | - Rodrigo Arancibia
- Cellus Medicina Regenerativa S.A., Santiago, Chile
- Cellus Biomédica, Parque Tecnológico de León, León, Spain
| | - Francisco Aulestia
- Cellus Medicina Regenerativa S.A., Santiago, Chile
- Cellus Biomédica, Parque Tecnológico de León, León, Spain
| | - Mauricio Vergara
- Escuela de Ingeniería Bioquímica, Facultad de Ingeniería, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile
| | - Flavio Carrión
- Cellus Medicina Regenerativa S.A., Santiago, Chile
- Departamento de Investigación, Postgrado y Educación Continua (DIPEC), Facultad de Ciencias de la Salud, Universidad del Alba, Santiago, Chile
| | - Nelson Osses
- Instituto de Química, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile
| | - Claudia Altamirano
- Escuela de Ingeniería Bioquímica, Facultad de Ingeniería, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile
- CREAS, Centro Regional de Estudios en Alimentos Saludables, Valparaíso, Chile
- *Correspondence: Claudia Altamirano,
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18
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Evaluation of a Novel Thiol–Norbornene-Functionalized Gelatin Hydrogel for Bioprinting of Mesenchymal Stem Cells. Int J Mol Sci 2022; 23:ijms23147939. [PMID: 35887286 PMCID: PMC9321464 DOI: 10.3390/ijms23147939] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/17/2022] [Revised: 07/13/2022] [Accepted: 07/16/2022] [Indexed: 12/04/2022] Open
Abstract
Introduction: Three-dimensional bioprinting can be considered as an advancement of the classical tissue engineering concept. For bioprinting, cells have to be dispersed in hydrogels. Recently, a novel semi-synthetic thiolene hydrogel system based on norbornene-functionalized gelatin (GelNB) and thiolated gelatin (GelS) was described that resulted in the photoclick hydrogel GelNB/GelS. In this study, we evaluated the printability and biocompatibility of this hydrogel system towards adipose-tissue-derived mesenchymal stem cells (ASCs). Methods: GelNB/GelS was synthesized with three different crosslinking densities (low, medium and high), resulting in different mechanical properties with moduli of elasticity between 206 Pa and 1383 Pa. These hydrogels were tested for their biocompatibility towards ASCs in terms of their viability, proliferation and differentiation. The extrusion-based bioprinting of ASCs in GelNB/GelS-high was performed to manufacture three-dimensional cubic constructs. Results: All three hydrogels supported the viability, proliferation and chondrogenic differentiation of ASCs to a similar extent. The adipogenic differentiation of ASCs was better supported by the softer hydrogel (GelNB/GelS-low), whereas the osteogenic differentiation was more pronounced in the harder hydrogel (GelNB/GelS-high), indicating that the differentiation fate of ASCs can be influenced via the adaption of the mechanical properties of the GelNB/GelS system. After the ex vivo chondrogenic differentiation and subcutaneous implantation of the bioprinted construct into immunocompromised mice, the production of negatively charged sulfated proteoglycans could be observed with only minimal inflammatory signs in the implanted material. Conclusions: Our results indicate that the GelNB/GelS hydrogels are very well suited for the bioprinting of ASCs and may represent attractive hydrogels for subsequent in vivo tissue engineering applications.
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De Belly H, Paluch EK, Chalut KJ. Interplay between mechanics and signalling in regulating cell fate. Nat Rev Mol Cell Biol 2022; 23:465-480. [PMID: 35365816 DOI: 10.1038/s41580-022-00472-z] [Citation(s) in RCA: 111] [Impact Index Per Article: 37.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 03/04/2022] [Indexed: 12/11/2022]
Abstract
Mechanical signalling affects multiple biological processes during development and in adult organisms, including cell fate transitions, cell migration, morphogenesis and immune responses. Here, we review recent insights into the mechanisms and functions of two main routes of mechanical signalling: outside-in mechanical signalling, such as mechanosensing of substrate properties or shear stresses; and mechanical signalling regulated by the physical properties of the cell surface itself. We discuss examples of how these two classes of mechanical signalling regulate stem cell function, as well as developmental processes in vivo. We also discuss how cell surface mechanics affects intracellular signalling and, in turn, how intracellular signalling controls cell surface mechanics, generating feedback into the regulation of mechanosensing. The cooperation between mechanosensing, intracellular signalling and cell surface mechanics has a profound impact on biological processes. We discuss here our understanding of how these three elements interact to regulate stem cell fate and development.
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Affiliation(s)
- Henry De Belly
- Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK
- Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA
- Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA, USA
| | - Ewa K Paluch
- Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK.
| | - Kevin J Chalut
- Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK.
- Wellcome/MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK.
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20
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Allahyari Z, Casillo SM, Perry SJ, Peredo AP, Gholizadeh S, Gaborski TR. Disrupted Surfaces of Porous Membranes Reduce Nuclear YAP Localization and Enhance Adipogenesis through Morphological Changes. ACS Biomater Sci Eng 2022; 8:1791-1798. [PMID: 35363465 DOI: 10.1021/acsbiomaterials.1c01472] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
The disrupted surface of porous membranes, commonly used in tissue-chip and cellular coculture systems, is known to weaken cell-substrate interactions. Here, we investigated whether disrupted surfaces of membranes with micron and submicron scale pores affect yes-associated protein (YAP) localization and differentiation of adipose-derived stem cells. We found that these substrates reduce YAP nuclear localization through decreased cell spreading, consistent with reduced cell-substrate interactions, and in turn enhance adipogenesis while decreasing osteogenesis.
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Affiliation(s)
- Zahra Allahyari
- Department of Microsystems Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States.,Department of Biomedical Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States
| | - Stephanie M Casillo
- Department of Biomedical Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States
| | - Spencer J Perry
- Department of Biomedical Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States
| | - Ana P Peredo
- Department of Biomedical Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States
| | - Shayan Gholizadeh
- Department of Microsystems Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States.,Department of Biomedical Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States
| | - Thomas R Gaborski
- Department of Biomedical Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States
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21
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Khalvandi A, Saber-Samandari S, Aghdam MM. Application of artificial neural networks to predict Young's moduli of cartilage scaffolds: An in-vitro and micromechanical study. BIOMATERIALS ADVANCES 2022; 136:212768. [PMID: 35929308 DOI: 10.1016/j.bioadv.2022.212768] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/22/2022] [Revised: 03/09/2022] [Accepted: 03/14/2022] [Indexed: 06/15/2023]
Abstract
In this study, four-phase Gelatin-Polypyrrole-Akermanite-Magnetite scaffolds were fabricated and analyzed using in-vitro tests and numerical simulations. Such scaffolds contained various amounts of Magnetite bioceramics as much as 0, 5, 10, and 15 wt% of Gelatin-Polypyrrole-Akermanite biocomposite. X-ray diffraction analysis and Fourier transform infrared spectroscopy were conducted. Swelling and degradation of the scaffolds were studied by immersing them in phosphate-buffered saline, PBS, solution. Magnetite bioceramics decreased the swelling percent and degradation duration. By immersing scaffolds in simulated body fluid, the highest formation rate of Apatite was observed in the 15 wt% Magnetite samples. The mean pore size was in an acceptable range to provide suitable conditions for cell proliferation. MG-63 cells were cultured on extracts of the scaffolds for 24, 48, and 72 h and their surfaces for 24 h. Cell viabilities and cell morphologies were assessed. Afterward, micromechanical models with spherical and polyhedral voids and artificial neural networks were employed to predict Young's moduli of the scaffolds. Based on the results of finite element analyses, spherical-shaped void models made the best predictions of elastic behavior in the 0, 5 wt% Magnetite scaffolds compared to the experimental data. Results of the simulations and experimental tests for the ten wt% Magnetite samples were well matched in both micromechanical models. In the 15 wt% Magnetite sample, models with polyhedral voids could precisely predict Young's modulus of such scaffolds.
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Affiliation(s)
- Ali Khalvandi
- Department of Mechanical Engineering, Amirkabir University of Technology, Tehran, Iran
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22
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Impact of Microenvironmental Changes during Degeneration on Intervertebral Disc Progenitor Cells: A Comparison with Mesenchymal Stem Cells. Bioengineering (Basel) 2022; 9:bioengineering9040148. [PMID: 35447707 PMCID: PMC9025850 DOI: 10.3390/bioengineering9040148] [Citation(s) in RCA: 10] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/23/2022] [Revised: 03/16/2022] [Accepted: 03/17/2022] [Indexed: 12/22/2022] Open
Abstract
Intervertebral disc (IVD) degeneration occurs with natural ageing and is linked to low back pain, a common disease. As an avascular tissue, the microenvironment inside the IVD is harsh. During degeneration, the condition becomes even more compromised, presenting a significant challenge to the survival and function of the resident cells, as well as to any regeneration attempts using cell implantation. Mesenchymal stem cells (MSCs) have been proposed as a candidate stem cell tool for IVD regeneration. Recently, endogenous IVD progenitor cells have been identified inside the IVD, highlighting their potential for self-repair. IVD progenitor cells have properties similar to MSCs, with minor differences in potency and surface marker expression. Currently, it is unclear how IVD progenitor cells react to microenvironmental factors and in what ways they possibly behave differently to MSCs. Here, we first summarized the microenvironmental factors presented in the IVD and their changes during degeneration. Then, we analyzed the available studies on the responses of IVD progenitor cells and MSCs to these factors, and made comparisons between these two types of cells, when possible, in an attempt to achieve a clear understanding of the characteristics of IVD progenitor cells when compared to MSCs; as well as, to provide possible clues to cell fate after implantation, which may facilitate future manipulation and design of IVD regeneration studies.
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23
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Janzekovic J, Hunt J, Peltz T, Wagels M, Brown T, Hutmacher DW. Biomechanical Principles of Breast Implants and Current State of Research in Soft Tissue Engineering for Cosmetic Breast Augmentation. Aesthetic Plast Surg 2022; 46:1-10. [PMID: 34494126 DOI: 10.1007/s00266-021-02559-y] [Citation(s) in RCA: 12] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/21/2021] [Accepted: 08/22/2021] [Indexed: 12/17/2022]
Abstract
Currently there are limited implant-based options for cosmetic breast augmentation, and problems associated with those have been increasingly appreciated, most commonly capsular contracture, which occurs due to a chronic foreign body reaction against non-degradable implant materials such as silicone and polyurethane leading to scar tissue formation, pain, and deformity. The underlying biomechanical concepts with implants create a reciprocal stress-strain relationship with local tissue, whilst acting as a deforming force. This means that with time, as the implant continues to have an effect on surrounding tissue the implant and host's biomechanical properties diverge, making malposition, asymmetry, and other complications more likely. Research directed towards development of alternative therapies based on tissue engineering and regenerative medicine seeks to optimize new tissue formation through modulation of tissue progenitors and facilitating tissue regeneration. Scaffolds can guide the process of new tissue formation by providing both an implant surface and a three-dimensional space that promotes the development of a microenvironment that guides attachment, migration, proliferation, and differentiation of connective tissue progenitors. Important to scaffold design are the architecture, surface chemistry, mechanical properties, and biomaterial used. Scaffolds provide a void in which vascularization, new tissue formation, and remodelling can sequentially occur. They provide a conduit for delivery of the different cell types required for tissue regeneration into a graft site, facilitating their retention and distribution. Whilst recent research from a small number of groups is promising, there are still ongoing challenges to achieving clinical translation. This article summarizes the biomechanical principles of breast implants, how these impact outcomes, and progress in scaffold-guided tissue engineering approaches to cosmetic breast augmentation. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
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Affiliation(s)
- Jan Janzekovic
- Centre for Regenerative Medicine, Institute of Health and Biomedical Innovation, Queensland University of Technology, 60 Musk Avenue, Brisbane, QLD, 4000, Australia
- Plastic and Reconstructive Surgery, Princess Alexandra Hospital, 199 Ipswich Rd, Woolloongabba, QLD, 4102, Australia
| | - Jeremy Hunt
- Surgical and Orthopaedic Research Laboratories, Prince of Wales Clinical School, University of New South Wales, Sydney, Australia
| | - Tim Peltz
- Centre for Regenerative Medicine, Institute of Health and Biomedical Innovation, Queensland University of Technology, 60 Musk Avenue, Brisbane, QLD, 4000, Australia
- Surgical and Orthopaedic Research Laboratories, Prince of Wales Clinical School, University of New South Wales, Sydney, Australia
- St Luke's and Prince of Wales Hospital Plastic Surgery Research Group, Potts Point, NSW, 2011, Australia
| | - Michael Wagels
- Plastic and Reconstructive Surgery, Princess Alexandra Hospital, 199 Ipswich Rd, Woolloongabba, QLD, 4102, Australia
- St Luke's and Prince of Wales Hospital Plastic Surgery Research Group, Potts Point, NSW, 2011, Australia
| | - Tim Brown
- Centre for Regenerative Medicine, Institute of Health and Biomedical Innovation, Queensland University of Technology, 60 Musk Avenue, Brisbane, QLD, 4000, Australia.
- Plastic Surgeon in Private Practice in Melbourne, 40-42 Clyde Road, Suite 2, Berwick, VIC, 3806, Australia.
- School of Mechanical, Medical and Process Engineering, Science and Engineering Faculty, Queensland University of Technology, Brisbane, QLD, 4000, Australia.
| | - Dietmar W Hutmacher
- Centre for Regenerative Medicine, Institute of Health and Biomedical Innovation, Queensland University of Technology, 60 Musk Avenue, Brisbane, QLD, 4000, Australia
- School of Mechanical, Medical and Process Engineering, Science and Engineering Faculty, Queensland University of Technology, Brisbane, QLD, 4000, Australia
- School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, QLD, 4000, Australia
- ARC ITTC in Additive Biomanufacturing, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, QLD, 4000, Australia
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24
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Mehta N, Shaik S, Prasad A, Chaichi A, Sahu SP, Liu Q, Hasan SMA, Sheikh E, Donnarumma F, Murray KK, Fu X, Devireddy R, Gartia MR. Multimodal Label-Free Monitoring of Adipogenic Stem Cell Differentiation Using Endogenous Optical Biomarkers. ADVANCED FUNCTIONAL MATERIALS 2021; 31:2103955. [PMID: 34924914 PMCID: PMC8680429 DOI: 10.1002/adfm.202103955] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/26/2021] [Indexed: 05/13/2023]
Abstract
Stem cell-based therapies carry significant promise for treating human diseases. However, clinical translation of stem cell transplants for effective treatment requires precise non-destructive evaluation of the purity of stem cells with high sensitivity (<0.001% of the number of cells). Here, a novel methodology using hyperspectral imaging (HSI) combined with spectral angle mapping-based machine learning analysis is reported to distinguish differentiating human adipose-derived stem cells (hASCs) from control stem cells. The spectral signature of adipogenesis generated by the HSI method enables identifying differentiated cells at single-cell resolution. The label-free HSI method is compared with the standard techniques such as Oil Red O staining, fluorescence microscopy, and qPCR that are routinely used to evaluate adipogenic differentiation of hASCs. HSI is successfully used to assess the abundance of adipocytes derived from transplanted cells in a transgenic mice model. Further, Raman microscopy and multiphoton-based metabolic imaging is performed to provide complementary information for the functional imaging of the hASCs. Finally, the HSI method is validated using matrix-assisted laser desorption/ionization-mass spectrometry imaging of the stem cells. The study presented here demonstrates that multimodal imaging methods enable label-free identification of stem cell differentiation with high spatial and chemical resolution.
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Affiliation(s)
- Nishir Mehta
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Shahensha Shaik
- Division of Basic Pharmaceutical Sciences, College of Pharmacy, Xavier University of Louisiana, New Orleans, LA 70125, USA
| | - Alisha Prasad
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Ardalan Chaichi
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Sushant P Sahu
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Qianglin Liu
- LSU AgCenter, School of Animal Sciences, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Syed Mohammad Abid Hasan
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Elnaz Sheikh
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Fabrizio Donnarumma
- Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Kermit K Murray
- Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Xing Fu
- LSU AgCenter, School of Animal Sciences, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Ram Devireddy
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
| | - Manas Ranjan Gartia
- Department of Mechanical and Industrial Engineering, Louisiana State University, Baton Rouge, LA 70803, USA
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25
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Strategies to address mesenchymal stem/stromal cell heterogeneity in immunomodulatory profiles to improve cell-based therapies. Acta Biomater 2021; 133:114-125. [PMID: 33857693 DOI: 10.1016/j.actbio.2021.03.069] [Citation(s) in RCA: 49] [Impact Index Per Article: 12.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/07/2020] [Revised: 03/15/2021] [Accepted: 03/31/2021] [Indexed: 02/06/2023]
Abstract
Mesenchymal stromal cells (MSCs) have gained immense attention over the past two decades due to their multipotent differentiation potential and pro-regenerative and immunomodulatory cytokine secretory profiles. Their ability to modulate the host immune system and promote tolerance has prompted several allogeneic and autologous hMSC-based clinical trials for the treatment of graft-versus-host disease and several other immune-induced disorders. However, clinical success beyond safety is still controversial and highly variable, with inconclusive therapeutic benefits and little mechanistic explanation. This clinical variability has been broadly attributed to inconsistent MSC sourcing, phenotypic characterization, variable potency, and non-standard isolation protocols, leading to functional heterogeneity among administered MSCs. Homogeneous MSC populations are proposed to yield more predictable, reliable biological responses and clinically meaningful properties relevant to cell-based therapies. Limited comparisons of heterogeneous MSCs with homogenous MSCs are reported. This review addresses this gap in the literature with a critical analysis of strategies aimed at decreasing MSC heterogeneity concerning their reported immunomodulatory profiles. STATEMENT OF SIGNIFICANCE: This review collates, summarizes, and critically analyzes published strategies that seek to improve homogeneity in immunomodulatory functioning MSC populations intended as cell therapies to treat immune-based disorders, such as graft-vs-host-disease. No such review for MSC therapies, immunomodulatory profiles and cell heterogeneity analysis is published. Since MSCs represent the most clinically studied experimental cell therapy platform globally for which there remains no US domestic marketing approval, insights into MSC challenges in therapeutic product development are imperative to providing solutions for immunomodulatory variabilities.
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26
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El-Rashidy AA, El Moshy S, Radwan IA, Rady D, Abbass MMS, Dörfer CE, Fawzy El-Sayed KM. Effect of Polymeric Matrix Stiffness on Osteogenic Differentiation of Mesenchymal Stem/Progenitor Cells: Concise Review. Polymers (Basel) 2021; 13:2950. [PMID: 34502988 PMCID: PMC8434088 DOI: 10.3390/polym13172950] [Citation(s) in RCA: 37] [Impact Index Per Article: 9.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/16/2021] [Revised: 08/04/2021] [Accepted: 08/05/2021] [Indexed: 01/23/2023] Open
Abstract
Mesenchymal stem/progenitor cells (MSCs) have a multi-differentiation potential into specialized cell types, with remarkable regenerative and therapeutic results. Several factors could trigger the differentiation of MSCs into specific lineages, among them the biophysical and chemical characteristics of the extracellular matrix (ECM), including its stiffness, composition, topography, and mechanical properties. MSCs can sense and assess the stiffness of extracellular substrates through the process of mechanotransduction. Through this process, the extracellular matrix can govern and direct MSCs' lineage commitment through complex intracellular pathways. Hence, various biomimetic natural and synthetic polymeric matrices of tunable stiffness were developed and further investigated to mimic the MSCs' native tissues. Customizing scaffold materials to mimic cells' natural environment is of utmost importance during the process of tissue engineering. This review aims to highlight the regulatory role of matrix stiffness in directing the osteogenic differentiation of MSCs, addressing how MSCs sense and respond to their ECM, in addition to listing different polymeric biomaterials and methods used to alter their stiffness to dictate MSCs' differentiation towards the osteogenic lineage.
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Affiliation(s)
- Aiah A. El-Rashidy
- Biomaterials Department, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt;
- Stem Cells and Tissue Engineering Research Group, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt; (S.E.M.); (I.A.R.); (D.R.); (M.M.S.A.)
| | - Sara El Moshy
- Stem Cells and Tissue Engineering Research Group, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt; (S.E.M.); (I.A.R.); (D.R.); (M.M.S.A.)
- Oral Biology Department, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt
| | - Israa Ahmed Radwan
- Stem Cells and Tissue Engineering Research Group, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt; (S.E.M.); (I.A.R.); (D.R.); (M.M.S.A.)
- Oral Biology Department, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt
| | - Dina Rady
- Stem Cells and Tissue Engineering Research Group, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt; (S.E.M.); (I.A.R.); (D.R.); (M.M.S.A.)
- Oral Biology Department, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt
| | - Marwa M. S. Abbass
- Stem Cells and Tissue Engineering Research Group, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt; (S.E.M.); (I.A.R.); (D.R.); (M.M.S.A.)
- Oral Biology Department, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt
| | - Christof E. Dörfer
- Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian Albrechts University, 24105 Kiel, Germany;
| | - Karim M. Fawzy El-Sayed
- Stem Cells and Tissue Engineering Research Group, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt; (S.E.M.); (I.A.R.); (D.R.); (M.M.S.A.)
- Clinic for Conservative Dentistry and Periodontology, School of Dental Medicine, Christian Albrechts University, 24105 Kiel, Germany;
- Oral Medicine and Periodontology Department, Faculty of Dentistry, Cairo University, Cairo 11562, Egypt
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27
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Stiffness Regulates the Morphology, Adhesion, Proliferation, and Osteogenic Differentiation of Maxillary Schneiderian Sinus Membrane-Derived Stem Cells. Stem Cells Int 2021; 2021:8868004. [PMID: 34306097 PMCID: PMC8285206 DOI: 10.1155/2021/8868004] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/20/2020] [Revised: 05/06/2021] [Accepted: 06/09/2021] [Indexed: 11/17/2022] Open
Abstract
Recent studies, which aim to optimize maxillary sinus augmentation, have paid significant attention exploring osteogenic potential of maxillary Schneiderian sinus membrane-derived cells (MSSM-derived cells). However, it remains unclear that how MSSM-derived cells could respond to niche's biomechanical properties. Herein, this study investigated the possible effects of substrate stiffness on rMSSM-derived stem cell fate. Initially, rMSSM-derived stem cells with multiple differentiation potential were successfully obtained. We then fabricated polyacrylamide substrates with varied stiffness ranging from 13 to 68 kPa to modulate the mechanical environment of rMSSM-derived stem cells. A larger cell spreading area and increased proliferation of rMSSM-derived stem cells were found on the stiffer substrates. Similarly, cells became more adhesive as their stiffness increased. Furthermore, the higher stiffness facilitated osteogenic differentiation of rMSSM-derived stem cells. Overall, our results indicated that increase in stiffness could mediate behaviors of rMSSM-derived stem cells, which may serve as a guide in future research to design novel biomaterials for maxillary sinus augmentation.
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28
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Chen H, Wang X, Wang J, Shi X, Li X, Wang J, Li D, Zhu Y, Tan W, Tan Z. In vitroadipogenesis and long-term adipocyte culture in adipose tissue-derived cell banks. Biofabrication 2021; 13. [PMID: 34044385 DOI: 10.1088/1758-5090/ac0610] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/19/2020] [Accepted: 05/27/2021] [Indexed: 11/12/2022]
Abstract
There is a critical need to developin vitroculture systems appropriate for the expansion of adipose tissue, in order to gain new insights into metabolic diseases and to assist in the restoration of tissue defects. Conventional two- or three-dimensional (2D or 3D)in vitromodels of adipocytes require a combination of supplements to induce adipocyte maturation that greatly increases the cost of large-scale industrial production. In the present study, a microporous, perforated bacterial cellulose (BC)-assisted culture system was developed that promoted the adhesion, proliferation, and adipogenic differentiation of preadipocytes. Additionally, the system maintained the cells as mature unilocular adipocytesex vivoin normal cell culture medium in long-term culture. All cells were derived from isolated adipose tissue without the use of expensive enzymes for tissue digestion. In contrast to culture in hard tissue culture plates, preadipocytes in the soft 3D environments formed multidimensional interlaced cell contacts, undergoing significant spontaneous lipid accumulation and could be cultured for up to threemonths in maintenance medium. More importantly, the cultured adipose tissue-derived cell bank created here was able to produce injury repair activators that promoted the proliferation of fibroblasts with little fibrosis and the functional differentiation of myoblasts, displaying the potential for use in adipose reconstruction. Thus, the present study demonstrates the potential of a mechanically flexible BC scaffold to generate volume tunable adipose constructs and provides a low-cost and user-friendly strategy for large-scale industrial production of adipose tissue.
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Affiliation(s)
- Haoxiang Chen
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China.,State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha, Hunan 410082, People's Republic of China
| | - Xiaocheng Wang
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China
| | - Jian Wang
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China.,State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha, Hunan 410082, People's Republic of China
| | - Xuelei Shi
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China
| | - Xinghuan Li
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China
| | - Jianlong Wang
- Department of Orthopedics, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, People's Republic of China
| | - Dan Li
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China
| | - Yonghua Zhu
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China
| | - Weihong Tan
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China
| | - Zhikai Tan
- College of Biology, Hunan University, Changsha, Hunan 410082, People's Republic of China.,State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha, Hunan 410082, People's Republic of China
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29
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Veernala I, Roopmani P, Singh R, Hasan U, Giri J. Cell encapsulated and microenvironment modulating microbeads containing alginate hydrogel system for bone tissue engineering. Prog Biomater 2021; 10:131-150. [PMID: 34224092 DOI: 10.1007/s40204-021-00158-3] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/09/2020] [Accepted: 05/22/2021] [Indexed: 11/28/2022] Open
Abstract
Functional tissue regeneration using synthetic biomaterials requires proliferation and heterotypic differentiation of stem/progenitor cells within a specialized heterogeneous (biophysical-biochemical) microenvironment. The current techniques have limitations to develop synthetic hydrogels, mimicking native extracellular matrix porosity along with heterogeneous microenvironmental cues of matrix mechanics, degradability, microstructure and cell-cell interactions. Here, we have developed a microenvironment modulating system to fabricate in situ porous hydrogel matrix with two or more distinct tailored microenvironmental niches within microbeads and the hydrogel matrix for multicellular tissue regeneration. Electrosprayed pectin-gelatin blended microbeads and crosslinked alginate hydrogel system help to tailor microenvironmental niches of encapsulated cells where two different cells are surrounded by a specific microenvironment. The effect of different microenvironmental parameters associated with the microbead/hydrogel matrix was evaluated using human umbilical-cord mesenchymal stem cells (hUCMSCs). The osteogenic differentiation of hUCMSCs in the hydrogel matrix was evaluated for bone tissue regeneration. This will be the first report on microenvironment modulating microbead-hydrogel system to encapsulate two/more types of cells in a hydrogel, where each cell is surrounded with distinct niches for heterogeneous tissue regeneration.
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Affiliation(s)
- Induvahi Veernala
- Department of Biomedical Engineering, Indian Institute of Technology, Hyderabad, Kandi, Telangana, India
| | - Purandhi Roopmani
- Department of Biomedical Engineering, Indian Institute of Technology, Hyderabad, Kandi, Telangana, India
| | - Ruby Singh
- Department of Biomedical Engineering, Indian Institute of Technology, Hyderabad, Kandi, Telangana, India
| | - Uzma Hasan
- Department of Biotechnology, Indian Institute of Technology, Hyderabad, Kandi, Telangana, India
| | - Jyotsnendu Giri
- Department of Biomedical Engineering, Indian Institute of Technology, Hyderabad, Kandi, Telangana, India.
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30
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Femtosecond laser-induced nanoporous layer for enhanced osteogenesis of titanium implants. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2021; 127:112247. [PMID: 34225886 DOI: 10.1016/j.msec.2021.112247] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/08/2021] [Revised: 05/31/2021] [Accepted: 06/07/2021] [Indexed: 11/22/2022]
Abstract
The osteogenic activity of medical metal can be improved by lowering its surface stiffness and elastic modulus. However, it is very difficult to directly reduce the elastic modulus of medical metal surfaces. In this paper, with selected parameters, the titanium surface was treated via femtosecond laser irradiation. Micro indentation revealed that the femtosecond laser ablation can effectively reduce the surface Young's modulus and Vickers hardness of titanium. Besides, In order to explain the mechanical properties of degradation of titanium surface, Large-scale Atomic/Molecular Massively Parallel Simulator (LAMMPS) was used to simulate the process of laser ablation process of titanium surface, and it was found that after the ablation of titanium surface, voids were produced in the subsurface layer. The simulation showed that the voids are formed by the cavitation of metastable liquid induced by high tensile stress and high temperature during femtosecond laser irradiation. Subsurface voids with a thickness of about 40 nm were observed under the oxide layer in the experiment. Cell experiments showed that the surface with low Young's modulus was more conducive to cell proliferation and osteogenic differentiation.
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31
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Lei R, Akins EA, Wong KCY, Repina NA, Wolf KJ, Dempsey GE, Schaffer DV, Stahl A, Kumar S. Multiwell Combinatorial Hydrogel Array for High-Throughput Analysis of Cell-ECM Interactions. ACS Biomater Sci Eng 2021; 7:2453-2465. [PMID: 34028263 DOI: 10.1021/acsbiomaterials.1c00065] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/29/2023]
Abstract
Biophysical cues in the extracellular matrix (ECM) regulate cell behavior in a complex, nonlinear, and interdependent manner. To quantify these important regulatory relationships and gain a comprehensive understanding of mechanotransduction, there is a need for high-throughput matrix platforms that enable parallel culture and analysis of cells in various matrix conditions. Here we describe a multiwell hyaluronic acid (HA) platform in which cells are cultured on combinatorial arrays of hydrogels spanning a range of elasticities and adhesivities. Our strategy utilizes orthogonal photopatterning of stiffness and adhesivity gradients, with the stiffness gradient implemented by a programmable light illumination system. The resulting platform allows individual treatment and analysis of each matrix environment while eliminating contributions of haptotaxis and durotaxis. In human mesenchymal stem cells, our platform recapitulates expected relationships between matrix stiffness, adhesivity, and cell mechanosensing. We further applied the platform to show that as integrin ligand density falls, cell adhesion and migration depend more strongly on CD44-mediated interactions with the HA backbone. We anticipate that our system could bear great value for mechanistic discovery and screening where matrix mechanics and adhesivity are expected to influence phenotype.
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Affiliation(s)
- Ruoxing Lei
- Department of Chemistry, Latimer Hall, University of California, Berkeley, Berkeley, California 94720, United States.,Department of Bioengineering, Stanley Hall, University of California, Berkeley, Berkeley, California 94720, United States
| | - Erin A Akins
- Department of Bioengineering, Stanley Hall, University of California, Berkeley, Berkeley, California 94720, United States.,University of California, Berkeley - University of California, San Francisco Graduate Program in Bioengineering, Stanley Hall, Berkeley, California 94720, United States
| | - Kelly C Y Wong
- Department of Bioengineering, Stanley Hall, University of California, Berkeley, Berkeley, California 94720, United States
| | - Nicole A Repina
- Department of Bioengineering, Stanley Hall, University of California, Berkeley, Berkeley, California 94720, United States.,University of California, Berkeley - University of California, San Francisco Graduate Program in Bioengineering, Stanley Hall, Berkeley, California 94720, United States
| | - Kayla J Wolf
- Department of Bioengineering, Stanley Hall, University of California, Berkeley, Berkeley, California 94720, United States.,University of California, Berkeley - University of California, San Francisco Graduate Program in Bioengineering, Stanley Hall, Berkeley, California 94720, United States
| | - Garrett E Dempsey
- Department of Nutritional Sciences and Toxicology, Morgan Hall, University of California, Berkeley, California 94720, United States
| | - David V Schaffer
- Department of Bioengineering, Stanley Hall, University of California, Berkeley, Berkeley, California 94720, United States.,University of California, Berkeley - University of California, San Francisco Graduate Program in Bioengineering, Stanley Hall, Berkeley, California 94720, United States.,Department of Molecular and Cell Biology, Life Sciences Addition, University of California, Berkeley, California 94720, United States.,Department of Chemical and Biomolecular Engineering, Gilman Hall, University of California, Berkeley, Berkeley, California 94720, United States
| | - Andreas Stahl
- University of California, Berkeley - University of California, San Francisco Graduate Program in Bioengineering, Stanley Hall, Berkeley, California 94720, United States.,Department of Nutritional Sciences and Toxicology, Morgan Hall, University of California, Berkeley, California 94720, United States
| | - Sanjay Kumar
- Department of Bioengineering, Stanley Hall, University of California, Berkeley, Berkeley, California 94720, United States.,University of California, Berkeley - University of California, San Francisco Graduate Program in Bioengineering, Stanley Hall, Berkeley, California 94720, United States.,Department of Chemical and Biomolecular Engineering, Gilman Hall, University of California, Berkeley, Berkeley, California 94720, United States.,Department of Bioengineering and Therapeutic Sciences, Byers Hall, University of California, San Francisco, San Francisco, California 94143, United States
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Prouvé E, Drouin B, Chevallier P, Rémy M, Durrieu MC, Laroche G. Evaluating Poly(Acrylamide-co-Acrylic Acid) Hydrogels Stress Relaxation to Direct the Osteogenic Differentiation of Mesenchymal Stem Cells. Macromol Biosci 2021; 21:e2100069. [PMID: 33870650 DOI: 10.1002/mabi.202100069] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/25/2021] [Revised: 03/30/2021] [Indexed: 11/09/2022]
Abstract
The aim of this study is to investigate polyacrylamide-based hydrogels stress relaxation and the subsequent impact on the osteogenic differentiation of human mesenchymal stem cells (hMSCs). Different hydrogels are synthesized by varying the amount of cross-linker and the ratio between the monomers (acrylamide and acrylic acid), and characterized by compression tests. It has been found that hydrogels containing 18% of acrylic acid exhibit an average relaxation of 70%, while pure polyacrylamide gels show an average relaxation of 15%. Subsequently, hMSCs are cultured on two different hydrogels functionalized with a mimetic peptide of the bone morphogenetic protein-2 to enable cell adhesion and favor their osteogenic differentiation. Phalloidin staining shows that for a constant stiffness of 55 kPa, a hydrogel with a low relaxation (15%) leads to star-shaped cells, which is typical of osteocytes, while a hydrogel with a high relaxation (70%) presents cells with a polygonal shape characteristic of osteoblasts. Immunofluorescence labeling of E11, strongly expressed in early osteocytes, also shows a dramatically higher expression for cells cultured on the hydrogel with low relaxation (15%). These results clearly demonstrate that, by fine-tuning hydrogels stress relaxation, hMSCs differentiation can be directed toward osteoblasts, and even osteocytes, which is particularly rare in vitro.
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Affiliation(s)
- Emilie Prouvé
- Department of mining, metallurgy, and materials engineering, Surface Engineering Laboratory, Research Center on Advanced Materials, Laval University, 1065 Avenue de la médecine, Québec, G1V 0A6, Canada.,Research Center of the University Hospital of Québec, Regenerative Medicine axis, St-François d'Assise Hospital, Laval University, 10 rue de l'Espinay, Québec, G1L 3L5, Canada.,Institute of Chemistry and Biology of Membranes and Nano-objects (UMR 5248 CBMN), Bordeaux University, Allée Geoffroy St Hilaire - Bât B14, Pessac, 33600, France.,CNRS, CBMN UMR5248, Allée Geoffroy Saint Hilaire - Bât B14, Pessac, 33600, France.,Bordeaux INP, CBMN UMR5248, Allée Geoffroy Saint Hilaire - Bât B14, Pessac, 33600, France
| | - Bernard Drouin
- Research Center of the University Hospital of Québec, Regenerative Medicine axis, St-François d'Assise Hospital, Laval University, 10 rue de l'Espinay, Québec, G1L 3L5, Canada
| | - Pascale Chevallier
- Department of mining, metallurgy, and materials engineering, Surface Engineering Laboratory, Research Center on Advanced Materials, Laval University, 1065 Avenue de la médecine, Québec, G1V 0A6, Canada.,Research Center of the University Hospital of Québec, Regenerative Medicine axis, St-François d'Assise Hospital, Laval University, 10 rue de l'Espinay, Québec, G1L 3L5, Canada
| | - Murielle Rémy
- Institute of Chemistry and Biology of Membranes and Nano-objects (UMR 5248 CBMN), Bordeaux University, Allée Geoffroy St Hilaire - Bât B14, Pessac, 33600, France.,CNRS, CBMN UMR5248, Allée Geoffroy Saint Hilaire - Bât B14, Pessac, 33600, France.,Bordeaux INP, CBMN UMR5248, Allée Geoffroy Saint Hilaire - Bât B14, Pessac, 33600, France
| | - Marie-Christine Durrieu
- Institute of Chemistry and Biology of Membranes and Nano-objects (UMR 5248 CBMN), Bordeaux University, Allée Geoffroy St Hilaire - Bât B14, Pessac, 33600, France.,CNRS, CBMN UMR5248, Allée Geoffroy Saint Hilaire - Bât B14, Pessac, 33600, France.,Bordeaux INP, CBMN UMR5248, Allée Geoffroy Saint Hilaire - Bât B14, Pessac, 33600, France
| | - Gaétan Laroche
- Department of mining, metallurgy, and materials engineering, Surface Engineering Laboratory, Research Center on Advanced Materials, Laval University, 1065 Avenue de la médecine, Québec, G1V 0A6, Canada.,Research Center of the University Hospital of Québec, Regenerative Medicine axis, St-François d'Assise Hospital, Laval University, 10 rue de l'Espinay, Québec, G1L 3L5, Canada
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Realizing tissue integration with supramolecular hydrogels. Acta Biomater 2021; 124:1-14. [PMID: 33508507 DOI: 10.1016/j.actbio.2021.01.034] [Citation(s) in RCA: 24] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/12/2020] [Revised: 01/04/2021] [Accepted: 01/21/2021] [Indexed: 12/17/2022]
Abstract
Biomaterial matrices must permit tissue growth and maturation for the success of tissue regeneration strategies. Naturally, this accommodation is achieved via the dynamic remodeling of a cell's extracellular matrix (ECM). Synthetically, hydrolytic or enzymatic degradation are often engineered into materials for this purpose. More recently, supramolecular interactions have been used to provide a biomimetic and tunable mechanism to facilitate tissue formation via their dynamic and reversible non-covalent interactions. By engineering the mechanical and bioactive properties of a material, supramolecular chemists are able to design permissivity into the construct and facilitate tissue integration in-vivo. Furthermore, via the reversibility of non-covalent interactions, injectability and responsiveness can be designed for enhanced delivery and spatio-temporal control. In this review, we delineate the basic considerations needed when designing permissive supramolecular hydrogels for tissue engineering with an eye toward tissue growth and integration. We highlight three archetypal hydrogel systems that have shown well-documented tissue integration in vivo, and provide avenues to assess tissue in-growth. Careful design and assessment of the biomedical potential of a supramolecular hydrogels can inspire the creation of robust and dynamic implants for new tissue engineering applications.
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Understanding of how the properties of medical grade lactide based copolymer scaffolds influence adipose tissue regeneration: Sterilization and a systematic in vitro assessment. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2021; 124:112020. [PMID: 33947531 DOI: 10.1016/j.msec.2021.112020] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/04/2020] [Revised: 02/24/2021] [Accepted: 02/27/2021] [Indexed: 01/22/2023]
Abstract
Aliphatic polyesters are the synthetic polymers most commonly used in the development of resorbable medical implants/devices. Various three-dimensional (3D) scaffolds have been fabricated from these polymers and used in adipose tissue engineering. However, their systematic evaluation altogether lacks, which makes it difficult to select a suitable degradable polymer to design 3D resorbable implants and/or devices able to effectively mimic the properties of adipose tissue. Additionally, the impact of sterilization methods on the medical devices, if any, must be taken into account. We evaluate and compare five different medical-grade resorbable polyesters with l-lactide content ranging from 50 to 100 mol% and exhibiting different physiochemical properties depending on the comonomer (d-lactide, ε-caprolactone, glycolide, and trimethylene carbonate). The salt-leaching technique was used to prepare 3D microporous scaffolds. A comprehensive assessment of physical, chemical, and mechanical properties of the scaffolds was carried out in PBS at 37 °C. The cell-material interactions and the ability of the scaffolds to promote adipogenesis of human adipose tissue-derived stem cells were assessed in vitro. The diverse physical and mechanical properties of the scaffolds, due to the different composition of the copolymers, influenced human adipose tissue-derived stem cells proliferation and differentiation. Scaffolds made from polymers which were above their glass transition temperature and with low degree of crystallinity showed better proliferation and adipogenic differentiation of stem cells. The effect of sterilization techniques (electron beam and ethylene oxide) on the polymer properties was also evaluated. Results showed that scaffolds sterilized with the ethylene oxide method better retained their physical and chemical properties. Overall, the presented research provides (i) a detailed understanding to select a degradable polymer that has relevant properties to augment adipose tissue regeneration and can be further used to fabricate medical devices/implants; (ii) directions to prefer a sterilization method that does not change polymer properties.
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Häussling V, Aspera-Werz RH, Rinderknecht H, Springer F, Arnscheidt C, Menger MM, Histing T, Nussler AK, Ehnert S. 3D Environment Is Required In Vitro to Demonstrate Altered Bone Metabolism Characteristic for Type 2 Diabetics. Int J Mol Sci 2021; 22:2925. [PMID: 33805833 PMCID: PMC8002142 DOI: 10.3390/ijms22062925] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/19/2021] [Revised: 03/09/2021] [Accepted: 03/10/2021] [Indexed: 12/19/2022] Open
Abstract
A large British study, with almost 3000 patients, identified diabetes as main risk factor for delayed and nonunion fracture healing, the treatment of which causes large costs for the health system. In the past years, much progress has been made to treat common complications in diabetics. However, there is still a lack of advanced strategies to treat diabetic bone diseases. To develop such therapeutic strategies, mechanisms leading to massive bone alterations in diabetics have to be well understood. We herein describe an in vitro model displaying bone metabolism frequently observed in diabetics. The model is based on osteoblastic SaOS-2 cells, which in direct coculture, stimulate THP-1 cells to form osteoclasts. While in conventional 2D cocultures formation of mineralized matrix is decreased under pre-/diabetic conditions, formation of mineralized matrix is increased in 3D cocultures. Furthermore, we demonstrate a matrix stability of the 3D carrier that is decreased under pre-/diabetic conditions, resembling the in vivo situation in type 2 diabetics. In summary, our results show that a 3D environment is required in this in vitro model to mimic alterations in bone metabolism characteristic for pre-/diabetes. The ability to measure both osteoblast and osteoclast function, and their effect on mineralization and stability of the 3D carrier offers the possibility to use this model also for other purposes, e.g., drug screenings.
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Affiliation(s)
- Victor Häussling
- Siegfried Weller Research Institute, BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany; (V.H.); (R.H.A.-W.); (H.R.); (C.A.); (M.M.M.); (T.H.); (S.E.)
| | - Romina H. Aspera-Werz
- Siegfried Weller Research Institute, BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany; (V.H.); (R.H.A.-W.); (H.R.); (C.A.); (M.M.M.); (T.H.); (S.E.)
| | - Helen Rinderknecht
- Siegfried Weller Research Institute, BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany; (V.H.); (R.H.A.-W.); (H.R.); (C.A.); (M.M.M.); (T.H.); (S.E.)
| | - Fabian Springer
- Department of Diagnostic and Interventional Radiology, University of Tübingen, Hoppe-Seyler-Str. 3, D-72076 Tübingen, Germany;
- Radiology Department, BG Trauma Center Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany
| | - Christian Arnscheidt
- Siegfried Weller Research Institute, BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany; (V.H.); (R.H.A.-W.); (H.R.); (C.A.); (M.M.M.); (T.H.); (S.E.)
| | - Maximilian M. Menger
- Siegfried Weller Research Institute, BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany; (V.H.); (R.H.A.-W.); (H.R.); (C.A.); (M.M.M.); (T.H.); (S.E.)
| | - Tina Histing
- Siegfried Weller Research Institute, BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany; (V.H.); (R.H.A.-W.); (H.R.); (C.A.); (M.M.M.); (T.H.); (S.E.)
| | - Andreas K. Nussler
- Siegfried Weller Research Institute, BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany; (V.H.); (R.H.A.-W.); (H.R.); (C.A.); (M.M.M.); (T.H.); (S.E.)
| | - Sabrina Ehnert
- Siegfried Weller Research Institute, BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Schnarrenbergstr. 95, D-72076 Tübingen, Germany; (V.H.); (R.H.A.-W.); (H.R.); (C.A.); (M.M.M.); (T.H.); (S.E.)
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López-Martínez S, Campo H, de Miguel-Gómez L, Faus A, Navarro AT, Díaz A, Pellicer A, Ferrero H, Cervelló I. A Natural Xenogeneic Endometrial Extracellular Matrix Hydrogel Toward Improving Current Human in vitro Models and Future in vivo Applications. Front Bioeng Biotechnol 2021; 9:639688. [PMID: 33748086 PMCID: PMC7973233 DOI: 10.3389/fbioe.2021.639688] [Citation(s) in RCA: 33] [Impact Index Per Article: 8.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/09/2020] [Accepted: 02/12/2021] [Indexed: 12/25/2022] Open
Abstract
Decellularization techniques support the creation of biocompatible extracellular matrix hydrogels, providing tissue-specific environments for both in vitro cell culture and in vivo tissue regeneration. We obtained endometrium derived from porcine decellularized uteri to create endometrial extracellular matrix (EndoECM) hydrogels. After decellularization and detergent removal, we investigated the physicochemical features of the EndoECM, including gelation kinetics, ultrastructure, and proteomic profile. The matrisome showed conservation of structural and tissue-specific components with low amounts of immunoreactive molecules. EndoECM supported in vitro culture of human endometrial cells in two- and three-dimensional conditions and improved proliferation of endometrial stem cells with respect to collagen and Matrigel. Further, we developed a three-dimensional endometrium-like co-culture system of epithelial and stromal cells from different origins. Endometrial co-cultures remained viable and showed significant remodeling. Finally, EndoECM was injected subcutaneously in immunocompetent mice in a preliminary study to test a possible hypoimmunogenic reaction. Biomimetic endometrial milieus offer new strategies in reproductive techniques and endometrial repair and our findings demonstrate that EndoECM has potential for in vitro endometrial culture and as treatment for endometrial pathologies.
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Affiliation(s)
- Sara López-Martínez
- Fundación Instituto Valenciano de Infertilidad, Instituto de Investigación Sanitaria La Fe, Valencia, Spain
| | - Hannes Campo
- Fundación Instituto Valenciano de Infertilidad, Instituto de Investigación Sanitaria La Fe, Valencia, Spain
| | - Lucía de Miguel-Gómez
- Fundación Instituto Valenciano de Infertilidad, Instituto de Investigación Sanitaria La Fe, Valencia, Spain.,University of Valencia, Valencia, Spain
| | - Amparo Faus
- Fundación Instituto Valenciano de Infertilidad, Instituto de Investigación Sanitaria La Fe, Valencia, Spain
| | - Alfredo T Navarro
- Fundación Instituto Valenciano de Infertilidad, Instituto de Investigación Sanitaria La Fe, Valencia, Spain
| | - Ana Díaz
- University of Valencia, Valencia, Spain
| | - Antonio Pellicer
- University of Valencia, Valencia, Spain.,IVIRMA Roma, Rome, Italy
| | - Hortensia Ferrero
- Fundación Instituto Valenciano de Infertilidad, Instituto de Investigación Sanitaria La Fe, Valencia, Spain.,IVIRMA Valencia, Valencia, Spain
| | - Irene Cervelló
- Fundación Instituto Valenciano de Infertilidad, Instituto de Investigación Sanitaria La Fe, Valencia, Spain
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37
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Pardo A, Gómez-Florit M, Barbosa S, Taboada P, Domingues RMA, Gomes ME. Magnetic Nanocomposite Hydrogels for Tissue Engineering: Design Concepts and Remote Actuation Strategies to Control Cell Fate. ACS NANO 2021; 15:175-209. [PMID: 33406360 DOI: 10.1021/acsnano.0c08253] [Citation(s) in RCA: 119] [Impact Index Per Article: 29.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/18/2023]
Abstract
Most tissues of the human body are characterized by highly anisotropic physical properties and biological organization. Hydrogels have been proposed as scaffolding materials to construct artificial tissues due to their water-rich composition, biocompatibility, and tunable properties. However, unmodified hydrogels are typically composed of randomly oriented polymer networks, resulting in homogeneous structures with isotropic properties different from those observed in biological systems. Magnetic materials have been proposed as potential agents to provide hydrogels with the anisotropy required for their use on tissue engineering. Moreover, the intrinsic properties of magnetic nanoparticles enable their use as magnetomechanic remote actuators to control the behavior of the cells encapsulated within the hydrogels under the application of external magnetic fields. In this review, we combine a detailed summary of the main strategies to prepare magnetic nanoparticles showing controlled properties with an analysis of the different approaches available to their incorporation into hydrogels. The application of magnetically responsive nanocomposite hydrogels in the engineering of different tissues is also reviewed.
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Affiliation(s)
- Alberto Pardo
- 3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciencia e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco-Guimarães, Portugal
- ICVS/3B's-PT Government Associate Laboratory, 4805-017 Braga/Guimarães, Portugal
| | - Manuel Gómez-Florit
- 3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciencia e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco-Guimarães, Portugal
- ICVS/3B's-PT Government Associate Laboratory, 4805-017 Braga/Guimarães, Portugal
| | - Silvia Barbosa
- Colloids and Polymers Physics Group, Condensed Matter Physics Area, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain
- Health Research Institute of Santiago de Compostela (IDIS), Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain
| | - Pablo Taboada
- Colloids and Polymers Physics Group, Condensed Matter Physics Area, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain
- Health Research Institute of Santiago de Compostela (IDIS), Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain
| | - Rui M A Domingues
- 3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciencia e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco-Guimarães, Portugal
- ICVS/3B's-PT Government Associate Laboratory, 4805-017 Braga/Guimarães, Portugal
| | - Manuela E Gomes
- 3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciencia e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco-Guimarães, Portugal
- ICVS/3B's-PT Government Associate Laboratory, 4805-017 Braga/Guimarães, Portugal
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38
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Yang F, Carmona A, Stojkova K, Garcia Huitron EI, Goddi A, Bhushan A, Cohen RN, Brey EM. A 3D human adipose tissue model within a microfluidic device. LAB ON A CHIP 2021; 21:435-446. [PMID: 33351023 PMCID: PMC7876365 DOI: 10.1039/d0lc00981d] [Citation(s) in RCA: 35] [Impact Index Per Article: 8.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/23/2023]
Abstract
An accurate in vitro model of human adipose tissue could assist in the study of adipocyte function and allow for better tools for screening new therapeutic compounds. Cell culture models on two-dimensional surfaces fall short of mimicking the three-dimensional in vivo adipose environment, while three-dimensional culture models are often unable to support long-term cell culture due, in part, to insufficient mass transport. Microfluidic systems have been explored for adipose tissue models. However, current systems have primarily focused on 2D cultured adipocytes. In this work, a 3D human adipose microtissue was engineered within a microfluidic system. Human adipose-derived stem cells (ADSCs) were used as the cell source for generating differentiated adipocytes. The ADSCs differentiated within the microfluidic system formed a dense lipid-loaded mass with the expression of adipose tissue genetic markers. Engineered adipose tissue showed a decreased adiponectin secretion and increased free fatty acid secretion with increasing shear stress. Adipogenesis markers were downregulated with increasing shear stress. Overall, this microfluidic system enables the on-chip differentiation and development of a functional 3D human adipose microtissue supported by the interstitial flow. This system could potentially serve as a platform for in vitro drug testing for adipose tissue-related diseases.
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Affiliation(s)
- Feipeng Yang
- Illinois Institute of Technology, Department of Biomedical Engineering, Chicago, 60616, USA
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39
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Toledano-Osorio M, Manzano-Moreno FJ, Ruiz C, Toledano M, Osorio R. Testing active membranes for bone regeneration: A review. J Dent 2021; 105:103580. [PMID: 33417978 DOI: 10.1016/j.jdent.2021.103580] [Citation(s) in RCA: 32] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2020] [Revised: 12/30/2020] [Accepted: 12/31/2020] [Indexed: 12/24/2022] Open
Abstract
OBJECTIVES Maxillofacial bone defects are the main hindering conditions for traditional dental implant strategies. Guided Bone Regeneration (GBR) is used to handle this situation. The principle of GBR is to use a membrane to prevent the colonization of soft tissue cells of the bone defect and favors the migration of osteogenic linages. Current membranes do not completely fulfill the requirements that an optimal membrane should have, sometimes resulting in non-predictable results. Thus, the need to develop an ideal membrane to perform this duty is clear. Recent developments in bio-manufacturing are driving innovations in membranes technology permitting the active participation of the membrane in the healing and regenerative process trough native tissue mimicking, drug-delivery and cells interaction, away from being a passive barrier. New membranes features need specific evaluation techniques, beyond the International Standard for membrane materials (last reviewed in 2004), being this the rationale for the present review. Nanotechnology application has completely shifted the way of analyzing structural characterization. New progresses on osteoimmmunomodulation have also switched the understanding of cells-membranes interaction. DATA AND SOURCES To propose an updated protocol for GBR membranes evaluation, critical reading of the relevant published literature was carried out after a MEDLINE/PubMed database search. CONCLUSIONS The main findings are that a potential active membrane should be assessed in its nanostructure, physicochemical and nanomechanical properties, bioactivity and antibacterial, osteoblasts proliferation, differentiation and mineralization. Immunomodulation testing for macrophages recruitment and M2 phenotype promotion in osteoblasts co-culture has to be achieved to completely analyze membranes/tissue interactions.
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Affiliation(s)
- Manuel Toledano-Osorio
- Biomaterials in Dentistry Research Group, Department of Stomatology, School of Dentistry, University of Granada, Spain; Medicina Clínica y Salud Pública PhD Programme, Spain
| | - Francisco Javier Manzano-Moreno
- Biomedical Group (BIO277), Department of Stomatology, School of Dentistry, University of Granada, Spain; Instituto Investigación Biosanitaria, ibs. Granada, Granada, Spain
| | - Concepción Ruiz
- Instituto Investigación Biosanitaria, ibs. Granada, Granada, Spain; Biomedical Group (BIO277), Department of Nursing, Faculty of Health Sciences. University of Granada, Spain; Institute of Neuroscience, University of Granada, Centro de Investigación Biomédica (CIBM), Parque Tecnológico de la Salud (PTS), Granada, Spain
| | - Manuel Toledano
- Biomaterials in Dentistry Research Group, Department of Stomatology, School of Dentistry, University of Granada, Spain.
| | - Raquel Osorio
- Biomaterials in Dentistry Research Group, Department of Stomatology, School of Dentistry, University of Granada, Spain
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40
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Ehnert S, Rinderknecht H, Aspera-Werz RH, Häussling V, Nussler AK. Use of in vitro bone models to screen for altered bone metabolism, osteopathies, and fracture healing: challenges of complex models. Arch Toxicol 2020; 94:3937-3958. [PMID: 32910238 PMCID: PMC7655582 DOI: 10.1007/s00204-020-02906-z] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/06/2020] [Accepted: 09/03/2020] [Indexed: 02/06/2023]
Abstract
Approx. every third hospitalized patient in Europe suffers from musculoskeletal injuries or diseases. Up to 20% of these patients need costly surgical revisions after delayed or impaired fracture healing. Reasons for this are the severity of the trauma, individual factors, e.g, the patients' age, individual lifestyle, chronic diseases, medication, and, over 70 diseases that negatively affect the bone quality. To investigate the various disease constellations and/or develop new treatment strategies, many in vivo, ex vivo, and in vitro models can be applied. Analyzing these various models more closely, it is obvious that many of them have limits and/or restrictions. Undoubtedly, in vivo models most completely represent the biological situation. Besides possible species-specific differences, ethical concerns may question the use of in vivo models especially for large screening approaches. Challenging whether ex vivo or in vitro bone models can be used as an adequate replacement for such screenings, we here summarize the advantages and challenges of frequently used ex vivo and in vitro bone models to study disturbed bone metabolism and fracture healing. Using own examples, we discuss the common challenge of cell-specific normalization of data obtained from more complex in vitro models as one example of the analytical limits which lower the full potential of these complex model systems.
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Affiliation(s)
- Sabrina Ehnert
- Siegfried Weller Research Institute at the BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Tübingen, Germany.
| | - Helen Rinderknecht
- Siegfried Weller Research Institute at the BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Tübingen, Germany
| | - Romina H Aspera-Werz
- Siegfried Weller Research Institute at the BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Tübingen, Germany
| | - Victor Häussling
- Siegfried Weller Research Institute at the BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Tübingen, Germany
| | - Andreas K Nussler
- Siegfried Weller Research Institute at the BG Trauma Center Tübingen, Department of Trauma and Reconstructive Surgery, University of Tübingen, Tübingen, Germany
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Biomimetic Aspects of Oral and Dentofacial Regeneration. Biomimetics (Basel) 2020; 5:biomimetics5040051. [PMID: 33053903 PMCID: PMC7709662 DOI: 10.3390/biomimetics5040051] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/15/2020] [Revised: 10/09/2020] [Accepted: 10/10/2020] [Indexed: 12/12/2022] Open
Abstract
Biomimetic materials for hard and soft tissues have advanced in the fields of tissue engineering and regenerative medicine in dentistry. To examine these recent advances, we searched Medline (OVID) with the key terms “biomimetics”, “biomaterials”, and “biomimicry” combined with MeSH terms for “dentistry” and limited the date of publication between 2010–2020. Over 500 articles were obtained under clinical trials, randomized clinical trials, metanalysis, and systematic reviews developed in the past 10 years in three major areas of dentistry: restorative, orofacial surgery, and periodontics. Clinical studies and systematic reviews along with hand-searched preclinical studies as potential therapies have been included. They support the proof-of-concept that novel treatments are in the pipeline towards ground-breaking clinical therapies for orofacial bone regeneration, tooth regeneration, repair of the oral mucosa, periodontal tissue engineering, and dental implants. Biomimicry enhances the clinical outcomes and calls for an interdisciplinary approach integrating medicine, bioengineering, biotechnology, and computational sciences to advance the current research to clinics. We conclude that dentistry has come a long way apropos of regenerative medicine; still, there are vast avenues to endeavour, seeking inspiration from other facets in biomedical research.
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Feng Q, Gao H, Wen H, Huang H, Li Q, Liang M, Liu Y, Dong H, Cao X. Engineering the cellular mechanical microenvironment to regulate stem cell chondrogenesis: Insights from a microgel model. Acta Biomater 2020; 113:393-406. [PMID: 32629189 DOI: 10.1016/j.actbio.2020.06.046] [Citation(s) in RCA: 39] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/07/2020] [Revised: 06/30/2020] [Accepted: 06/30/2020] [Indexed: 01/07/2023]
Abstract
Biophysical cues (especially mechanical cues) embedded in cellular microenvironments show a critical impact on stem cell fate. Despite the capability of traditional hydrogels to mimic the feature of extracellular matrix (ECM) and tune their physicochemical properties via diverse approaches, their relatively large size not only induces biased results, but also hinders high-throughput screening and analysis. In this paper, a microgel model is proposed to recapitulate the role of 3D mechanical microenvironment on stem cell behaviors especially chondrogenesis in vitro. The small diameter of microgels brings the high surface area to volume ratio and then the enlarged diffusion area and shortened diffusion distance of soluble molecules, leading to uniform distribution of nutrients and negligible biochemical gradient inside microgels. To construct ECM-like microenvironment with tunable mechanical strength, three gelatin/hyaluronic acid hybrid microgels with low, medium and high crosslinking densities, i.e., Gel-HA(L), Gel-HA(M) and Gel-HA(H), are fabricated in microfluidic devices by Michael addition reaction between thiolated gelatin (Gel-SH) and ethylsulfated hyaluronic acid (HA-VS) with different substitution degrees of vinyl sulfone groups. Our results show that mouse bone marrow mesenchymal stem cell (BMSC) proliferation, distribution and chondrogenesis are all closely dependent on mechanical microenvironments in microgels. Noteworthily, BMSCs show a clear trend of differentiating into hyaline cartilage in Gel-HA(L) and fibrocartilage in Gel-HA(M) and Gel-HA(H). Whole transcriptome RNA sequencing reveals that mechanical microenvironment of microgels affects BMSC differentiation via TGF-β/Smad signaling pathway, Hippo signaling pathway and Integrin/YAP/TAZ signaling pathway. We believe this microgel model provides a new way to further explore the interaction between cells and 3D microenvironment. STATEMENT OF SIGNIFICANCE: In recent years, hydrogels have been frequently used to construct 3D microenvironment for cells. However, their relatively large size not only brings biased experimental results, but also limits high-throughput screening and analysis. Herein we propose a gelatin/hyaluronic acid microgel model to explore the effects of 3D cellular mechanical microenvironment (biophysical cues) on BMSC behaviors especially chondrogenesis, which can minimize the interference of biochemical gradients. Our results reveal that BMSC differentiation into either hyaline cartilage or fibrocartilage can be regulated via tailoring the mechanical properties of microgels. Whole transcriptome RNA sequencing proves that "TGF-β/Smad signaling pathway", "Hippo signaling pathway" and "Integrins/YAP/ TAZ signaling pathway" are activated or inhibited in this process.
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Affiliation(s)
- Qi Feng
- Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou 510006, China; National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China
| | - Huichang Gao
- Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou 510006, China; National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China
| | - Hongji Wen
- Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou 510006, China; National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China
| | - Hanhao Huang
- Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou 510006, China; National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China
| | - Qingtao Li
- National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China; School of Medicine, South China University of Technology, Guangzhou 510006, China
| | - Minhua Liang
- Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou 510006, China; National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China
| | - Yang Liu
- Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou 510006, China; National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China
| | - Hua Dong
- Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou 510006, China; National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China; Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, South China University of Technology, Guangzhou 510006, P R China.
| | - Xiaodong Cao
- Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou 510006, China; National Engineering Research Center for Tissue Restoration and Reconstruction (NERC-TRR), Guangzhou 510006, China; Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, South China University of Technology, Guangzhou 510006, P R China; Guangdong Province Key Laboratory of Biomedical Engineering, South China University of Technology, Guangzhou 510641, China.
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Material-Dependent Formation and Degradation of Bone Matrix-Comparison of Two Cryogels. Bioengineering (Basel) 2020; 7:bioengineering7020052. [PMID: 32517006 PMCID: PMC7378764 DOI: 10.3390/bioengineering7020052] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/15/2020] [Revised: 06/02/2020] [Accepted: 06/03/2020] [Indexed: 01/19/2023] Open
Abstract
Cryogels represent ideal carriers for bone tissue engineering. We recently described the osteogenic potential of cryogels with different protein additives, e.g., platelet-rich plasma (PRP). However, these scaffolds raised concerns as different toxic substances are required for their preparation. Therefore, we developed another gelatin (GEL)-based cryogel. This study aimed to compare the two scaffolds regarding their physical characteristics and their influence on osteogenic and osteoclastic cells. Compared to the PRP scaffolds, GEL scaffolds had both larger pores and thicker walls, resulting in a lower connective density. PRP scaffolds, with crystalized calcium phosphates on the surface, were significantly stiffer but less mineralized than GEL scaffolds with hydroxyapatite incorporated within the matrix. The GEL scaffolds favored adherence and proliferation of the osteogenic SCP-1 and SaOS-2 cells. Macrophage colony-stimulating factor (M-CSF) and osteoprotegerin (OPG) levels seemed to be induced by GEL scaffolds. Levels of other osteoblast and osteoclast markers were comparable between the two scaffolds. After 14 days, mineral content and stiffness of the cryogels were increased by SCP-1 and SaOS-2 cells, especially of PRP scaffolds. THP-1 cell-derived osteoclastic cells only reduced mineral content and stiffness of PRP cryogels. In summary, both scaffolds present powerful advantages; however, the possibility to altered mineral content and stiffness may be decisive when it comes to using PRP or GEL scaffolds for bone tissue engineering.
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McColloch A, Liebman C, Liu H, Cho M. Alterted Adipogenesis of Human Mesenchymal Stem Cells by Photobiomodulation Using 1064 nm Laser Light. Lasers Surg Med 2020; 53:263-274. [PMID: 32495397 DOI: 10.1002/lsm.23278] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/26/2020] [Revised: 04/27/2020] [Accepted: 05/23/2020] [Indexed: 12/21/2022]
Abstract
BACKGROUND AND OBJECTIVES Photobiomodulation (PBM) describes the influence of light irradiation on biological tissues. Laser light in the near-infrared (NIR) spectrum has been shown to mitigate pain, reduce inflammation, and promote wound healing. The cellular mechanism that mediates PBM's effects is generally accepted to be at the site of the mitochondria, leading to an increased flux through the electron transport chain and adenosine triphosphate (ATP) production. Moreover, PBM has been demonstrated to reduce oxidative stress through an increased production of reactive oxygen species (ROS)-sequestering enzymes. The aim of the study is to determine whether these PBM-induced effects expedite or interfere with the intended stem cell differentiation to the adipogenic lineage. STUDY DESIGN/MATERIALS AND METHODS To determine the effects of 1064 nm laser irradiation (fluence of 8.8-26.4 J/cm2 ) on human mesenchymal stem cells (hMSCs) undergoing adipogenic differentiation, the ATP and ROS levels, and adipogenic markers were quantitatively measured. RESULTS At a low fluence (8.8 J/cm2 ) the ATP increase was essentially negligible, whereas a higher fluence induced a significant increase. In the laser-stimulated cells, PBM over time decreased the ROS level compared with the non-treated control group and significantly reduced the extent of adipogenesis. A reduction in the ROS level was correlated with a diminished lipid accumulation, reduced production of adipose-specific genetic markers, and delayed the chemically intended adipogenesis. CONCLUSION We characterized the use of NIR light exposure to modulate adipogenesis. Both the ATP and ROS levels in hMSCs responded to different energy densities. The current study is expected to contribute significantly to the growing field of PBM as well as stem cell tissue engineering by demonstrating the wavelength-dependent responses of hMSC differentiation. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.
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Affiliation(s)
- Andrew McColloch
- Department of Bioengineering, The University of Texas at Arlington, Arlington, Texas, 76019
| | - Caleb Liebman
- Department of Bioengineering, The University of Texas at Arlington, Arlington, Texas, 76019
| | - Hanli Liu
- Department of Bioengineering, The University of Texas at Arlington, Arlington, Texas, 76019
| | - Michael Cho
- Department of Bioengineering, The University of Texas at Arlington, Arlington, Texas, 76019
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Walter T, Gruenewald A, Detsch R, Boccaccini AR, Vogel N. Cell Interactions with Size-Controlled Colloidal Monolayers: Toward Improved Coatings in Bone Tissue Engineering. LANGMUIR : THE ACS JOURNAL OF SURFACES AND COLLOIDS 2020; 36:1793-1803. [PMID: 32017853 DOI: 10.1021/acs.langmuir.9b03308] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/10/2023]
Abstract
The surface structure of biomaterials is of key importance to control its interactions with biological environments. Industrial fabrication and coating processes often introduce particulate nanostructures at implant surfaces. Understanding the cellular interaction with particle-based surface topologies and feature sizes in the colloidal length scale therefore offers the possibility to improve the biological response of synthetic biomaterials. Here, surfaces with controlled topography and regular feature sizes covering the relevant length scale of particulate coatings (100-1000 nm) are fabricated by colloidal templating. Using fluorescent microscopy, WST assay, and morphology analysis, results show that adhesion and attachment of bone-marrow derived murine stromal cells (ST2) are strongly influenced by the surface feature size while geometric details play an insignificant role. Quantitative analysis shows enhanced cell adhesion, spreading, viability, and activity when surface feature size decreases below 200 nm compared to flat surfaces, while larger feature sizes are detrimental to cell adhesion. Kinetic studies reveal that most cells on surfaces with larger features lose contact with the substrate over time. This study identifies colloidal templating as a simple method for creating highly defined model systems to investigate complex cell functions and provides design criteria for the choice of particulate coatings on commercial implant materials.
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Affiliation(s)
- Teresa Walter
- Institute of Particle Technology , Friedrich-Alexander University Erlangen-Nürnberg , Cauerstrasse 4 , 91058 Erlangen , Germany
| | - Alina Gruenewald
- Institute of Biomaterials , Friedrich-Alexander University Erlangen-Nürnberg , Cauerstrasse 6 , 91058 Erlangen , Germany
| | - Rainer Detsch
- Institute of Biomaterials , Friedrich-Alexander University Erlangen-Nürnberg , Cauerstrasse 6 , 91058 Erlangen , Germany
| | - Aldo R Boccaccini
- Institute of Biomaterials , Friedrich-Alexander University Erlangen-Nürnberg , Cauerstrasse 6 , 91058 Erlangen , Germany
| | - Nicolas Vogel
- Institute of Particle Technology , Friedrich-Alexander University Erlangen-Nürnberg , Cauerstrasse 4 , 91058 Erlangen , Germany
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Castilla-Casadiego DA, Reyes-Ramos AM, Domenech M, Almodovar J. Effects of Physical, Chemical, and Biological Stimulus on h-MSC Expansion and Their Functional Characteristics. Ann Biomed Eng 2020; 48:519-535. [PMID: 31705365 PMCID: PMC6952531 DOI: 10.1007/s10439-019-02400-3] [Citation(s) in RCA: 27] [Impact Index Per Article: 5.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2019] [Accepted: 10/30/2019] [Indexed: 01/10/2023]
Abstract
Human adult mesenchymal stem or stromal cells (h-MSC) therapy has gained considerable attention due to the potential to treat or cure diseases given their immunosuppressive properties and tissue regeneration capabilities. Researchers have explored diverse strategies to promote high h-MSC production without losing functional characteristics or properties. Physical stimulus including stiffness, geometry, and topography, chemical stimulus, like varying the surface chemistry, and biochemical stimuli such as cytokines, hormones, small molecules, and herbal extracts have been studied but have yet to be translated to industrial manufacturing practice. In this review, we describe the role of those stimuli on h-MSC manufacturing, and how these stimuli positively promote h-MSC properties, impacting the cell manufacturing field for cell-based therapies. In addition, we discuss other process considerations such as bioreactor design, good manufacturing practice, and the importance of the cell donor and ethics factors for manufacturing potent h-MSC.
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Affiliation(s)
- David A Castilla-Casadiego
- Ralph E. Martin Department of Chemical Engineering, University of Arkansas, 3202 Bell Engineering Center, Fayetteville, AR, 72701, USA
| | - Ana M Reyes-Ramos
- Department of Chemical Engineering, University of Puerto Rico Mayagüez, Call Box 9000, Mayagüez, PR, 00681-9000, USA
| | - Maribella Domenech
- Department of Chemical Engineering, University of Puerto Rico Mayagüez, Call Box 9000, Mayagüez, PR, 00681-9000, USA
| | - Jorge Almodovar
- Ralph E. Martin Department of Chemical Engineering, University of Arkansas, 3202 Bell Engineering Center, Fayetteville, AR, 72701, USA.
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Zhong J, Yang Y, Liao L, Zhang C. Matrix stiffness-regulated cellular functions under different dimensionalities. Biomater Sci 2020; 8:2734-2755. [DOI: 10.1039/c9bm01809c] [Citation(s) in RCA: 21] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
The microenvironments that cells encounter with in vitro.
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Affiliation(s)
- Jiajun Zhong
- Guangdong Provincial Key Laboratory of Sensor Technology and Biomedical Instruments (Sun Yat-sen University)
- School of Biomedical Engineering
- Sun Yat-Sen University
- Guangzhou
- P. R. China
| | - Yuexiong Yang
- Guangdong Provincial Key Laboratory of Sensor Technology and Biomedical Instruments (Sun Yat-sen University)
- School of Biomedical Engineering
- Sun Yat-Sen University
- Guangzhou
- P. R. China
| | - Liqiong Liao
- Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering
- Biomaterials Research Center
- School of Biomedical Engineering
- Southern Medical University
- Guangzhou
| | - Chao Zhang
- Guangdong Provincial Key Laboratory of Sensor Technology and Biomedical Instruments (Sun Yat-sen University)
- School of Biomedical Engineering
- Sun Yat-Sen University
- Guangzhou
- P. R. China
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48
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McColloch A, Rabiei M, Rabbani P, Bowling A, Cho M. Correlation between Nuclear Morphology and Adipogenic Differentiation: Application of a Combined Experimental and Computational Modeling Approach. Sci Rep 2019; 9:16381. [PMID: 31705037 PMCID: PMC6842088 DOI: 10.1038/s41598-019-52926-8] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/01/2019] [Accepted: 10/22/2019] [Indexed: 12/18/2022] Open
Abstract
Stem cells undergo drastic morphological alterations during differentiation. While extensive studies have been performed to examine the cytoskeletal remodeling, there is a growing interest to determine the morphological, structural and functional changes of the nucleus. The current study is therefore aimed at quantifying the extent of remodeling of the nuclear morphology of human mesenchymal stem cells during biochemically-induced adipogenic differentiation. Results show the size of nuclei decreased exponentially over time as the lipid accumulation is up-regulated. Increases in the lipid accumulation appear to lag the nuclear reorganization, suggesting the nuclear deformation is a prerequisite to adipocyte maturation. Furthermore, the lamin A/C expression was increased and redistributed to the nuclear periphery along with a subsequent increase in the nuclear aspect ratio. To further assess the role of the nucleus, a nuclear morphology with a high aspect ratio was achieved using microcontact-printed substrate. The cells with an elongated nuclear shape did not efficiently undergo adipogenesis, suggesting the cellular and nuclear processes associated with stem cell differentiation at the early stage of adipogenesis cause a change in the nuclear morphology and cannot be abrogated by the morphological cues. In addition, a novel computational biomechanical model was generated to simulate the nuclear shape change during differentiation and predict the forces acting upon the nucleus. This effort led to the development of computational scaling approach to simulate the experimentally observed adipogenic differentiation processes over 15 days in less than 1.5 hours.
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Affiliation(s)
- Andrew McColloch
- University of Texas at Arlington, Department of Biomedical Engineering, Arlington, 76010, USA
| | - Manoochehr Rabiei
- University of Texas at Arlington, Department of Mechanical and Aerospace Engineering, Arlington, TX, 76010, USA
| | - Parisa Rabbani
- University of Texas at Arlington, Department of Biomedical Engineering, Arlington, 76010, USA
| | - Alan Bowling
- University of Texas at Arlington, Department of Mechanical and Aerospace Engineering, Arlington, TX, 76010, USA
| | - Michael Cho
- University of Texas at Arlington, Department of Biomedical Engineering, Arlington, 76010, USA.
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Yue D, Zhang M, Lu J, Zhou J, Bai Y, Pan J. The rate of fluid shear stress is a potent regulator for the differentiation of mesenchymal stem cells. J Cell Physiol 2019; 234:16312-16319. [PMID: 30784070 DOI: 10.1002/jcp.28296] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/13/2018] [Revised: 12/15/2018] [Accepted: 12/19/2018] [Indexed: 01/24/2023]
Abstract
We have previously demonstrated that the rate of fluid shear stress (ΔSS) can manipulate the fate of mesenchymal stem cells (MSCs) to osteogenic or chondrogenic cells. However, whether ΔSS is comparable to other two means of induction medium and substrate stiffness that have been proven to be potent in differentiation control is unknown. In this study, we subjected MSCs to 1-7 days of osteogenic or chondrogenic chemical induction, or 1-4 days of 37 or 86 kPa of substrate stiffness induction, followed by 20 min of Fast ΔSS (0-0') or Slow ΔSS (0-2'), which is a laminar FSS that linearly increased from 0 to 10 dyn/cm 2 in 0 (Fast) or 2 min (Slow) and maintained at 10 dyn/cm 2 for a total of 20 min. We found that 20 min of ΔSS could compete with 5 days' chemical and 2 days' substrate stiffness inductions. Our study confirmed that ΔSS is a powerful tool to control the differentiation of MSCs, which stressed the possible application in MSCs linage specification.
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Affiliation(s)
- Danyang Yue
- Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, College of Bioengineering, Chongqing University, Chongqing, China
| | - Mengxue Zhang
- Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, College of Bioengineering, Chongqing University, Chongqing, China
| | - Juan Lu
- Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, College of Bioengineering, Chongqing University, Chongqing, China
| | - Jin Zhou
- Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, College of Bioengineering, Chongqing University, Chongqing, China
| | - Yuying Bai
- Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, College of Bioengineering, Chongqing University, Chongqing, China
| | - Jun Pan
- Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, College of Bioengineering, Chongqing University, Chongqing, China
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Häussling V, Deninger S, Vidoni L, Rinderknecht H, Ruoß M, Arnscheidt C, Athanasopulu K, Kemkemer R, Nussler AK, Ehnert S. Impact of Four Protein Additives in Cryogels on Osteogenic Differentiation of Adipose-Derived Mesenchymal Stem Cells. Bioengineering (Basel) 2019; 6:67. [PMID: 31394780 PMCID: PMC6784125 DOI: 10.3390/bioengineering6030067] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/29/2019] [Revised: 07/30/2019] [Accepted: 08/05/2019] [Indexed: 12/21/2022] Open
Abstract
Human adipose-derived mesenchymal stem/stromal cells (Ad-MSCs) have great potential for bone tissue engineering. Cryogels, mimicking the three-dimensional structure of spongy bone, represent ideal carriers for these cells. We developed poly(2-hydroxyethyl methacrylate) cryogels, containing hydroxyapatite to mimic inorganic bone matrix. Cryogels were additionally supplemented with different types of proteins, namely collagen (Coll), platelet-rich plasma (PRP), immune cells-conditioned medium (CM), and RGD peptides (RGD). The different protein components did not affect scaffolds' porosity or water-uptake capacity, but altered pore size and stiffness. Stiffness was highest in scaffolds with PRP (82.3 kPa), followed by Coll (55.3 kPa), CM (45.6 kPa), and RGD (32.8 kPa). Scaffolds with PRP, CM, and Coll had the largest pore diameters (~60 µm). Ad-MSCs were osteogenically differentiated on these scaffolds for 14 days. Cell attachment and survival rates were comparable for all four scaffolds. Runx2 and osteocalcin levels only increased in Ad-MSCs on Coll, PRP and CM cryogels. Osterix levels increased slightly in Ad-MSCs differentiated on Coll and PRP cryogels. With differentiation alkaline phosphatase activity decreased under all four conditions. In summary, besides Coll cryogel our PRP cryogel constitutes as an especially suitable carrier for bone tissue engineering. This is of special interest, as this scaffold can be generated with patients' PRP.
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Affiliation(s)
- Victor Häussling
- Siegfried Weller Research Institute, BG Unfallklinik Tuebingen, Department of Trauma and Reconstructive Surgery, Eberhard Karls University Tübingen, 72074 Tübingen, Germany
| | - Sebastian Deninger
- Siegfried Weller Research Institute, BG Unfallklinik Tuebingen, Department of Trauma and Reconstructive Surgery, Eberhard Karls University Tübingen, 72074 Tübingen, Germany
| | - Laura Vidoni
- Siegfried Weller Research Institute, BG Unfallklinik Tuebingen, Department of Trauma and Reconstructive Surgery, Eberhard Karls University Tübingen, 72074 Tübingen, Germany
| | - Helen Rinderknecht
- Siegfried Weller Research Institute, BG Unfallklinik Tuebingen, Department of Trauma and Reconstructive Surgery, Eberhard Karls University Tübingen, 72074 Tübingen, Germany
| | - Marc Ruoß
- Siegfried Weller Research Institute, BG Unfallklinik Tuebingen, Department of Trauma and Reconstructive Surgery, Eberhard Karls University Tübingen, 72074 Tübingen, Germany
| | - Christian Arnscheidt
- Siegfried Weller Research Institute, BG Unfallklinik Tuebingen, Department of Trauma and Reconstructive Surgery, Eberhard Karls University Tübingen, 72074 Tübingen, Germany
| | - Kiriaki Athanasopulu
- Department of Applied Chemistry Reutlingen University, 72762 Reutlingen, Germany
| | - Ralf Kemkemer
- Department of Applied Chemistry Reutlingen University, 72762 Reutlingen, Germany
| | - Andreas K Nussler
- Siegfried Weller Research Institute, BG Unfallklinik Tuebingen, Department of Trauma and Reconstructive Surgery, Eberhard Karls University Tübingen, 72074 Tübingen, Germany.
| | - Sabrina Ehnert
- Siegfried Weller Research Institute, BG Unfallklinik Tuebingen, Department of Trauma and Reconstructive Surgery, Eberhard Karls University Tübingen, 72074 Tübingen, Germany
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