Induction of Tenogenic Differentiation Mediated by Extracellular Tendon Matrix and Short-Term Cyclic Stretching.
Abstract: Tendon and ligament pathologies are still a therapeutic challenge, due to the difficulty in restoring the complex extracellular matrix architecture and biomechanical strength. While progress is being made in cell-based therapies and tissue engineering approaches, comprehensive understanding of the fate of progenitor cells in tendon healing is still lacking. The aim of this study was to investigate the effect of decellularized tendon matrix and moderate cyclic stretching as natural stimuli which could potentially direct tenogenic fate. Equine adipose-derived mesenchymal stromal cells (MSC) were seeded on decellularized tendon matrix scaffolds. Mechanical stimulation was applied in a custom-made cyclic strain bioreactor. Assessment was performed 4 h, 8 h, and 24 h following mechanical stimulation. Scaffold culture induced cell alignment and changes in expression of tendon-related genes, although cell viability was decreased compared to monolayer culture. Short mechanical stimulation periods enhanced most of the scaffold-induced effects. Collagen 1A2 expression levels were decreased, while collagen 3A1 and decorin levels were increased. Tenascin-C and scleraxis expression showed an initial decrease but had increased 24 h after stimulation. The results obtained suggest that decellularized tendon matrix, supported by cyclic stretching, can induce tenogenic differentiation and the synthesis of tendon components important for matrix remodeling.
Publication Date: 2016-08-18 PubMed ID: 27630718PubMed Central: PMC5007347DOI: 10.1155/2016/7342379Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The study focuses on understanding how the application of a natural stimulus like decellularized tendon matrix and moderate cyclic stretching could potentially aid in tendon healing. The researchers discovered that decellularized tendon matrix with support from cyclic stretching can result in tenogenic differentiation and synthesis of tendon components crucial for matrix remodeling.
Objective and Need of the Study
- The main objective of this investigation is to understand the effect of natural stimuli including a decellularized tendon matrix and moderate cyclic stretching on tendon healing.
- Rationale behind the research is the challenge prevailing in treating tendon and ligament pathologies due to the difficulty in restoring the complex extracellular matrix architecture and biomechanical strength required for tendon healing.
- There is a significant lack of comprehensive understanding of how progenitor cells behave during tendon healing which this study attempts to analyze.
Methodology
- The research used Equine adipose-derived mesenchymal stromal cells (MSC) which were seeded on decellularized tendon matrix scaffolds.
- Cyclic strain was applied using a custom-made bioreactor. This mechanical stimulation was applied for 4 h, 8 h, and 24 h duration to evaluate the impact of the stimulus on cells.
Observations and Conclusions
- The scaffold culture process induced cell alignment as well as changes in the expression of tendon-associated genes. However, the cell viability noticed was decreased as compared to the traditional monolayer cell culture technique.
- Short periods of mechanical stimulation enhanced most of the effects caused by the scaffold. For instance, expression levels of Collagen 1A2 were decreased while Collagen 3A1 and decorin levels witnessed an increase.
- Expression of Tenascin-C and scleraxis showed an initial drop but exhibited an increase 24 h after stimulation.
- The results imply that utilizing decellularized tendon matrix, augmented by cyclic stretching, can encourage tenogenic differentiation. This further leads to the synthesis of significant tendon components that contribute to matrix remodeling, highlighting a prospective approach to tendon healing.
Cite This Article
APA
Burk J, Plenge A, Brehm W, Heller S, Pfeiffer B, Kasper C.
(2016).
Induction of Tenogenic Differentiation Mediated by Extracellular Tendon Matrix and Short-Term Cyclic Stretching.
Stem Cells Int, 2016, 7342379.
https://doi.org/10.1155/2016/7342379 Publication
Researcher Affiliations
- Translational Centre for Regenerative Medicine, University of Leipzig, Philipp-Rosenthal-Strasse 55, 04103 Leipzig, Germany; Saxon Incubator for Clinical Translation, University of Leipzig, Philipp-Rosenthal-Strasse 55, 04103 Leipzig, Germany; Institute of Veterinary Physiology, University of Leipzig, An den Tierkliniken 7, 04103 Leipzig, Germany.
- Large Animal Clinic for Surgery, University of Leipzig, An den Tierkliniken 21, 04103 Leipzig, Germany.
- Translational Centre for Regenerative Medicine, University of Leipzig, Philipp-Rosenthal-Strasse 55, 04103 Leipzig, Germany; Saxon Incubator for Clinical Translation, University of Leipzig, Philipp-Rosenthal-Strasse 55, 04103 Leipzig, Germany; Large Animal Clinic for Surgery, University of Leipzig, An den Tierkliniken 21, 04103 Leipzig, Germany.
- Translational Centre for Regenerative Medicine, University of Leipzig, Philipp-Rosenthal-Strasse 55, 04103 Leipzig, Germany; Department of Pathology and Laboratory Medicine, Tulane University, 1430 Tulane Avenue, New Orleans, LA 70112, USA.
- Large Animal Clinic for Surgery, University of Leipzig, An den Tierkliniken 21, 04103 Leipzig, Germany.
- Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Muthgasse 18, 1190 Vienna, Austria.
References
This article includes 43 references
- Dowling B. A., Dart A. J., Hodgson D. R., Smith R. K. W.. Superficial digital flexor tendonitis in the horse.. Equine Veterinary Journal 2000;32(5):369–378.
- Doroski D. M., Brink K. S., Temenoff J. S.. Techniques for biological characterization of tissue-engineered tendon and ligament.. Biomaterials 2007;28(2):187–202.
- Kader D., Saxena A., Movin T., Maffulli N.. Achilles tendinopathy: some aspects of basic science and clinical management.. British Journal of Sports Medicine 2002;36(4):239–249.
- Schnabel L. V., Lynch M. E., Van Der Meulen M. C. H., Yeager A. E., Kornatowski M. A., Nixon A. J.. Mesenchymal stem cells and insulin-like growth factor-I gene-enhanced mesenchymal stem cells improve structural aspects of healing in equine flexor digitorum superficialis tendons.. Journal of Orthopaedic Research 2009;27(10):1392–1398.
- Crovace A., Lacitignola L., Rossi G., Francioso E.. Histological and immunohistochemical evaluation of autologous cultured bone marrow mesenchymal stem cells and bone marrow mononucleated cells in collagenase-induced tendinitis of equine superficial digital flexor tendon.. Veterinary Medicine International 2010;2010:10.
- Smith R. K. W., Werling N. J., Dakin S. G., Alam R., Goodship A. E., Dudhia J.. Beneficial effects of autologous bone marrow-derived mesenchymal stem cells in naturally occurring tendinopathy.. PLoS ONE 2013;8(9).
- Guest D. J., Smith M. R. W., Allen W. R.. Monitoring the fate of autologous and allogeneic mesenchymal progenitor cells injected into the superficial digital flexor tendon of horses: preliminary study.. Equine Veterinary Journal 2008;40(2):178–181.
- Burk J., Gittel C., Heller S.. Gene expression of tendon markers in mesenchymal stromal cells derived from different sources.. BMC Research Notes 2014;7, article 826.
- Barsby T., Guest D.. Transforming growth factor beta3 promotes tendon differentiation of equine embryo-derived stem cells.. Tissue Engineering Part A 2013;19(19-20):2156–2165.
- Raabe O., Shell K., Fietz D.. Tenogenic differentiation of equine adipose-tissue-derived stem cells under the influence of tensile strain, growth differentiation factors and various oxygen tensions.. Cell and Tissue Research 2013;352(3):509–521.
- Brown J. P., Finley V. G., Kuo C. K.. Embryonic mechanical and soluble cues regulate tendon progenitor cell gene expression as a function of developmental stage and anatomical origin.. Journal of Biomechanics 2014;47(1):214–222.
- Alberton P., Popov C., Prägert M.. Conversion of human bone marrow-derived mesenchymal stem cells into tendon progenitor cells by ectopic expression of scleraxis.. Stem Cells and Development 2012;21(6):846–858.
- Chen X., Yin Z., Chen J.-L.. Scleraxis-overexpressed human embryonic stem cell-derived mesenchymal stem cells for tendon tissue engineering with knitted silk-collagen scaffold.. Tissue Engineering Part A 2014;20(11-12):1583–1592.
- Kishore V., Bullock W., Sun X., Van Dyke W. S., Akkus O.. Tenogenic differentiation of human MSCs induced by the topography of electrochemically aligned collagen threads.. Biomaterials 2012;33(7):2137–2144.
- Yin Z., Chen X., Song H.-X.. Electrospun scaffolds for multiple tissues regeneration invivo through topography dependent induction of lineage specific differentiation.. Biomaterials 2015;44:173–185.
- Zhang J., Li B., Wang J. H.-C.. The role of engineered tendon matrix in the stemness of tendon stem cells in vitro and the promotion of tendon-like tissue formation in vivo .. Biomaterials 2011;32(29):6972–6981.
- Yang G., Rothrauff B. B., Lin H., Gottardi R., Alexander P. G., Tuan R. S.. Enhancement of tenogenic differentiation of human adipose stem cells by tendon-derived extracellular matrix.. Biomaterials 2013;34(37):9295–9306.
- Durgam S. S., Stewart A. A., Pondenis H. C., Yates A. C., Evans R. B., Stewart M. C.. Responses of equine tendon- and bone marrow–derived cells to monolayer expansion with fibroblast growth factor-2 and sequential culture with pulverized tendon and insulin-like growth factor-I.. American Journal of Veterinary Research 2012;73(1):162–170.
- Little D., Guilak F., Ruch D. S.. Ligament-derived matrix stimulates a ligamentous phenotype in human adipose-derived stem cells.. Tissue Engineering—Part A 2010;16(7):2307–2319.
- Yin Z., Chen X., Zhu T.. The effect of decellularized matrices on human tendon stem/progenitor cell differentiation and tendon repair.. Acta Biomaterialia 2013;9(12):9317–9329.
- Altman G. H., Horan R. L., Martin I.. Cell differentiation by mechanical stress.. The FASEB Journal 2002;16(2):270–272.
- Doroski D. M., Levenston M. E., Temenoff J. S.. Cyclic tensile culture promotes fibroblastic differentiation of marrow stromal cells encapsulated in poly(ethylene glycol)-based hydrogels.. Tissue Engineering Part A 2010;16(11):3457–3466.
- Kreja L., Liedert A., Schlenker H.. Effects of mechanical strain on human mesenchymal stem cells and ligament fibroblasts in a textured poly(L-lactide) scaffold for ligament tissue engineering.. Journal of Materials Science: Materials in Medicine 2012;23(10):2575–2582.
- Kuo C. K., Tuan R. S.. Mechanoactive tenogenic differentiation of human mesenchymal stem cells.. Tissue Engineering Part A 2008;14(10):1615–1627.
- Scott A., Danielson P., Abraham T., Fong G., Sampaio A. V., Underhill T. M.. Mechanical force modulates scleraxis expression in bioartificial tendons.. Journal of Musculoskeletal Neuronal Interactions 2011;11(2):124–132.
- Joshi S. D., Webb K.. Variation of cyclic strain parameters regulates development of elastic modulus in fibroblast/substrate constructs.. Journal of Orthopaedic Research 2008;26(8):1105–1113.
- Rui Y. F., Lui P. P. Y., Ni M., Chan L. S., Lee Y. W., Chan K. M.. Mechanical loading increased BMP-2 expression which promoted osteogenic differentiation of tendon-derived stem cells.. Journal of Orthopaedic Research 2011;29(3):390–396.
- Chen Y.-J., Huang C.-H., Lee I.-C., Lee Y.-T., Chen M.-H., Young T.-H.. Effects of cyclic mechanical stretching on the mRNA expression of tendon/ligament-related and osteoblast-specific genes in human mesenchymal stem cells.. Connective Tissue Research 2008;49(1):7–14.
- Burk J., Erbe I., Berner D.. Freeze-thaw cycles enhance decellularization of large tendons.. Tissue Engineering Part C 2014;20(4):276–284.
- Diederichs S., Böhm S., Peterbauer A., Kasper C., Scheper T., van Griensven M.. Application of different strain regimes in two-dimensional and three-dimensional adipose tissue–derived stem cell cultures induces osteogenesis: implications for bone tissue engineering.. Journal of Biomedical Materials Research Part A 2010;94(3):927–936.
- Burk J., Brehm W., Aldag A.. Development of a new in vitro tenogenic differentiation model.. Journal of Tissue Engineering and Regenerative Medicine 2014;8(supplement 1):p. 475.
- Burk J., Ribitsch I., Gittel C.. Growth and differentiation characteristics of equine mesenchymal stromal cells derived from different sources.. Veterinary Journal 2013;195(1):98–106.
- Paebst F., Piehler D., Brehm W.. Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: an approach toward a standardized definition.. Cytometry Part A 2014;85(8):678–687.
- Pfaffl M. W.. A new mathematical model for relative quantification in real-time RT-PCR.. Nucleic Acids Research 2001;29, article e45.
- Bourin P., Bunnell B. A., Casteilla L.. Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT). Cytotherapy 2013;15(6):641–648.
- Smith R. K. W., Garvican E. R., Fortier L. A.. The current 'state of play' of regenerative medicine in horses: what the horse can tell the human.. Regenerative Medicine 2014;9(5):673–685.
- Xu B., Song G., Ju Y., Li X., Song Y., Watanabe S.. RhoA/ROCK, cytoskeletal dynamics, and focal adhesion kinase are required for mechanical stretch-induced tenogenic differentiation of human mesenchymal stem cells.. Journal of Cellular Physiology 2012;227(6):2722–2729.
- Dowling B. A., Dart A. J.. Mechanical and functional properties of the equine superficial digital flexor tendon.. Veterinary Journal 2005;170(2):184–192.
- Youngstrom D. W., Rajpar I., Kaplan D. L., Barrett J. G.. A bioreactor system for in vitro tendon differentiation and tendon tissue engineering.. Journal of Orthopaedic Research 2015;33(6):911–918.
- Gratzer P. F., Harrison R. D., Woods T.. Matrix alteration and not residual sodium dodecyl sulfate cytotoxicity affects the cellular repopulation of a decellularized matrix.. Tissue Engineering 2006;12(10):2975–2983.
- Scott A., Cook J. L., Hart D. A., Walker D. C., Duronio V., Khan K. M.. Tenocyte responses to mechanical loading in vivo: a role for local insulin-like growth factor 1 signaling in early tendinosis in rats.. Arthritis and Rheumatism 2007;56(3):871–881.
- Godwin E. E., Young N. J., Dudhia J., Beamish I. C., Smith R. K. W.. Implantation of bone marrow-derived mesenchymal stem cells demonstrates improved outcome in horses with overstrain injury of the superficial digital flexor tendon.. Equine Veterinary Journal 2012;44(1):25–32.
- Youngstrom D. W., LaDow J. E., Barrett J. G.. Tenogenesis of bone marrow-, adipose-, and tendon-derived stem cells in a dynamic bioreactor.. Connective Tissue Research 2016.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists