Muscle Tissue Engineering Using Gingival Mesenchymal Stem Cells Encapsulated in Alginate Hydrogels Containing Multiple Growth Factors
- 864 Downloads
Repair and regeneration of muscle tissue following traumatic injuries or muscle diseases often presents a challenging clinical situation. If a significant amount of tissue is lost the native regenerative potential of skeletal muscle will not be able to grow to fill the defect site completely. Dental-derived mesenchymal stem cells (MSCs) in combination with appropriate scaffold material, present an advantageous alternative therapeutic option for muscle tissue engineering in comparison to current treatment modalities available. To date, there has been no report on application of gingival mesenchymal stem cells (GMSCs) in three-dimensional scaffolds for muscle tissue engineering. The objectives of the current study were to develop an injectable 3D RGD-coupled alginate scaffold with multiple growth factor delivery capacity for encapsulating GMSCs, and to evaluate the capacity of encapsulated GMSCs to differentiate into myogenic tissue in vitro and in vivo where encapsulated GMSCs were transplanted subcutaneously into immunocompromised mice. The results demonstrate that after 4 weeks of differentiation in vitro, GMSCs as well as the positive control human bone marrow mesenchymal stem cells (hBMMSCs) exhibited muscle cell-like morphology with high levels of mRNA expression for gene markers related to muscle regeneration (MyoD, Myf5, and MyoG) via qPCR measurement. Our quantitative PCR analyzes revealed that the stiffness of the RGD-coupled alginate regulates the myogenic differentiation of encapsulated GMSCs. Histological and immunohistochemical/fluorescence staining for protein markers specific for myogenic tissue confirmed muscle regeneration in subcutaneous transplantation in our in vivo animal model. GMSCs showed significantly greater capacity for myogenic regeneration in comparison to hBMMSCs (p < 0.05). Altogether, our findings confirmed that GMSCs encapsulated in RGD-modified alginate hydrogel with multiple growth factor delivery capacity is a promising candidate for muscle tissue engineering.
KeywordsTissue engineering Muscle regeneration Dental mesenchymal stem cells RGD-coupled alginate hydrogel
This work was supported by grants from the National Institute of Dental, Craniofacial Research (K08DE023825 to A.M. and R01 DE017449 to S.S.). The authors declare no potential conflicts of interest with respect to the authorship and/or publication of this article.
- 7.Borselli, C., H. Storrie, F. Benesch-Lee, D. Shvartsman, C. Cezar, J. W. Lichtman, H. H. Vandenburgh, and D. J. Mooney. Functional muscle regeneration with combined delivery of angiogenesis and myogenesis factors. Proc. Natl. Acad. Sci. USA 107:3287–3292, 2010.CrossRefPubMedPubMedCentralGoogle Scholar
- 29.Morosetti, R., M. Mirabella, C. Gliubizzi, A. Broccolini, L. De Angelis, E. Tagliafico, et al. MyoD expression restores defective myogenic differentiation of human mesoangioblasts from inclusion-body myositis muscle. Proc. Natl. Acad. Sci. USA 103:16995–17000, 2006.CrossRefPubMedPubMedCentralGoogle Scholar
- 31.Moshaverinia, A., C. Chen, K. Akiyama, S. Ansari, X. Xu, W. W. Chee, et al. Alginate hydrogel as a promising scaffold for dental-derived stem cells: an in vitro study. J. Mater. Sci: Mater. Med. 23:3041–3051, 2012.Google Scholar
- 32.Moshaverinia, A., C. Chen, K. Akiyama, X. Xu, W. W. Chee, S. R. Schricker, and S. Shi. Encapsulated dental-derived stem cells in an injectable and biodegradable scaffold for applications in bone tissue engineering. J. Biomed. Mater. Res. Part. A. 101:3285–3294, 2013.Google Scholar
- 33.Moshaverinia, A., C. Chen, X. Xu, K. Akiyama, S. Ansari, H. H. Zadeh, and S. Shi. Bone regeneration potential of stem cells derived from periodontal ligament or gingival tissue sources encapsulated in RGD-modified alginate scaffold. Tissue Eng. Part A. 20:611–621, 2013.PubMedPubMedCentralGoogle Scholar
- 34.Moshaverinia, A., C. Chen, X. Xu, S. Ansari, H. H. Zadeh, S. R. Schricker, et al. Regulation of the stem cell-host immune system interplay using hydrogel coencapsulation system with an anti-inflammatory drug. Adv Funct Mater. 15:2296–2307, 2015.Google Scholar
- 41.Sachlos, E., and J. T. Czernuszka. Making tissue engineering scaffolds work. Review on the application of solid freeform fabrication technology to the production of tissue engineering scaffold. Euro. Cell. Mater. 5:29–40, 2003.Google Scholar
- 42.Salani, S., C. Donadoni, F. Rizzo, N. Bresolin, G. P. Comi, and S. Corti. Generation of skeletal muscle cells from embryonic and induced pluripotent stem cells as an in vitro model and for therapy of muscular dystrophies. J. Cell Mol. Med. 16:1353–1364, 2012.CrossRefPubMedPubMedCentralGoogle Scholar
- 54.Zhang, Q., S. Shi, Y. Liu, J. Uyanne, Y. Shi, S. Shi, et al. mesenchymal stem cells derived from human gingiva are capable of immunomodulatory functions and ameliorate inflammation-related tissue destruction in experimental colitis. J. Immunol. 183:7787–7798, 2009.CrossRefPubMedPubMedCentralGoogle Scholar