Muscle Tissue Engineering Using Gingival Mesenchymal Stem Cells Encapsulated in Alginate Hydrogels Containing Multiple Growth Factors

Abstract

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 analyses 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.

Dental and orofacial mesenchymal stem cells in craniofacial regeneration: The prosthodontist’s point of view

“…Our group and others 64–70 have confirmed that alginate hydrogel scaffold can be considered as a promising encapsulating biomaterial for dental- and orofacial-derived MSCs, including GMSCs and PDLSCs (Fig. 2).…”

“…More recently, Ansari et al 70 confirmed that GMSCs, when encapsulated in an alginate hydrogel microencapsulation system with multiple growth factor delivery capacity (loaded with myogenic growth factors), can be used in the repair and regeneration of muscular tissues.…”

Hydrogel biomaterials and their therapeutic potential for muscle injuries and muscular dystrophies

“…More interestingly, the expression level of myogenic-related genes (MyoG, MyoD and Myf5) in GMSCs encapsulated in alginate hydrogel was higher than in GMSCs that were not encapsulated. Most importantly, during in vivo trails, the formation of small islands of muscle-like structures was observed eight weeks following subcutaneous transplantation of GMSCs in microspheres into immunocompromised mice figure 4b [74].…”

“…For example, alginate is a naturally occurring linear polysaccharide typically derived from brown seaweed that has been widely used as a hydrogel for tissue engineering and cell encapsulation applications due to its favourable properties including low toxicity, high biocompatibility and ease of gelation [107]. Possessing a porous microstructure allowing the diffusion of oxygen, nutrients and growth factors makes this material highly suitable as a scaffold for the growth and differentiation of myogenic cells in mass culture and for myoblast engraftment in skeletal muscle regeneration [33,74]. Among the natural hydrogels, those derived from decellularized extracellular matrix (ECM) have gained much attention due to their ability to conform to irregular shapes and the inherent bioactivity of the matrix source.…”

Regenerative Medicine Weapons to Fight the Degenerative Defect of Musculoskeletal System Disorders

“…The concept of combining the stem cells, growth factors and scaffold was demonstrated to be efficient for regeneration of bone [1,2] , cartilage [3] , muscle [4] , tendon [5] and nerve [6] . Most of these studies was conducted at in vitro and in vivo level, supposing that will perform the same efficiency when be applied in the clinical cases.…”