The Application of Three‐Dimensional Collagen‐Scaffolds Seeded with Myoblasts to Repair Skeletal Muscle Defects

Abstract

Three-dimensional (3D) engineered tissue constructs are a novel and promising approach to tissue repair and regeneration. 3D tissue constructs have the ability to restore form and function to damaged soft tissue unlike previous methods, such as plastic surgery, which are able to restore only form, leaving the function of the soft tissue often compromised. In this study, we seeded murine myoblasts (C2C12) into a collagen composite scaffold and cultured the scaffold in a roller bottle cell culture system in order to create a 3D tissue graft in vitro. The 3D graft created in vitro was then utilized to investigate muscle tissue repair in vivo. The 3D muscle grafts were implanted into defect sites created in the skeletal muscles in mice. We detected that the scaffolds degraded slowly over time, and muscle healing was improved which was shown by an increased quantity of innervated and vascularized regenerated muscle fibers. Our results suggest that the collagen composite scaffold seeded with myoblasts can create a 3D muscle graft in vitro that can be employed for defect muscle tissue repair in vivo.

Therapeutic angiogenesis by a myoblast layer harvested by tissue transfer printing from cell-adhesive, thermosensitive hydrogels

“…It has been reported that myoblasts have the morphological and functional capacity to regenerate muscle [35,36]. Although myoblasts cannot directly participate in neovascularization, the paracrine signaling effects of myoblasts after transplantation have been reported and these secreted factors may contribute indirectly to therapeutic angiogenesis [37].…”

Biomaterials based strategies for skeletal muscle tissue engineering: Existing technologies and future trends

“…One of the issues associated with protein-based scaffolds is immune rejection and the onset of a foreign body response, leading to many in vivo studies being carried out in immunodeficient animal models. For example, Ma et al used a porous collagen scaffold seeded with murine myoblasts for the treatment of skeletal muscle defects, and reported that although vascularization, innervation, and the generation of myofibers were observed, successful integration of the scaffold-tissue graft was only evident in immunecompromised animals [88].…”

Engineering Muscle Constructs for the Creation of Functional Engineered Musculoskeletal Tissue

“…These microfabricated hydrogel substrates allow for finely tuned special features, such as grooves and wells with specific placement of proteins to direct cell attachment, proliferation and fate [53,54]. Finally, gel-based scaffolds of fibrin, collagen, and other ECM proteins are seeded with MPCs in their commercially available gel form [3,17,19,46], or can be suspended into matrices with a determined pore size by cryogelation [18,55]. …”

“…Scaffold-free technologies utilize fibroblasts obtained from skeletal muscle co-cultured with muscle satellite cells to create cell monolayers that self-assemble into 3D constructs [17,19,21,26,43,44]. Our scaffoldless tissue engineered muscle constructs are unique in that they are co-cultured with engineered tendon constructs and thus are fabricated with functional tendon ends [11,19,23,45,46]. …”