Laminin-Coated Poly(Methyl Methacrylate) (PMMA) Nanofiber Scaffold Facilitates the Enrichment of Skeletal Muscle Myoblast Population

Abstract

Myoblasts, the contractile cells of skeletal muscle, have been invaluable for fundamental studies of muscle development and clinical applications for muscle loss. A major limitation to the myoblast-based therapeutic approach is contamination with non-contractile fibroblasts, which overgrow during cell expansion. To overcome these limitations, this study was carried out to establish a 3D culture environment using nanofiber scaffolds to enrich the myoblast population during construct formation. Poly(methyl methacrylate) (PMMA) nanofiber (PM) scaffolds were fabricated using electrospinning techniques and coated with extracellular matrix (ECM) proteins, such as collagen or laminin, in the presence or absence of genipin. A mixed population of myoblasts and fibroblasts was isolated from human skeletal muscle tissues and cultured on plain surfaces, as well as coated and non-coated PM scaffolds. PMMA can produce smooth fibers with an average diameter of 360 ± 50 nm. Adsorption of collagen and laminin on PM scaffolds is significantly enhanced in the presence of genipin, which introduces roughness to the nanofiber surface without affecting fiber diameter and mechanical properties. It was also demonstrated that laminin-coated PM scaffolds significantly enhance myoblast proliferation (0.0081 ± 0.0007 h−1) and migration (0.26 ± 0.04 μm/min), while collagen-coated PM scaffolds favors fibroblasts proliferation (0.0097 ± 0.0009 h−1) and migration (0.23 ± 0.03 μm/min). Consequently, the myoblast population was enriched on laminin-coated PM scaffolds throughout the culture process. Therefore, laminin coating of nanofiber scaffolds could be a potential scaffold for the development of a tissue-engineered muscle substitute.

Electrospun Nanometer to Micrometer Scale Biomimetic Synthetic Membrane Scaffolds in Drug Delivery and Tissue Engineering: A Review

“…Hence, currently, a blend of both natural and synthetic polymers is largely preferred to fabricate electrospun nanofibers to exploit their respective strengths [60]. He et al co-electrospun collagen type I with poly (L-lactic acid)-co-poly(ε-caprolactone) (PLLA-CL) to produce nanofiber scaffolds, which enhanced the growth of human artery endothelial cells [63]. Another method to use multiple polymers is by layering individual fiber meshes one on top of another by sequential electrospinning.…”

Tailoring the Interface of Biomaterials to Design Effective Scaffolds

“…Interesting results on the application of nanofibers as scaffolds for TE come also from skeletal muscle and skin TE. Laminin-coated poly(methyl methacrylate) nanofibers showed to promote primary myoblasts proliferation, while nanofiber silk fibroin scaffold revealed to have great potential in wounds healing of skin in vivo [ 57 , 58 ].…”

Mechanophysical Cues in Extracellular Matrix Regulation of Cell Behavior

“…Physical factors, such as mechanical forces exerted on cells, play an important role in the adhesion and formation of cells and tissue . Biomechanics is an interdisciplinary field that studies such interactions .…”

“…The ECM has been studied as a scaffold for cell culture for over 40 years . Laminin, fibronectin, collagen, and vitronectin have been adopted as cell culturing scaffolds, substrates or components. These macromolecules facilitate cell culturing and improve cell survival rates .…”

“…Laminin, fibronectin, collagen, and vitronectin have been adopted as cell culturing scaffolds, substrates or components. These macromolecules facilitate cell culturing and improve cell survival rates . Cell mechanical support by the ECM relates mainly to enhancing cell survival rate, while high cell adhesiveness to the ECM results in a high degree of cell spreading.…”