title: *Design of an In Vitro Model of Cell Recruitment for Skeletal Muscle Regeneration Using Hepatocyte Growth Factor-Loaded Fibrin Microthreads.
abstract: Large skeletal muscle defects that result in volumetric muscle loss (VML) result in the destruction of the basal lamina, which removes key signaling molecules such as hepatocyte growth factor (HGF) from the wound site, eliminating the endogenous capacity of these injuries to regenerate. We recently showed that HGF-loaded fibrin microthreads increased the force production in muscle tissues after 60 days in a mouse VML model. In this study, we created an in vitro, three-dimensional (3D) microscale outgrowth assay system designed to mimic cell recruitment in vivo, and investigated the effect of HGF-loaded, cross-linked fibrin microthreads on myoblast recruitment to predict the results observed in vivo. This outgrowth assay discretely separated the cellular and molecular functions (migration, proliferation, and chemotaxis) that direct outgrowth from the wound margin, creating a powerful platform to model cell recruitment in axially aligned tissues, such as skeletal muscle. The degree of cross-linking was controlled by pH and microthreads cross-linked using physiologically neutral pH (EDCn) facilitated the release of active HGF; increasing the two-dimensional migration and 3D outgrowth of myoblasts twofold. While HGF adsorbed to uncross-linked microthreads, it did not enhance myoblast migration, possibly due to the low concentrations that were adsorbed. Regardless of the amount of HGF adsorbed on the microthreads, myoblast proliferation increased significantly on stiffer, cross-linked microthreads. Together, the results of these studies show that HGF loaded onto EDCn microthreads supported enhanced myoblast migration and recruitment and suggest that our novel outgrowth assay system is a robust in vitro screening tool that predicts the performance of fibrin microthreads in vivo.
authors.affiliation: 1 Department of Biomedical Engineering, Worcester Polytechnic Institute , Worcester, Massachusetts.|2 Bioengineering Institute, Worcester Polytechnic Institute , Worcester, Massachusetts.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Grasman
authors.affiliation: 1 Department of Biomedical Engineering, Worcester Polytechnic Institute , Worcester, Massachusetts.|2 Bioengineering Institute, Worcester Polytechnic Institute , Worcester, Massachusetts.
authors.forename: Raymond L
authors.initials: RL
authors.lastname: Page
authors.affiliation: 1 Department of Biomedical Engineering, Worcester Polytechnic Institute , Worcester, Massachusetts.|2 Bioengineering Institute, Worcester Polytechnic Institute , Worcester, Massachusetts.
authors.forename: George D
authors.initials: GD
authors.lastname: Pins
mesh_terms: D000327:Adsorption; D000818:Animals; D002417:Cattle; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000494:pharmacology; D005337:Fibrin / Q000494:pharmacology*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D051379:Mice; D018482:Muscle, Skeletal / Q000187:drug effects / Q000502:physiology*; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D003432:Cross-Linking Reagents; D012710:Serum Albumin, Bovine; D017228:Hepatocyte Growth Factor; D005337:Fibrin
keywords: 3D in vitro model system; biomaterials; cell migration; fibrin microthreads; hepatocyte growth factor; skeletal muscle regeneration; tissue engineering