Induction of mesenchymal stem cell differentiation in the absence of soluble inducer for cutaneous wound regeneration by a chitin nanofiber-based hydrogel

J Tissue Eng Regen Med. 2018 Feb;12(2):e867-e880. doi: 10.1002/term.2400. Epub 2017 May 23.

Abstract

Transplantation of bone marrow mesenchymal stem cells (BMSCs) has been considered to be a promising strategy for wound healing. However, poor viability of engrafted BMSCs and limited capabilities of differentiation into the desired cell types in wounds often hinder its application. Few studies report the induction of BMSC differentiation into the skin regeneration-related cell types using natural biopolymer, e.g. chitin and its derivative. Here we utilized a chitin nanofiber (CNF) hydrogel as a directive cue to induce BMSC differentiation for enhancing cutaneous wound regeneration in the absence of cell-differentiating factors. First, a 'green' fabrication of CNF hydrogels encapsulating green fluorescence protein (GFP)-transfected rat BMSCs was performed via in-situ physical gelation without chemical cross-linking. Without soluble differentiation inducers, CNF hydrogels decreased the expression of BMSC transcription factors (Oct4 and Klf4) and concomitantly induced their differentiation into the angiogenic cells and fibroblasts, which are indispensable for wound regeneration. In vivo, rat full-thickness cutaneous wounds treated with BMSC hydrogel exhibited better viability of the cells than did local BMSC injection-treated wounds. Similar to that of the in vitro result, CNF hydrogels induced BMSCs to differentiate into beneficial cell types, resulting in accelerated wound repair characterized by granulation tissue formation. Our data suggest that three-dimensional CNF hydrogel may not only serve as a 'protection' to improve the viability of exogenous BMSCs, but also provide a functional scaffold capable of enhancing BMSC regenerative potential to promote wound healing. This may help to overcome the current limitations to stem cell therapy that are faced in the field of wound regeneration. Copyright © 2017 John Wiley & Sons, Ltd.

Keywords: biomaterial scaffold; chitin nanofibers hydrogel; cutaneous wound regeneration; differentiation; mesenchymal stem cells; three-dimensional culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biocompatible Materials / pharmacology
  • Cell Differentiation* / drug effects
  • Cell Survival / drug effects
  • Chitin / pharmacology*
  • Collagen / biosynthesis
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Granulation Tissue / drug effects
  • Granulation Tissue / pathology
  • Hydrogels / pharmacology*
  • Kruppel-Like Factor 4
  • Male
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects
  • Mice
  • Nanofibers / chemistry*
  • Neovascularization, Physiologic / drug effects
  • Rats, Sprague-Dawley
  • Skin / drug effects
  • Skin / pathology*
  • Wound Healing* / drug effects

Substances

  • Biocompatible Materials
  • Hydrogels
  • Klf4 protein, mouse
  • Klf4 protein, rat
  • Kruppel-Like Factor 4
  • Chitin
  • Collagen