Chitosan-hyaluronic acid polyelectrolyte multi-layers films as a platform for the chondrogenic differentiation of Wharton’s Jelly mesenchymal stem cells for tissue engineering in osteoarthritis

Abstract

Recently, the use of layer-by-layer (L-b-L) assembly of natural polyelectrolytes has been suggested as a new technique of surface modification aimed to mimic cells natural environment. The ease of harvesting mesenchymal stem cells from human umbilical cords Wharton’s Jelly (hWJ-MSCs), their great differentiation potential and low immunogenicity make them a suitable tool for cartilage tissue engineering. Our L-b-L was based on alternated deposition of 10 layers of the polycation: Chitosan (CHI) and 10 layers of the polyanion: Hyaluronic acid (HA). In this study, we have evaluated the chondrogenic differentiation of hWJ-MSCs (3000 cells/cm2, at 4th passage) seeded on (CHI-HA)10 films. hWJ-MSCs stimulated with TGF-Beta were used as positive control for chondrogenic differentiation, and cells seeded on glass slides without growth factor were used as negative control. mRNA and protein expression of the main chondrogenic markers (Sox9 and Aggrecan) along with Chondroitin sulfate staining were performed. The isolated hWJ-MSCs showed typical fibroblastic-like morphology and were positive for MSCs markers (CD44, CD73, CD90, CD105) and negative for hematopoietic markers (CD34 and CD45). We have shown that hWJ-MSCs highly express chondrogenic markers at genic and proteic levels after 21 days of culture in DMEM compared to our positive and negative markers. We report here a new biocompatible coating allowing chondrogenic differentiation of hWJ-MSCs. This substrate opens new routes in cartilage tissue engineering to design allogenic cartilage graft.