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

Abstract

Osteoarthritis, a condition that develops from arthritic cartilage abnormalities, impairs the quality of life for millions of individuals globally and causes significant healthcare inflation. Efforts in the field of tissue engineering to treat osteoarthritis have primarily focused on enhancing the growth and differentiation of stem cells, along with the utilization of various scaffolding methods. Layer-by-layer assembly of naturally occurring polyelectrolytes has recently been proposed as an innovative approach to surface modification that attempts to imitate the environment in which cells normally exist. For cartilage regeneration, Wharton’s jelly mesenchymal stem cells from the human umbilical cord show promise due to the fact that they are easily accessible, have significant differentiation potential, and have minimal immunogenicity. This study evaluated the feasibility of a novel layer-by-layer matrix consisting of cross-linked 10 layers of polyanionic hyaluronic acid with 10 layers of polycationic chitosan, along with Wharton’s jelly mesenchymal stem cells, in promoting chondrogenic differentiation. Wharton’s jelly mesenchymal stem cells were seeded onto the chitosan-hyaluronic acid polyelectrolyte multilayer film at a density of 3000 cells/cm² with normal growth medium. The positive control involved Wharton’s jelly mesenchymal stem cells onto the polyelectrolyte multilayer films with transforming growth factor beta, while the negative control consisted of cells on glass slides with normal growth medium and no growth factors. A number of elements, including cellular growth, matrix development, as well as crucial gene expressions, were evaluated during in vitro culture. Based on ISCT guidelines, the cells had fibroblastic shape, plastic adherence capacity, and the required MSC markers (CD73, CD90, and CD44), but not thehematopoietic markers (CD34 and CD45). In addition, these cells have preserved their multipotency, as shown by osteogenic induction (confirmed by quantitative Real-time Polymerase Chain Reaction, immunofluorescence staining, and alizarin red staining). As a result, a favorable environment for mesenchymal stem cellspontaneous development into chondrocytes capable of forming cartilage matrix was created by the chitosan-hyaluronic acid polyelectrolyte multilayers films; this was confirmed at the genic level (Sox9 and Aggrecan) and by the use of chondroitin sulfate staining. The use of this type of polyelectrolyte multilayer films might thereby pave the way for novel approaches to cartilage tissue engineering