Hyaluronic acid and chondrogenesis of murine bone marrow mesenchymal stem cells in chitosan sponges

Abstract

To establish the dose-dependent effects of high-molecular-weight hyaluronic acid (HA) supplementation on chondrogenesis by mesenchymal stem cells (MSCs) cultured on chitosan sponges and to determine the extent to which MSC matrix production (chondrogenesis) can be influenced by incorporation of high-molecular-weight HA into chitosan scaffolds. Murine MSCs derived from a multipotent bone marrow stromal precursor. MSCs were seeded on chitosan and chitosan-HA scaffolds in chondrogenic medium with various HA concentrations. Scanning electron microscopy, fluorescence microscopy (viability assay), and DNA quantification were used to assess cell attachment, distribution, and viability 48 hours after seeding. Constructs were cultured for 3 weeks prior to evaluation of cell distribution and chondrogenic differentiation via histologic evaluation and quantification of DNA, glycosaminoglycan, and collagen II. 48 hours after MSC seeding, cell viability and DNA content were similar among groups. Three weeks after seeding, HA supplementation of the culture medium improved matrix production in a dose-dependent manner, as indicated by matrix glycosaminoglycan and collagen II concentrations. The scaffold composition, however, had no significant effect on matrix production. High-molecular-weight HA supplementation in culture medium had a dose-dependent effect on matrix production and thus chondrogenic differentiation of MSCs cultured on chitosan sponges. The addition of HA in the surrounding fluid during chondrogenesis should improve cartilage production and may be useful for producing engineered cartilage tissues.