Growth and Chondrogenic Differentiation of Mesenchymal Stem Cells Derived from Human Adipose Tissue on Chitosan Scaffolds

Treatment of cartilage damage for any reason is associated with temporary relief of joint pain. Providing the possibility of differentiating various stem cells into adult tissues can contribute to recovery and treatment of damaged cartilage tissue in skeletal system. In this study, chondrogenic potential of chitosan scaffold, CH-β-GP-HEC, with stem cells derived from human adipose tissue.

In this cross-sectional study, adipose tissue-derived stem cells were separated from abdomen of 15 patients who underwent inguinal hernia repair. 6-7×105 cells were cultured in plate one-dimensionally and on chitosan scaffold three-dimensionally for 21 days. MTT assay was run to evaluate the toxic effect of scaffold on cell viability. Proliferation and differentiation of cells were studied in the two types of culture after toluidine blue staining. To confirm the formation of cartilage, expression of collagen type II was assessed by immunohistochemistry.
FINDING: In MTT assay, the average OD for cells cultured on scaffold is higher than 0.8 compared with control group, which confirms the nontoxicity of scaffold for culturing stem cells (p>0.05). Chondrogenic differentiation of cells on scaffold shows more glycosaminoglycan deposition in the extracellular matrix compared with one-layer culture. Moreover, in group with three-dimensional culture system, cells were spherical and the morphology of nucleus was different from one-layer culture. Regarding immunohistochemistry results, increased synthesis was observed in collagen type II as chondrogenesis markers in three-dimensional culture system compared with one-layer culture.

Results of the study revealed that hydrogel scaffold, CH-β-GP-HEC, with porous structure provides a better environment for the growth of mesenchymal stem cells and their differentiation into cartilage tissue.