Investigation on the Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells Cultured on a Non-woven Carboxymethyl Cellulose Scaffold Into Chondrocyte-Like Cells

Abstract

Background: Trauma can significantly impact cartilage tissue and human quality of life. Limited cartilage regeneration capability has led to medical interventions for tissue repair. Carboxymethyl cellulose (CMC) is an anionic cellulose with suitable price and mechanical properties. Carboxymethyl cellulose is used as a lubricant or in constructing hydrogel scaffolds for cell culture. Objectives: Since cartilage cells differentiate from bone marrow-derived mesenchymal stem cells, this study aimed to investigate their chondrogenic differentiation potential on a non-woven CMC scaffold in a monolayer system. Methods: Non-woven CMC was synthesized and evaluated using Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and mechanical tests. Human bone marrow-derived mesenchymal stem cells were isolated and identified using flow cytometry. After chondrogenic differentiation induction, cell attachment and expression of chondrogenic-related genes were examined using dimethyl thiazolyl diphenyl tetrazolium (MTT) and Quantitative real-time polymerase chain reaction (Q-PCR). Results: dimethyl thiazolyl diphenyl tetrazolium assay showed that cells adhered well to the scaffold surface. There was no significant difference in cell viability between the control group (cells without scaffold) and the experimental group. The SEM images showed that the cells were successfully attached to the membrane. Fourier-transform infrared spectroscopy spectra showed the singes of carbonyl, methyl, and hydroxyl groups. The tensile strength was as much as 0.224 ± 0.155 and 0.306 ± 0.164 in dry and wet forms, respectively. The tensile strength was measured as 0.155 ± 0.224 and 0.164 ± 0.306 MPa in the scaffold's dry and wet forms, which was insignificant. Young's modulus was equal to 0.077 and 0.079 MPa in dry and wet conditions, respectively. After two weeks of differentiation culture, a significant increase in Sox-9 and Collagen II expression was observed. No significant increase in the expression of Collagen X and Aggrecan was detected.Conclusion: Human bone marrow-derived mesenchymal stem cells successfully adhered to the non-woven CMC scaffold and could express early chondrogenic pathway genes in the presence of inducing factors. However, expression of more specific cartilage-related genes morphological changes toward a chondrogenic phenotype and lacuna formation were not observed.