Abstract
Articular cartilage defects are common, causing significant pain and morbidities. To date, tissue engineering techniques using adipose-derived mesenchymal stem cells have offered a promising modality for cartilage repair. In this study, we investigated the effects of Eucomis autumnalis aqueous crude extracts on porcine adipose-derived mesenchymal stem cells (pADMSCs) to determine whether they can differentiate pADMSCs into chondrocytes. pADMSCs were prepared and treated with different concentrations of E. autumnalis aqueous crude extracts and transforming growth factor (TGFβ3) was used as a positive control in vitro. The cells were then stained with safranin O and toluidine blue to observe change in morphology. Micromass culture was used to assess chondrogenic differentiation. The deposition of glycosaminoglycan (GAG) chondroitin sulfate was assessed as well with dimethylmethylene blue dye assay. Microscopic observations showed that pADMSCs were adherent with fibroblastic morphology features and trypan blue staining showed that 96% of pADMSCs were viable. Micromass sections stained with alcian blue and safranin O showed blue and orange color deposition in between cells on the outer side of the sections, suggesting the presence of cartilage GAGs and collagens, respectively. We also noticed that E. autumnalis extracts at 5 and 15 μg/mL concentrations were able to induce chondrogenesis of pADMSCs, as observed in the morphology of the resultant cells. Treatment of cells with E. autumnalis extracts at all concentrations upregulated gene expression of SRY-type high-mobility group box 9 (SOX9) and downregulated gene expression of collagen type II and X. Increased GAG content was noticed on cells treated with E. autumnalis extracts at 5 μg/mL concentration compared to other treatment groups. These findings suggest that E. autumnalis crude extracts of at least 5 μg/mL concentration could possibly differentiate pADMSCs into chondrocyte-like cells. These results show the potential role that E. autumnalis could play in the differentiation factor.
Impact Statement
Eucomis autumnalis is one plant that is used by various traditional healers to alleviate the signs and symptoms associated with osteoarthritis. Although the exact mechanisms remain unknown, we hypothesized that this plant can induce chondrogenesis. In this work, we explored the potential for an aqueous crude extract from E. autumnalis to induce chondrogenesis in porcine adipose-derived mesenchymal stem cells (MSCs). The results reported in our article indicate that the aqueous crude extract from E. autumnalis was able to indeed induce chondrogenesis. Our research is relevant to communities that rely on plant-based remedies for their well-being.
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