Therapeutic potential of hAMSCs in ligament regeneration: insights from the UCA1-miR-16-5p-FGF2 pathway.

BACKGROUND: With the growing demand for physical activity, there is a large population of patients suffering from ligament injuries, characterized by a high incidence and a need for early return to sports. A critical issue to address is how to promote the repair of damaged ligaments at the microscopic level and facilitate tendon-bone healing post-surgery. Among various studies, hAMSCs ( human amniotic mesenchymal stem cells ) have shown strong differentiation capabilities, and by inducing their differentiation into ligament cells, they may promote tendon-bone healing. METHODS: Human amniotic mesenchymal stem cells were extracted and cultured from human amniotic tissues. The cells were identified as mesenchymal stem cells through toluidine blue, safranin O, and oil red O staining. The differentiation of these cells into ligament cells was assessed by detecting relevant markers, including protein expression via Western blot and mRNA levels through qPCR(Quantitative polymerase chain reaction). Additionally, we used dual-luciferase reporter assays to verify the targeted interactions between lncRNA UCA1 and miR-16-5p, as well as between miR-16-5p and mRNA FGF2. RESULTS: Overexpression of lncRNA UCA1 and mRNA FGF2 increased the expression levels of Type I collagen (COL 1), Type III collagen (COL 3), and mohawk homeobox (MKX), key markers of ligament differentiation. LncRNA UCA1 was found to bind with miR-16-5p. The miR-16-5p can target mRNA FGF2, forming a regulatory axis that controls the differentiation of hAMSCs into ligament cells. Furthermore, we discovered that upregulation of miR-16-5p could inhibit the regulatory effects of overexpressed lncRNA UCA1 and mRNA FGF2 on mesenchymal stem cells. CONCLUSION: In this study, we identified the presence of a ceRNA network involving the UCA1-miR-16-5p-FGF2 axis in hAMSCs. By regulating the key components of this axis, we can modulate the differentiation of hAMSCs into ligament cells, providing a potential therapeutic strategy for promoting tendon-bone healing.