Enhanced tenogenic differentiation and tendon-like tissue formation by Scleraxis overexpression in human amniotic mesenchymal stem cells

Tendon and ligament injuries are not uncommon in clinics and have poor self-healing capacity due to their bloodless and slow-proliferative nature. Promoting the repair or reconstruction of an injured structure is an urgent problem. While Scleraxis (Scx) is a highly specific tendon cell marker, its function has not been explored to a large extent. Hence, Recombinant adenovirus was used to study the influence of Scx overexpression on directional differentiation of human amniotic mesenchymal stem cells (hMSCAs). hAMSCs modified with Scx could dramatically enhance the gene expression of tendon-related molecules, containing Scx, collagens I and III, Tenascin-C, fibronectin, matrix metalloproteinase-2 (MMP-2), lysyl oxidase-1 (LOX-1) and Tenomodulin at all-time points (P < 0.05), and the secretion of collagen I and III, fibronectin and Tenascin-C over time (P < 0.05) but did not impact the cell proliferation capacity (P > 0.05). Immunofluorescence staining showed the cobweb-like fusion of collagen I and fibronectin in the AdScx group on day 7, with higher average fluorescence intensity than the control (P < 0.05). After mixing with Matrigel, transplants were subcutaneously implanted in nude mice, obvious inflammation and rejection of immune response were not observed and HE staining showed a histological feature of swirl of fibers is closely linked in parallel in hAMSCs modified with Scx. On the contrary, in the control group, an unorganized connective structure with cell distributed randomly was spotted. The results of promoted directional differentiation of stem cells and the spatial structure of the normal tendon tissue in three-dimensional space manifested that Scx can be used as a specific marker for tendon cells, and as a positive regulator for directional differentiation of hAMSCs, which is possible to be applied to novel therapeutics for clinical tendon and ligament injury by hAMSCs modified with Scx.