Synergistic effects of enamel matrix derivatives and surface morphology of anodized titanium on osteogenic differentiation of bone marrow mesenchymal stem cells sheet.

Objective: To explore the potential osteogenic induction mechanism of enamel matrix derivatives (EMDs) on bone marrow mesenchymal stem cell (BMSC) sheets with different titanium surface morphologies.

Methods: The BMSCs were inoculated on the surfaces of titanium alloys with different morphologies: anodic oxidation (AO), sand-blasted, large grit and acid-etched, and no treatment (control). The proliferation and osteogenic differentiation of BMSCs on the different surface morphologies were observed with the same concentration of EMDs. To further understand the osteogenic mechanism of EMDs on BMSC sheets with different morphologies, a real-time RT-PCR and a western blot were used to detect the overall levels of osteogenic genes and osteogenic proteins. Finally, to verify the osteogenic effect of BMSC sheets stimulated by EMDs in vivo, BMSC sheets with different morphologies were implanted into the subcutaneous tissue of the back of nude mice, and the bone formation was detected by HE staining.

Results: The EMDs and surface morphology in the AO group synergically increased the expression levels of osteogenic active factors (RUNX2, OSX and OCN) and enhanced the osteogenic differentiation effect of BMSCs. The in vivo experiments showed that the BMSC sheets in the AO group were rich in osteogenic active factors, and promoted the formation of ectopic bone tissue after implantation into the subcutaneous tissue of the back of nude mice.

Conclusion: EMDs and AO morphology synergically enhance the secretion of bone osteogenic active factors of BMSCs and promote the formation of heterotopic bone.