Biphasic calcium phosphate loaded with advanced platelet rich fibrin can promote the activity of rabbit bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Adding growth factor to scaffold material can promote the proliferation and osteogenic differentiation of cultured stem cells in vitro and promote bone tissue regeneration. Advanced platelet rich fibrin (A-PRF) contains a variety of growth factors, which can promote the proliferation activity of a variety of cells and the expression of related functional proteins. OBJECTIVE: To observe the effects of the sole and combined usage regarding A-PRF and biphasic calcium phosphate (BCP) on the growth of bone marrow mesenchymal stem cells. METHODS: The third generation of rabbit bone marrow mesenchymal stem cells was cultured in four groups, including basic medium (blank group), A-PRF condition medium (A-PRF group), BCP (BCP group), A-PRF condition medium and BCP (A-PRF+BCP group). Cell proliferation activity was detected by CCK-8 assay. Cell adhesion was observed based on methylrhodamine-phalloidin fluorescence staining. Intracellular alkaline phosphatase expression and mineralized nodules were quantitatively measured. RESULTS AND CONCLUSION: (1) The absorbance value of proliferation of the A-PRF+BCP group was higher than that of the other three groups at 3, 5, 7, 11 and 14 days after culture. The absorbance value of proliferation of cells cultured in the A-PRF group was higher than that of the BCP group and the blank group at 1, 3 and 5 days after culture. The absorbance value of proliferation of BCP group was higher than that of the A-PRF group and the blank group at 7, 11 and 14 days after culture. (2) Methylrhodamine-phalloidin fluorescence staining showed that bone marrow mesenchymal stem cells in the BCP group and the A-PRF+BCP group adhered to the surface of BCP and the number of cells and microfilaments in the A-PRF+BCP group was higher than that in BCP group. (3) The synthesis of mineralized nodules was A-PRF+BCP group > BCP group > A-PRF group > blank group at 1, 21 and 28 days after surgery. (4) The expression of alkaline phosphatase was A-PRF+BCP group > BCP group > A-PRF group > blank group at 5, 7 and 9 days. There was significant difference between the two groups (P < 0.05). (5) The results showed that BCP exerted a weak influence on promoting cell proliferation in the early stage, but its effect of scaffold was apparent. A-PRF had a significant effect on promoting cell proliferation with considerable influence on promoting cell proliferation and differentiation when combined BCP.