The Effect Of Wnt3a Factor And Bone Morphogenetic Protein-2 Co-regulating Bone Marrow Mesenchymal Stem Cell Proliferation And Osteogenic Differentiation

Objective: To observe the impact of classical Wnt/β-catenin and BMP signaling pathway effect on rat bone marrow-derived mesenchymal stem cell proliferation and osteogenic differentiation. Methods: Bone marrow mesenchymal stem cells were isolated by density gradient centrifugation combined adherence separation methods and divided into four groups: control group, Wnt group, BMP group, Wnt3a and BMP-2 combination group. Cell proliferation activity and alkaline phosphatase (ALP) activity were analyzed quantitatively by CCK8 protocol. Cellular osteogenic differentiation and matrix mineralization were decided by Von Kossa staining. The expression of osteoblast-specific markers were detected by RT-PCR. Results: At culture day 7, CKK-8 activity was 0.845 in combination group, 0.738 in Wnt3a group and significantly higher than 0.409 of control group (P <0.05). At culture day 6 and day12, cellular ALP activity was 63.8 and 144.3 of combination group, 40.8 and 104.1 of BMP-2 group, higher than 7.3 and 18.9 of control group (P <0.05). Cultured for 14 days, of combination group osteocalcin mRNA expression was significantly higher, while the expression of Runx2 and Osterix was not significant compared with other groups. Culture for 3 weeks, Von Kossa staining showed the number and size of calcium nodules of combination group were higher than of control group. Conclusion: Combined induction of Wnt3a factor and bone morphogenetic protein2 can effectively promote mesenchymal stem cell proliferation and osteogenic differentiation.