Objective
The purpose was to observe whether systemic administration with silibinin(SIL) have an positive effect on bone defect regeneration through HIF-1α/VEGF and Notch signaling pathway in an ovariectomized(OVX) rat model.
Methods
The MC3T3-E1 cells were co-cultured with lower SIL and higher SIL and induced to osteogenesis, and the cell viability, osteogenic activity were observed by Cell Count Kit-8(CCK-8), Alkaline phosphatase (ALP) staining, Alizarin Red(RES) staining and Western blotting(WB). After the drilling defect model was established, the OVX rats were treated with SIL for 12 weeks. Micro-CT, histology and Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis were used to observe the therapeutic effect and explore the possible mechanism.
Results
CCK-8, ALP and ARS staining results show that the cell mineralization and osteogenic activity of LSIL and HSIL group is significantly higher than the Con group. Protein expressions show that related regulatory proteins such as ALP, OPN, RUNX-2, OC, VEGFA, HIF-1α, Notch 1, JAG 1, HEY 1 and HES 1 of LSIL and HSIL group are significantly higher than Con group. Micro-CT and Histological analysis evaluation show that group SIL + OVX presented the stronger effect on bone regeneration, bone mineralization, higher expression of VEGFA and HIF-1α, when compared with OVX group. RT-qPCR analysis shows that SIL + OVX group showed increased Notch 1, HES1, HEY1 and JAG1 than the OVX group(p < 0.05).
Conclusions
Our current study demonstrated that systemic administration with SIL is a scheme for rapid repair of femoral condylar defects, and these effects may be achieved by activating HIF-1α/VEGF and Notch signaling pathway.