Abstract: BACKGROUND: Compared with the traditional autogenous bone graft methods, the bone tissue engineering repair strategies have their advantages of less trauma, free immunogenicity, and promotion of bone regeneration among the bone defect repair modes in the implant area. The scheme that applies the traditional Chinese medicine ingredients with less adverse reactions than chemical synthetic drugs to the bone defect area of osteoporosis patients constitutes an important part of the bone tissue engineering scaffold to deliver drugs.
OBJECTIVE: To compare effects of puerarin and icariin on the proliferation, differentiation and mineralization of mouse preosteoblasts (MC3T3-E1) and select effective osteogenic drugs.
METHODS:  The optimal concentration of puerarin and icariin to promote the proliferation and differentiation of MC3T3-E1 was determined. Cultured cells were divided into control group, 17β-estradiol group, icariin group, and puerarin group. Cell proliferation and viability were detected at 1, 4, and 7 days after drug intervention. Alkaline phosphatase activity was detected at 1, 7, and 14 days after osteogenic induction. Calcified nodules after alizarin red staining were measured at 21 days after osteogenic induction. Real time-qPCR was used to detect the expression of osteoprotegerin and Runt-associated transcription factor 2 mRNA at 7 days after osteogenic induction. The morphology of the cytoskeleton after 24 hours was observed using phalloidin staining.
RESULTS AND CONCLUSION: Puerarin and icariin exhibited the best effect on proliferation and osteogenic differentiation at concentration of 10-7 mol/L and 10-6 mol/L. At 4 days of culture, compared with the icariin group, the ability of puerarin to promote cell proliferation was stronger, and the difference was significant (P < 0.05). At 7 days of osteogenic induction, compared with the puerarin group, the alkaline phosphatase activity of the icariin group was stronger; the expression levels of osteoprotegerin and Runt-related transcription factor 2 mRNA were significantly up-regulated; at 21 days of induction, the number and area of calcified nodules increased significantly, and the difference was significant (P < 0.05). After 24 hours of culture, the cytoskeletal morphology was grid-like spreading. Compared with the control group, the actin ran clearly in the estrogen group, the puerarin group and the icariin group. According to the results, puerarin can better accelerate the proliferation of MC3T3-E1 cells, and icariin can better promote the differentiation and mineralization of MC3T3-E1 cells.

Key words: puerarin, icariin, cell proliferation, cell differentiation, cell mineralization, osteoblast