Combination treatment of bisphosphonate (pamidronate) and Quercus infectoria semi-purified fraction promotes proliferation and differentiation of osteoblast cell via expression of Osterix and Runx2 marker

ABSTRACT

Objective:To understand the effects of combination treatment of pamidronate with isolated Quercus infectoria semi-purified fraction (QIsm-F) on human foetal osteoblast cel model (hFOB 1.19 cel line) through assessment of Runt related transcription fraction-2 (Runx2) and Osterix (Osx). Methods:The isolation and purification of QIsm-F were conducted by chromatographic technique. In order to assess relative efficacy of QIsm-F to the osteoblast model, the determination of half maximal effective concentration (EC50) was performed by MTT assay. hFOB 1.19 cel s were cultured in DMEM F-12 and supplemented with 10％ fetal bovine serum along with 1％ penicil in-streptomycin incubated in 5％ CO2 at 37 ℃. Expression of Runx2 and Osx was assessed through western blotting and confirmed with immunofluorescence staining. Results: Results of western blot analysis and immunofluorescence staining demonstrated that compared to hFOB 1.19 cells treated with single individual treatment of QIsm-F and control groups, levels of Runx2 and Osx were elevated with higher fluorescence intensity and more rapid proliferation in hFOB 1.19 cells treated with combined treatment of QIsm-F and pamidronate. Conclusions: The finding demonstrates the synergistic effect between osteoporotic drug pamidronate and established QIsm-F. The combination treatment helps increase the efficiency of pamidronate acting on osteoblast cells by stimulating osteoblast proliferation and differentiation via expression of Runx2 and Osx.