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 cell model (hFOB 1.19 cell 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 cells were cultured in DMEM F-12 and supplemented with 10% fetal bovine serum along with 1% penicillin-streptomycin incubated in 5% CO2 at 37 °C. 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.

Keywords

• pamidronate
• runx2
• osterix
• osteoblast
• quercus infectoria semi-purified fraction