摘要
Objective:To understand the effects of combination treatment of pamidronate with isolated Quercus infectoria semi-purified fraction(Qlsm-F)on human foetal osteoblast cell model(hFOB1.19 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(EC_(50))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%CO_2 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.
Objective:To understand the effects of combination treatment of pamidronate with isolated Quercus infectoria semi-purified fraction(Qlsm-F)on human foetal osteoblast cell model(hFOB1.19 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(EC_(50))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%CO_2 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.
基金
supported by Ministry of Higher Education,Malaysia under the Fundamental Research Grant Scheme grant number 203/PPSK/6171169
