Effects and optimum concentration of zoledronate on RAW264.7

doi:10.3877/cma.j.issn.2096-0263.2018.01.002

Abstract

Abstract:

Objective

To further observe the concentration range of zoledronate on RAW264.7 cell toxic effect and confirm the optimum concentration in inhibition of RAW264.7 differentiation of osteoclast.

Methods

Mouse leukemia macro phage RAW264.7 was cultured as research object. RAW264.7 osteoclast toxic effect of zoledronate was detected by MTT method. Inhibition of osteoclast differentiation by different concentration of zoledronate were detected by tartrate-resistant acid phosphatase staining method.

Results

After 24 h vitro culture, Enzyme-linked immunosorbent assay absorbance indicated that 10^-3 mol/L (0.511±0.920), 10^-4 mol/L (0.615±0.577) concentrations of zoledronate had significantly toxic effects on cells compared with the blank control group (0.789±0.061)(F=5.880, P<0.01). TRAP staining osteoclast count indicated that 10^-5 mol/L (8.333±0.817), 10^-6 mol/L (10.400±1.817), 10^-7 mol/L (11.250±2.750) and 10^-8 mol/L (11.143±1.864) concentrations of zoledronate all significantly inhibited RAW264.7 differentiation compared with the blank control group (F=27.972, P<0.05), showing concentration dependence, 10^-5 mol/L zoledronate had the highest inhabitation effect (P<0.01).

Conclusion

Zoledronate can effectively inhibit RAW264.7 differentiation, and the optimum concentration in vitro is 10^-5 mol/L.

Key words: Zoledronic acid, Osteoclast, Cell differentiation, Cytotoxicity, RAW264.7

Lukai Zhang, Jianxiong Ma, Jie Zhao, Mingjie Kuang, Bin Lu, Ying Wang, Lei Sun, Xinlong Ma. Effects and optimum concentration of zoledronate on RAW264.7[J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2018, 04(01): 4-8.

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试剂及仪器	来源
DMEM培养基、胰蛋白酶、胎牛血清、	Sigma公司，美国
磷酸缓冲液D-Hank’s	自制
二甲基亚砜	美国sigma公司
青链霉素溶液	美国Invitrogen公司
MTT试剂	美国sigma公司
碱性磷酸酶试剂盒	南京建成生物工程研究所
细胞培养皿	美国Corning公司
倒置显微镜	德国Leica公司
细胞培养箱	德国Heraeus公司
唑来膦酸	美国sigma公司

试剂及仪器	来源
DMEM培养基、胰蛋白酶、胎牛血清、	Sigma公司，美国
磷酸缓冲液D-Hank’s	自制
二甲基亚砜	美国sigma公司
青链霉素溶液	美国Invitrogen公司
MTT试剂	美国sigma公司
碱性磷酸酶试剂盒	南京建成生物工程研究所
细胞培养皿	美国Corning公司
倒置显微镜	德国Leica公司
细胞培养箱	德国Heraeus公司
唑来膦酸	美国sigma公司

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