Objective To investigate the effect of cigarette smoke extract(CSE)on osteoclast size and gene expression of integrin αvβ3 in vitro.Methods 7-week-old C57BL/6J mouse bone marrow mononuclear cells were added with M-CSF and RANKL to induce osteoclasts.5%CSE was added to the experimental group for osteoclasts for 3 days,while no CSE was added in the control group.The osteoclasts in the control and experimental groups were identified using TRAP staining and images collected under light microscopy.The bone lacunae formed by osteoclasts on bone slices was identified using toluidine blue staining and images were taken under light microscopy.NIH imageJ was used to calculate the quantity and area.Real-time fluorescence quantitative polymerase chain reaction was used with GAPDH as the internal reference.The were detected by Trap,CTR,and integrin αvβ3 mRNA expression levels,and integrin αvβ3 protein expression was detected by 2-ΔΔCT calculation.The integrin αvβ3 protein expression was detected by Western blot.Results The induced osteoclasts in the experimental group were significantly larger than those in the control group(P<0.01).The number and area of absorption lacunae on the bone slices of the experimental group was significantly higher than that of the control group(P<0.05).The TRAP mRNA level in the CSE treatment group was 1.16±0.13,which was significantly higher than that of the control group(P<0.05).The CTR mRNA level was 0.38±0.04,which was significantly lower than that of the control group(P<0.01).Integrins αvβ3 mRNA level in the CSE treatment group was 1.75±0.05,significantly higher than the control group(P<0.01),and the results of Western blot analysis were also significantly higher than the control group(P<0.05).Conclusions CSE can significantly increase the size of osteoclasts and upregulate the gene expression of integrins αvβ3,which may be involved in the promotion of bone resorption.
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