低浓度唑来膦酸对成骨细胞和破骨细胞影响的体外实验

Effects of low concentration of zoledronic acid on osteoclasts and osteoblasts in vitro

目的观察低浓度(10-6 mol/L)唑来膦酸(zoledronate acid,ZA)对体外大鼠破骨细胞及成骨细胞的影响。方法体外分别培养大鼠来源的成骨细胞和破骨细胞,将两种细胞各分为两组:空白对照组及低浓度(10-6 mol/L)ZA组。应用抗酒石酸酸性磷酸酶染色、图像分析计算骨吸收陷窝面积,检测破骨细胞形态及骨吸收情况。碱性磷酸酶(alkaline phosphatase,ALP)染色、四甲基偶氮唑盐比色法了解成骨细胞的形态及增殖情况。结果培养1周后破骨细胞具有典型的形态特征,并在骨片上形成了吸收陷窝;ZA组与对照组相比,破骨细胞数量及生成吸收陷窝的数目和面积减少,差异有统计学意义(P<0.05)。成骨细胞有典型的梭形、ALP染色阳性特征,培养至第7天ZA组成骨细胞光吸收值(3.37±0.11)高于对照组(2.87±0.12),差异有统计学意义(P<0.05)。结论低浓度(10-6 mol/L)的ZA能够抑制破骨细胞的增殖和活性,促进成骨细胞的增殖,选择恰当给药方式和剂量能够在抑制破骨的同时促进成骨。

Objective To explore the effects of low concentration （10^-6 mol/L） of zoledronic acid （ZA） on osteoclasts and osteoblasts. Methods Osteoclasts and osteoblasts were obtained from neonatal rats and cultured in vitro, and they were divided into control group and ZA group. TRAP staining and image analysis were used to detect the proliferation and activity of osteoclasts. ALP staining and MTT were used to detect the morphological characteristics and proliferation of osteoblasts.Results After a week, osteoclasts showed typical morphological characteristics, and bone resorption lacunas were found in vitro. Compared with control group, the number of osteoclasts and lacunae （P〈0.05）, and the area of lacunae （P〈0.05） in ZA group were significantly decreased. Osteoblasts showed typical fusiform and ALP showed positive staining characteristics. The optical density of osteoblasts was dramatically higher in ZA group （3.37±0.11） than control group （2.87±0.12） on day 7 after being cultured, and it had significant difference （P〈0.05）.Conclusion The low concentration （10^-6 mol/L） of ZA can inhibit proliferation and activity of osteoclasts and promote the proliferation of osteoblasts. Therefore, it is important to choose right administration and dose of ZA to enhance the osteogenesis and inhibit the bone resorption.