结合状态伊班膦酸盐对破骨细胞骨吸收作用的影响

 目的 观察伊班膦酸盐（ibandronate，Iba）与羟基磷灰石（hydroxyapatite，HA）结合后对破骨细胞骨吸收作用的影响。方法 将Iba与HA复合成伊班膦酸盐-羟基磷灰石（Iba-HA）作为实验组，未结合Iba的HA作为对照组。分离、培养、纯化大鼠破骨细胞，Giemsa染色和抗酒石酸酸性磷酸酶(TRAP)染色显示破骨细胞及其细胞核的形态和数目。用两组材料与破骨细胞复合培养，透射电镜观察细胞内结构变化，SNAFL染色测定破骨细胞内氢离子浓度，骨吸收陷窝面积测定并辅以上清液内胶原C端肽片段（CTx）的浓度测定观察破骨细胞骨吸收功能。结果 Gimsa染色标本可见多核的破骨细胞、细胞核染成蓝紫色，TRAP染色标本可见TRAP酶活性的部位被染成红色，分布于破骨细胞胞浆内。透射电镜观察发现，破骨细胞与Iba-HA复合培养后，较之与HA复合培养的破骨细胞生物活性下降。SNAFL染色显示Iba-HA组破骨细胞荧光强度小于HA组，统计学有差异（P<0.05）；骨吸收检测时面积显示，Iba-HA组小于HA组，统计学有差异（P<0.05），上清液CTx浓度，Iba-HA组小于HA组，统计学有差异（P<0.05）。结论 与ＨＡ处于结合状态的Iba对破骨细胞的骨吸收作用仍有显著影响。

Effect of Binding Ibandronate on Bone Resorption through the Mediation of Osteoclasts

OBJECTIVE To investigate the effects of binding ibandronate with hydroxyapatite on bone resorption through the mediation of osteoclasts. METHODS Ibandronate was integrated with hydroxyapatite to form compound materials and was incabated with osteoclasts in the experimental group， while hydroxyapatite without ibandronate was incubated with osteoclasts in the control group. Osteoclasts of the rats were separated and cultured， and Giemsa staining and TRAP staining were used to determined their nuclear morpha and the account. Samples in two groups were co-cultured with osteoclasts. Then electron microscope were used to investigate the structural change of the osteoclasts. And hydrogen ion concentration in the cells were acquired by seminaphthofluoresceins. The classic bone reseption trap area estimation was used to investigate the function of absorption， CTx concentration in the supernatant was detected complementarity. RESULTS Compared with the osteoclasts cultured without ibandronate-HA， the co-cultured cells demonstrated less Endoplasmic Reticulum and lysosomes in their cytoplasm， and nucleus crimpled， nucleolus disappeared. There were differences between the two groups in the hydrogen ion concentration (P<0.05) and bone reseption trap area estimation (P<0.05). The difference of CTx concentration in the supernatant was significant (P<0.05). CONCLUSION The binding ibandronate with hydroxyapatite shows significant effect on the bone resorption of the osteoclasts.