目的采用先天性成骨不全(0I)小鼠,oim/oim为动物模型,应用破骨细胞一颅骨联合培养体系研究OB和OC两种细胞在0I骨再建过程中的功能改变和相互作用。方法实验采用小鼠颅骨(CAL)组织培养,实验设两组:WTCAL.WTOC组:联合培养对照组颅(WTCAL)与对照破骨细胞(WTOC);OICAL—OIOC组:联合培养OI颅骨(OICAL)与OI破骨细胞(OIOC)。以免疫组化染色方法-TRAP识别破骨细胞,ALP免疫组化染色方法识别成骨细胞。破骨细胞骨吸收活性为骨吸收陷窝占颅骨表面百分比。单位OC吸收面积为总骨吸收陷窝除以破骨细胞数。结果于7d,OICAL、OIOC组破骨细胞数低于WTCAL-WTOC组;OICAL—OIOC组的OC/OB比例低WTCAL—WTOC组;OICAL—OIOC组单位破骨细胞吸收能力高于WTCAL—WTOC组。结论OI的小鼠模型骨再建中骨量丢失一方面南于其成骨细胞功能异常,另一方面也可能因为其破骨细胞的代偿性功能活跃。
Objective To evaluate the effects of osteoblast (OB) and osteoclast (OC) on the bone remodeling of osteogenesis imperfecta (OI) adopting osteoclast-calvaria co-culture system in vitro on oim/oim (OI) mouse model. Methods Wild (WT) and OI mice were used and compared in this study. OC cells were cultured in calvaria (CAL) in vitro for WT (WTCAL-WTOC group) and OI mice (OICAL-OIOC group) respectively, Tartrate-resistant acid phosphatase (TRAP) staining and alkaline phosphatase (ALP) staining were used to identify OCs and OBs respectively. Bone resorption of OCs was assessed by the area percentage of absorption lacunam, which is the rate of OCs number to the whole calvarial surfaces. Results At the culturing time of d7, the number of OCs and OC/OB rate of group O1CAL-OIOC were significantly lower than that of group WTCAL-WTOC. However, the OCs number normalized to resorption pit number was significantly greater in group OICAI-OIOC compared to that of WTCAI-WTOC group. Conclusions The mechanism of the increased OCs function is partially due to the increased bone turnover in OI mice model for the compensation of decreased OBs function.