钛颗粒负荷下成骨细胞对破骨细胞功能的调节作用

Stimulative and Regulative Functions of Osteoblasts Loaded under the Titanium Particles on Osteoclasts

前期研究的结果表明,磨损颗粒可通过假体-界面膜进入假体周围间隙与成骨细胞(OBs)直接相互作用,影响OBs功能、促进OBs分泌活性因子。为进一步探讨磨损颗粒与OBs作用后是否会影响破骨细胞(OCs)的骨吸收功能,本研究通过结合形态学特征(TRAP染色、OCs扫描电镜观察、F-actin荧光标记、骨吸收陷窝染色和扫描电镜观察、钙离子荧光追踪)和生化学评价(ABC-ELISA法测定I型胶原交联羧基末端肽浓度),综合分析了3种直径(6.9、2.7和0.9μm)钛颗粒与OBs共孵育后的的条件培养基(CM)对OCs形态和功能的影响。结果表明,较小的0.9μm组CM可促进OCs分化、发育、成熟,部分活化OCs,影响OCs的存活;2.7μm和6.9μm组的CM,尤其是6.9μm组可明显促进OCs分化、发育、成熟、活化,维持OCs的存活;这与OCs细胞内钙离子浓度升高有关。以上结果提示OCs的活化、存活以及分化在假体松动中起着重要的作用;本研究为假体无菌性松动的防治、人工关节设计的材料优化选择提供了新的实验依据。

Our previous studies on the function of the osteoblasts（OBs） have shown that worn titanium particles decrease osteoblast＇ function and promot secretion of bone resorption cytokines of OBs surrounding the synovium-like interface membrane of loosening implants.The current study was aimed to test the hypothesis that osteoclasts（OCs） bone absorption function is induced by conditioned media（CM） prepared from OBs loaded in the presence or absence of titanium particles（with three mean diameters 6.9μm,2.7μm,and 0.9μm,respectively）.The effects of CM on OCs fuction were examined using a combination of the morphological characteristics tests,i.e.,TRAP dyeing,scanning electron microscopy,F-actin immunofluorescence protocol for confocal microscopy,bone resorption lacunae assay,osteoclastic calcium tracking,with biochemical evaluation,i.e.,C-terminal cross-linked telopeptides of type I collagen evaluated with ABC-ELISA method.The results showed that CM from 0.9μm titanium particles could induce osteoclastic differentiation and formation,could partially influence the survival of the OCs;while CM of 2.7μm and 6.9μm titanium particles,especially the latter,could obviously augmented osteoclastic activity,survival,or differentiation.The stimulation of osteoclast function may be due to a parallel increase in the intracellular free calcium concentration.The present study provides strong support for the hypothesis that osteoclastic activity,survival,or differentiation are very important in the development of aseptic loosening.The development of therapeutic interventions to reduce osteoclastic function and optimization of biomaterials may be useful approaches for improving the performance of orthopaedic implants.