羟基磷灰石形貌对RAW264.7细胞破骨分化影响研究

Effect of different hydroxyapatite shapes on osteoclastogenesis of RAW264.7 cells in vitro

目的分析羟基磷灰石(HAp)不同形貌对小鼠单核RAW264.7细胞破骨分化性能的影响。方法采用水热合成及pH调节的方法获得不同形貌羟基磷灰石微米颗粒,通过扫描电子显微镜(SEM)表征材料形貌。将不同形貌HAp与RAW264.7细胞共培养,通过Live/Dead活性染色检测HAp对细胞活性有无影响;并在RANKL破骨诱导因子的作用下通过抗酒石酸酸性磷酸酶(TRAP)染色、实时荧光定量(qRT-PCR)分析不同形貌HAp对RAW264.7细胞破骨分化的影响。结果 24h Live/Dead活性染色证实微棒与微球HAp均具有良好的生物相容性。qRT-PCR结果表明,两种形貌HAp均抑制RAW264.7细胞的破骨分化,而相对于微球HAp,微棒HAp抑制作用较显著。结论羟基磷灰石形貌的差异能够引起RAW264.7破骨分化性能的差异表达。

Objective To investigate the effect of different hydroxyapatite shapes on osteoclastogenesis of RAW264.7cells. Methods Hydroxyapatite with different shapes was synthesized through a hydrothermal treatment with further pH adjustments, and characterized by filed scanning electron microscope（SEM）.Live/Dead staining was performed to evaluate the cell viability of different hydroxyapatites co-cultured with RAW264.7 cells. Tartrate-resistant acid phosphatase （TRAP） staining and quantitative RT- PCR （qRT-PCR） analysis were used to identify osteoclastogenesis and investigate transcriptional changes of osteoclastogenesis-related genes, under the stimulation of the receptor activator of nuclear factor-r,B ligand （RANKL） in vitro. Results Live/Dead staining validated that the microrod and microsphere HAp particles were biocompatible and no difference of cell viability was detected between two particles after 24h co-cultttre. The qRT-PCR results showed that the two shaped HAp particles significantly inhibited the RANKL-induced osteoclastogenesis of RAW264.7 cells, especially for micrord particles compared to that of microsphere ones. Conclusions Hydroxyapatites with different shapes could initiate different degree on osteoclastogenesis of RAW264.7cells.