钛颗粒对成骨细胞增殖分化及细胞形态的影响

发布时间：2018-03-29 14:19

本文选题：钛颗粒　切入点：成骨细胞　出处：《中国修复重建外科杂志》2014年05期

【摘要】：目的通过观察钛颗粒对成骨细胞增殖分化和细胞形态的影响,探讨其可能的内在联系和作用机制。方法取10只新生24 h SD大鼠颅骨,采用多次酶消化法体外分离培养成骨细胞,ALP和茜素红染色进行细胞鉴定。取生长良好的第3代细胞,分别采用浓度为0.01、0.05、0.1、0.5、1 mg/mL的钛颗粒培养基(0.01、0.05、0.1、0.5、1 mg/mL组)培养,7 d后用细胞计数试剂盒8检测吸光度(A)值,比较不同浓度钛颗粒对成骨细胞增殖能力的影响,并筛选半数致死浓度;ELISA法检测细胞Ⅰ型胶原表达,比较不同浓度钛颗粒对成骨细胞分化能力的影响。另取以半数致死浓度钛颗粒培养7 d的成骨细胞(实验组)进行FITC-鬼笔环肽和碘化丙啶双重荧光染色,激光共聚焦显微镜下观察吞噬钛颗粒后细胞形态学的改变。以上观察指标均以正常第3代成骨细胞作为对照组。结果 ALP及茜素红染色鉴定提示培养的细胞为成骨细胞。培养7 d,各浓度组成骨细胞增殖、分化能力与对照组比较均有不同程度下降,差异均有统计学意义(P0.05)。同时随浓度增加,各浓度组对成骨细胞增殖的抑制作用逐渐增强,成骨细胞Ⅰ型胶原表达逐渐降低;除0.01、0.05 mg/mL组间差异无统计学意义(P0.05)外,其余各浓度组间比较差异均有统计学意义(P0.05)。钛颗粒的半数致死浓度为0.5 mg/mL。激光共聚焦显微镜下示,与对照组比较,实验组吞噬了钛颗粒的成骨细胞皱缩变形,伪足收缩变短,微丝排列紊乱。结论钛颗粒可抑制成骨细胞的增殖和分化能力,这种作用可能与吞噬钛颗粒后细胞形态学的改变有关。
[Abstract]:Objective to observe the effect of titanium granule on proliferation, differentiation and cell morphology of osteoblasts, and to explore its possible internal relationship and mechanism. Methods the cranial bones of 10 newborn 24 h SD rats were taken out. Osteoblasts were isolated and cultured with ALP and alizarin red staining by multiple enzyme digestion in vitro. The cell count kit 8 was used to detect the absorbance of osteoblasts in 0. 01% 0. 05 and 0. 5 mg/mL titanium granule culture medium containing 0. 01 0. 05 and 0. 5 mg/mL for 7 days. The effects of different concentrations of titanium particles on the proliferation of osteoblasts were compared, and the effects of different concentrations of titanium particles on the proliferation of osteoblasts were compared. The expression of type I collagen was detected by Elisa. The effects of titanium particles at different concentrations on the differentiation of osteoblasts were compared. The osteoblasts cultured with titanium particles with LD50 for 7 days (experimental group) were stained with FITC- phloropylinin and promadime iodide double fluorescence staining. The morphologic changes of the cells after phagocytosis of titanium granules were observed by laser confocal microscopy. All the above indexes were normal third passage osteoblasts as control group. Results ALP and alizarin red staining indicated that the cultured cells were formed. Osteoblasts were cultured for 7 days. Compared with the control group, the differentiation ability decreased to some extent, and the difference was statistically significant (P 0.05). At the same time, with the increase of concentration, the inhibition of osteoblast proliferation was gradually enhanced, and the expression of type I collagen in osteoblasts was gradually decreased. There was no significant difference between 0. 01 mg/mL and 0. 01 mg/mL groups (P 0. 05), but there were significant differences among the other groups. The LD50 of titanium particles was 0. 5 mg / mL. Laser confocal microscope was used to compare the results with that of the control group, and compared with the control group, the LD50 of titanium particles was 0. 5 mg 路mL ~ (-1) 路ml ~ (-1) 路L ~ (-1). In the experimental group, the osteoblasts of the experimental group were engulfed with titanium particles, the contraction of pseudopodia became shorter and the arrangement of microfilaments was disordered. Conclusion Titanium granules can inhibit the proliferation and differentiation of osteoblasts. This effect may be related to the changes of cell morphology after phagocytosis of titanium particles.
【作者单位】：深圳市龙岗中心医院骨关节科;
【基金】：深圳市龙岗区科技发展基金资助项目(YS2012163)~~
【分类号】：R318.08

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