不锈钢微粒对人成骨样细胞毒性及护骨素及其配体基因表达的影响

Effects of Stainless on Osteoblasts:Cytotoxicity and the Releasion of OPG and RANKL

目的评价两种不锈钢微粒(镍铬和钴铬合金)对人成骨样细胞MG-63的细胞毒性及对人成骨样细胞分泌护骨素(OPG)、核因子kB配体受体激活子(RANKL)的影响。方法选择人成骨样细胞系MG-63作为细胞毒性的实验对象,干预材料选用镍铬、钴铬合金微粒,使二者均以0.005%、0.01%和0.05%三个终浓度作用于细胞48 h;并设立对照组。MMT法检测细胞活力,评价不锈钢种植体的细胞毒性;用半定量RT-PCR检测人成骨样细胞内OPG/RANKL mRNA的表达量,分析微粒对破骨细胞重要信号传导通路OPG/RANKL/RANK系统的影响。结果 (1)与对照组比较,当镍铬粒子浓度>0.01%时,对细胞生长产生一定的抑制作用;而钴铬对人成骨样细胞的生长无明显的抑制作用。(2)与对照组比较,镍铬粒子浓度<0.01%时,对造骨细胞OPG的表达无明显影响,但可以诱导人成骨样细胞中RANKL的表达;钴铬粒子对细胞OPG表达无明显的影响,只有当粒子浓度达到0.05%的时候,才对人成骨样细胞中RANKL有一定的诱导效果。结论不锈钢微粒对人成骨样细胞无细胞毒性;低浓度不会促进人成骨样细胞的增殖、分化及抑制破骨细胞的形成。

Objective To evaluate the cytotoxicity of two kinds of stainless microparticle on human osteoblas-like cellst（MG-63） and the effects on the secretion of OPG / RANKL from osteoblast.Methods Human osteoblast-like cells MG-63 was chosen to be the cytotoxicity of the object.The proliferation of osteoblast was detected by MTT method.The differentiation of osteoblast was detected by the activity of ALP mRNA expression of osteoprotegerin（OPG） and RANKL（receptor activator nuclear factor-kappa B ligand,RANK） were semiquantified by RT-PCR,and the ratio of OPG/RANKL mRNA was used to evaluated the effect of diosgenin on osteoclast signal transduction pathway of OPG/RANKL/RANK system.Results Compared to the control,MTT test demonstrated that NiCr can obviously decrease the cell viability of osteoblast when the concentration is exceed 0.01%.CoCr can not decrease the cell viability of osteoblast.Compared to the control,the expression of OPG has no effect when the the concentration of NiCr is less than 0.01%.But the expression of osteoblast RANKL can be induced.CoCr has no obviously effect in the expression of OPG.Only when the concentration is exceed 0.05%,the osteoblast RANKL can be induced.Conclusion The stainless microparticle has no cell cytotoxicity and can not promote the proliferation,differentiation of osteoblast and inhibit the formation of osteoclast inlow-grade concentration.