Ti6A14V,PMMA,UHMWPE和Co-Cr颗粒对体外培养人外周血单核细胞影响的实验研究

发布时间：2018-02-26 03:18

本文关键词： 无菌性松动骨溶解磨损颗粒单核细胞细胞因子　出处：《滨州医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的：目前公认的人工关节假体无菌性松动主要是由于磨损颗粒诱导骨溶解导致。为更好地研究临床常见磨损颗粒诱发骨溶解反应初始阶段的特征,我们观察分析Ti6A14V, PMMA, UHMWPE和 Co-Cr等四种磨损颗粒对体外培养人外周血单核细胞的影响并了解其功能生物学活性的差异,进一步探讨各种磨损颗粒诱导骨溶解反应引起无菌性松动可能的作用机理。方法：选取15例健康志愿者,每人采集外周循环血40m1。采用密度梯度离心法分离单核细胞,分别同Ti6A14V, PMMA, UHMWPE和Co-Cr等四种磨损颗粒进行共培养。采用台盼蓝染液检测细胞存活率。根据添加磨损颗粒种类的不同分为四组,A组：单核细胞+Ti6A14V颗粒组；B组：单核细胞+PMMA颗粒组；C组：单核细胞+JHMWPE颗粒组；D组：单核细胞+Co-Cr颗粒组。每组根据颗粒/细胞比值为0、0.1、1、10、100、1000等不同比例添加磨损颗粒,培养72小时后提取细胞上清液。采用ELISA法检测上清液中细胞因子TNF-α、IL-1和IL-6的表达水平。各组组内比较观察磨损颗粒浓度变化对单核细胞的影响；组间比较采用颗粒/细胞比值为10、100、1000时进行观察磨损颗粒理化性质对单核细胞的影响。结果：我们发现所有组在加入不同浓度的颗粒前单核细胞存活率均大于95%。共培养72h后发现仅有D组颗粒/细胞比值为1000时单核细胞的存活率降至87%,其它各组单核细胞存活率仍大于95%。四种磨损颗粒均可激活单核细胞,促使其合成分泌不同水平的TNF-α、IL-1和IL-6等细胞因子。当颗粒/细胞比值10、100或1000时,活化的单核细胞分泌TNF-α、IL-1和IL-6的水平与原始幼稚的单核细胞(颗粒/细胞比值为0时)相比,差异具有统计学意义(P0.05)。各组组内比较显示A、B、C和D组中四种磨损颗粒不同的浓度对人外周血单核细胞分泌TNF-α、IL-1和IL-6具有不同的影响。A组、B组和C组中磨损颗粒对单核细胞分泌TNF-α、IL-1和IL-6的影响,伴随颗粒浓度的增加出现不同程度地增强。TNF-α、IL-1和IL-6的表达水平与颗粒浓度呈正相关。但D组中当颗粒/细胞比值达到1000时却出现TNF-α、IL-1和IL-6的表达水平降低。组间比较四种磨损颗粒对TNF-α、IL-1和IL-6表达水平的影响显示在骨溶解反应的初始阶段中Ti6Al4V颗粒的功能生物学活性最强,其次是Co-Cr颗粒和PMMA颗粒,UHMWPE颗粒的功能生物学活性最低。结论：临床常见四种磨损颗粒具有不同功能生物学活性,在骨溶解反应初始阶段对单核细胞的影响存在剂量依赖性。Ti6A14V颗粒较强功能生物学活性和Co-Cr颗粒存在一定的细胞毒性提示可能是影响骨溶解反应初始阶段的关键因素,日益积聚难以降解的PMMA和UHMWPE颗粒可能在假体周围骨溶解反应终末阶段和形成假性肉芽肿过程中具有重要的作用。
[Abstract]:Objective: at present, the aseptic loosening of artificial joint prosthesis is mainly caused by wear particles inducing osteolysis. We observed and analyzed the effects of Ti6A14V, PMMA, UHMWPE and Co-Cr on human peripheral blood mononuclear cells (PBMC) cultured in vitro. To further explore the possible mechanism of aseptic loosening induced by various wear particles. Methods: 15 healthy volunteers were selected to collect peripheral circulating blood 40m1.Monocytes were isolated by density gradient centrifugation. Four kinds of wear particles were co-cultured with Ti6A14V, PMMA, UHMWPE and Co-Cr, respectively. The cell survival rate was measured by trypan blue staining. According to the different types of wear particles added, they were divided into four groups: monocyte Ti6A14V particle group: monocyte Ti6A14V particle group: monocyte group: mononuclear cells. Group C of cell PMMA granules: monocyte JHMWPE granules group D: monocyte Co-Cr granule group. Wear particles were added to each group according to the ratio of 0 0. After 72 hours of culture, the supernatant was extracted. The expression of cytokine TNF- 伪 -1 and IL-6 in the supernatant was detected by ELISA assay. The effect of the concentration of wear particles on monocytes was observed comparatively in each group. The effects of physical and chemical properties of wear particles on monocytes were observed at 1000. Results: we found that the survival rate of monocytes was higher than 95% in all groups before adding different concentrations of particles. After 72 hours of culture, it was found that the survival rate of monocytes in group D was reduced to 87 when the ratio of particles / cells was 1000, and the survival rate of monocytes in other groups was still greater than 95.The four kinds of wear particles could activate monocytes. When the granulosa / cell ratio was 10,100 or 1000, the activated monocytes secreted TNF- 伪 IL-6 and IL-1 compared with the original immature monocytes (the granulosa / cell ratio was 0:00). The difference was statistically significant (P 0.05). The results showed that the different concentrations of four kinds of wear particles in group C and D had different effects on the secretion of TNF- 伪伪 -1 and IL-6 by human peripheral blood monocytes. Group A, group B and group C had different effects of wear particles on mononuclear cells. The effect of TNF- 伪 on the secretion of IL-1 and IL-6, With the increase of particle concentration, the expression levels of IL-1 and IL-6 were positively correlated with the particle concentration, but the expression level of IL-1 and IL-6 decreased when the ratio of granule to cell reached 1000 in group D. The effects of four kinds of wear particles on the expression of IL-1 and IL-6 of TNF- 伪 showed that the functional biological activity of Ti6Al4V particles was the highest in the initial stage of osteolysis. Secondly, the functional biological activities of Co-Cr and PMMA particles were the lowest. Conclusion: four kinds of wear particles have different functional biological activities. In the initial stage of osteolysis reaction, there were dose-dependent effects of Ti6A14V particles on monocytes and cytotoxicity of Co-Cr particles in the initial stage of osteolysis, suggesting that these factors may be the key factors affecting the initial stage of osteolysis reaction. The accumulation of undegradable PMMA and UHMWPE particles may play an important role in the end stage of osteolysis and the formation of pseudogranulomatous granuloma around the prosthesis.
【学位授予单位】：滨州医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R687.4

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