磷酸钙骨水泥/聚乳酸羟基乙酸降解产物促进小鼠单核细胞破骨向分化  

Calcium phosphate cement/poly(lactic-co-glycolic acid)degradation products promote osteoclast differentiation of mouse monocytes

作　　者：龙桂月 李冬冬 廖红兵[2] Long Guiyue;Li Dongdong;Liao Hongbing(Guangxi Key Laboratory of Oral and Maxillofacial Restoration and Reconstruction,Guangxi Cranio-Maxillofacial Malformation Clinical Research Center,Guangxi Health Commission Key laboratory of prevention and treatment for oral infectious diseases,School of Stomatology,Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Department of Prosthetics,School of Stomatology,Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学附属口腔医学院广西口腔颌面修复与重建研究重点实验室,广西颅颌面畸形临床医学研究中心,广西壮族自治区卫生健康委员会口腔感染性疾病防治重点实验室,广西壮族自治区南宁市530021 [2]广西医科大学附属口腔医学院口腔修复科,广西壮族自治区南宁市530021

出　　处：《中国组织工程研究》2023年第8期1193-1198,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81860201,81560190),项目负责人:廖红兵。

摘　　要：背景:前期研究发现磷酸钙骨水泥/聚乳酸羟基乙酸复合材料可加速骨改建过程,其降解产物对单核细胞破骨向分化的影响有待进一步研究。目的:观察磷酸钙骨水泥/聚乳酸羟基乙酸降解产物对小鼠RAW264.7单核细胞破骨向分化的影响。方法:实验分为空白对照组、羟基乙酸组、磷酸钙骨水泥/聚乳酸羟基乙酸组,按照实验分组,将配制好的溶液和50μg/L核因子κB受体活化因子配体添加到完全培养基中制备成不同条件的培养基,培养小鼠RAW264.7细胞5 d,诱导其分化。应用CCK-8法分析各组培养液对细胞增殖作用的影响;RT-PCR和Western blot检测小鼠RAW264.7细胞破骨向分化相关因子活化T细胞核因子c1、基质金属蛋白酶9、核因子κB受体活化因子、抗酒石酸酸性磷酸酶基因和蛋白表达水平的变化;使用抗酒石酸酸性磷酸酶染色法鉴定破骨样细胞。结果与结论:①CCK-8结果显示,细胞在前72 h处于快速生长期,96 h后开始下降;②RT-PCR、Western blot结果显示磷酸钙骨水泥/聚乳酸羟基乙酸组的活化T细胞核因子c1、核因子κB受体活化因子、抗酒石酸酸性磷酸酶mRNA的表达水平及活化T细胞核因子c1、核因子κB受体活化因子的蛋白表达水平显著高于羟基乙酸组、对照组(P<0.05);③羟基乙酸组和磷酸钙骨水泥/聚乳酸羟基乙酸组基质金属蛋白酶9mRNA和蛋白的表达水平差异无显著性意义,但高于对照组(P<0.01);④抗酒石酸酸性磷酸酶染色可见磷酸钙骨水泥/聚乳酸羟基乙酸组破骨样细胞数高于对照组和羟基乙酸组,差异有显著性意义(P<0.05);⑤结果表明磷酸钙骨水泥/聚乳酸羟基乙酸组降解产物形成的微环境可促进小鼠RAW264.7细胞破骨向分化。BACKGROUND:Preliminary studies found that calcium phosphate cement/poly(lactic-co-glycolic acid)composite can accelerate the process of bone remodeling,and the effect of its degradation products on the differentiation of mononuclear osteoclasts needs further study.OBJECTIVE:To observe the effect of calcium phosphate cement/poly(lactic-co-glycolic acid)degradation products on the differentiation of mouse RAW264.7monocytes and osteoclasts.METHODS:The experiment was divided into blank control group,glycolic acid group,and calcium phosphate cement/poly(lactic-co-glycolic acid)group.According to the experimental group assignment,the prepared solution and 50μg/L ligand of nuclear factorκB receptor activator were added to complete medium to prepare media with different conditions.Mouse RAW264.7 cells were cultured for 5 days to induce their differentiation.Cell counting kit-8 assay was used to analyze the effect of different culture media on cell proliferation.RT-PCR and western blot assay were used to detect the changes of gene and protein expression levels of nuclear factor of activated T-cells cytoplasmic 1,matrix metalloproteinase-9,nuclear factorκB receptor activator and tartrate-resistant acid phosphatase activated by osteoclast differentiation-related factors in mouse RAW264.7 monocytes.The osteoclast-like cells were identified by tartrate-resistant acid phosphatase staining.RESULTS AND CONCLUSION:Cell counting kit-8 assay results showed that the cells were in a rapid growth period within 72 hours,and then began to decline after 96 hours.The results of RT-PCR and western blot assay showed that the expression levels of nuclear factor of activated T-cells cytoplasmic 1,nuclear factor-κB receptor activator and tartrate-resistant acid phosphatase mRNA and protein expression levels of nuclear factor-1 and nuclear factor-κB receptor activator in activated T cells in calcium phosphate cement/poly(lactic-co-glycolic acid)group were significantly higher than those in glycolic acid group and control group(P<0.05).Ther

关 键 词：磷酸钙骨水泥/聚乳酸羟基乙酸 单核细胞 破骨分化 基质金属蛋白酶9 活化T细胞核因子c1 核因子ΚB受体活化因子 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R394.2