纳米二氧化锆对鼻黏膜外胚层间充质干细胞成骨分化的影响  

Effects of nano-zirconium dioxide on osteogenic differentiation of ectomesenchymal stem cells in nasal mucosa

作　　者：卞璐 夏丹丹 钱源[2] 史雯 阙云端 吕龙 徐爱花 史文涛 Bian Lu;Xia Dandan;Qian Yuan;Shi Wen;Que Yunduan;Lyu Long;Xu Aihua;Shi Wentao(Central Laboratory of Gaochun Hospital Affiliated to Jiangsu University,Nanjing 211300,Jiangsu Province,China;Department of Orthopedics,Affiliated Hospital of Jiangnan University,Nanjing 211300,Jiangsu Province,China)

机构地区：[1]江苏大学附属高淳医院中心实验室,江苏省南京市211300 [2]江南大学附属医院骨科,江苏省南京市211300

出　　处：《中国组织工程研究》2024年第15期2346-2350,共5页Chinese Journal of Tissue Engineering Research

基　　金：南京市卫生科技发展专项资金项目(重点项目:ZKX21063),项目负责人:阙云端,史文涛;南京市卫生科技发展专项资金项目(YKK19129),项目负责人:史文涛。

摘　　要：背景:纳米二氧化锆在骨组织修复领域表现出良好的应用潜力,研究纳米二氧化锆对成骨分化的影响将有助于推广纳米二氧化锆治疗骨缺损的临床应用。目的:探究纳米二氧化锆对鼻黏膜外胚层间充质干细胞成骨分化的影响。方法:分离培养大鼠鼻黏膜来源外胚层间充质干细胞,使用CCK-8法检测纳米二氧化锆的细胞毒性,根据细胞毒性选择生物安全浓度,然后将细胞随机分为对照组、纳米二氧化锆组和纳米羟基磷灰石组,定向诱导各组细胞成骨分化。在诱导分化第7天进行碱性磷酸酶染色,采用qRT-PCR和Western blot检测早期成骨标志物(Runx2和Osx)的表达水平,在诱导分化第21天进行茜素红染色,采用qRT-PCR和Western blot检测中晚期成骨标志物(OPN和OCN)的表达水平。结果与结论:①纳米二氧化锆对鼻黏膜外胚层间充质干细胞的半数致死质量浓度为0.6 mg/mL,该实验选择质量浓度为200μg/mL进行干预,二氧化锆对细胞增殖无明显影响;②与对照组相比,纳米二氧化锆组细胞碱性磷酸酶染色更明显且细胞矿化水平更高,但与纳米羟基磷灰石组对比无明显差异;③与对照组相比,纳米二氧化锆组成骨相关基因和蛋白的表达均明显增多,但与纳米羟基磷灰石组对比无明显差异;④结果表明:纳米二氧化锆具有良好的生物安全性,且能对鼻黏膜外胚层间充质干细胞的成骨分化产生促进作用,这种促进效果与纳米羟基磷灰石相当。BACKGROUND:Nano-zirconium dioxide has good application potential in the field of bone tissue repair.Studying the effect of nano-zirconium dioxide on osteogenic differentiation will help to promote the clinical application of nano-zirconium dioxide in the treatment of bone defects.OBJECTIVE:To explore the effect of nano-zirconium dioxide on the osteogenic differentiation of ectomesenchymal stem cells in the nasal mucosa.METHODS:Ectomesenchymal stem cells derived from rat nasal mucosa were isolated and cultured,and the biotoxicity of nano-zirconium dioxide to the cells was detected by CCK-8 assay.The biosafety concentration was selected according to the cytotoxicity,and the cells were randomly divided into a control group,a nano-zirconium dioxide group,and a nano-hydroxyapatite group.Osteogenic differentiation of cells was directionally induced in each group.On day 7 of induced differentiation,alkaline phosphatase staining was performed.qRT-PCR and western blot assay were used to detect the expression of early osteogenic markers(Runx2 and Osx).On day 21 of induced differentiation,alizarin red staining was conducted.qRT-PCR and western blot assay were utilized to determine the expression levels of late osteogenic markers(OPN and OCN).RESULTS AND CONCLUSION:(1)The median lethal concentration of nano-zirconium dioxide on ectomesenchymal stem cells in nasal mucosa was 0.6 mg/mL.In the experiment,the mass concentration of 200μg/mL was selected for intervention.Zirconium dioxide had no significant effect on the proliferation of the cells.(2)Compared with the control group,the alkaline phosphatase staining of the cells in the nano-zirconium dioxide group was more obvious and the level of cell mineralization was higher,but there was no significant difference compared with the nano-hydroxyapatite.(3)Compared with the control group,the expression of bone-related genes and proteins increased significantly,but there was no significant difference compared with nano-hydroxyapatite.(4)The results show that nano-zirconium dioxide

关 键 词：鼻黏膜 外胚层间充质干细胞 二氧化锆 羟基磷灰石 成骨分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R683