载纳米银磷酸三钙的体外细胞毒性及兔体内毒性  

Silver nanoparticle-doped tricalcium phosphate: in vitro and in vivo toxicity in rabbits

作　　者：程坚 张军[1,3] 管捷[1] 曾俊凯 赵鑫[1] 谢幼专 Cheng Jian;Zhang Jun;Guan Jie;Zeng Junkai;Zhao Xin;Xie Youzhuan(Department of Orthopedics, Ninth People’s Hospital of Shanghai Jiaotong University Medical School, Shangahi 200011, China;Fifth Department of Surgery, Yuyao People’s Hospital, Yuyao 315400, Zhejiang Province, China;Department of Orthopedics, Qingpu Central Hospital of Shanghai, Shanghai 200011, China)

机构地区：[1]上海交通大学医学院附属第九人民医院骨科上海市内植物重点实验室,上海市200011 [2]余姚市人民医院外五科,浙江省余姚市315400 [3]上海青浦区中心医院骨科,上海市200011

出　　处：《中国组织工程研究》2019年第6期917-923,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81071453);项目负责人:谢幼专;上海市科委基金(1441901700);项目负责人:谢幼专~~

摘　　要：背景:课题组前期研究发现,负载5%纳米银颗粒β-磷酸三钙材料具有抗菌性和良好的生物相容性。目的:10%纳米银颗粒β-磷酸三钙材料的体内外毒性。方法:(1)体外细胞毒性:分别采用含质量分数10%纳米银颗粒β-磷酸三钙材料浸提液、多孔β-磷酸三钙材料浸提液、正常组织培养液培养SD大鼠(上海西普尔-必凯实验动物有限公司提供)骨髓间充质干细胞,培养1,3,5,7 d,CCK-8法检测细胞增殖;(2)动物体内毒性:取20只新西兰大白兔(上海市甲干生物科技有限公司提供),在左侧股骨外髁制作直径6 mm、深度13 mm的骨缺损,随机分2组,实验组缺损处植入含质量分数10%纳米银颗粒的β-磷酸三钙材料,对照组植入β-磷酸三钙材料。植入后12周,采用电感耦合等离子体质谱法检测血液和各组织器官内银离子浓度;苏木精-伊红染色切片观察各组织器官病理状态;Van Gieson染色切片观察材料植入区组织生长情况。结果与结论:(1)β-磷酸三钙浸提液组、正常组织培养液组、含纳米银颗β-磷酸三钙材料浸提液组培养不同时间点的细胞增殖比较无差异(P <0.05);(2)实验组和对照组兔血液、心、肝、脾、肺、肾各脏器中的银离子浓度均小于0.1×10^(-6),小于WHO规定的人体内银离子的最高浓度0.5×10^(-6);(3)实验组与对照组兔心、肝、脾、肺、肾组织切片均未见明显病理性改变;(4)实验组与对照组材料轮廓可分辨,材料周围可见被染成红色的新骨组织,材料内部多孔结构内可见大量红染的新骨组织,两组材料多孔结构内的新骨组织分布未见明显差异;(5)结果表明,含质量分数10%纳米银颗粒的β-磷酸三钙材料无明显细胞毒性与动物体内毒性,具有良好的生物相容性。BACKGROUND:Previous studies by the research team found that5%silver nanoparticle-dopedβ-tricalcium phosphate material has antibacterial properties and good biocompatibility.OBJECTIVE:To study the biotoxicity of porousβ-tricalcium phosphate containing10%mass fraction of silver nanoparticles(10%SNPs-TCP)in vitro and in vivo.METHODS:(1)In vitro cytotoxicity:Bone marrow mesenchymal stem cells from Sprague-Dawley rats(provided by Shanghai Xipuer-Beikai Animal Co.,Ltd.,China)were cultured with the extract containing10%SNPs-TCP,porous TCP extract and normal tissue culture medium for1,3,5,7days.Cell counting kit-8method was used to detect cell proliferation.(2)In vivo toxicity:Twenty New Zealand white rabbits(provided by Shanghai Jiagan Biotechnology Co.,Ltd.,China)were used to make bone defects of the left femoral condyle(6mm in diameter and13mm in depth),which were randomly divided into two groups.The experimental group was implanted with10%SNPs-TCP,and the control group was implanted with single TCP.Twelve weeks after implantation,inductively coupled plasma mass spectrometry was used to detect the concentration of silver ions in the blood and tissues.Hematoxylin-eosin staining sections were made to observe the pathological changes of various tissues and organs.Van Gieson staining sections were used to observe the tissue growth in the implantation area.RESULTS AND CONCLUSION:(1)There was no difference in cell proliferation between the TCP group,normal culture group and10%SNPs-TCP group at different time points(P<0.05).(2)The concentration of silver ions in the blood,heart,liver,spleen,lung and kidney of the experimental and control groups was less than0.1×10^-6,which was less than the highest concentration of silver ions(0.5×10^-6)in the human body specified by the WHO.(3)There were no obvious pathological changes in the heart,liver,spleen,lung and kidney sections of the experimental and control groups.(4)The contours of the implantation materials in the experimental and control groups could be distinguished.New bo

关 键 词：纳米银颗粒 Β-磷酸三钙 生物毒性 骨缺损 载纳米银磷酸三钙 银 磷酸钙类 毒性试验 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318.06[医药卫生—临床医学]