新型三元复合材料纳米羟基磷灰石/聚酰胺66/氧化锆的制备及体外生物相容性  

Preparation and in vitro biocompatibility of a novel ternary biomaterial, yttria-stabilized zirconia reinforced nano-hydroxyapatite/polyamide 66

作　　者：李毓灵 蒋科[1] 陈路[1] 蔚芃[1] 陈骞[1] 谯波[2] 蒋电明 Li Yuling;Jiang Ke;Chen Lu;Yu Peng;Chen Qian;Qiao Bo;Jiang Dianming(Department of Orthopedics, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China;the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China;Bone and Trauma Centre, the Third Affiliated Hospital of Chongqing Medical University, Chongqing 401120, China)

机构地区：[1]川北医学院附属医院骨科,四川省南充市637000 [2]重庆医科大学附属第一医院,重庆市400016 [3]重庆医科大学附属第三医院骨与创伤中心,重庆市401120

出　　处：《中国组织工程研究》2019年第6期930-935,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金青年基金(NSFC81501876);项目负责人:谯波;四川省科技厅应用基础项目(2018JY0250);项目负责人:李毓灵~~

摘　　要：背景:纳米羟基磷灰石/聚酰胺66复合材料具有高仿生特性,可通过与宿主骨直接结合来发挥生物活性作用,但其缺乏足够的力学强度。目的:制备新型三元复合材料纳米羟基磷灰石/聚酰胺66/氧化锆(nano-hydroxyapatite/polyamide66/yttria-stabilized tetragonal zirconia,nH A/PA66/YTZ),验证其力学特性及体外生物相容性。方法:采用两步法制备三元复合材料n HA/PA66/YTZ,其中纳米羟基磷灰石与氧化锆的质量比分别100∶0、90∶10、80∶20、60∶40。扫描电镜观察复合材料表征,力学测试仪测试其抗弯强度、抗张强度、抗压强度、弹性模量、断裂伸长率等力学参数,评价其力学性能,筛选最佳质量比复合材料,用于以下实验。分别采用细胞培养液(空白对照组)、纳米羟基磷灰石/聚酰胺66材料浸提液(对照组)、nHA/PA66/YTZ材料浸提液(实验组)培养小鼠成骨细胞MC3T3-E1,CCK-8法检测细胞增殖;将纳米羟基磷灰石/聚酰胺66材料(对照组)、nHA/PA66/YTZ材料(实验组)分别与小鼠成骨细胞MC3T3-E1共培养,24h后采用激光共聚焦显微镜观察MC3T3-E1细胞在复合材料表面的黏附、增殖情况。结果与结论:(1)扫描电镜显示,氧化锆晶粒填充了原本纳米羟基磷灰石晶粒之间的空隙,纳米羟基磷灰石/氧化锆均匀分散在聚酰胺66基体中;(2)生物力学测试显示,纳米羟基磷灰石与氧化锆质量比为60∶40nHA/PA66/YTZ材料的抗压强度、抗弯强度、抗张强度、断裂伸长率及弹性模量最高,力学性能最优,选择其进行细胞相容性实验;(3)CCK-8检测显示,随着时间的延长,3组细胞数量逐渐增加,3组间细胞增殖比较无差异;(4)激光共聚焦显微镜显示,实验组复合材料上的细胞呈现融合、团聚及分层现象,细胞内肌动蛋白丝更多;对照组复合材料上的细胞呈现单层及分散现象,细胞数量与细胞内的肌动蛋白丝较实验组少;(5)结果表明,三元复合材料nH A/PA66/YTZ在体外�BACKGROUND:Nano-hydroxyapatite/polyamide66(nHA/PA66)composite materials possess high bionic properties and exert biological activity by directly combining with host bone,but it lacks sufficient mechanical strength.OBJECTIVE:To prepare a novel ternary biomaterial composed of nHA/PA66/yttria-stabilized tetragonal zirconia(YTZ),and to investigate its mechanical properties and biocompatibility.METHODS:The biomaterial was prepared by two-step approach,and the mass ratio of nano-hydroxyapatite to yttria-stabilized tetragonal zirconia was100:0,90:10,80:20,and60:40,respectively.The characterization of nHA/PA66/YTZ was observed by scanning electron microscope.The mechanical parameters of nHA/PA66/YTZ including bending strength,tensile strength,compressive strength,elastic modulus and breaking elongation were tested to evaluate its mechanical properties.The mechanical properties were evaluated by the mechanical tester to select composite materials with the optimum mass ratio applied to the following experiments.The MC3T3-E1cells were cultured by the cell-culture medium(blank control group),nHA/PA66material extract(control group),nHA/PA66/YTZ material extract(experimental group).The cell proliferation was detected by cell counting kit-8assay.The nHA/PA66material(control group)and nHA/PA66/YTZ material(experimental group)were respectively co-cultured with MC3T3-E1cells.The adhesion and proliferation of MC3T3-E1cells on the surface of composite materials were observed by the laser scanning confocal microscope after24hours.RESULTS AND CONCLUSION:The scanning electron microscope showed that YTZ grains filled the gaps between the original nano-hydroxyapatite grains and the nHA/YTZ was evenly dispersed in the matrix of polyamide66.The biomechanical test revealed that the compressive strength,bending strength,tensile strength,elongation at break and elastic modulus were the highest at60:40of the mass ratio of nHA and YTZ,while the mechanical properties were optimal,which could be chosen for the cell compatibility experiments.Cell

关 键 词：口腔材料 纳米羟基磷灰石 氧化钇稳定型氧化锆 聚酰胺66 三元复合材料 肌动蛋白丝 羟基磷灰石类 尼龙 材料试验 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]