纳米羟基磷灰石-氧化锌复合支架生物性能及对MC3T3-E1成骨细胞行为的影响  

Biological properties of nano-hydroxyapatite-zinc oxide composite scaffolds and their effects on the behavior of MC3T3-E1 osteoblasts

作　　者：刘子璇 李岩 伋琳 夏德林 Liu Zixuan;Li Yan;Ji Lin;Xia Delin(Department of Oral and Maxillofacial Surgery,Affiliated Stomatological Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Taizhou Polytechnic College,Bone Tissue Engineering Research Center of Taizhou,Taizhou 225300,Jiangsu Province,China;Department of Plastic and Maxillofacial Surgery,Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]西南医科大学附属口腔医院口腔颌面外科,四川省泸州市646000 [2]泰州职业技术学院药学院,泰州市骨组织工程研究中心,江苏省泰州市225300 [3]重庆医科大学附属第二医院整形与颌面外科,重庆市400010

出　　处：《中国组织工程研究》2023年第34期5441-5447,共7页Chinese Journal of Tissue Engineering Research

基　　金：江苏省高等学校自然科学面上项目(21KJB180001),项目负责人:李岩;江苏省“青蓝工程”优秀青年骨干教师项目(2021年度),项目负责人:李岩。

摘　　要：背景:有研究发现,在纳米羟基磷灰石中掺杂金属离子锌可改善纳米羟基磷灰石的力学性能。目的:探讨含不同质量分数氧化锌的纳米羟基磷灰石陶瓷支架的生物性能,以及对MC3T3-E1成骨细胞的生物学行为的影响。方法:应用甲壳素凝胶体系制备含有不同质量分数(0%,0.5%,1.5%,2.5%,3.5%)氧化锌的纳米羟基磷灰石陶瓷支架,表征支架的物理性能。将5组纳米羟基磷灰石-氧化锌复合支架分别与MC3T3-E1成骨细胞共培养,分别进行Calcein-AM/PI活死细胞染色、CCK-8细胞增殖实验、CellTiter-LumiTM发光实验、碱性磷酸酶活性检测、茜素红染色及钙离子定量分析。结果与结论:(1)扫描电镜下可见纳米羟基磷灰石-氧化锌复合支架表面具有三维贯通的孔隙,复合支架的孔径大小均匀,孔径范围在300-400μm,孔隙率均在70%以上;随着氧化锌质量分数的增加,复合支架的抗压强度逐渐升高。(2)Calcein-AM/PI活死细胞染色显示,5组支架均无明显的细胞毒性,其中2.5%组细胞存活率高于0%组(P<0.05)。CCK-8细胞增殖与CellTiter-LumiTM发光实验实验显示,各组细胞增殖及活性由强到弱的顺序为:2.5%组>1.5%组>0.5%组>0%组>3.5%组。各组细胞碱性磷酸酶活性由高到低的顺序为:2.5%组>1.5%组>3.5%组>0.5%组>0%组。茜素红染色与钙离子定量分析显示,各组细胞矿化结节形成能力由强到弱的顺序为:2.5%组>1.5%组>0.5%组>3.5%组>0%组。(3)结果表明,氧化锌的掺杂提高了纳米羟基磷灰石支架的力学性能,可促进成骨细胞的增殖与分化,其中掺杂质量分数2.5%氧化锌的复合支架促成骨细胞增殖和分化能力最佳。BACKGROUND:It has been found that the mechanical properties of nano-hydroxyapatite can be improved by doping zinc ions in nano-hydroxyapatite.OBJECTIVE:To investigate the biological properties of nano-hydroxyapatite ceramic scaffold mixed with different concentrations of zinc oxide,and to study the effects of the scaffold on the biological behavior of MC3T3-E1 osteoblasts.METHODS:Nano-hydroxyapatite ceramic scaffolds containing different mass fractions(0%,0.5%,1.5%,2.5%,3.5%)of zinc oxide were prepared by a chitin gel system to characterize the physical properties of scaffolds.The five groups of nano-hydroxyapatite-zinc oxide composite scaffolds were co-cultured with MC3T3-E1 osteoblasts.Calcein AM/PI staining,CCK-8 assay,CellTiter LumiTM Luminescent assay,alkaline phosphatase activity detection,alizarin red staining,and quantitative analysis of calcium ions were detected.RESULTS AND CONCLUSION:(1)Under a scanning electron microscope,the surface of the nano-hydroxyapatite-zinc oxide composite scaffolds had threedimensional perforated pores.The pore size of the composite scaffolds was uniform,ranging from 300 to 400μm,and the porosity was above 70%.With the increase of the mass fraction of zinc oxide,the compressive strength of the composite scaffold increased gradually.(2)Calcein-AM/PI staining showed no obvious cytotoxicity in the five groups,and the survival rate of cells in the 2.5%group was higher than that in the 0%group(P<0.05).CCK-8 assay and CellTiter-LumiTM luminescent assay showed that cell proliferation and activity of each group were in the order from strong to weak:2.5%group>1.5%group>0.5%group>0%group>3.5%group.The order of alkaline phosphatase activity from high to low was 2.5%group>1.5%group>3.5%group>0.5%group>0%group.Alizarin red staining and quantitative analysis of calcium ions showed that the formation ability of cell mineralized nodules was in the order of 2.5%group>1.5%group>0.5%group>3.5%group>0%group from strong to weak.(3)The results showed that zinc oxide doping improved the mechanical

关 键 词：羟基磷灰石 氧化锌 骨组织工程 支架 骨再生 甲壳素凝胶 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R-331