亚微米拓扑结构磷酸三钙陶瓷的研制及骨诱导性能  

Preparation and osteoinductive properties of tricalcium phosphate ceramics with submicron topology

作　　者：芦笛 万欣宇 杨金鑫 丁珂欣 张成 段荣泉 刘宗响 Lu Di;Wan Xinyu;Yang Jinxin;Ding Kexin;Zhang Cheng;Duan Rongquan;Liu Zongxiang(School of Stomatology,Xuzhou Medical University,Xuzhou 221004,Jiangsu Province,China;Affiliated Stomatological Hospital of Xuzhou Medical University,Xuzhou 221002,Jiangsu Province,China)

机构地区：[1]徐州医科大学口腔医学院,江苏省徐州市221004 [2]徐州医科大学附属口腔医院,江苏省徐州市221002

出　　处：《中国组织工程研究》2023年第16期2473-2479,共7页Chinese Journal of Tissue Engineering Research

基　　金：四川省科技厅转化项目(2019ZYZF0081),课题名称:复合球粒堆积支架异位成骨和原位修复机制技术研发,项目负责人:段荣泉;徐州医科大学优秀人才科研启动基金(D2020005),课题名称:多孔磷酸钙陶瓷的异位成骨机制及其临床转化,项目负责人:段荣泉。

摘　　要：背景:人工合成的磷酸钙陶瓷材料与天然骨组织无机成分相似,通过表面形貌和化学组成进行功能化设计可赋予其优异的骨传导和骨诱导性能,研发具有骨诱导性能的磷酸钙陶瓷材料是目前的研究热点。目的:通过材料形貌调控和功能化设计赋予亚微米拓扑结构磷酸三钙陶瓷骨诱导性能,检测其理化性能及骨诱导性能。方法:采用高温烧结法制备亚微米拓扑结构的磷酸三钙陶瓷,以市场可供商品化的骨修复材料Bio-Oss骨粉为对照组,表征两种材料的表面形貌、蛋白吸附能力及体外矿化性能。将第3代人牙周膜干细胞与两种材料浸提液共培养,采用CCK-8法检测细胞增殖,茜素红染色检测细胞矿化性能;将第3代人牙周膜干细胞分别接种至两种材料表面,采用碱性磷酸酶染色检测早期成骨,qR T-PCR检测成骨相关因子的表达。结果与结论:(1)扫描电镜下可见两种材料均具有颗粒状纹理的微孔表面,Bio-Oss颗粒明显小于磷酸三钙陶瓷,两种材料的总孔隙度、大孔隙度和微孔隙度相似,磷酸三钙陶瓷主要为亚微米级孔隙,晶粒粒径100 nm-1.0μm,Bio-Oss骨粉主要为纳米级孔隙;体外矿化实验显示,磷酸三钙陶瓷表面诱导骨磷灰石沉积的能力强于Bio-Oss骨粉;与Bio-Oss骨粉相比,磷酸三钙陶瓷可从胎牛血清、牛血清白蛋白溶液中吸附更多的蛋白质(P<0.05);(2)CCK-8实验显示,两种材料均促进细胞增殖,两组间比较差异无显著性意义(P>0.05);(3)磷酸三钙陶瓷组培养4,7 d的碱性磷酸酶活性高于Bio-Oss组(P<0.05),培养21 d的矿化结节数量多于Bio-Oss组;培养7,14 d的碱性磷酸酶、骨钙素、骨桥蛋白及Runx-2的mRNA表达均高于Bio-Oss组(P<0.05);(4)结果表明,新型磷酸钙陶瓷具有优越的体外骨诱导性能。BACKGROUND:Synthetic calcium phosphate ceramic materials are similar to the inorganic components of natural bone tissue.Functional design through surfa ce morphology and chemical composition can endow them with excellent osteoconductive and osteoinductive properties.The development of calcium phosphate ceramic materials with osteoinductive properties is the current research hotspot.OBJECTIVE:To endow the submicron topology tricalcium phosphate ceramics with osteoinductive properties through material morphology regulation and functional design,and test their physicochemical prope rties and osteoinductive properties.METHODS:Novel tricalcium phosphate ceramics were prepared by high tempe rature sintering method.Clinical available Bio-oss bone meal was set as the control group.The sample surface morphology,the protein adsorption capacity,and the in vitro mineralizability of the two materials were characte rized.Human periodontal membrane stem cells at passage 3 were co-cultured with the extra cts of the two mate rials.Cell prolife ration was detected by CCK-8 assay.Alizarin red staining was used to detect cell mineralization performance.The human periodontal ligament stem cells at passage 3 were seeded on the surfaces of the two materials.Early osteogenesis was detected by alkaline phosphatase staining.The expression of osteogenesis-related factors was detected by q RT-PCR.RESULTS AND CONCLUSION:(1)Scanning electron microscope exhibited that both materials had microporous surfa ces with granular texture.Bio-Oss particles were significantly smaller than that of tricalcium phosphate ceramics.The total porosity,macroporosity,and microporosity were similar for both mate rials.Tricalcium phosphate ceramics were mainly submicron pores,with a grain size of 100 nm-1.0μm.Bio-Oss bone meal was mainly nano-scale pores.In vitro mineralization experiments displayed that the ability of the surfa ce of tricalcium phosphate ceramics to induce bone apatite deposition was stronger than that of Bio-Oss bone meal.Compared with Bio-Oss

关 键 词：骨缺损 磷酸三钙 骨修复 骨诱导 骨引导 表面形貌 人牙周膜干细胞 因子 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R-332