3D打印聚己内酯/氧化铜支架促成骨及抗菌性能的体外研究  

Study on the Osteogenic and Antibacterial Properties of 3D Printed PCL/CuO Scaffolds:An In Vitro Study

作　　者：赵双 谢凯[1] 郭煜[1] 谭嘉 吴钧翔 杨阳子 王磊[1] 姜闻博[1,2] 郝永强[1,2] ZHAO Shuang;XIE Kai;GUO Yu;TAN Jia;WU Junxiang;YANG Yangzi;WANG Lei;JIANG Wenbo;HAO Yongqiang(Shanghai Key Laboratory of Orthopaedic Implants,Department of Orthopaedic Surgery,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,200011 Shanghai,China;Clinical and Translational Research Center for 3D Printing Technology,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,200011 Shanghai,China)

机构地区：[1]上海交通大学医学院附属第九人民医院,上海市骨科内植物重点实验室,上海200011 [2]上海交通大学医学3D打印创新与临床转化中心

出　　处：《组织工程与重建外科杂志》2020年第4期263-269,共7页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：十三五国家科技部重点研发计划项目(2016YFC1100600);国家自然科学基金(81972058);上海市卫生健康委员会先进适宜技术推广项目(2019SY033);上海市卫生健康委员会临床重点专科建设项目-生物医用材料;上海交通大学医工交叉青年基金(YG2017QN09)。

摘　　要：目的探索聚己内酯/氧化铜(PCL/CuO)支架的促成骨性能和抗菌性能。方法采用熔融沉积成型(FDM)方法分别制备含有0%、5%、10%CuO的3D打印多孔PCL支架(PCL、PCL/5%CuO和PCL/10%CuO)支架。用ICP离子浓度检测仪测量各组支架在浸泡液中的累积铜离子(Cu^2+)浓度。将支架及浸提液与大鼠骨髓间充质干细胞(rBMSCs)共培养,通过CCK-8法评估细胞黏附和增殖情况,通过碱性磷酸酶(ALP)染色及活力测定、茜素红染色及半定量分析和聚合酶链式反应(PCR)检测成骨分化能力,通过划痕实验分析细胞迁移情况。将浸提液与耐甲氧西林金黄色葡萄球菌(MRSA)共培养,通过光密度值检测及细菌活死染色观察抗细菌生物膜形成性能。结果PCL/CuO复合支架可以持续释放Cu^2+。PCL/5%CuO和PCL/10%CuO支架均可增加rBMSCs的黏附、增殖与成骨分化,其中PCL/10%CuO支架组ALP活性更高、钙结节数量更多、成骨分化相关基因表达更高。此外,PCL/10%CuO支架还可以促进rBMSCs迁移,抑制MRSA细菌活性和细菌生物膜形成。结论PCL/CuO多孔支架可以用于骨缺损修复,其中PCL/10%CuO支架具备最佳的促成骨与抗菌性能。Objective To explore the osteogenic and antibacterial properties of polycaprolactone/cupric oxide(PCL/CuO)scaffolds.Methods Using fused deposition modeling(FDM)method to manufacture three-dimensional(3D)printed porous PCL scaffolds,contained different mass fractions(wt.0%,5%,10%)of CuO:PCL,PCL/5%CuO and PCL/10%CuO scaffolds.The concentration of copper ions(Cu^2+)accumulated in the extracts was measured by ICP.After the scaffolds and extracts co-cultured with rat bone marrow mesenchymal stem cells(rBMSCs),CCK-8 method was used to evaluate cell adhesion and proliferation,alkaline phosphatase(ALP)staining and activity determination,alizarin red staining,semi quantitative analysis and polymerase chain reaction(PCR)were used to detect osteogenic differentiation ability,and wound healing assay was used to analyze cell migration.After the extracts co-cultured with methicillin-resistant Staphylococcus aureus(MRSA),optical density(OD)value and live/dead staining were used to observe the performance of anti-bacterial biofilm.Results PCL/CuO scaffolds could continuously release Cu^2+.Both PCL/5%CuO and PCL/10%CuO scaffolds could increase rBMSCs adhesion,proliferation and osteogenic differentiation.And PCL/10%CuO group showed higher ALP activity,more calcium nodules and higher expression of osteogenic differentiation related genes.Moreover,PCL/10%CuO group promoted the migration of rBMSCs,and inhibited the performance of anti-bacterial biofilm.Conclusion PCL/CuO porous scaffolds could be used for repairing bone defect and PCL/10%CuO scaffold has the most optimal performance.

关 键 词：三维打印 聚己内酯 铜 骨缺损 骨支架 

分 类 号：R318.08[医药卫生—生物医学工程]