镀膜工艺结合3D打印制备Mg-F膜/淫羊藿素膜/β-磷酸三钙支架的表征及成骨能力  

Characterization and osteogenic ability of Mg-F membrane/icariin membrane/beta-tricalcium phosphate scaffolds fabricated by coating process combined with 3D printing

作　　者：薛鹏 杜斌[1] 刘锌[1] 孙光权[1] 程童飞 陈浩 何帅 Xue Peng;Du Bin;Liu Xin;Sun Guangquan;Cheng Tongfei;Chen Hao;He Shuai(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,Jiangsu Province,China;Institute of Chemical Industry,Nanjing University of Science and Technology,Nanjing 210094,Jiangsu Province,China)

机构地区：[1]南京中医药大学附属医院,江苏省南京市210029 [2]南京理工大学化工学院,江苏省南京市210094

出　　处：《中国组织工程研究》2023年第16期2480-2487,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(82074471),项目负责人:杜斌;江苏省卫生健康委科研项目(NK2019027),项目负责人:杜斌;江苏省研究生实践创新计划项目(SJCX22_0769),项目负责人:薛鹏。

摘　　要：背景:修饰生长因子的骨组织工程支架在骨修复材料中具有广阔的应用前景,但生长因子释放过快导致复合支架仅能在早期促进骨修复,镀膜工艺为解决该问题提供了新思路。目的:制备Mg-F膜/淫羊藿素/β-磷酸三钙支架,表征该支架的生物学特性。方法:应用3D打印技术制备β-磷酸三钙支架,通过低能电子束沉积技术制备淫羊藿素膜/β-磷酸三钙支架,再通过脉冲激光沉积技术制备Mg-F膜/淫羊藿素膜/β-磷酸三钙支架,检测支架的微观结构、孔隙直径、孔隙率、丝径、抗压强度及元素组成,并分析淫羊藿素的结合力和缓释性能。将兔骨髓间充质干细胞分别与β-磷酸三钙支架、淫羊藿素/β-磷酸三钙支架与Mg-F膜/淫羊藿素膜/β-磷酸三钙支架浸提液共培养,利用CCK8法检测细胞增殖;将兔骨髓间充质干细胞分别与上述3种支架共培养,加入成骨诱导培养基,利用茜素红染色观察成骨分化能力。结果与结论:(1)扫描电镜下可见,β-磷酸三钙支架结构较为规则,孔隙连通率好;淫羊藿素膜/β-磷酸三钙支架表面有较为致密的淫羊藿素膜覆盖原微观孔隙;Mg-F膜在淫羊藿素膜表面沉积,留下较为粗糙、疏松的表面,存在微观孔隙;3种支架的孔隙直径、孔隙率、丝径、抗压强度比较差异均无显著性意义(P>0.05);(2)Mg-F膜/淫羊藿素膜/β-磷酸三钙支架的淫羊藿素结合力优于淫羊藿素膜/β-磷酸三钙支架(P<0.05),且淫羊藿素缓释更持久;(3)CCK8检测结果显示,3种支架浸提液不影响骨髓间充质干细胞的生长;茜素红染色显示,成骨诱导21 d时,Mg-F膜/淫羊藿素膜/β-磷酸三钙支架组和淫羊藿素膜/β-磷酸三钙支架架组钙结节数及钙结节成熟度优于β-磷酸三钙支架组;(4)结果表明,Mg-F膜/淫羊藿素膜/β-磷酸三钙支架具有良好的细胞相容性、促成骨能力、药物结合力及缓释性能。BACKGROUND:Bone tissue engineering scaffolds with modified growth factors have promising applications in bone repair mate rials,but the rapid release of growth factors leads to composite scaffolds that can promote bone repair only at an early stage,and the coating process provides a new idea to solve this problem.OBJECTIVE:To prepare Mg-F membrane/icaritin/β-tricalcium phosphate scaffold to chara cterize the biological properties of the new scaffold.METHODS:Theβ-tricalcium phosphate scaffold was prepared by applying 3 D printing technology,and the icaritin/β-tricalcium phosphate scaffold was prepared by low energy electron beam deposition technology.Mg-F membrane/icaritin/β-tricalcium phosphate scaffolds were fabricated by pulsed laser deposition technology.The microstructure,pore diameter,porosity,wire diamete r,compressive strength,and elemental composition of the scaffolds were examined,and the icaritin binding force and slow-release properties were analyzed.Rabbit bone marrow mesenchymal stem cells were co-cultured withβ-tricalcium phosphate scaffold,icariin/β-tricalcium phosphate scaffold and Mg-F membrane/icariin mem b rane/β-tricalcium phosphate scaffold extra ct.Cell proliferation was measured by CCK8 assay.Rabbit bone marrow mesenchymal stem cells were co-cultured with the above three scaffolds,separately,and osteogenic induction medium was added,and the osteogenic differentiation ability was observed by alizarin red staining.RESULTS AND CONCLUSION:(1)Scanning electron microscope showed that the structure ofβ-tricalcium phosphate scaffold was relatively regular and the pore connectivity rate was good;the surface of the icariin membrane/β-tricalcium phosphate scaffold had relatively dense icariin membrane covering the original microscopic pores;Mg-F membrane was deposited on the surfa ce of the icariin membrane,leaving a relatively rough and loose surfa ce with microscopic pores.The diffe rences in pore diameter,porosity,wire diameter,and compressive strength of three kinds of scaffolds were not

关 键 词：脉冲激光沉积 低能电子束沉积 缓释膜 Β-磷酸三钙 骨组织工程支架 淫羊藿素 

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