具有开放孔结构的左旋聚乳酸/卵磷脂多孔支架的制备及成骨性能研究  

Preparation and osteogenic properties of poly(L-lactic acid)/lecithin porous scaffolds with open pore structure

作　　者：谌斯 杜昶 CHEN Si;DU Chang(Department of Biomedical Engineering,School of Materials Science and Engineering,South China University of Technology,Guangzhou Guan gdong,510641,P.R.China;National Engineering Research Center for Tissue Restoration and Reconstruction,South China University of Technology,Guangzhou Guangdong,510006,P.R.China;Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education,South China University of Technology,Guangzhou Guangdong,510006,P.R.China)

机构地区：[1]华南理工大学材料科学与工程学院生物医学工程系,广州510641 [2]华南理工大学国家人体组织功能重建工程技术研究中心,广州510006 [3]华南理工大学生物医用材料与工程教育部重点实验室,广州510006

出　　处：《中国修复重建外科杂志》2018年第9期1123-1130,共8页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家重点研发计划资助项目(2017YFC1105000);国家自然科学基金资助项目(51572087)~~

摘　　要：目的探讨具有开放孔结构的左旋聚乳酸[poly(L-lactic acid),PLLA]/卵磷脂多孔支架的制备方法及成骨性能。方法采用热致相分离法制备含不同比例卵磷脂(0、5%、10%、20%、30%、40%、50%)的PLLA/卵磷脂多孔支架(分别对应为A、B、C、D、E、F、G组)。扫描电镜观察各支架表面形貌;广角X射线衍射(X-ray diffraction,XRD)和差示扫描量热法(differential scanning calorimetry,DSC)检测各支架结晶度;测量各组支架吸水率;采用细胞计数试剂盒8(cell counting kit 8,CCK-8)法检测小鼠BMSCs在各支架表面的增殖情况;通过检测ALP活性观察小鼠BMSCs在各组支架上的成骨分化能力。最后使用SD大鼠颅骨缺损模型观察含不同比例卵磷脂的支架在动物体内的骨修复情况,采用Micro-CT对缺损处进行三维模型重建,并对缺损处的新生骨体积和骨矿物密度进行定量分析。结果扫描电镜观察示,卵磷脂会导致支架孔径略微缩小;当卵磷脂含量为50%时,支架表面会出现片状结构。广角XRD和DSC检测示,支架结晶度随卵磷脂含量的升高而逐渐降低。亲水性测试示,随着卵磷脂含量增加,支架的亲水性逐渐增强。CCK-8法检测示,随培养时间增加,BMSCs在各组支架上均有明显增殖;培养7 d时,C、D、E、F组吸光度(A)值显著高于A、B、G组(P<0.05),C、D、E、F组间比较差异无统计学意义(P>0.05)。成骨诱导14 d时,随卵磷脂含量增加,各组ALP活性出现了显著差异,D、E、F、G组ALP活性显著高于A、B、C组(P<0.05)。大鼠颅骨缺损修复实验中,Micro-CT检测及新生骨体积和骨矿物密度结果均显示,含30%卵磷脂支架修复效果最佳。结论通过热致相分离法制备的PLLA/卵磷脂多孔支架的细胞相容性、成骨分化性能及骨修复能力显著强于单纯PLLA支架;卵磷脂含量适宜的PLLA/卵磷脂多孔支架有望作为骨组织工程支架材料。Objective To investigate the preparation and osteogenic properties of poly（L-lactic acid）（PLLA）/lecithin porous scaffolds with open pore structure. Methods PLLA/lecithin porous scaffolds with different lecithin contents（0, 5%, 10%, 20%, 30%, 40%, 50%） were prepared by thermally induced phase separation（groups A, B, C, D, E, F,and G, respectively）. Scanning electron microscopy（SEM） was used to observe the surface morphology of the scaffolds.Wide-angle X-ray diffraction（XRD） and differential scanning calorimetry（DSC） were used to detect the crystallinity ofthe scaffolds. The water uptake ability of the scaffolds was measured. The cell growth and viability of bone marrow mesenchymal stem cells（BMSCs） of mouse on each scaffold was assessed by cell counting kit 8（CCK-8） method. The osteogenic differentiation ability of BMSCs on each scaffold was evaluated by alkaline phosphatase（ALP） activity. Finally,a critical-size rat calvarial bone defect model was used to evaluate the osteogenesis of the scaffolds in vivo. Micro-CT was used to reconstruct the three-dimensional model of the defect area, and the bone volume and bone mineral density were quantitatively analyzed. Results SEM results showed that the lecithin could slightly reduce the pore size; when lecithin content was 50%, platelet-like structure could be observed on the scaffolds. Wide angle XRD and DSC showed that the crystallinity of scaffolds gradually decreased with the increase of lecithin content. The water uptake ability test showed that the hydrophilicity of scaffolds increased with the increase of lecithin content. CCK-8 assay showed that cell activity gradually increased with the increase of culture time. After 7 days of culture, the absorbance（A） value of groups C, D, E,and F were significantly higher than that of groups A, B, and G（P〈0.05）, but no significant difference was found among groups C, D, E, and F（P〉0.05）. After 14 days of osteogenic induction, with the increase of lecithin content, there

关 键 词：热致相分离 左旋聚乳酸 卵磷脂 多孔支架 骨组织工程 

分 类 号：R318.08[医药卫生—生物医学工程]