中国修复重建外科杂志

中国修复重建外科杂志

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

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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。

ObjectiveTo investigate the preparation and osteogenic properties of poly (L-lactic acid)(PLLA)/lecithin porous scaffolds with open pore structure.MethodsPLLA/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 of the 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.ResultsSEM 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 was a significant difference in ALP activity of each group. The ALP activity in groups D, E, F, and G were significantly higher than that in groups A, B, and C (P<0.05).In vivo, the results of Micro-CT examination and bone volume and bone mineral density showed that the scaffolds with 30% lecithin had the best repairing effect.ConclusionPrepared by thermally induced phase separation, the cytocompatibility, osteogenic differentiation, and bone repair ability of the PLLA/lecithin porous scaffold is obviously better than that of pure PLLA scaffold. PLLA/lecithin porous scaffold with suitable lecithin content is a promising scaffold material for bone tissue engineering.

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

Key words: Thermally induced phase separation; poly (L-lactic acid); lecithin; porous scaffold; bone tissue engineering

引用本文: 谌斯, 杜昶. 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. 中国修复重建外科杂志, 2018, 32(9): 1123-1130. doi: 10.7507/1002-1892.201804127 复制

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