搭载淫羊藿苷聚乳酸-羟基乙酸共聚物(PLGA-ICA)复合支架对兔软骨细胞的影响

Effect of patchouli glycoside polylactic acid-hydroxyacetic acid copolymer(PLGA-ICA)composite scaffold on rabbit chondrocytes

目的 本课题探究搭载不同浓度淫羊藿苷的聚乳酸-羟基乙酸共聚物(PLGA-ICA)复合支架对兔软骨细胞的影响,阐明了其作为骨关节炎潜在治疗手段的可能性。方法 本研究将淫羊藿苷和PLGA以不同比例混合,利用静电纺丝技术构建PLGA-ICA复合支架。利用扫描电镜(SEM)观察PLGA-ICA复合支架的表面形貌,测量支架及复合支架直径和接触角;与兔软骨细胞共培养,将细胞分为:空白对照组、0.01%ICA组、0.1%ICA组、1%ICA组、PLGA组、0.01%ICA+PLGA组、0.1%ICA+PLGA组、1%ICA+PLGA组,MTT法检测支架对兔软骨细胞的细胞相容性,并在培养24h后,荧光显微镜下观察兔软骨细胞的细胞黏附与扩散区域面积。结果 扫描电镜表明PLGA支架平均直径为1.58±0.16μm;复合纤维支架的平均直径分别为1.01μm±0.59μm、1.75μm±0.32μm、1.89μm±0.43μm。纯PLGA组的水接触角为88.32±4.57°,随着ICA的浓度增加,其接触角变化范围为82.58°±8.03°-71.25°±0.51°。MTT法检测表明与空白对照组相比,随着ICA的浓度自0.01%-1.00%上升,兔软骨细胞增殖的效果差异有统计学意义(P<0.05),与未进行任何处理的空白PLGA支架组相比,搭载了ICA的复合支架的兔软骨细胞随ICA浓度增加(0.01%-1%)兔软骨细胞增殖的效果差异有统计学意义(P<0.05)。24h后荧光显微镜观察发现与空白对照组相比,随着ICA的浓度自0.01%-1%上升,兔软骨细胞粘附数量明显增加(P<0.05);随着ICA的浓度自0.01%-1%上升,兔软骨细胞复合支架中培养的细胞其扩散区域比单纯PLGA支架的细胞扩散区域增加(P<0.05)。结论 PLGA-ICA复合支架能够促进兔软骨细胞的增殖,在骨关节炎的治疗中有潜在应用价值。

Objective This study investigated the effects of poly(lactic acid)-hydroxy acetic acid copolymer(PLGAICA)composite scaffolds carrying different concentrations of epimedium on rabbit chondrocytes and elucidated its possibility as a potential therapeutic tool for osteoarthritis.Methods In this study,patchouli glycosides and PLGA were mixed in different ratios and PLGA-ICA composite scaffolds were constructed using electrostatic spinning technique.The surface morphology of the PLGA-ICA composite scaffold was observed using scanning electron microscopy(SEM),and the diameter and contact angle of the scaffold and composite scaffold was measured;the cells were co-cultured with rabbit chondrocytes and divided into blank control group,0.01%ICA group,0.1%ICA group,1%ICA group,PLGA group,0.01%ICA+PLGA group,0.1%ICA+PLGA group,and 1%ICA+PLGA group,MTT method was used to detect the cytocompatibility of the scaffold on rabbit chondrocytes,and the area of cell adhesion and spreading area of rabbit chondrocytes was observed under a fluorescence microscope after 24hof culture.Results Scanning electron microscopy showed that the average diameter of PLGA scaffold was 1.58±0.16μm;the average diameter of the composite fiber scaffold was 1.01μm±0.59μm,1.75μm±0.32μm and 1.89μm±0.43μm,respectively.the water contact angle of the pure PLGA group was 88.32±4.57°,and with the increase of ICA concentration,the contact angle change The MTT assay showed a statistically significant difference in the effect of rabbit chondrocyte proliferation as the concentration of ICA increased from 0.01%to 1.00% compared with the blank control group(P<0.05),and compared with the blank PLGA scaffold group without any treatment,the rabbit chondrocytes in the composite scaffold equipped with ICA After 24h,fluorescence microscopy revealed that the number of rabbit chondrocyte adhesions increased significantly(P<0.05)as the concentration of ICA increased from 0.01% to 1% compared with the blank control group;as the concentration of ICA increased from 0.01%to 1%,the number of rabbit chondrocyte composite scaffolds increased from 0.01%to 1%(P<0.05).rose,the cell spreading area of the cells cultured in the rabbit chondrocyte composite scaffold increased than that of the PLGA scaffold alone(P<0.05).Conclusion The PLGA-ICA composite scaffold can promote the proliferation of rabbit chondrocytes and has potential application in the treatment of osteoarthritis.