新型活性修饰聚乳酸组织工程骨的体外成骨研究Osteogenic ability of poly-D,I-lactide acid(PDLLA) scaffolds affected by bioactive modification in vitro
许子星;陈建庭;张鑫鑫;姜晓锐;李方国;李涛;查丁胜;刘富强;巫松辉;朱青安;
摘要(Abstract):
[目的]探讨氨等离子体改性、酰胺键接枝甘氨酸-精氨酸-甘氨酸-天冬氨酸-丝氨酸(GRGDS)短肽的活性修饰方法对消旋聚乳酸(PDLLA)组织工程骨体外成骨能力的影响。[方法]制备圆片状直径8 mm、厚1 mm的PDLLA三维多孔支架,分为三组:表面氨基化PDLLA(aminated PDLLA,A/PDLLA,A组),接枝肽A/PDLLA(peptides conjugated A/PDLLA,PA/PDLLA,PA组),以未经处理的PDLLA(P组)作为对照组。实时荧光定量聚合酶链式反应(RT-qPCR)检测各组材料上骨髓间充质干细胞(BMSCs)、骨钙素(OCN)、Ⅰ型胶原(Col-Ⅰ)、碱性磷酸酶(ALP)、骨形态发生蛋白-2(Bmp-2)和骨桥蛋白(OPN)mRNA的表达变化并进行ALP活性测定和钙黄绿素矿化荧光染色。[结果]qPCR结果示:第3 d,PA组各基因表达倍数均较P组高;除OCN外的各基因表达也均较A组高。第7 d,A组OCN(13.13±1.28)、Col-Ⅰ(23.71±6.51)和OPN(27.4±7.17)mRNA表达倍数为最高,PA组次之。第14 d,多数基因表达上调,A组和PA组表达倍数较对照组明显增高。PA组OCN(49.21±7.03)、ALP(24.26±3.41)和BMP-2(11.82±2.38)mRNA的表达较A组高,差异有统计学意义(P<0.05)。ALP活性检测结果示:A组和PA组ALP活性持续升高,P组活性第21 d较第14 d时略有下降。第7 d,A组和PA组之间ALP活性比较无统计学意义(P>0.05),但均高于P组(P<0.01);第14 d和第21 d,三组间ALP活性两两比较均有统计学意义,ALP活性PA组>A组>P组。钙黄绿素矿化荧光染色观察示,骨支架上钙盐沉积与ALP的活性变化趋势相平行。[结论]氨等离子体改性能够促进聚乳酸组织工程骨支架上BMSCs早期向成骨细胞分化,改性后接枝GRGDS肽的新型活性修饰PDLLA具有更好的促BMSCs体外成骨能力。
关键词(KeyWords): 消旋聚乳酸;活性修饰;仿生材料;骨组织工程;基因表达
基金项目(Foundation): 广东省科技计划项目(编号:2007B031003005)
作者(Author): 许子星;陈建庭;张鑫鑫;姜晓锐;李方国;李涛;查丁胜;刘富强;巫松辉;朱青安;
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