聚己内酯胶原支架纳米纤维方向对小鼠MC3T3-E1成骨前体细胞的形态,增值和分化的影响.docVIP

PAGE \* MERGEFORMAT 3 聚己内酯/胶原支架纳米纤维方向对小鼠MC3T3-E1成骨前体细胞的形态、增值和分化的影响 路 菊[通迅作者] xxx，电话:(023 [通迅作者] xxx，电话:(023E—mail： [摘要] 目的：探讨聚己内酯/胶原（PCL/COL）纳米纤维方向对小鼠成骨前体细胞（MC3T3-E1）的形态，增殖，分化的影响。方法：通过扫描电子显微镜（SEM）对非取向和取向的PCL/COL纳米纤维支架的形貌进行观察;将MC3T3-E1细胞接种在非取向与取向的PCL/COL纳米纤维支架上，通过SEM及激光共聚焦显微镜(LSCM)观察细胞在两组材料上的生长形态，CCK-8法检测细胞在两组材料上的增殖情况，Real time RT-PCR检测细胞在两组材料上成骨分化的功能性基因Ⅰ型胶原（COLⅠ）、碱性磷酸酶（ALP）、骨钙素（OCN）的表达情况。结果：非取向PCL/COL纳米纤维支架的纤维方向无规则，取向PCL/COL纳米纤维支架的纤维方向相同,两者纤维直径差异无统计学意义(p 0.05)，分别为586±239 nm和625±155 nm); SEM及LSCM检测结果显示MC3T3-E1细胞在非取向的纳米纤维上成多边形生长，细胞骨架呈不规则形态，而在取向纳米纤维上成纺锤形生长，且细胞骨架沿纳米纤维方向整齐排列；CCK-8检测结果显示在1、3、5和7d MC3T3-E1细胞在非取向与取向PCL/COL纳米纤维支架上的增殖率比较差异无统计学意义(p 0.05); Real time RT-PCR检测结果显示MC3T3-E1细胞在两组材料上COLⅠ表达水平比较差异无统计学意义(p 0.05)，而细胞在非取向组较在取向组上能表达更多的ALP和OCN，比较差异均有统计学意义(p 0.05)。结论：非取向和取向的PCL/COL纳米纤维支架均能提供了一个良好的微环境利于MC3T3-E1细胞的生长、增值，同时非取向PCL/COL纳米纤维支架较取向PCL/COL纳米纤维支架更有利于MC3T3-E1细胞的成骨分化，是一种有应用前景的骨组织工程支架材料。 [关键词] 聚己内酯/胶原；纳米纤维；MC3T3-E1细胞；方向 The effect of poly(ε-caprolactone)/collagen nanofiber orientation on MC3T3-E1 preosteoblast morphology, proliferation, and differentiation Lu Ju (Biomedical Analysis Center，Third Military Medical University，Chongqing 400038，China) [Abstract] Objective: To explore the effect of poly(ε-caprolactone) (PCL)/collagen (COL) nanofiber orientation on mouse preosteoblast cells ( MC3T3-E1 ) morphology, proliferation, and differentiation. Methods: The morphologies of the random and aligned PCL/COL nanofiber scaffolds were observed by scanning electron microscopy ( SEM ); MC3T3-E1 cells were cultured on the random and aligned PCL/COL nanofibers scaffolds to study the effects of fiber orientation on cell morphology through SEM and laser confocal microscope ( LSCM ), To evaluate proliferation, numbers of viable cells were measured using the Cell Counting Kit-8 (CCK-8), To explore how MC3T3-E1 sense and respond to different scaffolds, real-time RT-PCR was used to testify the expression of osteogenic marker genes (including collagen type Ⅰ( COL