聚己内酯/胶原支架纳米纤维方向对小鼠成骨前体细胞形态、增殖和分化的影响  

Effect of poly( ε-caprolactone ) /collagen nanofiber orientation on morphology,proliferation and differentiation in mouse preosteoblast MC3T3-E1 cells

作　　者：路菊[1] 

机构地区：[1]第三军医大学科研部生物医学分析测试中心,重庆400038

出　　处：《第三军医大学学报》2013年第19期2069-2072,共4页Journal of Third Military Medical University

摘　　要：目的探讨聚己内酯/胶原(PCL/COL)支架纳米纤维方向对小鼠成骨前体细胞(MC3T3-E1)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呈星形生长,细胞骨架呈不规则形态,而在取向纳米纤维上呈扁长形生长,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 d和7 d 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取向组能表达更多的ALP和OCN,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一个良好的微环境,利于MC3T3-E1细胞的生长、增殖,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。Objective To determine the effect of the direction of poly （s-caprolactone） （PCL）/ collagen （COL） nanofiber orientation on the morphology, proliferation, and differentiation in mouse preosteo- blast cells （MC3T3-E1）. Methods The morphologies of the random （bad-oriented） and aligned （well- oriented） PCL,/COL nanofiber scaffolds were observed by scanning electron microscopy （SEM）. MC3T3-E1 cells were cultured on the 2 kinds of nanofiber scaffolds to study the effects of fiber orientation on cell morpholo- gy through SEM and laser confocal microscopy （LSCM）. The cell proliferation and numbers of viable cells were observed by cell counting kit-8 （ CCK-8 ）. Real-time RT-PCR was used to testify the mRNA expression of osteo- genie markers, such as collagen type I （COL I ）, alkaline phosphatase （ALP） and osteocalein （OCN） to study the sense and response of MC3T3-E1 cells to different oriented scaffolds. Results The random PCL/ COL nanofibers showed isotropic fiber alignments, the aligned nanofibers exhibited anisotropic fiber alignments. The difference of the fibers diameter was not statistically significant between the 2 scaffolds （586 ± 239 vs 625 ± 155 nm, P 〉0.05）. SEM and LSCM showed that MC3T3-E1 cells exhibited a star-like shape and irregular cytoskeletons on the random nanofibers, while these cells on the aligned nanofibers had a prolate shape and elongated cytoskeletons along the aligned fibers. CCK-8 results showed that there was no significant difference in the proliferation rates of the cells on the random and aligned nanofiber scaffolds at 1, 3, 5 and 7 d （P 〉 0. 05）. Real-time PCR demonstrated the differences in expression profiles of osteogenie markers in MC3T3-E1 cells cultured on 2 scaffolds. The expression levels of COL I on random and aligned scaffolds were not signifi-cantly different （P 〉 0.05 ）, while ALP and OCN expression levels were scaffold compared with the aligned scaffolds （ P 〈 0.05 ）. Conclusion significantly elevated

关 键 词：聚己内酯 胶原 纳米纤维 MC3T3-E1细胞 方向 

分 类 号：R318.08[医药卫生—生物医学工程] R329.28[医药卫生—基础医学]