摘要
背景:目前,骨组织工程的研究主要集中在成骨细胞特性研究和骨修复材料研制两大领域,而对细胞与细胞外材料的相互作用,尤其是调节和促进两者间相互作用的相关研究尚未给予足够的重视。目的:在低聚乳酸接枝纳米羟基磷灰石/聚丙交酯-乙交酯(Hydroxyapatite nanocrystals surface-grafted with L-lactic acid oligomer/Poly(lactide-co-glycolide),op-HA/PLGA)复合材料上接种成骨细胞,观察脉冲电场刺激对成骨细胞增殖和成骨相关基因表达的影响。设计、时间及地点:对比观察实验,于2008-05/2009-01在中国科学院长春应用化学研究所高分子物理与化学国家重点实验室完成。材料:op-HA和PLGA由中科院长春应用化学研究所制备。方法:实验分为PLGA无电刺激组、op-HA/PLGA无电刺激组、PLGA电刺激组和op-HA/PLGA电刺激组。将op-HA/PLGA和PLGA氯仿溶液分别于盖玻片表面铺成薄膜,并接种兔成骨细胞,体外培养7d。电刺激组刺激电压为2V,频率为1Hz,占空比为20%,每天刺激1h。主要观察指标:流式细胞仪检测细胞周期,Real-time PCR检测骨形态发生蛋白2、Ⅰ型胶原蛋白、骨连接蛋白的表达。结果:脉冲电刺激组op-HA/PLGA上生长的成骨细胞S期细胞百分比明显高于其他组(P<0.05)。op-HA/PLGA与PLGA比较能上调骨形态发生蛋白2、Ⅰ型胶原蛋白、骨连接蛋白3种基因表达量,并且施加脉冲电刺激组骨形态发生蛋白2、Ⅰ型胶原蛋白、骨连接蛋白3种基因表达量高于未加电刺激组。结论:op-HA/PLGA复合材料是一种很好的骨修复材料,脉冲电刺激能促进成骨细胞在材料上的增殖能力,提高成骨活性相关基因的表达水平。
BACKGROUND： Recently, bone tissue engineering is focus on researching characteristics of osteoblasts and bone repairing materials; however, there is less attention to interaction between cells and extracellular materials, in particular, regulating and promoting the interaction. OBJECTIVE： To explore the effect of pulsed electrical stimulation on esteoblast proliferation and osteeblast-related gene expression cultured on Hydroxyapatite nanocrystals surface-grafted with L-lactic acid oligomer/poly （lactide-co-glycolide） （op-HA/PLGA） composite material. DESIGN, TIME AND SETTING： A contrast study was provided by State Key Laboratory of Polymer Physics and Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences from May 2008 to January 2009. MATERIALS： op-HA and PLGA were provided by Changchun Institute of Applied Chemistry, Chinese Academy of Sciences. METHODS： The osteoblasts were assigned into PLGA group, op-HA/PLGA group, PLGA electrical stimulation group, and op-HNPLGA electrical stimulation group, op-HA/PLGA and PLGA chloroform solution were covered on the surface of coverslip of which rabbit osteoblasts were inoculated and cultured in vitro for 7 days. Electrical stimulation was performed as follows： voltage 2V, frequency 1 Hz, duty cycle 20%, and 1 hour each day. MAIN OUTCOME MEASURES： Cell cycle was detected using flow cytometry; while, expressions of bone morphogenetic protein-2 （BMP2）, collagen I, and osteonectin were measured using Real-time PCR. RESULTS： Percentage of osteoblasts in the op-HA/PLGA electrical stimulation group was significantly higher than other groups in the S phase （P 〈 0.05）. op-HA/PLGA could up-regulate the expressions of BMP-2, collagen I, and osteonectin compared to PLGA. In addition, electrical stimulation could also up-regulate the expressions of BMP-2, collagen I, and osteonectin. CONCLUSION： Op-HA/PLGA composite is a good bone repair material. Pulse electrical stimulation can promote osteoblast proliferation and increase related gene expression of osteogenic capacity.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第16期3065-3069,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然科学基金资助项目(50673090)
国家“八六三”计划(2007AA03Z320)~~
