掺锶聚磷酸钙对内皮细胞来源促血管化因子基质金属蛋白酶2表达的影响  

Effect of strontium-doped calcium polyphosphate on endothelial cells-derived angiogenic factor matrix metalloproteinase-2 expression

作　　者：任大伟[1] 刘菲[1] 余喜讯[1] 邓晓薇[1] 张小华[1] 万昌秀[1] 

机构地区：[1]四川大学高分子科学与工程学院,四川省成都市610065

出　　处：《中国组织工程研究与临床康复》2009年第38期7433-7436,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30870616)~~

摘　　要：背景:课题组研究的掺锶聚磷酸钙作为一种新型骨修复材料,具有良好的生物相容性和生物可降解性,经过前期的研究已证明有促血管化作用,但机制尚不清楚。目的:内皮细胞与掺锶聚磷酸钙支架于体外复合培养,观察细胞的增殖和促血管化因子基质金属蛋白酶2的分泌情况。设计、时间及地点:体外细胞学对比观察实验,于2008-09/2009-04在四川大学组织工程研究室完成。材料:在制备聚磷酸钙过程中掺入锶制备掺锶量为1%,2%,5%,8%,10%的掺锶聚磷酸钙骨组织工程支架材料。方法:①材料置于24孔培养板中,将浓度为3×107L-1的内皮细胞悬液300μL接种于24孔培养板上,补加200μLRPMI1640全培养液。分别于1,3,5,7d通过MTT法观察内皮细胞的增殖情况。②聚磷酸钙和8%掺锶聚磷酸钙多孔支架放于24孔板中,将浓度为1×108L-1的内皮细胞悬液300μL接种于支架材料上,补加600μLRPMI1640全培养液。培养5d后取出材料,扫描电镜观察内皮细胞的形貌。③内皮细胞与不同掺锶量的掺锶聚磷酸钙支架材料复合培养5d,至细胞汇合后,离心取上清液,采用酶联免疫吸附试验检测内皮细胞来源的基质金属蛋白酶2蛋白的分泌量。主要观察指标:观察掺锶聚磷酸钙和聚磷酸钙材料上内皮细胞的增殖及形貌,内皮细胞来源的基质金属蛋白酶2蛋白的分泌量。结果:MTT法实验结果表明,与聚磷酸钙组及其他掺锶量的掺锶聚磷酸钙比较,8%掺锶聚磷酸钙拥有更高的内皮细胞增殖度;扫描电镜表明在8%掺锶聚磷酸钙材料表面生长的内皮细胞具有更好的生长状态和生物活性;酶联免疫吸附试验法结果表明,与聚磷酸钙组相比,掺锶聚磷酸钙能上调基质金属蛋白酶2的蛋白分泌量,其中以8%掺锶聚磷酸钙最为明显(P<0.05)。结论:掺锶聚磷酸钙与内皮细胞具有良好的生物相容性,明显上调促血管化因子基质金属蛋白酶2的分泌,具有促血管BACKGROUND： Strontium-doped calcium polyphosphate （SCPP）, as a new repairing material, has been demonstrated to have favorable biocompatibility and biodegradability and some effects on promoting self-angiogenesis. However, the mechanism remains still unknown. OBJECTIVE： Endothelial cells were cultured with SCPP scaffolds in vitro, as well as the cell proliferation and angiogenic factor matrix metalloproteinase-2 （MMP-2） secretion were observed. DESIGN, TIME AND SETTING： A contrast study was performed at the Laboratory of Tissue Engineering of Sichuan University from September 2008 to April 2009. MATERIALS： A series of calcium polyphosphate （CPP） respectively containing 1%, 2%, 5%, 8%, and 10% Sr^2＋ were prepared. METHODS： ① Materials were plated on 24-well culture plate, and endothelial cell suspension （300 μL） were seeded on 24-well culture plate at the concentration of 3×10^7/L and cultured with 200 μL RPMI1640 culture media. Endothelial cell proliferation was observed using MTT method at days 1,3, 5, and 7 after culture. ~ CPP and 8% SCPP were plated on 24-well culture plate, and endothelial cell suspension （300 μL） was then incubated in 24-well culture plate at the concentration of 1 ×10^8/L and cultured with 600 μL RPMI1640 culture media. The morphology of endothelial cells was observed by scanning electron microscopy （SEM） at day 5 after culture. ③ Endothelial cells were co-cultured with SCPP of various Sr^2＋ contents for 5 days. After confluence, cells were centrifuged to obtain supernatant. Angiogenic factor MMP-2 secretion was evaluated by ELISA assay. MAIN OUTCOME MEASURES： The proliferation and morphology of endothelial cells on SCPP and CPP were observed. The amount of endothelial cells-derived MMP-2 protein secretion was detected. RESULTS： MTT method demonstrated that the proliferation of endothelial cells on the 8% SCPP scaffold showed a higher level compared to CPP, and other SCPP groups. Scanning electron microscope results suggested that endothe

关 键 词：掺锶聚磷酸钙 内皮细胞 血管化 基质金属蛋白酶2 

分 类 号：R318[医药卫生—生物医学工程]