CAJ | 학술논문

背景: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可控降解性.课题组前期研究表明其具有促进血管牛成的作用,但其机制尚不清楚.目的:将ROS17/2.8成骨细胞与掺锶聚磷酸钙支架在体外共培养,观察细胞增殖情况和促血管生成因子血管内皮生长因子的分泌情况.设计、时间及地点:对比观察实验,于2008-10/2009-06在四川大学组织工程研究室完成.材料:制备掺锶量为1%,2%,5%,8%,10%的掺锶聚磷酸钙和未掺锶聚磷酸钙骨组织工程支架材料.ROS17/2.8成骨细胞株购自于四川大学华西医院移植免疫与移植工程实验室. 方法:①制备细胞支架复合物:将材料置于24孔培养板中,每孔接种300 μL细胞悬液(细胞浓度为2×10~7L~(-1)),培养14 d,隔天换液.②分别于1,3,5,7,10,14 d行MTT法观察成骨细胞的增殖情况.③将培养7 d的细胞支架复合物,离心取上清液,用夹心ELISA法检测血管内皮生长因子的分泌情况.主要观察指标:观察掺锶聚磷酸钙和未掺锶聚磷酸钙支架上成骨细胞的增殖情况及血管内皮生长因子的分泌情况.结果:MTT法实验结果表明,与未掺锶聚磷酸钙组相比,掺锶聚磷酸钙组能促进成骨细胞的增殖,且8%掺锶聚磷酸钙效果最好;ELISA结果表明,与未掺锶聚磷酸钙组相比,掺锶聚磷酸钙组能上调血管内皮生长因子的蛋白分泌量,其中以8%掺锶聚磷酸钙促进作用最为明显(P<0.05).结论:锶能有效促进成骨细胞增殖,并能明显上调血管内皮生长因子的分泌,且以8%掺锶聚磷酸钙效果最好,在一定程度上揭示了其具有促血管化作用的机制.
배경:참송취린산개작위일충신형골수복재료,구유량호적생물상용성화가공강해성.과제조전기연구표명기구유촉진혈관우성적작용,단기궤제상불청초.목적:장ROS17/2.8성골세포여참송취린산개지가재체외공배양,관찰세포증식정황화촉혈관생성인자혈관내피생장인자적분비정황.설계、시간급지점:대비관찰실험,우2008-10/2009-06재사천대학조직공정연구실완성.재료:제비참송량위1%,2%,5%,8%,10%적참송취린산개화미참송취린산개골조직공정지가재료.ROS17/2.8성골세포주구자우사천대학화서의원이식면역여이식공정실험실. 방법:①제비세포지가복합물:장재료치우24공배양판중,매공접충300 μL세포현액(세포농도위2×10~7L~(-1)),배양14 d,격천환액.②분별우1,3,5,7,10,14 d행MTT법관찰성골세포적증식정황.③장배양7 d적세포지가복합물,리심취상청액,용협심ELISA법검측혈관내피생장인자적분비정황.주요관찰지표:관찰참송취린산개화미참송취린산개지가상성골세포적증식정황급혈관내피생장인자적분비정황.결과:MTT법실험결과표명,여미참송취린산개조상비,참송취린산개조능촉진성골세포적증식,차8%참송취린산개효과최호;ELISA결과표명,여미참송취린산개조상비,참송취린산개조능상조혈관내피생장인자적단백분비량,기중이8%참송취린산개촉진작용최위명현(P<0.05).결론:송능유효촉진성골세포증식,병능명현상조혈관내피생장인자적분비,차이8%참송취린산개효과최호,재일정정도상게시료기구유촉혈관화작용적궤제.
BACKGROUND: Strontium-doped calcium polyphosphate (SCPP) is a new type of bone repair materials with good biocompatibility and controlled degradation. The preliminary studies of our group indicate their role in promoting angiogenesis,but its mechanism is unclear.OBJECTIVE: By co-culturing osteoblasts ROS17/2.8 with SCPP in vitro to observe cell proliferation and the secretion of vascular endothelial growth factor (VEGF).DESIGN, TIME AND SETTING: A contrast study was performed at the Laboratory of Tissue Engineering of Sichuan University from October 2008 to June 2009.MATERIALS: A series of calcium polyphosphate (CPP) respectively containing 0%, 1 %, 2%, 5%, 8%, and 10% Sr~(2+) were prepared. ROS17/2.8 osteoblastic cell strain was provided by Laboratory of Transplantation Immunity and Transplantation Engineering, West China Hospital, Sichuan University.METHODS: ①Preparation of cell scaffold complexes: The materials were placed in 24-well plates, then 300 μL cell suspension with a concentration of 2×10~7 cells/Lwas inoculated into each hole. These complexes were cultured for 14 days and the liquid was changed every two days. ②These complexes were measured by MTT assay to observe the proliferation of osteoblasts on the 1~(st), 3~(rd), 5~(th), 7~(th), 10~(th) and 14~(th) days, respectively. ③ The centrifugal supernatant of the complex cultured for seven days was measured by ELISA assay to check the secretion of VEGF.MAIN OUTCOME MEASURES: The proliferation of osteoblastic cells on SCPP and CPP was observed. The amount of VEGF protein secreting from osteoblastic cells was detected.RESULTS: The results of MTT showed that, compared with the CPP group, SCPP groups could promote the proliferation of osteoblasts, and 8% SCPP group was the best; ELISA results showed that, compared with the CPP group, SCPP groups could increase the amount of VEGF protein secretion, of which the promoting role of 8% SCPP was the most obvious (P < 0.05).CONCLUSION: When cultured with osteoblasts, SCPP can promote cell proliferation, and can significantly increase the secretion of VEGF; moreover, 8% SCPP is the best, which reveals a certain mechanism of its promoting angiogenesis.