摘要
背景:如何使生长因子安全的在体内稳定持久的表达,并获得长期的促血管生成效应是亟待解决的问题,这就需要一个良好的药物缓释系统,即可控释的生物模拟系统。目的:探讨不同配伍的生物膜对鸡胚绒毛尿囊膜血管生成的影响,寻找最佳的组合方式。设计、时间及地点:生物材料学体外观察,于2008-06/2009-05在江苏省中医院中心实验室完成。材料:通过交联剂将肝素、中药与Ⅰ型胶原发生共价结合,然后rhVEGF165、碱性成纤维细胞生长因子与肝素发生生理性结合,将血管生成素1制成微球均匀植入生物膜内,即为实验所需生物膜。方法:将制备的生物膜置于正对观察窗的鸡胚绒毛尿囊膜表面的血管最少处,然后贴在鸡胚绒毛尿囊膜表面上,封口膜封闭假气室,继续孵育。实验组在给予生物膜基础上,依次加入0,10,50,100ng碱性成纤维细胞生长因子,从中筛选出最显著促进鸡胚绒毛尿囊膜血管新生的组合作为生物膜1;再依次加入100μL丹参注射液、黄芪注射液、三七总甙注射液、川芎嗪注射液,同法筛选出生物膜2;再依次加入10,50ng血管生成素Ⅰ,同法筛选出生物膜3作为实验最终结果。同时设立空白对照组,鸡胚绒毛尿囊膜表面未贴生物膜,仅加入PBS。加药7d后取绒毛尿囊膜,拍摄给药部位,采用Image Pro Plus 5.0.2进行数据收集。主要观察指标:不同配伍的生物膜对血管新生面积的影响。结果:①与空白对照组比较,生物膜+0,10,50,100ng碱性成纤维细胞生长因子各组新生血管面积均明显增加,其中生物膜+100ng碱性成纤维细胞生长因子组增加幅度最高(P<0.01),为此将其选为生物膜1。②与生物膜1组比较,生物膜1+三七总甙注射液组、生物膜1+川芎嗪注射液组无明显变化,生物膜1+丹参注射液组、生物膜1+黄芪注射液组新生血管面积分别增加25.48%,63.21%(P均<0.01),因此将生物膜1+黄芪注射液组作为生物膜2的最佳组成。③与生物膜2组比较,生物膜2+10ng血管生成素Ⅰ组新生血管面积无明显差异;生物膜2+50ng血管生成素Ⅰ组新生血管面积增加21.49%(P<0.05),因此将其作为生物膜3。结论:在实验制备的生物膜基础上,加入100ng碱性成纤维细胞生长因子、100μL黄芪注射液组、50ng血管生成素Ⅰ的组合,具有明显促进鸡胚绒毛尿囊膜血管生成的作用。
BACKGROUND： A well drug sustained release system, i.e., controllable biosimulated system, is needed to investigate how to stably and permanently express growth factors in vivo and promote angiogenesis. OBJECTIVE： To investigate the effect of different combinations of the biomembrane on the chick embryo chorioallantoic membrane angiogenesis, and to find the best combination. DESIGN, TIME AND SETTING： An in vitro biomaterials study was performed at Central Laboratory of Jiangsu Hospital of Traditional Chinese Medicine from June 2008 to May 2009. MATERIALS： Heparin, Chinese herb, and type I collagen were covalently bonded through crosslinking agent. Additionally, rhVEGF165 and basic fibroblast growth factor were bonded with heparin. Angiogenin I was made into microsphere which was implanted into biomembrane. METHODS： The different combinations of the biofilm were put on the surface of chick embryo chorioallantoic membrane （CAM）. In the experimental group, basic fibroblast growth factors （0, 10, 50, and 100 ng） were added respectively, and the biomembrane which remarkably promote angiogenesis was considered as membrane 1. In addition, Danshen injection （100 μL）, Huangqi injection, Panaxnotoginseng saponins injection, and Ligustrazine injection were used to screen biomembrane 2, while angiogenin I （10 and 50 ng） was used to screen biomembrane 3. Moreover, blank control group was set, i.e., biomembrane was not attached to surface of chick embryo chorioallantoic membrane but PBS was performed on samples. Seven days later, chorioallantoic membrane was photographed and Image Pro Plus5.0.2 software was used for data collection. MAIN OUTCOME MEASURES： Effect of different combinations of the biomembrane on angiogenesis area. RESULTS： ① As compared with blank control group, the new vascular area of the biomembrane 1, the biomembrane 1 ＋ 10 ng, 50 ng, and 100 ng basic fibroblast growth factors group was increased, in particular, the increase was the highest in the biomembrane 1 ＋ 100 ng basic fibroblast growth factors group （P 〈 0.01）. ② As compared with biomembrane 1 group, the new vascular area of biomembrane 1 ＋ Panaxnotoginseng saponins injection and biomembrane 1 ＋ Ligustrazine injection groups was not changed; however, the vascular area of biomembrane 1 ＋ Danshen injection and biomembrane 1 ＋ Huangqi injection groups was increased 25.48% and 63.21%, respectively （P 〈 0.01）. Therefore, biomembrane 1 ＋ Huangqi injection was recorded as the best combination of biomembrane 2. ③ As compared with biomembrane 2 group, the vascular area of biomembrane 2 ＋ 10 ng angiogenin I group was not changed, but the area of biomembrane 2 ＋ 50 ng angiogenin I group was increased 21.49% （P 〈 0.05）, which was considered as the best combination of biomembrane 3. CONCLUSION： The biomembrane combined with 100 ng basic fibroblast growth factors, 100 μL Huangqi injection, and 50 ng ngiogenin I can remarkably promote the chick embryo chorioallantoic membrane angiogenesis.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第42期8216-8220,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
江苏省人事厅"六大人才高峰"第三批资助项目(06-B-0222006)~~
