负载可递送内皮抑制素基因的胶原基复合角膜替代物在兔角膜移植中的研究  

Application of Coll/Glyco IPN loading endostatin in rabbit corneal transplantation

作　　者：李雪[1,2] 王海燕[1,2] 郝永娜[1,2] 赵少贞[1,2] 刘文广[3] 李亚楠[1,2] 边领斋[1,2] 粘红[1,2] 魏瑞华[1,2] 

机构地区：[1]天津医科大学眼科医院 [2]天津医科大学眼科研究所,天津市300384 [3]天津大学材料科学与工程学院,天津市300072

出　　处：《眼科新进展》2014年第2期108-113,共6页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:50973082;81100646);天津市自然科学基金资助(编号:10JCZDJC17400);天津市科委自然科学基金(编号:11JCYBJC26000)~~

摘　　要：目的观察负载可递送内皮抑制素基因的胶原基复合角膜替代物,在行板层角膜移植术后角膜的愈合情况,评价其作为角膜移植替代物的可行性。方法选用健康成年大耳白兔105只,随机分为Ⅰ组、Ⅱ组、Ⅲ组,每组各30只,Ⅰ组、Ⅱ组分别行板层植入胶原基复合角膜替代物、负载可递送内皮抑制素基因的胶原基复合角膜替代物;Ⅲ组为同种异体对照组。各组根据不同处死时间随机分为3 d、7 d、15 d、1个月、3个月、6个月组,每组各5只。其余15只(30眼)作为Ⅲ组角膜移植供体来源。术后行裂隙灯显微镜观察角膜透明度与角膜新生血管(corneal neovascularization,CNV)评分情况,行HE染色以及上皮细胞标志蛋白K3和血管内皮生长因子(vascular endothelial growth factor,VEGF)的免疫组织化学检测。采用Friedman秩和检验行组间比较,组内两两比较采用Wilcoxon符号秩和检验。结果Ⅰ组、Ⅱ组、Ⅲ组术后角膜透明度逐渐升高(P1=0.000、P2=0.000、P3=0.006)。术后3 d、7 d、3个月,Ⅰ组和Ⅲ组角膜透明度评分差异均有统计学意义(P=0.000、0.000、0.022);术后7 d,Ⅰ组和Ⅱ组角膜透明度评分差异有统计学意义(P=0.037)。术后三组角膜CNV评分随时间延长逐渐升高,至术后15 d达最高,后逐渐降低(P1=0.001、P2=0.001、P3=0.000)。术后3 d、7 d、15 d,Ⅰ组和Ⅲ组CNV评分差异有统计学意义(P=0.041、0.000、0.024);术后7 d,Ⅰ组和Ⅱ组CNV评分差异有统计学意义(P=0.011)。组织学观察结果显示:Ⅰ组、Ⅱ组术后1个月移植材料几乎完全降解,角膜基质胶原排列趋于规则,Ⅲ组胶原纤维排列局部欠规则;术后6个月三组胶原纤维排列基本规则。免疫组织化学检测结果显示:术后6个月,三组角膜上皮细胞特异性标志蛋白K3阳性表达。术后6个月VEGF表达较术后15 d减少,Ⅲ组较Ⅰ组、Ⅱ组表达减少。结论胶原基复合角膜替代物能支持角膜上皮的生长,促进角膜�Objective To investigate corneal wound healing in rabbit after lamellar keratoplasty with Collagen-Glyco Semi-Inter penetrating Ploymer Network （ ColV Glyco IPN） loading endostatin delivered, and evaluate the feasibility of it as a corneal graft. Methods One hundred and five Chinese white rabbits, ninety rabbits were randomly divided into group Ⅰ （transplanted with Coll/Glyco IPN）, groupⅡ （transplanted with Coll/Glyco IPN loading endostatin delivered） and group Ⅲ（ transplanted with al- lograft） ,and the rest 15 rabbits＇ eyes（30 eyes）were used as the source of allograft for groupⅢ. The corneal transparency and corneal neovascularization （CNV）were observed by slit lamp at postoperative 3 days ,7 days, 15 days, 1 month,3 months ,6 months （ each group five eyes）. The corneas were stained by HE, and the epithelial cells marker protein K3 and vascular endothelial growth factor （VEGF）were detected by immunohis- tochemisty. The results were analyzed by Rank sum test. Results The corneal trans- parency was increased gradually in all three groups （ P = 0. 000,0. 000,0. 006 ）. There were statistical differences in corneal transparency at postoperative 3 days,7 days and 3 months between group Ⅰ and group Ⅲ（P = 0.000,0. 000,0.022） ; There was statistical difference between group Ⅰand group Ⅱ at postoperative 7 days（P =0. 037）. The score of CNV was increased gradually in all groups, and reached the peak at postoperative 15 days ,then decreased gradually （P = 0. 001,0. 001,0. 000 ）. There were statistical differ- ences in scores of CNV at postoperative 3 days,7 days, 15 days between group Ⅰ and group Ⅲ（P = 0. 041,0. 000,0. 024） ;There was statistical difference between group Ⅰ and group Ⅱ at postoperative 7 days （P =0.011 ）. The result of HE staining showed the graft material of group Ⅰ and group Ⅱ almost all disappeared at postoperative 1 month, and the arrangement of collagen tended to be regular; At postoperative 5 months, ar- rayed collage

关 键 词：角膜移植 兔 伤口愈合 支架材料 

分 类 号：R735.2[医药卫生—肿瘤]