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,促进角膜基质的改建,抑制CNV生成,进一步改进后有望成为新型的板层角膜移植支架材料。
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 collagen was regular in general, but disordered partly. Immunohistochemisty showed the epithelial cells marker protein K3 was positive at postoperative 5 months in all three groups. The expression of VEGF at postoperative 5 months was lower than that at postoperative 15 days,and the expression of VEGF in group Ⅲ was less than that of group Ⅰ and group Ⅱ. Conclusion Collagen-Glyco Semi-Inter penetrating Ploymer Network （ Coll/Glyco IPN） can support the growth of corneal epithelium, promote the reconstruct of corneal stroma, and inhibit the growth of CNV. But it needs more study to be a new corneal substitute of lamellar keratoplasty.
出处
《眼科新进展》
CAS
北大核心
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)~~
关键词
角膜移植
兔
伤口愈合
支架材料
corneal transplantation
rabbit
wound healing
support material
