兔眼角膜去端肽猪皮Ⅰ型胶原膜移植的研究

目的 探讨去端肽猪皮I型胶原行板层角膜移植术后的角膜愈合情况,及其作为角膜移植替代物的可行性.方法 实验研究.选用中国白兔100只,其中80只兔根据角膜植片来源不同随机分为胶原材料实验组(A组)及同种异体对照组(B组),两组均行单眼前板层角膜移植术.各组根据不同处死时间,随机分为3 d、14 d、1个月、3个月和6个月亚组,每组8只.余20只兔(40只眼)作为B组角膜移植供体来源.术后行大体、裂隙灯观察角膜愈合情况并记录角膜透明度与角膜新生血管(CNV)评分情况,HE染色组织学观察以及上皮细胞标志蛋白K3的免疫组化检测.结果 采用Friedman秩和检验行组内比较,两两之间比较行Wilcoxon符号秩和检验.结果 两组术后角膜透明度逐渐升高(A组:x2=31.250,P=0.000;B组:x2=32.566,P=0.000),1、3及6个月两组角膜透明度评分(A组:2.50、1.94、1.94;B组:2.88、2.25、1.63)比较差异无统计学意义(Z=-1.414,0.000,-0.743;P=0.157,1.000,0.458).术后7 d两组均有CNV长入,A组CNV评分逐渐增高,14 d最高,术后1个月变化不明显,后逐渐减轻(x2=20.727,P=0.001);B组CNV评分逐渐增高,14 d达高峰,后逐渐减轻(x2=25.562,P=0.000);术后6个月两组CNV比较差异有统计学意义(Z=-2.070,P=0.038).组织学观察A组术后1个月胶原材料几乎完全降解,胶原排列趋于规则,术后6个月胶原排列基本规则,但两组局部均有不同程度胶原紊乱区.结论 去端肽猪皮I型胶原可促进角膜上皮细胞与基质细胞重建,但完全取代同种异体角膜植片还需进一步研究.

Abstract：

Objective To investigate corneal wound healing in rabbit after lamellar keratoplasty with porcine type I atelocollagen. Methods One hundred Chinese white rabbits, 80 rabbits were randomly divided into collagen materials group (group A) and allograft group (group B) , and group A and B were operated for the lamellar keratoplasty in one eye. Rest 20 rabbits'eyes (40 eyes) were used as the source of allograft for group B. After operation,the rabbits'eyes were observed by naked eyes and slit lamp, and recorded the score of the transparency and neovascularization of the cornea. The rabbits were sacrificed at 3rd, 14th day, the 1st, 3rd, and 6th month (each group eight eyes). The corneas were observed by histopathology, and the epithelial cells marker protein K3 were detected by immunohistochemistry. The results were analyzed by Rank sum test. Results The transparency of the cornea was increased gradually in A and B groups, and reached the peak on the 6th month(group A; x2 =31.250, P=0.000; group B: x2= 32.566, P = 0.000). The difference had not statistics significance in the comparison of the transparency from the 1st to 6th month(Z= -1.414, 0.000, -0.743;P=0. 157, 1.000, 0.458). The two groups had corneal neovascularization after 7 days, the intensity of corneal neovascularization in A group increased gradually, reached the peak on day 14, then decreased gradually (x2 = 20.727, P = 0.001); the intensity of corneal neovascularization in B group increased gradually, reached the peak on day 14, and then decreased gradually (x2 =25. 562, P =0. 000). The difference had statistics significance in the comparison of the neovascularzation of the cornea on the 6th month between A and B group(Z=-2.070,P=0.038).Histopathology showed the collagen material nearly disappeared after 1 month of surgery and arrangement of collagen tends to be regular.On the 6th month,arrayed collagen was regular in general,but disordered partly.Conclusions Porcine type I atelocollagen can promote corneal cells regeneration.But it needs more study to be a cornea substitute that can completely replace the corneal allogaft.

Transplantation of porcine type I atelocollagen into rabbit cornea

Objective To investigate corneal wound healing in rabbit after lamellar keratoplasty with porcine type I atelocollagen. Methods One hundred Chinese white rabbits, 80 rabbits were randomly divided into collagen materials group (group A) and allograft group (group B) , and group A and B were operated for the lamellar keratoplasty in one eye. Rest 20 rabbits'eyes (40 eyes) were used as the source of allograft for group B. After operation,the rabbits'eyes were observed by naked eyes and slit lamp, and recorded the score of the transparency and neovascularization of the cornea. The rabbits were sacrificed at 3rd, 14th day, the 1st, 3rd, and 6th month (each group eight eyes). The corneas were observed by histopathology, and the epithelial cells marker protein K3 were detected by immunohistochemistry. The results were analyzed by Rank sum test. Results The transparency of the cornea was increased gradually in A and B groups, and reached the peak on the 6th month(group A; x2 =31.250, P=0.000; group B: x2= 32.566, P = 0.000). The difference had not statistics significance in the comparison of the transparency from the 1st to 6th month(Z= -1.414, 0.000, -0.743;P=0. 157, 1.000, 0.458). The two groups had corneal neovascularization after 7 days, the intensity of corneal neovascularization in A group increased gradually, reached the peak on day 14, then decreased gradually (x2 = 20.727, P = 0.001); the intensity of corneal neovascularization in B group increased gradually, reached the peak on day 14, and then decreased gradually (x2 =25. 562, P =0. 000). The difference had statistics significance in the comparison of the neovascularzation of the cornea on the 6th month between A and B group(Z=-2.070,P=0.038).Histopathology showed the collagen material nearly disappeared after 1 month of surgery and arrangement of collagen tends to be regular.On the 6th month,arrayed collagen was regular in general,but disordered partly.Conclusions Porcine type I atelocollagen can promote corneal cells regeneration.But it needs more study to be a cornea substitute that can completely replace the corneal allogaft.