国产猪源纤维蛋白黏合剂玻璃体腔注射后对兔眼视网膜组织的生物安全性研究

Safety of porcine fibrin sealant kit on ocular tissue following intravitreal injection

背景猪源纤维蛋白黏合剂已广泛应用于临床手术中，能够起到止血和闭合伤口的作用。目前眼科学者认为猪源纤维蛋白黏合剂在玻璃体切割术中可替代长效气体或硅油作为填充物，但国产猪源纤维蛋白黏合剂是否对视网膜有毒性作用仍在研究中。目的研究国产猪源纤维蛋白黏合剂在眼内应用后对视网膜组织的生物安全性。方法15只青紫兰兔任选一眼作为实验眼，对侧眼作为对照眼。兔眼行玻璃体切割术，实验眼术毕于玻璃体腔内注射猪源纤维蛋白胶0．5ml，对照眼注射等量平衡盐溶液。术后l、3、7、15、30d用裂隙灯及直接／间接检眼镜检查眼前后节的炎症反应，术后1、3、7d用SchiStz眼压计测量眼压，分别于术前和术后30d进行视网膜电图（ERG）检查，评估视网膜的功能变化，术后30d行眼科B型超声检查。玻璃体腔内注射后第30天摘除眼球并制备视网膜切片，光学显微镜下检查视网膜的组织形态学变化，透射电子显微镜下观察视网膜的超微结构变化。结果实验组15只兔眼中出现并发症者7只眼，其中晶状体损伤后发生白内障者3只眼，术后增生性玻璃体病变和视网膜脱离者5只眼，眼内炎者1只眼。对照组15只兔眼晶状体损伤后发生白内障者2只眼，术中发生视网膜损伤者2只眼。眼内注射后30d实验组未出现并发症的8只眼及对照组的11只眼均未见明显的眼内炎症反应，术后眼压均正常，2个组间及手术前后各时间点间的眼压变化差异均无统计学意义（F分组=0．008，P=0，929；F时间=3．600，P=0．075）。实验组及对照组间兔眼注药前后ERGa波、b波振幅的总体差异均无统计学意义（a波：F分组=0．728，P=0．405；b波：F分组=0．222，P=0．644）；各组兔眼手术前后ERGa波、b波振幅的总体差异均无统计学意义（a波：F时间=0．516，P=0．482；b波：F时间=0．057，P=0．814）。眼内注射后30d，实验眼和对照眼视网膜组织层次清晰，无水肿及阳性细胞浸润，各层视网膜的细胞及视网膜光感受器、视网膜色素上皮的色素细胞形态均正常，亚细胞器结构清晰，细胞膜和核膜完整，线粒体嵴排列整齐。结论国产猪源纤维蛋白黏合剂玻璃体腔注射后对兔眼视网膜无毒性作用。

Background Porcine fibrin sealant kit has been widely used in surgery for clotting and wounds- sealing. It is thought to be a substitution for inert gas and silicone oil in vitrectomy. But whether it produce toxic effect on retina or not after intravitreal injection is still studying. Objective This study aimed to investigate the retinal toxicity of porcine fibrin sealant kit following intraocular application. Methods ~itrectomy was performed on bilateral eyes of 15 pigmented rabbits. Porcine fibrin sealant kit of O. 5 ml was intravitreally injected in the lateral eyes of the rabbits as the experimental group, and equal volume of BSS was used at the same way in contra]ateral eye as control group. The inflammatory response of eye was observed under the slit lamp and ophthalmoscopy in 1 day,3,7, 15,30 days after injection. Schicitz tonometer was used to measure intraocular pressure （IOP） in 1 day, 3,7 days, and retinal function was evaluated by electroretinogram （ERG） before operation and 30 days after operation. B-ultrasonic examination was carried out 30 days after surgery. Animals were sacrificed and eyeballs were obtained for retinal histopathological and ultrastructural examination in 30 days after operation. Results Complications appeared in 7 eyes of the experimental group,including cataract in 3 eyes, proliferation vitreoretinopathy and retinal detachment in 5 eyes,endophthalmitis in 1 eye, however, the cataract in 2 eyes and retina injury in 2 eyes were found in 15 control eyes. On the 30 days after injection,no inflammation was found in the 8 eyes without complication in the experimental group and 11 eyes in the control group. The IOP change was insignificant in different groups and various time points （ Fgroup = 0. 008, P = 0. 929 ; Ftime = 3. 600 P = 0. 075 ） ~ No significant difference was found in a-wave amplitude between groups and various time points（ Fgroup=0. 728 ,P= 0. 405 ;Ftime= 0. 516 ,P = 0. 482 ） , and so was the b-wave amplitude （ Fgroup = 0. 222, P = 0. 644 ; Ftime = 0. 057, P = 0.814 ）. On the 30th day after operation, arrangement of the retina was regular and no tissue edema and inflammatory cell infiltration were seen in both groups under the light microscope. Transmission electron microscope revealed that the ultrastructures of retinal cells, photoreceptor and retinal pigment cell were normal with the clear subcellular organelle, intact cellular membrane structure and mitochondria in both groups. Conclusions Domestic porcine fibrin sealant kit does not produce toxic effect on retina after intravitreal injection in rabbit.