玻璃体腔注射人PRP联合巩膜外冷冻建立PVR动物模型的研究

Human PRP intravitreal injection combining cryotherapy for establishing animal models with proliferative vitreoretinopathy

目的探讨玻璃体腔内注射人富含血小板血浆(platelet-richplasma,PRP)联合巩膜外冷冻建立增生性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR)动物模型的效果。方法选取青紫蓝兔52只,随机分成实验组及对照组,每组各26只,每只兔随机选取一眼作为实验眼。取健康体检者静脉血制备PRP。暴露实验眼颞上方巩膜,于角膜缘后3.5mm行巩膜切开,剪除玻璃体约0.5mL,实验组玻璃体腔注射PRP0.4mL,同时联合巩膜外冷冻;对照组单纯行玻璃体腔注射PRP0.4mL。分别于造模后1d、3d、7d、14d、28d行裂隙灯、间接眼底镜及B超观察玻璃体增殖及视网膜脱离情况,并进行PVR分级。分别于造模后7d、14d及28d每组随机选取2只兔摘除眼球行组织病理学检查。结果实验组和对照组造模后28d视网膜脱离的发生率分别为95%和65%,2组成模率比较,差异有统计学意义(χ2=3.906,P=0.048)。临床和B超观察:实验组造模后1d出现明显玻璃体混浊、3d出现玻璃体增殖条索、7d出现局限性视网膜脱离、14d广泛视网膜脱离、28d全视网膜脱离;对照组造模后1d玻璃体轻度混浊、3d出现明显玻璃体混浊、7d可见少量玻璃体增殖条索、14d出现局限性视网膜脱离、28d有广泛视网膜脱离发生。病理组织学观察:实验组造模后7d视网膜表面炎性细胞聚集、成纤维细胞增生,14d视网膜前增殖条索、视网膜皱褶,28d视网膜固定皱褶形成、呈花节样外观;对照组造模后7d视网膜表面见炎性细胞及渗出,14d视网膜表面增殖膜,28d视网膜表面增殖条索出现。结论玻璃体内注射人PRP联合巩膜外冷冻建立PVR动物模型简便有效、成模率高,可作为PVR相关基础研究的建模方法。

Objective To investigate the efficiency of animal model for proliferative vitreoretinopathy（PVR） by using human platelet-rich plasma（PRP） injection combined with cryotherapy.Methods Fifty two pigmented rabbits were divided into experimental group and control group with 26 rabbits in each group.One eye of each rabbit was chosen for experimental eye.Venous blood was taken from healthy examination rabbit for PRP.Sclera at upper temple for experimental eye was exposed,and cut open at 3.5 mm behind limbus cornea.About 0.5 mL vetreous body was taken for experimental eye.Rabbits in experimental group were taken intravitreous injection of 0.4 mL PRP,and cryotherapy out of sclera;Rabbits in control group were just taken intravitreous injection of 0.4 mL PRP.All rabbits were taken slit lamp examination,indirect ophthalmoscopy,B scan for observing vitreous proliferation and retinal detachment,and PVR ranking at 1 day,3 days,7 days,14 days and 28 days after modeling.Two rabbits were randomly chosen in each group,and eyeballs were taken out for histopathological examination.Results Ninety five percent of rabbit eyes in experimental group were observed with retinal detachment at 28 day after operation,and only 65% in control group.The difference was statistically significant（χ2=3.906,P=0.048）.Clinical observation and B-scan ultrasound showed vitreous cloudy at 1 day,epiretinal tissue at 3 days,localized retinal detachment at 7 days,widely retinal detachment at 14 days,and total retinal detachment at 28 days in experimental group,and performed mild vitreous cloudy at 1 day,vitreous cloudy at 3 days,epiretinal tissue at 7 days,localized retinal detachment at 14 days,and widely retinal detachment at 28 days in control group.Histopathological examination revealed epiretinal inflammatory cell and fibroblast concentrating at 7 days,epiretinal tissue and retinal folds at 14 days,flower appearance fixed retinal folds at 28 days in experimental group,and presented epiretinal inflammatory cells and exudation at 7 days,epiretinal proliferative member at 14 days,and epiretinal tissues at 28 days in control group.Conclusions The method of human PRP intravitreal injection combining cryotherapy is efficient and convenient to build the animal model with PVR,which could produce reliable experimental animal models for basic scientific research.