摘要目的:评价离子导入角膜胶原交联法治疗兔眼棘阿米巴性角膜炎的效果.方法:实验研究.采用随机数字表法将40只(80眼)实验兔平均分为模型组(A组)、药物组(B组)、交联组(C组)、药物联合交联组(D组),每组各10只.均选取右眼为实验眼,左眼为对照眼.A组不治疗,B组给予药物治疗,C组给予交联治疗,D组给予药物联合交联治疗.治疗90 d后行裂隙灯显微镜、共聚焦显微镜和角膜组织病理切片检查.共聚焦显微镜选取病变区深度分别为100、200、300μm的图片,对图像中棘阿米巴包囊的密度使用Image J图像处理软件进行测定.所得实验数据采用Kruskal-Wallis H检验和2×2的析因设计方差分析进行分析.结果:治疗90 d后,裂隙灯显微镜示各组角膜病变不同分级的眼数不同,差异具有统计学意义(χ2=19.738,P<0.001),药物组与模型组、交联组与模型组差异均有统计学意义(P<0.05),药物组与交联组差异无统计学意义(P=0.189).共聚焦显微镜显示药物组、交联组、联合组棘阿米巴原虫密度值均低于模型组,差异具有统计学意义(均P<0.05),药物组与交联组在治疗上存在交互作用,这种交互作用表现为协同作用.组织病理切片HE染色显示药物组、交联组、联合组棘阿米巴原虫密度值均低于模型组,差异具有统计学意义(均P<0.05),药物组与交联组在治疗上存在交互作用,这种交互作用表现为协同作用.结论:离子导入角膜胶原交联法治疗兔眼棘阿米巴性角膜炎有效,且药物联合交联的效果明显优于单纯药物疗法和单纯交联疗法.

abstractsObjective: To evaluate the efficacy of drug treatment and iontophoretic collagen crosslinking alone and in combination in the treatment for acanthamoeba keratitis in rabbits. Methods: Rabbit corneas were scraped with circular blades and 0.1 ml acanthamoeba suspension was injected into the central corneal stroma. The injection caused a~3.0-mm diameter white edematous area. A soft corneal contact lens was then applied. Forty-eight hours after acanthamoeba were injected into the corneal stroma, the rabbits were randomly divided into four groups: Group A, untreated (n=10). Group B, treated 7 times/day with eyedrops containing 0.02% chlorhexidine combined with 2% metronidazole (n=10). Group C, stromal crosslinking with 0.1% riboflavin penetration by 5 min of iontophoresis, followed with 370 nm ultraviolet irradiation (10 mW/cm2, 9-mm aperture diameter, 9 min, total energy 5.4 J) (n=10). Group D, drug+crosslinked treatment (n=10). The right eye was selected as the experimental eye, and the left eye was the control eye. After treatment for 90 days, slit-lamp microscopy, confocal microscopy, and corneal histopathology were performed. Confocal microscopy was used to select the images with lesion depths of 100, 200, and 300 μm. The density of the acanthamoeba in each image was measured by Image J software. The experimental data were analyzed by Kruskal-Wallis H tests and 2×2 analysis of variance analysis. Results: After treatment for 90 days, the number of corneal lesions was significantly different in four groups (χ2=19.738, P<0.001). The classification of corneal lesions in the Groups B and C was significantly lower than that in the Group A, and the difference was statistically significant (P<0.05). The large reduction in the number of corneal lesions in Group D showed that the drug treatment and crosslinking acted synergistically to control the infection. Confocal microscopy measurements of the acanthamoeba density were significantly lower in the Groups B, C and D compared with the Group A (P<0.05). The combination of drug and crosslinking treatment synergistically lowered the density of the parasites. Histopathological sections stained by hematoxylin and eosin confirmed the reduced density of acanthamoeba in the three treated groups compared to the Group A (P<0.05). The combination of drug and crosslinking treatment synergistically lowered the density of the parasites. Conclusions: Iontophoretic corneal collagen crosslinking of acanthamoeba keratitis in rabbit eyes is effective. The combination of drug therapy and crosslinking is superior to local drug therapy alone or simple crosslinking alone.