Rudraprasad Dhanwini, Gandhi Jaishree, Naik Poonam, N Naik Milind, Naidu Chenchu, Kumar Mishra Dilip, Joseph Joveeta
Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India.
Manipal Academy of Higher Education, Manipal, Karnataka, India.
J Ophthalmic Vis Res. 2023 Jul 28;18(3):272-282. doi: 10.18502/jovr.v18i3.13775. eCollection 2023 Jul-Sep.
Animal models are necessary in understanding the pathogenesis of endophthalmitis and are also necessary to assist the development of new therapeutics for this sight-threatening ocular inflammation. Hamilton syringes are usually preferred to inject pathogens when performing experiments on test subjects, however, this method has technical and financial disadvantages. In this study, we report the findings and assess the related benefits of applying a novel low-cost intravitreal injection technique to initiate endophthalmitis in a mouse model while using the Eppendorf tip and a 26G needle.
The 18-hr culture of clinical isolates of bacteria ( and ) and fungus ( and ) were resuspended to a final concentration of 10,000 colony forming units (CFU)/1 µL which were separately injected intravitreally into C57BL/6 mice (6-8 weeks) using a 0.1-2.5µL pipette attached to the modified Eppendorf tip with a 26G needle. The contralateral eye served as vehicle/uninjected control. Disease progression was determined by assessing the corneal haze, opacity, bacterial burden, and retinal histology of the eyes used in the model. Following euthanization, bacteria-infected mice were enucleated after 24 hr of the initial injection, and fungus-infected mice after 72 hr.
Of the 50 mice injected, the modified technique was successful in 48 mice. Two mice were excluded due to cataract formed by accidental injury to the lens. The experimental endophthalmitis mice model successfully mimicked the natural clinical course. Clinical assessment and histopathology confirmed the influx of inflammatory cells into the posterior segment of the eye along with dissolution of retinal architecture.
Our novel method of injection using a modified Eppendorf tip and 26G needle yielded a cost-effective mouse model of clinical endophthalmitis, resulting in reproducible infection for understanding various aspects of its pathobiology.
动物模型对于理解眼内炎的发病机制是必要的，对于辅助开发针对这种威胁视力的眼部炎症的新疗法也是必要的。在对实验对象进行实验时，通常首选汉密尔顿注射器来注射病原体，然而，这种方法存在技术和经济方面的缺点。在本研究中，我们报告了使用新型低成本玻璃体内注射技术在小鼠模型中引发眼内炎的研究结果，并评估了相关益处，该技术使用的是艾本德移液器吸头和26G针头。
将细菌（ 和 ）和真菌（ 和 ）临床分离株的18小时培养物重悬至终浓度为10,000菌落形成单位（CFU）/1微升，使用连接有26G针头的改良艾本德移液器吸头的0.1 - 2.5微升移液器分别将其玻璃体内注射到C57BL / 6小鼠（6 - 8周龄）体内。对侧眼作为载体/未注射对照。通过评估模型中所用眼睛的角膜混浊、不透明度、细菌负荷和视网膜组织学来确定疾病进展。安乐死后，细菌感染的小鼠在初次注射后24小时摘除眼球，真菌感染的小鼠在72小时后摘除眼球。
在注射的50只小鼠中，改良技术在48只小鼠中成功。两只小鼠因晶状体意外受伤形成白内障而被排除。实验性眼内炎小鼠模型成功模拟了自然临床病程。临床评估和组织病理学证实炎症细胞流入眼后段，同时视网膜结构溶解。
我们使用改良艾本德移液器吸头和26G针头的新型注射方法产生了一种具有成本效益的临床眼内炎小鼠模型，可实现可重复的感染，以了解其病理生物学的各个方面。
