An Optimized Porcine Corneal Ex Vivo Model Using Plumbing Rings and Medical Adhesive for Controlled Localized Infection with Topical Treatments v1

• DOI: 10.17504/protocols.io.eq2ly6nkmgx9/v1
• Publication Date: 2025-02-02T13:09:16Z
• Authors: Daniel Foulkes; Keri Mclean; Tarunima Sharma; David Fernig; Stephen Kaye
• Keywords: Ex vivo

Infection models are essential tools for understanding host-pathogen interactions and developing noveltherapies. Ex vivo infection offer significant ethical practical advantages, enabling more accurate physiologically relevant insights compared to traditional cell culture systems. The versatility of porcine corneal ex has made them valuable studying a range ofpathogens, including Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae, Candida albicans, Acanthamoeba, viruses such as Herpes Simplex adenovirus. Here, we present anoptimized protocol that addresses several limitations, using plumbing rings medical adhesive enhance experimental accuracy reproducibility. application 10 mm ring the centre cornea allows localized inoculation, preventing "run-off" topically added aqueous solutions ensuring pathogens remain confined site infection. Additionally, this setup enables precise maintenance therapeutic agents on surface, allowing reliable assessments antimicrobial effects. For analysis, describe methods colony-forming unit (CFU) quantification within cornea, evaluation opacity, histological analysis hematoxylin eosin staining assess progression over 48 hours. This improved, accessible, cost-effective model provides standardized platform evaluating potential treatments, ultimately supporting clinical translation reducing burden microbial keratitis.