Exploring interspecies differences in ex vivo models of Pseudomonas aeruginosa keratitis: a comparative study of human, pig and sheep corneas.

Katarzyna Okurowska, Sheila MacNeil, Sanhita Roy, Prashant Garg, Peter N Monk, Esther Karunakaran
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Abstract

Introduction. Interspecies differences in human, pig and sheep corneal thickness may affect the Pseudomonas aeruginosa colonization. Currently, there is no research investigating the impact of these differences, along with variable storage and culture conditions on infection in ex vivo cornea models. These factors could significantly influence utilizing ex vivo models for drug testing research.Aim. In this study, we aim to compare the relevance of sheep and pig cornea infection models to human.Methodology. The corneas were stored in McCarey-Kaufman medium or Eagle's Minimum Essential Medium or Dulbecco's Modified Eagle's Medium/Mixture F-12 Ham medium (incubator) and then infected after varying storage durations. The effect of added foetal bovine serum (FBS) to media and continuous shaking mimicking rinsing with tears on infection was also investigated. The infection outcome was evaluated by comparing c.f.u. between conditions.Results. The study found that storage conditions, culture media, FBS and continuous rinsing of corneas with media had no significant effect on infection progression in ex vivo keratitis models across selected species.Conclusions. Pig and sheep models yield results comparable to human corneas. These findings support the interchangeability of ex vivo human, pig and sheep keratitis models for P. aeruginosa infection studies, emphasizing their relevance and reliability in research contexts. This interchangeability is particularly useful for research groups where one particular animal model may not be available. The media in this ex vivo keratitis model can be free of animal components by the removal of FBS, which reduces the reliance on animal-derived products, aligning with ethical considerations and promoting more sustainable and humane scientific practices. This study advances the understanding of ex vivo keratitis models, demonstrating their robustness and potential for broader application in ophthalmic research and drug testing.

介绍。人、猪和羊角膜厚度的种间差异可能会影响铜绿假单胞菌的定植。目前,还没有研究调查这些差异以及不同的储存和培养条件对体外角膜模型感染的影响。这些因素可能会严重影响利用体外模型进行药物测试研究。本研究旨在比较绵羊和猪角膜感染模型与人类角膜感染模型的相关性。将角膜保存在麦卡雷-考夫曼培养基或老鹰最低限度基本培养基或杜贝科改良老鹰培养基/F-12 Ham 混合培养基(培养箱)中,然后在不同的保存时间后进行感染。此外,还研究了在培养基中添加胎牛血清(FBS)和模拟用眼泪冲洗连续振荡对感染的影响。感染结果通过比较不同条件下的 c.f.u. 进行评估。研究发现,在所选物种的体外角膜炎模型中,储存条件、培养基、FBS 和用培养基持续冲洗角膜对感染进展没有显著影响。猪和绵羊模型的结果与人类角膜相当。这些研究结果支持在铜绿假单胞菌感染研究中使用体外人、猪和绵羊角膜炎模型的互换性,强调了它们在研究中的相关性和可靠性。这种互换性对于没有特定动物模型的研究小组尤其有用。这种体外角膜炎模型中的培养基不含动物成分,去除了 FBS,从而减少了对动物衍生产品的依赖,符合伦理要求,促进了更可持续和人道的科学实践。这项研究加深了人们对体外角膜炎模型的了解,证明了其在眼科研究和药物测试中的稳健性和广泛应用潜力。
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