Department of Eye and Vision Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool, United Kingdom.
SpheriTech Ltd, The Heath Business and Technical Park, Runcorn, Cheshire, United Kingdom.
Invest Ophthalmol Vis Sci. 2022 Jan 3;63(1):11. doi: 10.1167/iovs.63.1.11.
To determine the amoebicidal activity of functionalized poly-epsilon-lysine hydrogels (pɛK+) against Acanthamoeba castellanii.
A. castellanii trophozoites and cysts were grown in the presence of pɛK solution (0-2.17 mM), pɛK or pɛK+ hydrogels, or commercial hydrogel contact lens (CL) for 24 hours or 7 days in PBS or Peptone-Yeast-Glucose (PYG) media (nutrient-deplete or nutrient-replete cultures, respectively). Toxicity was determined using propidium iodide and imaged using fluorescence microscopy. Ex vivo porcine corneas were inoculated with A. castellanii trophozoites ± pɛK, pɛK+ hydrogels or commercial hydrogel CL for 7 days. Corneal infection was assessed by periodic acid-Schiff staining and histologic analysis. Regrowth of A. castellanii from hydrogel lenses and corneal discs at 7 days was assessed using microscopy and enumeration.
The toxicity of pɛK+ hydrogels resulted in the death of 98.52% or 83.31% of the trophozoites at 24 hours or 7 days, respectively. The toxicity of pɛK+ hydrogels resulted in the death of 70.59% or 82.32% of the cysts in PBS at 24 hours or 7 days, respectively. Cysts exposed to pɛK+ hydrogels in PYG medium resulted in 75.37% and 87.14% death at 24 hours and 7 days. Ex vivo corneas infected with trophozoites and incubated with pɛK+ hydrogels showed the absence of A. castellanii in the stroma, with no regrowth from corneas or pɛK+ hydrogel, compared with infected-only corneas and those incubated in presence of commercial hydrogel CL.
pɛK+ hydrogels demonstrated pronounced amoebicidal and cysticidal activity against A. castellanii. pɛK+ hydrogels have the potential for use as CLs that could minimize the risk of CL-associated Acanthamoeba keratitis.
确定功能化聚赖氨酸水凝胶（pɛK+）对棘阿米巴Castellanii的杀阿米巴活性。
在 PBS 或 Peptone-Yeast-Glucose（PYG）培养基（分别为营养缺乏或营养丰富的培养物）中，将棘阿米巴滋养体和包囊在 pɛK 溶液（0-2.17mM）、pɛK 或 pɛK+水凝胶或商业水凝胶隐形眼镜（CL）存在下生长 24 小时或 7 天。使用碘化丙啶测定毒性，并通过荧光显微镜成像。将猪眼角膜外植体接种棘阿米巴滋养体±pɛK、pɛK+水凝胶或商业水凝胶 CL 7 天。通过过碘酸希夫染色和组织学分析评估角膜感染。在第 7 天，通过显微镜和计数评估从水凝胶镜片和角膜片上棘阿米巴的再生。
pɛK+水凝胶的毒性导致 98.52%或 83.31%的滋养体在 24 小时或 7 天内死亡。pɛK+水凝胶的毒性导致 PBS 中 70.59%或 82.32%的包囊在 24 小时或 7 天内死亡。在 PYG 培养基中暴露于 pɛK+水凝胶的包囊在 24 小时和 7 天时分别导致 75.37%和 87.14%的死亡。与仅感染的角膜和用商业水凝胶 CL 孵育的角膜相比，用 pɛK+水凝胶孵育的感染滋养体的外植体角膜在基质中没有发现棘阿米巴，并且从角膜或 pɛK+水凝胶中没有再生。
pɛK+水凝胶对棘阿米巴Castellanii表现出明显的杀阿米巴和杀包囊活性。pɛK+水凝胶有可能用作隐形眼镜，可最大程度地降低隐形眼镜相关棘阿米巴角膜炎的风险。
