Comparative cytotoxicity of Acanthamoeba castellanii-derived conditioned medium on human corneal epithelial and stromal cells

Alhazmi, Abdullah; Sidney, Laura E.; Hopkinson, Andy; Elsheikha, Hany M.

Abstract

Soluble factors in the secretome of Acanthamoeba castellanii play crucial roles in the pathogenesis of Acanthamoeba keratitis (AK). Investigating the pathological effects of A. castellanii-derived conditioned medium (ACCM) on ocular cells can provide insights into the damage inflicted during AK. This study examined ACCM-induced cytotoxicity in primary human corneal stromal cells (CSCs) and a human SV40 immortalized corneal epithelial cell line (ihCECs) at varying ACCM concentrations (25 %, 50 %, 75 %, and 100 %). MTT, AlamarBlue, Sulforhodamine B, lactate dehydrogenase, and Caspase-3/7 activation assays were used to assess the impact of ACCM on the cell viability, proliferation and apoptosis. Additionally, fluorescent staining was used to reveal actin cytoskeleton changes. ACCM exposure significantly decreased cell viability, increased apoptosis, and disrupted the actin cytoskeleton, particularly at higher concentrations and longer exposures. Proteases were found to mediate these cytopathogenic effects, highlighting the need for characterization of A. castellanii proteases as key virulence factors in AK pathogenesis.

Citation

Alhazmi, A., Sidney, L. E., Hopkinson, A., & Elsheikha, H. M. (2024). Comparative cytotoxicity of Acanthamoeba castellanii-derived conditioned medium on human corneal epithelial and stromal cells. Acta Tropica, 257, Article 107288. https://doi.org/10.1016/j.actatropica.2024.107288

• Journal Article Type: Article
• Acceptance Date: Jun 11, 2024
• Online Publication Date: Jun 18, 2024
• Publication Date: Sep 1, 2024
• Deposit Date: Jul 7, 2024
• Publicly Available Date: Jul 8, 2024
• Journal: Acta Tropica
• Print ISSN: 0001-706X
• Electronic ISSN: 1873-6254
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 257
• Article Number: 107288
• DOI: https://doi.org/10.1016/j.actatropica.2024.107288
• Keywords: Acanthamoeba castellanii, Corneal stromal cells, Corneal epithelial cells, Cytotoxicity, Metabolic activity, Actin cytoskeleton
• Public URL: https://nottingham-repository.worktribe.com/output/36306968
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0001706X24001700?via%3Dihub