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Savitri Sharma1, Lakshminarayanan Gowtham2, and Bhupesh Bagga3
1Jhaveri Microbiology Centre, 2Department of Ophthalmic Pharmacology, 3Shanthilal Shanghvi Cornea Institute; LV Prasad Eye Institute, Hyderabad, India
Purpose: To assess the safety of miltefosine to corneal cells and tissues and to evaluate its efficacy to eliminate Acanthamoeba cysts and trophozoites.
Methods: Clinical isolates of Acanthamoeba spp. from corneal scrapings of patients with keratitis (n=17) were grown axenically, subjected to genotyping and tested against miltefosine for minimum cysticidal and trophozoiticidal concentrations (MCC, MTC). Cytotoxicity of miltefosine was assessed on human corneal epithelial cells (HCEC) at four incubation time points at different concentrations. Confluent monolayers of HCEC and ex-vivo human corneal models were challenged with A. castellanii (T4) trophozoites and cysts in the presence and absence of miltefosine for 24hrs. Cytopathic effects and changes in the corneal morphology were evaluated using microscopy and histopathology analysis.
Results: Majority of Acanthamoeba isolates tested were T4 genotype (82.3%). MTC90 and MCC90 values of miltefosine were 125 μg/mL and 4 mg/mL respectively. Miltefosine was found to be safe at 62.5 μg/mL and 125 μg/mL on HCE for 4 hours and 15 minutes, respectively. Without miltefosine, A.castellani trophozoites and cysts destroyed the cellular structures and corneal architecture within 24 hours while miltefosine pre-treatment completely prevented the infection at both the tested concentrations (62.5μg/mL, 125μg/mL).
Conclusions: Miltefosine was effective against Acanthamoeba cysts and trophozoites in vitro and ex vivo, however, cidal concentration was higher by more than 20 fold for cysts compared to trophozoites. At the effective concentration the drug was safe for corneal epithelial cells and ex vivo corneal tissue.
Disclosure: N
Support: Hyderabad Eye Research Foundation
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