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Abstract Timely diagnosis and treatment of Acanthamoeba keratitis (AK) are crucial for improving the visual outcome of patients. New medications with lethal effects on the resistant cyst stage are required to eliminate the infection. The present study aimed to evaluate the therapeutic effect of Nigella sativa aqueous extract, Chitosan nanoparticles (CS NPs) and their combination in comparison to chlorhexidine 0.02% on experimentally induced AK. Experimental induction of AK was performed in 64 rats. The study medications; N. sativa (Ns) aqueous extract [30 and 60 mg/ ml], Chitosan nanoparticles (nCs) [50 and 100 g/ ml], Ns-nCs combinations [Ns 30 mg/ml + nCs 50 g/ml and Ns 60 mg/ml + nCs 100 g/ml] and Chlorhexidine (Ch) 0.02% as a drug control were applied, starting from the 5th day post-inoculation, and were evaluated by clinical examination of the cornea, direct microscopic examination, cultivation and PCR testing of corneal scraps on the 10th , 15th and 20th days post-treatment. A 100% cure of corneal opacity was obtained with CS NPs 100 g/ ml and combined doses of Ns 60 mg/ ml plus nCs 100 g/ ml on the 10th day post-treatment, as well as with N. sativa aqueous extract 60 mg/ml on the 15th day post-treatment. Meanwhile, on the 20th day post-treatment N. sativa (30 mg/ml) gave 87.5% cure, CS NPs (50 ug/ml) and combined doses (Ns 30 mg/ml + nCs 50 g/ml) produced 75% cure, while Ch 0.02% resulted in only 25% cure. The clinical outcome was in accordance with the results of the plate culture method, therefore the culture sensitivity was 100%. The sensitivities of direct microscopic examination and PCR were 47.1% and 58.8% respectively compared to cultivation. The results of the present study hold the promise of finding new therapeutic alternatives, N. sativa aqueous extract and CS NPs, for AK treatment with no tissue side effects. Also, the study spots the light on their utilization alone or in combination for successful treatment of AK. |