Abstract
BackgroundThe use of contact lenses has increased in recent years as has the incidence of Dry Eye Syndrome, partly due to their use. Artificial tears are the most common treatment option. Since these changes can facilitate Acanthamoeba infection, the present study has been designed to evaluate the effect of three artificial tears treatments in the viability of Acanthamoeba genotype T4 trophozoites. Optava Fusion™, Oculotect®, and Artelac® Splash were selected due to their formulation.MethodsViability was assessed using two staining methods, Trypan Blue stain and CTC stain at different time intervals (2, 4, 6, 8 and 24 h). Trypan Blue viability was obtained by manual count with light microscopy while the CTC stain was determined using flow cytometry.ResultsTrypan Blue staining results demonstrated a decrease in viability for Optava Fusion™ and Artelac® Splash during the first 4 h of incubation. After, this effect seems to lose strength. In the case of Oculotect®, complete cell death was observed after 2 h. Using flow cytometry analysis, Optava Fusion™ and Oculotect® exhibited the same effect observed with Trypan Blue staining. However, Artelac® Splash revealed decreasing cell respiratory activity after four hours, with no damage to the cell membrane.ConclusionsThe present study uses, for the first time, CTC stain analyzed by flow cytometry to establish Acanthamoeba viability demonstrating its usefulness and complementarity with the traditional stain, Trypan Blue. Artelac® Splash, with no preservatives, and Optava Fusion TM, with Purite®, have not shown any useful amoebicidal activity. On the contrary, promising results presented by Ocultect®, with BAK, open up a new possibility for Acanthamoeba keratitis prophylaxis and treatment although in vivo studies should be carried out.
Highlights
The use of contact lenses has increased in recent years as has the incidence of Dry Eye Syndrome, partly due to their use
Viability tests Trypan blue stain Viability studied using Trypan Blue showed a decrease in the case of Optava FusionTM and Artelac® Splash after the first 4 h
In the present study, three artificial tear brands were selected according to the preservative agents or the lack of them to study their possible amoebicidal activity against trophozoites, as this is the initial state of the infection
Summary
The use of contact lenses has increased in recent years as has the incidence of Dry Eye Syndrome, partly due to their use. Acanthamoeba spp. are one of the most commonly isolated amoebae in environmental samples.They have a cosmopolitan distribution and can act as both opportunistic and non-opportunistic pathogens [1] These amoebae have been isolated from soil, dust, air, seawater, swimming pools, sewage, sediments, airconditioning units, domestic tap water, bottled water, dental treatment units, hospitals, dialysis apparatus, eyewash stations, contact lenses and their cases and. These free-living amoebae are typically harmless to humans, but in rare instances can cause severe infections. It is associated with symptoms of ocular discomfort, and contact lens dry eye is considered a sub-classification of this syndrome [13, 14]
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