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. 2018 Jan 22;11(1):50.
doi: 10.1186/s13071-018-2639-5.

Can artificial tears prevent Acanthamoeba keratitis? An in vitro approach

Angela Magnet  1 , Thiago Santos Gomes  1   2 , Carmen Pardinas  1 , Natalia Garcia de Blas  1 , Cruz Sadaba  3 , Eugenia Carrillo  4 , Fernando Izquierdo  1 , José Manuel Benítez Del Castillo  5 , Carolina Hurtado  1 , Carmen Del Aguila  1 , Soledad Fenoy  6
Affiliations

Can artificial tears prevent Acanthamoeba keratitis? An in vitro approach

Angela Magnet et al. Parasit Vectors. .

Abstract

Background: The 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.

Methods: Viability 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.

Results: Trypan 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.

Conclusions: The 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.

Keywords: Acanthamoeba; Amoebicidal effect; Artificial tears; Preservatives; Prophylaxis.

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Competing interests

The authors declare that they have no competing interests.

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Figures

Fig. 1
Fig. 1
Acanthamoeba culture appearance at 4, 6 and 8 h of incubation with the studied artificial tears. Scale-bars: 100 μm
Fig. 2
Fig. 2
Histogram statistic of the CTC viability assay of Artelac® Splash, Optava Fusion™, Oculotect® and NEFF by flow cytometry at 2 h, 4 h, 6 h and 8 h incubation. R1: normal amoebae population; R2: altered smaller amoebae trophozoites
See this image and copyright information in PMC

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