Antimicrobial Action of Biguanides on the Viability of Acanthamoeba Cysts and Assessment of Cell Toxicity

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dc.contributor.authorMafra, Cecilia Sales Pires [UNIFESP]
dc.contributor.authorCarrijo-Carvalho, Linda Christian [UNIFESP]
dc.contributor.authorChudzinski-Tavassi, Ana Marisa
dc.contributor.authorTaguchi, Felipe Marques de Carvalho [UNIFESP]
dc.contributor.authorForonda, Annette Silva [UNIFESP]
dc.contributor.authorCarvalho, Fabio Ramos de Souza [UNIFESP]
dc.contributor.authorFreitas, Denise de [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionButantan Inst
dc.date.accessioned2016-01-24T14:34:19Z
dc.date.available2016-01-24T14:34:19Z
dc.date.issued2013-09-01
dc.description.abstractPURPOSE. To assess dose- and concentration-dependent rates of biguanides on the viability of Acanthamoeba cysts isolated from severe ulcerative keratitis, and to correlate cysticidal activites with cytotoxic profiles in corneal and endothelial cells.METHODS. Cysticidal activities of polyhexamethylene biguanide and chlorhexidine digluconate were evaluated in the Acanthamoeba castellanii strain and clinical isolates of Acanthamoeba spp obtained from two severe and recurrent cases of ulcerative keratitis. the molecular characterization of protozoa used in the experimental assays was performed by sequencing reactions of the 18S rDNA gene. Acanthamoeba cysts were exposed at different dosages and concentrations of both biguanides; the application of double-biguanides was also evaluated. Automated cell viability assessment of cysts was performed using the trypan blue dye exclusion method. Cytotoxicity assays of biguanides were conducted using primary cultures of endothelial cells alone or in coculture with Acanthamoeba cysts. Human corneal epithelial cells were used as a comparative pattern to assess the toxicity of biguanide compounds. Cell viability was measured using both quantitative and qualitative methods. Statistical analyses were applied to the data.RESULTS. the in vitro study showed that all dosages, concentrations, and combinations of biguanides tested had a cysticidal effect on Acanthamoeba spp strains tested compared with control cultures not exposed to any antimicrobials; the difference in response was statistically significant. the use of both biguanides in combination demonstrated the best cysticidal effect. the use of isolated biguanides was associated with greater cytotoxic effects than with biguanides used in combination. Chlorhexidine digluconate used alone tended to have greater cytotoxicity than polyhexamethylene biguanide. Furthermore, the double-biguanide application had a statistically significant decrease in the deleterious effect on endothelial cells at higher dosage and concentration. Quantitative and qualitative analyses demonstrated the toxic effect of biguanide compounds on the viability of corneal epithelial cells, under single or in combination usage.CONCLUSIONS. We demonstrated that the combined use of biguanides had greater cysticidal activity than individual drug application as well as a possible protective effect on endothelial cells. the biguanide compounds tested were able to induce corneal epithelial cell death in time and concentration-independent fashions. Findings support the hypothesis concerning the cysticidal effect and the differential patterns of toxicity expressed by polyhexamethylene biguanide and chlorhexidine digluconate on the endothelial and corneal cells.en
dc.description.affiliationUniversidade Federal de São Paulo, Paulista Sch Med, Dept Ophthalmol, São Paulo, Brazil
dc.description.affiliationButantan Inst, Lab Biochem & Biophys, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Paulista Sch Med, Dept Ophthalmol, São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipIDFAPESP: 11/51626-1
dc.description.sponsorshipIDFAPESP: 08/53969-0
dc.description.sponsorshipIDCNPq: 311612/2012-1
dc.description.sponsorshipIDFAPESP: 2012/15603-0
dc.format.extent6363-6372
dc.identifierhttp://dx.doi.org/10.1167/iovs.13-11990
dc.identifier.citationInvestigative Ophthalmology & Visual Science. Rockville: Assoc Research Vision Ophthalmology Inc, v. 54, n. 9, p. 6363-6372, 2013.
dc.identifier.doi10.1167/iovs.13-11990
dc.identifier.issn0146-0404
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/36680
dc.identifier.wosWOS:000325169500053
dc.language.isoeng
dc.publisherAssoc Research Vision Ophthalmology Inc
dc.relation.ispartofInvestigative Ophthalmology & Visual Science
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectAcanthamoebaen
dc.subjectchlorhexidineen
dc.subjectpolyhexamethylene biguanideen
dc.subjectendothelial cellen
dc.titleAntimicrobial Action of Biguanides on the Viability of Acanthamoeba Cysts and Assessment of Cell Toxicityen
dc.typeinfo:eu-repo/semantics/article
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