Influência da rugosidade de superfície e ação de diferentes desinfetantes na formação de biofilme em resina acrílica para prótese ocular e sua biocompatibilidade em contato com células da conjuntiva humana
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Data
2017-08-31
Autores
Andreotti, Agda Marobo [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O controle da formação de biofilme é essencial para manter a saúde da cavidade anoftálmica dos portadores de prótese ocular. As propriedades físicas da resina acrílica, como a sua rugosidade, podem interferir na adesão de microrganismos nas próteses e na formação de biofilmes. Sendo assim, é necessário utilizar soluções de limpeza eficazes e que não causem irritação tecidual na conjuntiva ocular. O objetivo desse trabalho foi verificar a influência da rugosidade de superfície da resina acrílica utilizada na confecção de próteses oculares na formação de biofilme bacteriano, bem como avaliar a eficácia antimicrobiana e citotoxicidade de diferentes soluções para desinfecção de próteses oculares. Para a análise da rugosidade, corpos de prova de resina acrílica foram confeccionados e divididos em 5 grupos de acordo com a granulação de lixa de polimento utilizada: 120, 400, 800, 1200 e 1200S (lixa + sílica). A rugosidade dos corpos de prova foi mensurada por meio de perfilômetro e imagens de Microscopia de Força Atômica foram obtidas para cada grupo. O crescimento das espécies Staphylococcus aureus e Staphylococcus epidermidis sobre os corpos de prova foi avaliado após 4 e 24h, por meio da contagem das Unidades Formadoras de Colônia (UFC)/mL. O efeito dos seguintes tratamentos: soro fisiológico, sabão neutro, clorexidina 4%, pastilhas efervescentes Efferdent®, triclosan 1% e citronela sobre biofilme formado sobre os corpos de prova de resina acrílica também foi avaliado por meio da determinação do número de UFC/mL. A citotoxicidade dos agentes antimicrobianos foi avaliada por meio dos testes de MTT e Neutral Red, após tratamento dos corpos de prova, diariamente, por 1, 7, 15, 30, 60 e 90 dias com as soluções já citadas, sendo avaliados ao fim de cada período. Os dados obtidos foram submetidos à análise estatística adequada, considerando se provinham de dados paramétricos ou não paramétricos. Os resultados demonstraram que os menores valores de rugosidade foram encontrados no grupo 800, 1200 e 1200S. A contagem microbiana foi menor no grupo 1200S e esta diferiu estatisticamente (p>0,05) do controle para ambos os tempos de avaliação e bactérias. Na análise da ação antimicrobiana das soluções para desinfecção, todos os protocolos promoveram redução do crescimento dos microrganismos testados comparados ao controle (crescimento sem a presença do antimicrobiano), sendo que os grupos submetidos ao tratamento com clorexidina 4%, pastilhas efervescentes Efferdent® e triclosan 1% eliminaram o biofilme (p>0,01). Ainda nos ensaios de citotoxicidade, em todos os grupos foi observada proliferação celular das células da conjuntiva acima de 84% (p>0,05). Dessa forma, conclui-se que a rugosidade não interfere significativamente na adesão bacteriana e formação de biofilme, exceto para o grupo 1200S, e que os protocolos de limpeza com imersão em clorexidina 4%, pastilhas efervescentes Efferdent® e triclosan 1% são totalmente eficazes na desinfecção e não apresentam citotoxicidade para com as células da conjuntiva humana, sugerindo sua indicação para desinfecção de próteses oculares.
The control of biofilm formation is essential to maintain the health of anophthalmic cavity of ocular prosthesis wearers. The physical properties of the acrylic resin, such as surface roughness, may interfere with the adhesion of microorganisms to the prostheses. Therefore, it is necessary to use cleaning solutions that are effective and do not cause tissue irritation in ocular conjunctiva. The aim of this study was to verify the influence of surface roughness of the acrylic resin used in the preparation of ocular prosthesis in the formation of bacterial biofilm, as well as to evaluate the antimicrobial efficacy and cytotoxicity of different disinfectants in ocular prostheses. For roughness analysis, acrylic resin samples were prepared and divided into 5 groups according to the granulation of polishing sandpaper used: 120, 400, 800, 1200 and 1200S (sandpaper + silica). The roughness of the samples was measured using a profilometer and Atomic Force Microscopy images were obtained for each group. The microbial growth for Staphylococcus aureus and Staphylococcus epidermidis on the samples was evaluated after 4 and 24 h by counting the Colony Forming Units (CFU)/mL. The effect of the following solutions was also evaluated by CFU/mL: non treated group (immersed in saline solution), neutral soap, 4% chlorhexidine, Efferdent® effervescent tablets, 1% triclosan and citronella. The cytotoxicity of the solutions was evaluated by MTT and Neutral Red after treating the samples daily for 1, 7, 15, 30, 60 and 90 days with the solutions already mentioned, being evaluated at the end of each period. Data were submitted to proper statistical analysis, considering whether data were parametric or non-parametric. The results showed that the lowest roughness values were found in groups 800, 1200 and 1200S. 1200S group also presented the lowest values of microbial growth statistically different from control group (p>0,05), for both periods and bacteria evaluated. In the analysis of the solutions, compared to the control group (microbial growth without the presence of the antimicrobial), all protocols promoted a reduction in the growth of the microorganisms, and the groups submitted to disinfection with 4% chlorhexidine, Efferdent® effervescent tablets and 1% triclosan eliminated the biofilm (p>0,01). In addition, all groups showed cell proliferation of conjunctival cells above 84% in the cytotoxicity assays (p>0,05). Thus, it was concluded that the roughness does not significantly interfere in the adhesion of the microorganisms, except for group 1200S, and that the cleaning protocols with 4% chlorhexidine, Efferdent® effervescent tablets and 1% triclosan are totally effective in the disinfection and do not present cytoxicitiy with human conjunctive cells, what suggests their indication for cleaning of ocular prosthesis.
The control of biofilm formation is essential to maintain the health of anophthalmic cavity of ocular prosthesis wearers. The physical properties of the acrylic resin, such as surface roughness, may interfere with the adhesion of microorganisms to the prostheses. Therefore, it is necessary to use cleaning solutions that are effective and do not cause tissue irritation in ocular conjunctiva. The aim of this study was to verify the influence of surface roughness of the acrylic resin used in the preparation of ocular prosthesis in the formation of bacterial biofilm, as well as to evaluate the antimicrobial efficacy and cytotoxicity of different disinfectants in ocular prostheses. For roughness analysis, acrylic resin samples were prepared and divided into 5 groups according to the granulation of polishing sandpaper used: 120, 400, 800, 1200 and 1200S (sandpaper + silica). The roughness of the samples was measured using a profilometer and Atomic Force Microscopy images were obtained for each group. The microbial growth for Staphylococcus aureus and Staphylococcus epidermidis on the samples was evaluated after 4 and 24 h by counting the Colony Forming Units (CFU)/mL. The effect of the following solutions was also evaluated by CFU/mL: non treated group (immersed in saline solution), neutral soap, 4% chlorhexidine, Efferdent® effervescent tablets, 1% triclosan and citronella. The cytotoxicity of the solutions was evaluated by MTT and Neutral Red after treating the samples daily for 1, 7, 15, 30, 60 and 90 days with the solutions already mentioned, being evaluated at the end of each period. Data were submitted to proper statistical analysis, considering whether data were parametric or non-parametric. The results showed that the lowest roughness values were found in groups 800, 1200 and 1200S. 1200S group also presented the lowest values of microbial growth statistically different from control group (p>0,05), for both periods and bacteria evaluated. In the analysis of the solutions, compared to the control group (microbial growth without the presence of the antimicrobial), all protocols promoted a reduction in the growth of the microorganisms, and the groups submitted to disinfection with 4% chlorhexidine, Efferdent® effervescent tablets and 1% triclosan eliminated the biofilm (p>0,01). In addition, all groups showed cell proliferation of conjunctival cells above 84% in the cytotoxicity assays (p>0,05). Thus, it was concluded that the roughness does not significantly interfere in the adhesion of the microorganisms, except for group 1200S, and that the cleaning protocols with 4% chlorhexidine, Efferdent® effervescent tablets and 1% triclosan are totally effective in the disinfection and do not present cytoxicitiy with human conjunctive cells, what suggests their indication for cleaning of ocular prosthesis.
Descrição
Palavras-chave
Olho artificial, Resinas acrílicas, Propriedades de superfície, Biofilmes, Desinfecção, Toxicidade, Eye, Artificial, Acrylic resins, Surface properties, Biofilms, Disinfection, Toxicity