Revista de Odontologia da UNESP
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Revista de Odontologia da UNESP
Original Article

Antifungal activity of chlorhexidine on Candida spp. biofilm

Atividade antifúngica da clorexidina em biofilmes de Candida spp.

Machado, Fernanda Campos; Portela, Marista Barbosa; Cunha, Amanda Carneiro; Souza, Ivete Pomarico Ribeiro de; Soares, Rosângela Maria de Araújo; Castro, Gloria Fernanda Barbosa de Araújo

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Abstract

Oral candidiasis is the most common opportunistic infection in immunocompromised patients and chlorhexidine is an important antimicrobial for its treatment. The antifungal effect of different CHX solutions (Periogard®, NoPlak Max®, Noplak®, Perioxidin®, Chlorhexidine 0.06%, Paradontax® and Chlorhexidine 1%) was evaluated on artificial biofilms of Candida spps: C. albicans (ATCC36801), C. parapsilosis (ATCC22019), C. krusei (ATCC6258), C. glabrata (ATCC2001) and Candida tropicalis (ATCC750). The strains were grown, in a BHI agar medium on bovine teeth enamel for 72 hours at 37 °C. After growth, the fragments were immersed in the CHX solutions for 3 minutes. Nystatin and saline solutions were used as positive and negative controls respectively. To remove the non-adhered cells, the fragments were inoculated in saline solution for 10 minutes, transferred to Falcon tubes containing saline solution and mixed in a vortex. Aliquots of 100 µL were inoculated on BHI agar for 24 hours at 37 °C to count the number of colony forming units (CFU). We observed that the number of (CFU) of C. albicans and C. parapsilosis, showed a reduction rate ranging from 79 to 99% with the use of different solutions (p < 0.001), except for NoPlak Max® (2.94 and 1.3%, respectively). For C. krusei and C. glabrata, nystatin was the least effective solution (23 and 3.4%, respectively); and for C. tropicalis, all the substances presented a high reduction percentage (99-100%). The chlorhexidine solutions were able to reduce the colony forming units of Candida biofilm.

Keywords

Chlorhexidine, candida, biofilms

Resumo

Candidíase oral é a infecção oportunista mais comum em pacientes imunocomprometidos, sendo a clorexidina um importante antimicrobiano auxiliar no seu tratamento. O objetivo do presente estudo foi avaliar o efeito antifúngico de diferentes soluções de clorexidina (Periogard®, NoPlak Max®, Noplak®, Perioxidin®, Clorexidina 0.06%, Paradontax® e Clorexidina 1%) sobre biofilmes artificiais de Candida spp.: C. albicans (ATCC36801); Cparapsilosis (ATCC22019); C. krusei (ATCC6258); C. glabrata (ATCC2001) e C. tropicalis (ATCC750). As cepas foram cultivadas em meio de cultura BHI ágar sobre fragmentos de esmalte bovino por 72 horas a 37 °C. Após o crescimento, cada fragmento de esmalte bovino foi imerso nas diferentes soluções de Clorexidina por 3 minutos. Nistatina e solução salina foram utilizadas como controle negativo e positivo, respectivamente. Para remoção das células não aderidas, os fragmentos foram então imersos em solução salina por 10 minutos e agitados em vortex. Alíquotas de 100 µL foram inoculadas em placas contendo BHI ágar por 24 horas a 37 °C para contagem de unidades formadoras de colônias (UFC). Observamos que o número de UFC de C. albicans e C. parapsilosis, apresentou um percentual de redução variando de 79 a 99% quando do uso das diferentes soluções (p < 0,001), o mesmo não foi observado para o NoPlak Max® (2,94 e 1,3%, respectivamente); Para C. krusei e C. glabrata, a solução menos efetiva foi a Nistatina (23 e 3,4%, respectivamente) enquanto que para C. tropicalis, todas as soluções apresentaram um alto percentual de redução (99 a 100%). As soluções de clorexidina foram capazes de reduzir significativamente o número de UFC provenientes de biofilme de Candida spp. in vitro.

Palavras-chave

Clorexidina, cândida, biofilmes

References



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