Revista de Odontologia da UNESP
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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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.


Chlorhexidine, candida, biofilms


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.


Clorexidina, cândida, biofilmes


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