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
Rev. odontol. UNESP, vol.39, n5, p.271-275, 2010
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); C. parapsilosis (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
1. Kuriyama T, Williams DW, Bagg J, Coulter WA, Ready D, Lewis MA. In vitro susceptibility of oral Candida to seven antifungal agents. Oral Microbiol Immunol. 2005; 20: 349-53.
2. Greenspan D. Treatment of oral candidiasis in HIV infection Oral Surg Oral Med Oral Pathol. 1994; 78: 211-5.
3. Chandra J, Kuhn DM, Mukherjee PK, Hoyer LL, McCormick T, Ghannoum MA. Biofilm formation by the fungal pathogen Candida albicans: Development, architecture and drug resistance. J Bacteriol. 2001; 183: 5385-94.
4. Lamfon H, Porter SR, McCullought M, Pratten J. Susceptibility of Candida albicans biofilms grown in a constant depth film fermentor to chlorhexidine, fluconazole and miconazole: a longitudinal study. J Antimicrobial Chemother. 2004; 53: 383-5.
5. Suci PA, Tyler BJ. Action of chlorhexidine digluconate against yeast and filamentous forms in early-stage Candida albicans biofilm. Antimicrob Agents Chemother. 2002; 46: 3522-31.
6. Ellepola ANB, Samaranayake LP. Adjunctive use of chlorhexidine in oral candidosis: a review. Oral Dis. 2001; 7: 11-7.
7. Torres SR, Peixoto CB, Caldas DM, Akiti T, Barreiros MGC, Uzeda M, et al. A prospective randomized trial to reduce Candida spp. colonization in patients with hyposalivation. Braz Oral Res. 2007; 21: 182-7.
8. Bobichon H, Bouchet P. Action of chlorhexidine on budding Candida albicans: screening and transmission electron microscopic study. Mycopathologia. 1987; 100: 27-35.
9. Alviano WS, Mendonça-Filho RR, Alviano DS, Bizzo HR, Souto-Padrón T, Rodrigues ML, et al. Antimicrobial activity of Croton cajucara Benth linalool-rich essential oil on artificial biofilms and planktonic microorganisms. Oral Microbiol Immunol. 2003; 20: 101-5.
10. Nakamichi I, Iwaku M, Fusayama T. Bovine teeth as possible substitutes in the adhesion test. J Dent Res. 1983; 62: 1076-81.
11. Tagami J, Tao L, Pashley DH, Horner JA. The permeability of dentine from bovine incisors in vitro. Arch Oral Biol. 1989; 34: 773-7.
12. Abuabara A, Santos AJ, Aguiar FH, Lovadino JR. Evaluation of microleakage in human, bovine and swine enamels. Braz Oral Res. 2004; 18: 312-6.
13. Pratten J, Smith AW, Wilson M. Response of single biofilm and microcosm dental plaques to pulsing with chlorhexidine. J Antimocrobial Chemother. 1998; 42: 253-9.
14. Makihira S, Nikawa H, Tamagami M, Hamada t, Samaranayake LP. Differences in Candida albicans adhesion to intact and denature type I collagen in vitro. Oral Microbiol Immunol. 2002; 17: 129-31.
15. Meiller TF, Kelley JI, Jabra-Rizk MA, DePaola LG, Baqui A, Falker WA. In vitro studies of the efficacy of antimicrobials against fungi. Oral Surg Oral Med Oral Pahol Oral Radiol Endod. 2001; 91: 663-70.
16. Vaahtoniemi LH. Surface ultrastructure of intact and in situ chlorhexidine treated human buccal cells. A method for scanning electron microscopy. Acta Odontol Scand. 1997; 55: 266-81.
17. Franco Neto CA, Parolo CCF, Rösing CK, Maltz M. Comparative analysis of the effect of two chlorhexidine mouthrinses on plaque accumulation and gingival bleeding. Braz Oral Res. 2008; 22: 139-44.
18. MacNeill S, Rindler E, Walker A, Brown AR, Cobb CM. Effects of tetracycline hydrochloride and chlorhexidine gluconate on Candida albicans. An in vitro study. J Clin Periodontol. 1997; 24: 753-60.
19. Lamfon H, Al-Karaawi Z, McCullough M, Porter SR, Pratten J. Composition of in vitro denture plaque biofilms and susceptibility to antifungals. FEMS Microbiol Lett. 2005; 242: 343-51.
20. Guiuliana G, Pizzo G, Milici ME, Musotto GC, Giangreco R. In vitro antifungal properties of mouthrinses containing antimicrobial agents. J Periodontol. 1997; 68: 729-33.
21. Patton AJ, Shugars DA. A systematic review of antifungal drugs for the prevention and treatment of oropharyngeal candidiasis in HIV‑positive patients. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2000; 92: 170-9.
22. Abrams DI. Potential interventions for HIV/AIDS wasting: an overview. J Acquir Immune Defic Syndr. 2000; 25: 74-80.
23. Muzyka BC. Oral fungal infections. Dent Clin North Am. 2005; 49: 49-65.
24. Mariné M, Serena C, Pastor F, Guarro J. Combined antifungal therapy in a murine infection by Candida glabtata. J Antimicrobial Chemother. 2006; 58: 1295-8.
25. Posteraro B, Tumbarello M, La Sorda ML, Spanu T, Trecarichi EM, Bernardis F, et al. Azole resistance of Candida glabrata in a case of recurrent fungaemia. J Clin Microbiol. 2006; 44: 3046-7.
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