Impact of Brief Exposure to Lysozyme and Lactoferrin on Pathogenic Attributes of Oral Candida.

INTRODUCTION AND AIMS: Adhesion to buccal epithelial cells (BEC) and denture acrylic surfaces (DAS), germ tube (GT) formation, cell surface hydrophobicity (CSH), and haemolysin production are attributes associated with pathogenicity of Candida. Candida albicans and Candida dubliniensis are allied in causing oral candidosis. Lysozyme and lactoferrin exert antimicrobial activity on a range of oral microorganisms, including Candida. There is no information on the impact of brief exposure to lysozyme and lactoferrin on adhesion-related attributes and haemolysin production of aforementioned oral Candida isolates. Thus, we investigated the impact of lysozyme and lactoferrin on adhesion to BEC and DAS, GT formation, CSH, and haemolysin production of these isolates. METHODS: After exposure to lysozyme and lactoferrin for 1 hour, susceptibility to lysozyme and lactoferrin of 20 isolates each of C albicans and C dubliniensis isolates was determined following a 48-hour period of incubation. Candida cell suspensions, obtained from colony-forming units after this period, were assessed for adhesion to BEC and DAS, GT formation, CSH, and haemolysin production using in vitro assays. RESULTS: Exposure to lysozyme and lactoferrin significantly suppressed the ability of C albicans and C dubliniensis isolates to adhere to BEC and DAS, GT formation, CSH, and haemolysin production (P < 0.01 for all virulent attributes tested). CONCLUSIONS: These data provide a tantalising glimpse into the possibility that exposure to either lysozyme or lactoferrin, even for a brief period, would induce a sustainable antifungal effect by suppressing adhesion-related attributes and haemolysin production of these oral Candida species in vitro. Resistance to conventional antifungal agents has been reported in clinical isolates of Candida. The presence of such resistance indicates the need for possible alternative therapies to facilitate the management of oral candidosis. Further research on the pharmacodynamics of lysozyme and lactoferrin and their effects on candidal pathogenic attributes should be fostered, with the vision of developing novel topical antifungal drugs.

Impact of Cigarette Smoke Condensate on Adhesion-Related Traits and Hemolysin Production of Oral Candida dubliniensis Isolates.

BACKGROUND: Cigarette smoke is associated with higher oral Candida carriage and possible predisposition and increased susceptibility to oral candidal infection. Candida dubliniensis is associated with oral candidosis. Candidal adherence to buccal epithelial cells (BEC) and denture acrylic surfaces (DAS), germ tube (GT) formation, cell surface hydrophobicity (CSH) and hemolysin production are pathogenic traits of Candida. OBJECTIVES: The impact of exposure to cigarette smoke on the aforementioned pathogenic attributes of oral C. dubliniensis has not been studied. Hence, the impact of cigarette smoke condensate (CSC) on adhesion to BEC and DAS, GT formation, CSH and hemolysin production of 20 oral C. dubliniensis isolates after exposure to CSC for 24, 48 and 72 h was ascertained. METHODS: After preparation of the CSC, using an in-house smoking device, the Candida isolates were exposed to the CSC for 24, 48 and 72 h, by a previously described in vitro method. Thereafter, the adhesion to BEC and DAS, GT formation, CSH and hemolysin production of C. dubliniensis isolates was investigated by hitherto described in vitro assays. RESULTS: Exposure to CSC significantly increased the ability of C. dubliniensis oral isolates to adhere to BEC, DAS, GT formation, CSH and produce hemolysin following 24-h, 48-h and 72-h exposure periods to CSC (P < 0.001 for all attributes tested). CONCLUSIONS: Exposure of oral C. dubliniensis isolates to CSC may significantly promote in vitro adhesion traits and hemolysin production of these isolates, thereby augmenting its pathogenicity in vitro in the presence of cigarette smoke.

In vitro Post-Antifungal Effect of Posaconazole and Its Impact on Adhesion-Related Traits and Hemolysin Production of Oral Candida dubliniensis Isolates.

OBJECTIVE: Candidal adherence to denture acrylic surfaces (DAS) and oral buccal epithelial cells (BEC), formation of candidal germ tubes (GT), candidal cell surface hydrophobicity (CSH), and hemolysin production are important pathogenic traits of Candida. The antifungal drug-induced post-antifungal effect (PAFE) also impacts the virulence of Candida. Candida dubliniensis isolates are associated with the causation of oral candidiasis which could be managed with posaconazole. Thus far there is no evidence on posaconazole-induced PAFE and its impact on adhesion-related attributes and production of hemolysin by C. dubliniensis isolates. Hence, the PAFE, adhesion to DAS and BEC, formation of GT, CSH, and hemolysin production of 20 oral C. dubliniensis isolates after brief exposure to posaconazole was ascertained. MATERIALS AND METHODS: The PAFE, adherence to DAS and BEC, formation of GT, candidal CSH, and hemolysin production were investigated by hitherto described in vitro assays. RESULTS: The mean PAFE (h) induced by posaconazole on C. dubliniensis isolates was 1.66. Exposure to posaconazole suppressed the ability of C. dubliniensis to adhere to DAS, BEC, formation of candidal GT, candidal CSH and to produce hemolysin by a reduction of 44, 33, 34, 36, and 15% (p < 0.005 to p < 0.001), respectively. CONCLUSION: Exposure of C. dubliniensis isolates to posaconazole for a brief period induced an antimycotic impact by subduing its growth in addition to suppressing pathogenic adherence-associated attributes, as well as production of hemolysin.

Caspofungin-induced in-vitro post-antifungal effect and its impact on adhesion related traits of oral Candida dubliniensis and Candida albicans isolates.

Adhesion to buccal epithelial cells (BEC) and denture acrylic surfaces (DAS), germ tube (GT) formation and cell surface hydrophobicity (CSH) are all virulence traits involved in the pathogenicity of Candida. Post-antifungal effect (PAFE) also have a bearing on pathogenicity and virulence of Candida. Candida dubliniensis is associated with oral and systemic candidosis, which can be managed with caspofungin. There is no published information on caspofungin-induced PAFE and its impact on adhesion traits of C. dubliniensis isolates. Thus, the purpose of this investigation was to determine the in vitro duration of PAFE on 20 C. dubliniensis isolates following transient exposure to caspofungin. Furthermore the impacts of caspofungin-induced PAFE on adhesion to BEC and DAS, GT formation and CSH of these isolates were also determined. After establishing the minimum inhibitory concentration (MIC) of caspofungin, C. dubliniensis isolates were exposed to sub-lethal concentrations (×3 MIC) of caspofungin for 1 hr. Thereafter the duration of PAFE, adhesion to BEC and DAS, GT formation and CSH were determined by previously described in-vitro assays. MIC (µg/mL) of C. dubliniensis isolates to caspofungin ranged from 0.004 to 0.19. Caspofungin-induced mean PAFE on C. dubliniensis isolates was 2.17 hr. Exposure to caspofungin suppressed the ability of C. dubliniensis isolates to adhere to BEC and DAS, form GT and CSH by 69.97%, 71.95%, 90.06% and 32.29% (P < 0.001 for all), respectively. Thus, transient exposure of C. dubliniensis isolates to caspofungin produces an antifungal effect not only by suppressing its growth but also by altering its adhesion traits.