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Journal of Clinical Microbiology, February 2005, p. 818-825, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.818-825.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

In Vitro Method To Study Antifungal Perfusion in Candida Biofilms

Y. H. Samaranayake, J. Ye, J. Y. Y. Yau, B. P. K. Cheung, and L. P. Samaranayake*

Oral Biosciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong, Special Administrative Region, People's Republic of China

Received 20 October 2003/ Returned for modification 11 February 2004/ Accepted 5 August 2004

Antimycotic perfusion through Candida biofilms was demonstrated by a modification of a simple in vitro diffusion cell bioassay system. Using this model, the perfusion of three commonly used antifungal agents, amphotericin B, fluconazole, and flucytosine, was investigated in biofilms of three different Candida species (i.e., Candida albicans, Candida parapsilosis, and Candida krusei) that were developed on microporous filters. Scanning electron microscopy revealed that C. albicans formed a contiguous biofilm with tightly packed blastospores and occasional hyphae compared with C. parapsilosis and C. krusei, which developed confluent biofilms displaying structural heterogeneity and a lesser cell density, after 48 h of incubation on nutrient agar. Minor structural changes were also perceptible on the superficial layers of the biofilm after antifungal perfusion. The transport of antifungals to the distal biofilm-substratum interface was most impeded by C. albicans biofilms in comparison to C. parapsilosis and C. krusei. Fluconazole and flucytosine demonstrated similar levels of perfusion, while amphotericin B was the least penetrant through all three biofilms, although the latter appeared to cause the most structural damage to the superficial cells of the biofilm compared with the other antifungals. These results suggest that the antifungal perfusion through biofilm mode of growth in Candida is dependent both on the antimycotic and the Candida species in question, and in clinical terms, these phenomena could contribute to the failure of Candida biofilm-associated infections. Finally, the in vitro model we have described should serve as a useful system to investigate the complex interactions that appear to operate in vivo within the biofilm-antifungal interphase.

* Corresponding author. Mailing address: Dean's Office, Faculty of Dentistry, University of Hong Kong, 34 Hospital Rd., Hong Kong, SAR, People's Republic of China. Phone: 852 2859 0480. Fax: 852 2547 6133. E-mail: lakshman{at}hku.hk.

Journal of Clinical Microbiology, February 2005, p. 818-825, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.818-825.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Perumal, P., Mekala, S., Chaffin, W. L. (2007). Role for Cell Density in Antifungal Drug Resistance in Candida albicans Biofilms. Antimicrob. Agents Chemother. 51: 2454-2463 [Abstract] [Full Text]  
  • El-Azizi, M. (2007). Enhancement of the in vitro activity of amphotericin B against the biofilms of non-albicans Candida spp. by rifampicin and doxycycline. J Med Microbiol 56: 645-649 [Abstract] [Full Text]  
  • LaFleur, M. D., Kumamoto, C. A., Lewis, K. (2006). Candida albicans Biofilms Produce Antifungal-Tolerant Persister Cells. Antimicrob. Agents Chemother. 50: 3839-3846 [Abstract] [Full Text]  


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