Effects of aspirin on lipase production and biofilm formation by candida species recovered from oral lesion.

CASTILLO G; BARENBAUM, S; SCATENA, G, ; , LEHNER; SOTOMAYOR C; ASCURRA A

Rosario

Congreso; 2da Reunión Internacional de Ciencias Farmacéuticas; 2012

Sociedad Argentina de Ciencias Farmaceuticas

  Introduction The emergence and opportunistic nature of Candida species has attracted interest in the inhibition of virulence factors, such as exoenzymes lipases (LIP) and their ability to form biofilm (BP). There is in vitro evidence of the inhibitory role of aspirin (ASP) on LIP to concentrations that can be used in therapeutics, suggesting a potential clinical use for combination therapy against biofilm-associated infections. The objective of this study was to evaluate the ability of ASP to inhibit the LIP activity and BP formation by Candida isolated recovered from patients with different stomatological lesions. Materials and methods Strains were isolated from patients with oral lesions with clinical, mycological and/or pathological diagnostic of oral candidiasis (ORC, n=20), lichens (LIC, n=10) and oral cancer (OCA, n=13). Isolates were identified in chromogenic medium (CHROMagar, France) resulting 23 C.albicans, 7 C. tropicalis, 6 C. krusei and 7 mixed cultures. BP was assessed by crystal violet method (1). To semi-quantify the LIP activity, Rhodamine-B assay was used and the Pz ratio as halo diameter/colony diameter was measured (2). Inhibition of LIP activity with ASP (ICLIP) (Sigma, China) was assessed by successive dilution (concentrations between 0.313 mM and 10 mM in minimal medium) (3). Determinations were performed in triplicate. Data were analyzed using the t test, Wilcoxon and multivariate correspondence test. Strains were isolated from patients with oral lesions with clinical, mycological and/or pathological diagnostic of oral candidiasis (ORC, n=20), lichens (LIC, n=10) and oral cancer (OCA, n=13). Isolates were identified in chromogenic medium (CHROMagar, France) resulting 23 C.albicans, 7 C. tropicalis, 6 C. krusei and 7 mixed cultures. BP was assessed by crystal violet method (1). To semi-quantify the LIP activity, Rhodamine-B assay was used and the Pz ratio as halo diameter/colony diameter was measured (2). Inhibition of LIP activity with ASP (ICLIP) (Sigma, China) was assessed by successive dilution (concentrations between 0.313 mM and 10 mM in minimal medium) (3). Determinations were performed in triplicate. Data were analyzed using the t test, Wilcoxon and multivariate correspondence test. Results All isolates showed LIP activity and capacity of BP. ASP did not produce a growth inhibition in any of the concentration studied. Mean ICLIP of ASP was of 1.25 mM for 43 isolates studied, and only one (C.tropicalis, LIC lesion) showed no ICLIP of ASP. The higher values of ICLIP were observed in yeast recovered from OCA (1.88 mM, p<0.0001) and species C.krusei and C.tropicalis (1.88 mM, p<0.04). Isolates with greater BP capacity showed the highest activity LIP (p=0.0005). Multivariate analysis showed an association between higher values of BP capacity, ICLIP and LIP (inertia = 56.7%). When considering the type of oral lesion, association among the higher values of ICLIP, LIC and LIP activity was observed, and also the higher capacity of BP with OCA (inertia = 46.88%). All isolates showed LIP activity and capacity of BP. ASP did not produce a growth inhibition in any of the concentration studied. Mean ICLIP of ASP was of 1.25 mM for 43 isolates studied, and only one (C.tropicalis, LIC lesion) showed no ICLIP of ASP. The higher values of ICLIP were observed in yeast recovered from OCA (1.88 mM, p<0.0001) and species C.krusei and C.tropicalis (1.88 mM, p<0.04). Isolates with greater BP capacity showed the highest activity LIP (p=0.0005). Multivariate analysis showed an association between higher values of BP capacity, ICLIP and LIP (inertia = 56.7%). When considering the type of oral lesion, association among the higher values of ICLIP, LIC and LIP activity was observed, and also the higher capacity of BP with OCA (inertia = 46.88%). Conclusions Although ASP could not be considered a cure for Candida biofilm associated to stomatological lesions, decreasing its contribution to pathogenicity through the inhibition of LIP has clinical implications. This study would contribute to the knowledge about ASP as a LIP inhibitor, especially in combination with antifungal agents commonly employed in clinic therapeutics. Although ASP could not be considered a cure for Candida biofilm associated to stomatological lesions, decreasing its contribution to pathogenicity through the inhibition of LIP has clinical implications. This study would contribute to the knowledge about ASP as a LIP inhibitor, especially in combination with antifungal agents commonly employed in clinic therapeutics.