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Waltimo
Yeasts in apical periodontitis.Waltimo TM, Sen BH, Meurman JH, Orstavik D, Haapasalo MP. Crit Rev Oral Biol Med. 2003;14(2):128-37. Review.
"Microbiological reports of apical periodontitis have revealed that yeasts can be isolated from approximately 5-20% of infected root canals. They occur either in pure cultures or together with bacteria. Almost all isolated yeasts belong to the genus Candida, and the predominant species is C. albicans. Pheno- and genotypic profiles of C. albicans isolates show heterogeneity comparable with those of isolates from other oral sites. C. albicans expresses several virulence factors that are capable of infecting the dentin-pulp complex, including dentinal tubules. This causes, consequentially, an inflammatory response around the root apex, which suggests a pathogenic role for this organism in apical periodontitis. Yeasts are particularly associated with persistent root canal infections that do not respond favorably to conservative root canal therapy. This may be due to the resistance of all oral Candida species against a commonly used topical medicament, calcium hydroxide. However, other antimicrobial agents may offer alternative therapeutic approaches and improve the treatment of these persistent cases of apical periodontitis."
In vitro yeast infection of human dentin.Waltimo TM, Orstavik D, Siren EK, Haapasalo MP. J Endod. 2000 Apr;26(4):207-9. [abstract only]
"An in vitro model was developed for investigation of Candida albicans penetration into human dentinal tubules. The model consisted of a dentin disc mounted between two cuvettes that each had a circular opening facing the disc. The cuvettes were filled with Tryptic-Soy-Broth, and the pulpal side cuvette was inoculated with C. albicans and incubated at 37 degrees C in air until growth occurred in the uninoculated cuvette or up to 30 days. The system was also used with Enterococcus faecalis. Completely glue-covered dentin specimens served as negative controls. Brown & Brenn-stained histological preparations of the specimens were examined with light microscopy. The time needed before growth occurred in the uninoculated cuvette showed great variation with C. albicans, whereas E. faecalis penetrated within 1 to 5 days of incubation. Slight penetration both by hyphae and yeast cells was observed in specimens inoculated with C. albicans, whereas specimens inoculated with E. faecalis showed deep and effective penetration. This study demonstrates the penetration of dentin as a possible pathway of infection by C. albicans. However, dentin penetration by C. albicans was slow and limited in comparison with E. faecalis."
In vitro susceptibility of Candida albicans isolates from apical and marginal periodontitis to common antifungal agents.Waltimo TM, Orstavik D, Meurman JH, Samaranayake LP, Haapasalo MP. Oral Microbiol Immunol. 2000 Aug;15(4):245-8. [abstract only]
"The susceptibility of a total of 70 Candida albicans strains to five common antifungal agents was determined. Thirty-five of the strains were isolated from persistent cases of apical periodontitis and 35 from cases of marginal periodontitis. The susceptibility of the strains to amphotericin B, 5-fluorocytosine and three azoles: fluconazole, miconazole and clotrimazole, was tested. The antifungal agents and yeast inoculums were prepared according to the NCCLS (National Committee for Clinical Laboratory Standards) recommendations. The yeasts were incubated with ten different concentrations of antifungal agents at 35 degrees C for 48 h. Yeast growth was measured spectrophotometrically. All strains from both isolation sources were susceptible to low concentrations of amphotericin B and 5-fluorocytosine, whereas the susceptibility to the three azoles varied, and three of the strains showed azole cross-resistance. These findings are in agreement with recent reports of increased azole resistance in Candida species in general and suggest the possibility that the oral cavity may act as a reservoir of resistant yeast isolates in systemic infections."
In vitro susceptibility of Candida albicans to four disinfectants and their combinations.Waltimo TM, Orstavik D, Siren EK, Haapasalo MP. Int Endod J. 1999 Nov;32(6):421-9.
"AIM: The aim of this study was to evaluate the susceptibility of seven strains of Candida albicans to four disinfectants: iodine potassium iodide, chlorhexidine acetate, sodium hypochlorite and calcium hydroxide. In addition, all possible pairs of the disinfectants were tested in order to compare the effect of the combination and its components.
METHODOLOGY: Filter paper discs were immersed in standardized yeast suspensions and then transferred to disinfectant solutions of different concentrations and incubated at 37 degrees C for 30 s, 5 min, 1 h and 24 h. After incubation the filter paper discs were transferred to vials with PBS and glass beads that were then vigorously shaken for dispersal of the yeast cells. PBS with resuspended yeasts was serially diluted 10-fold. Droplets of 25 microL from each dilution were inoculated on TSB agar plates and incubated in air at 37 degrees C for 24 h. The number of colony-forming units was then calculated from appropriate dilutions.
RESULTS: C. albicans cells were highly resistant to calcium hydroxide. Sodium hypochlorite (5% and 0.5%) and iodine (2%) potassium iodide (4%) killed all yeast cells within 30 s, whilst chlorhexidine acetate (0.5%) showed complete killing after 5 min. Combinations of disinfectants were equally or less effective than the more effective component. All C. albicans strains tested showed similar susceptibility to the medicaments tested.
CONCLUSIONS: This study indicates that sodium hypochlorite, iodine potassium iodide and chlorhexidine acetate are more effective than calcium hydroxide against C. albicans in vitro. However, combining calcium hydroxide with sodium hypochlorite or chlorhexidine may provide a wide-spectrum antimicrobial preparation with a long-lasting effect."
Susceptibility of oral Candida species to calcium hydroxide in vitro.Waltimo TM, Siren EK, Orstavik D, Haapasalo MP. Int Endod J. 1999 Mar;32(2):94-8. [abstract only]
"AIM: The susceptibility of common oral Candida species to saturated aqueous calcium hydroxide solution was studied.
METHODOLOGY: The yeast species tested were C. albicans (16 strains). C. glabrata (three strains), C. guilliermondii (three strains), C. krusei (two strains), and C. tropicalis (two strains). At least one reference strain of each species was used; the others were clinical isolates either from persistent apical periodontitis or from marginal periodontitis. The susceptibility of Enterococcus faecalis (ATCC 29212) was studied for comparative purposes. Standardized inocula of the strains were incubated in aqueous calcium hydroxide solution, pH 12.4, for time-periods ranging from 5 min to 6 h. Volumes of 0.1 mL of the test suspension were cultured directly on Brucella blood agar and incubated in air at 37C. The plates were inspected for growth at 24 and 48 h and the colonies were counted. The time required to reduce the number of colony-forming units to less than 0.1% of the initial number was determined for each strain.
RESULTS: The sensitivity of the C. albicans strains was generally low, with 16 h of incubation required to kill 99.9% of the colony-forming units. No differences in susceptibility between C. albicans strains isolated from root-canal infections and from periodontitis were found. Both strains of C. tropicalis were killed between 3 and 6 h of incubation, whilst strains of C. guilliermondii were killed after only 1020 min of incubation. All strains of C. glabrata survived 20 min, but not 1 h, of incubation, whilst 13 h were required to kill C. krusei. Compared with E. faecalis, all Candida spp. showed either equally high or higher resistance to aqueous calcium hydroxide.
CONCLUSIONS: This study indicates that Candida spp. are resistant to calcium hydroxide in vitro, which may explain the isolation of yeasts from cases of persistent apical periodontitis."
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