universidade de são paulo - Faculdade de Odontologia - Unesp

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pooled and clarified by centrifugation at 10000 g for 5 min at 4 ºC 18 and then
sterilized using membrane filter with a 0.22 μm pore size 10,13,27,28 . The resulting
saliva was immediately stored at -70 ºC until use. The study was approved by the
Ethics Committee of Araraquara Dental School, and all subjects volunteered to
participate and signed an informed consent form.
Test specimen sterilization
Before the microbiological tests, the test specimens were kept in distilled
water at ambient temperature for 48 hours, in order to eliminate residual
monomers 18 . After this, they received an ultrasound bath for 20 minutes and were
exposed to ultraviolet light in the laminar flow chamber for 20 minutes on each
side 29 .
Preconditioning with saliva
The 90 test specimens were divided into five groups (n=18), as follows: 1
control (without preconditioning in saliva) and 4 experimental groups that were
conditioned with saliva for periods of: 30 minutes (30 min); 1 hour (60 min); 3
hours (180 min); or 12 hours (720 min). The specimens of the four experimental
groups were incubated in 12-well microtiter plates with 3 ml of the saliva
preparation at room temperature 17 prior to the adherence assay.
Adherence assay
The stock culture of Candida albicans strain ATCC 90028 was maintained
in YEPD medium (1% yeast extract, 2% peptone, 2% dextrose, 2% agar) stored at