The presence Tanespimycin of five different Candida species (C. albicans, C. dubliniensis, C. glabrata, C. krusei and C. tropicalis) was simultaneously investigated using the DNA checkerboard hybridisation method. Additionally, we have correlated these findings to differences in surface roughness and total amount of formed biofilm. The null hypotheses were as follows: (I) there are no significant differences in terms of cell counts between target species for tested materials and (II) there is a positive correlation between count and surface roughness and the total amount of formed biofilm. Six healthy men aged between 21 and 27 years (mean age: 24 years) were enrolled in the study. The subjects

selected had no clinical signs of diseases in the oral mucosa and the gingival sulci were <3 mm deep without clinical signs of inflammation. Additional exclusion criteria were pregnancy, lactation, periodontal or antibiotic treatment in the earlier 3 months, current smokers or any systemic disease that could influence the periodontal status. The sample size was determined by means of sample size estimation for comparison of means considering estimated standard deviations (SDs). The study was approved by the local ethics committee (Ethical

Committee of the Faculty of Dentistry of Ribeirão Preto) and all the experiments were undertaken with the understanding and written consent of each subject according to the ethical principles (Process number: 2011.1.371.583). For this microbiological study, two different types of titanium and one type of Fulvestrant in vivo ceramic specimen (10 mm in diameter and 2 mm in thickness) were used to evaluate the oral biofilm formation and composition after oral cavity exposure.

Three individual removable intraoral acrylic upper jaw splints for mounting test disc samples were fabricated for each subject, one for each type of evaluated substrate. Four disc specimens of the same material were fixed in the buccal region of each GBA3 splint, two positioned in the anterior region (incisive) and two in the posterior region (premolar). The entire tested surface of each material was totally exposed in the oral cavity after mounting. Before contamination test, all the splints containing mounted specimens were sterilised with hydrogen peroxide plasma for 60 min. Subjects were advised to use each splint for 24 h, removing it only for food consumption and tooth brushing. A period of 1 week was stated as ‘washout’ between each tested splint. After enrolment, participants randomly received the following three splint interventions, according to a ‘crossover’ design: (1) machined pure titanium (MPT; n = 24) specimens, (2) zirconia (Zc; n = 24) specimens and (3) cast and polished titanium (CPT; n = 24) specimens. MPT and Zc discs were obtained from Neodent® (Neodent, Curitiba, PR, Brazil).