RESUMO
The present study investigated, for the first time, enzymes in the in-situ pellicle of children. Furthermore, it was purposed to detect glucosyltransferase (GTF) isoforms in each child's pellicle. Twenty-four children (5-9 yr of age) participated in the study. Twelve were caries free with no decayed, missing, and filled teeth (dmft), whereas 12 had active caries (dmft ≥ 2, indicating at least two untreated carious lesions). Bovine enamel slabs, fixed on individual upper jaw splints, were utilized for pellicle formation in situ. After 3 and 30 min, samples were tested for amylase, lysozyme, and peroxidase activities; total GTF activity was examined only in the 30-min pellicle. Gold-immunolabelling was used to quantify the GTF B, C, and D isoforms in the pellicle by transmission electron microscopy (TEM). All enzymes tested were detected in the children's in-situ pellicle in an active conformation, and there were no significant differences in their levels of activity between caries-free and caries-active children. All GTF isoforms were found to be randomly distributed within all pellicle layers, althoug GTF B was only detected very sporadically. A significantly higher amount of GTF D was detected in the pellicle of caries-active children. Pellicle formation in children is characterized by uniformity and selectivity. Glucosyltransferase D might represent a possible biomarker for high caries risk in children.
RESUMO
The present in situ study investigated the influence of a preparation containing CPP/ACP (caseinphosphopeptide-amorphous calcium phosphate) (GC Tooth mousse) on initial bacterial colonization of enamel and dentin. Therefore, pellicle formation was performed in situ on bovine enamel and dentin specimens fixed to individual upper jaw splints worn by 8 subjects. After 1 min of pellicle formation GC Tooth mousse was used according to manufacturer's recommendations. Rinses with chlorhexidine served as positive controls. Specimens carried without any rinse served as negative controls. After 8 h overnight exposure of the splints, bacterial colonization was quantified by fluorescence microscopy (DAPI and BacLight live/dead staining). Additionally, the colony forming units (CFU) were determined after desorption. Furthermore, the effects on Streptococcus mutans bacteria were tested in vitro (BacLight). There was no significant impact of CPP/ACP on initial bacterial colonization proved with DAPI and BacLight. Determination of CFU showed statistical significance for CPP/ACP to reduce bacterial adherence on enamel. The in vitro investigation indicated no antimicrobial effects for CPP/ACP on Streptococcus mutans suspension. Under the chosen conditions, CPP/ACP (GC Tooth mousse) had no significant impact on initial biofilm formation on dental hard tissues. The tested preparation cannot be recommended for biofilm management.