Investigation of the effect of the coronal restoration quality on the composition of the root canal microflora in teeth with apical periodontitis by means of T-RFLP analysis

Aim To investigate the effect of the radiographic and clinical quality of coronal restorations on the composition of the root canal flora of teeth with necrotic pulps and teeth with root fillings associated with apical periodontitis. Methodology Twenty-eight necrotic pulps and 35 root filled canals with signs of apical periodontitis were studied. Both the coronal filling (presence of radiographically or clinically deficient margins and/or secondary caries) and the root filling (homogeneity and length) were scored. Bacterial root canal samples were taken with sterile paper points under rubber dam and using measures to prevent contamination. A DNA-based nonculture bacterial identification technique was used, namely terminal restriction fragment length polymorphism (T-RFLP) analysis. Results Twelve samples were negative for bacterial DNA. A total of 33 different terminal restriction fragments (TRFs) were detected. The Fusobacterium nucleatum/Streptococcus mitis group was the most frequently encountered TRF. The mean number of TRFs per necrotic pulp was 6.2 and 5.8 for the groups with acceptable and unacceptable coronal restorations, respectively. This difference was not significant. In the root filled group, these values (respectively, 5.2 and 8.6) were statistically significantly different (P<0.05). The following parameters in root filled teeth had no significant influence on the mean numbers of TRFs detected: the length and homogeneity of the root filling and the type of tooth (anterior-premolar-molar). Conclusion T-RFLP allowed the rapid assessment of bacterial biodiversity in root canal samples. The technique revealed the presence of bacteria that have rarely been described in the root canals of teeth with apical periodontitis. Biodiversity in the root filled group was high, as compared with culture-dependent studies where monoinfections were more frequently reported. Only in root filled teeth did defective coronal restorations have a statistically significant influence on the mean numbers of detected TRFs per sample.