The effect of cross-linker on endogenous enzymatic activity within radicular dentin and bond strength between intraradicular post and radicular dentin

The primary aim was to evaluate the effect of 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide (EDC) on endogenous enzymatic activity within radicular dentin and push-out bond strength of adhesively luted fiber posts, at baseline and after artificial aging. Additionally, the effect of different cementation strategies on endogenous enzymatic activity and fiber post retention was evaluated. The experiment was carried out on extracted human teeth, following endodontic treatment and fiber post cementation. Three cementation strategies were performed: resin cement in combination with etch-and-rinse (EAR) adhesive system, with self-etch (SE) system and self-adhesive (SE) cement. Each of the mentioned strategies had a control and experimental (EDC) group in which root canal was irrigated with 0.3M EDC for 1 minute. The push-out bond strength test was performed 24h after cementation and after 40.000 thermocycles. In order to investigate the effect of EDC and different cementation strategies, in situ zymography analyses of the resin-dentin interfaces were conducted. Statistical analyses were conducted with the software Stata 12.0 (Stata Corp, College Station, Texas, USA) and the significance was set for p<0.05. The results of statistical analysis (ANOVA) showed that the variables “EDC”, “root region” and “artificial aging” significantly influenced fiber posts’ retention to root canal (p<0.05). The highest values were observed in coronal third. The mean values observed after artificial aging were lower when compared to baseline, however EDC was effective in preserving bond strength. The level of enzymatic activity varied between the groups and EDC had a beneficial effect on silencing the enzymatic activity. Within the limitations of the study, it was concluded that the choice of cementation strategy did not influence posts’ retention, while EDC contributed to the preservation of bond strength after artificial aging and reduced enzymatic activity within radicular dentin. In vivo trials are necessary to confirm the results of this in vitro study.

Development of new collagen cross-linkers for adhesive bonding stabilization

Aims: This thesis aimed to investigate the influence of different collagen cross-linkers, as separate primers or contained within desensitizing agents, on the longevity of dental restorations and on the dentinal enzymatic activity immediately, or after aging in vitro. Methods: A series of studies was conducted using several different cross-linking molecules and several adhesive systems. Four studies investigated the longevity of the hybrid layer by means of microtensile bond strength test, and the enzymatic activity using gelatin and in situ zymography, immediately or after 1 year of aging in the artificial saliva. The first study tested samples bonded with or without a cross-linking agent, that were previously aged for 5 years. The degradation of the hybrid layer was observed using transmission electron microscopy, the enzymatic activity in the hybrid layer using in situ zymography. Raman spectroscopy was used to investigate whether the active substance was still within the hybrid layer after 5 years. Results: The results of the studies showed that collagen cross-linkers were efficient in preserving bond strength after aging in vitro when used as separate primers on demineralized or partially demineralized dentin. In the cases when the cross-linker was utilized on mineralized dentin, bond strength results were higher than in the control groups immediately and after aging, however, no difference in enzymatic activity was detected after aging. Conclusions: The tested cross-linker molecules used as separate primers in etch-and-rinse and self-etch adhesives seem to be clinically applicable, since the procedure is not overly time-consuming and seems to preserve the hybrid layer over time. As for the cross-linkers contained in the desensitizing agent, when utilized before the adhesive procedures, it has shown to increase the bond strength of self-etch adhesives, but further studies are needed to better understand its effect on the enzymatic activity and crosslinking effects on mineralized dentin.