Chitosan-Based Extrafibrillar Demineralization for Dentin Bonding.

Instability of resin-dentin bonds is the Achilles' heel of adhesive dentistry. To address this problem, a chelate-and-rinse extrafibrillar dentin demineralization strategy has been developed that keeps intrafibrillar minerals within collagen fibrils intact to prevent activation of endogenous proteases that are responsible for collagen degradation within hybrid layers. The objective of the present study was to evaluate the potential of using chitosan >40 kDa as an antimicrobial extrafibrillar dentin-chelating agent to enhance bond durability. Transmission electron microscopy provided evidence for retention of intrafibrillar minerals and smear plugs in dentin conditioned with 1 wt% chitosan. Analyzed by Kruskal-Wallis analysis of variance, Dunn's statistic, and separate Mann-Whitney tests, tensile bond strengths to wet- and dry-bonded dentin indicated that chelating dentin with chitosan for 60 s prior to bonding did not result in a significant decline in resin-dentin bond strength when compared with that of phosphoric acid etching ( P > 0.05). Gelatinolytic activity within the hybrid layers was examined via in situ zymography after 24-h storage or after thermomechanical cycling and analyzed with 3-factor analysis of variance. After 24 h, enzymatic activity was detected only within completely demineralized phosphoric acid-etched dentin, with values derived from dry bonding significantly higher than those derived from wet bonding ( P < 0.05). Negligible fluorescence was detected within hybrid layers when dentin was conditioned with chitosan, even after thermomechanical cycling, as compared with the controls. Reduction in water permeability in chitosan-conditioned dentin, attributed to smear plug retention, also fostered long-term bond stability. Antibacterial testing performed with live/dead staining indicated that the acetic acid-solubilized chitosan possessed antibacterial activities against 3 single-species biofilms: Streptococcus mutans, Actinomyces naeslundii, and Enterococcus faecalis. Taken together, the new chitosan-based extrafibrillar demineralization strategy retains intrafibrillar minerals, reduces endogenous protease-initiated collagen degradation, prevents water permeation within hybrid layers, and kills bacteria on dentin surfaces, which are crucial factors for enhancing resin-dentin bond durability.

[Effects of exogenous enzymes on the degradation of adhesive-dentin interfaces].

OBJECTIVE: To compare the effects of exogenous enzymes on the degradation of adhesive-dentin interface. METHODS: Forty molars were sectioned to expose the middle-coronal dentin surface and randomly divided into two adhesive systems: an etch-and-rinse adhesive Adper Single Bond 2 and a self-etching adhesive G-Bond. After composite building up, the specimens were then randomly assigned to four groups(n=5 for each group)as follows: group 1, 24 h of water storage(the control group); group 2, six months of water storage; group 3, twelve weeks storage in artificial saliva containing clostridium histolyticum collagenase; group 4, twelve weeks storage in artificial saliva containing cholesterolesterase. The microtensile bond strengths(MTBS)were then tested. The failure modes and nanoleakage were analyzed. RESULTS: After aging treatments, the three aging groups showed significantly lower MTBS compared with the control group in both adhesive systems(P<0.05). For etch-and-rinse adhesive Adper Single Bond 2, the MTBS of group 3([19.6±3.5]MPa)was lower than that of group 2([23.4±4.2]MPa)and group 4([24.2±4.2]MPa)(P<0.05). For self-etching adhesive G-Bond, there was no difference on MTBS among different aging groups(P>0.05). SEM observation showed that, compared with the control group, water storage(group 2)and the exogenous enzymes(group 3 and 4)increased the nanoleakage expression(silver deposition)of both adhesive systems. Adhesive failure was the predominant fracture modes in all groups. CONCLUSIONS: Storage in artificial saliva containing clostridium histolyticum collagenase or cholesterol esterase could be used to accelerate the degradation process of adhesive-dentine interface.