M.C. Giacomini

Role of Proteolytic Enzyme Inhibitors on Carious and Eroded Dentin Associated With a Universal Bonding System

OBJECTIVES The aim of this study was to evaluate the effect of proteolytic inhibitors on the bond strength of a universal adhesive system (etch-and-rinse mode) applied to artificial carious and eroded dentin. METHODS Ninety molars were prepared and randomly divided into three groups according to the substrate: N, no challenges; ACD, artificial carious dentin simulation and ERO, artificial erosion simulation with orange juice. All groups were redivided into three subgroups according to the dentin pretreatment: W, water; CHX, 2% digluconate chlorhexidine; and E-64 (trans-epoxysuccinyl-L-leucylamido-[4-guanidino] butane), 5 μM E-64 inhibitor. They constituted a total of nine groups (n=10): N-W, N-CHX, N-E64, ACD-W, ACD-CHX, ACD-E64, ERO-W, ERO-CHX, and ERO-E64. All specimens were restored with Adper Single Bond Universal/Filtek Z250. Beams (0.64 mm2) were obtained and subjected to the microtensile test (μTBS) in a universal testing machine at 0.5 mm/min. The failure mode of the interfaces was determined by optical microscopy (40× magnification). Data were statistically analyzed by three-way analysis of variance and Tukey tests (p<0.05). RESULTS All individual factors (p<0.0001) and the interaction between substrate and treatment (p=0.0011) and between substrate and time (p=0.0003) were statistically significant. The caries substrate contributed negatively to bond strength. Chlorhexidine reduced bond strength for normal and eroded conditions. Only the normal substrate was negatively affected by time despite the pretreatment. CONCLUSIONS The universal bonding system appears to be a promising bonding strategy for the maintenance of bond strength to affected dentin. E-64 did not affect bonding to the dentin in contrast to the use of chlorhexidine, which, when associated with the universal system, did affect the microtensile bond strength for artificial carious dentin.

Effect of ethanol-dissolved rhodamine B marker on mechanical properties of non-simplified adhesives

OBJECTIVES Rhodamine B (RB) is commonly used to evaluate dental polymers, including dental bonding systems (DBS). For reliability assessments, its effect should not only allow visualization of the dentin-polymer interface but also must not interfere with the bonding of the DBS to dentin as measured by the microtensile bond strength and hardness tests. MATERIAL AND METHODS Flat human dentin surfaces were prepared and randomly distributed (n = 10) into six groups: Adper Scotchbond Multi-Purpose (MP) or Clearfil SE Bond (SE) in concentrations of none/control, 0.02 or 0.1 mg/mL. These combinations were prepared through ethanol dissolution to improve their penetration into the dentin. All specimens were fabricated with Filtek Z250 (n = 10) and prepared for a microtensile bond test (μTBS) (0.5 mm/min) after 7 days and 6 months. The failure modes were determined using a stereomicroscope (×40). For the hardness test, flat human dentin blocks were prepared and treated as previously described (n = 6). The specimens were stored at 37 °C/48 h and were tested (Knoop indenter - 25 gF/10 s). Data were analyzed with two-way ANOVA and Tukey tests for multiple comparisons (α = 0.05). The effect of time was evaluated using the Student t-test. RESULTS For 7-day μTBS, both the DBS and RB concentrations were significant factors (p < 0.01). After 6 months, only the RB concentration was significantly different. Adhesive failures were prevalent for all groups. Regarding hardness, the DBS differed only with the use of 0.10 mg/mL of RB. CONCLUSIONS Ethanol-dissolved rhodamine B in concentrations of 0.02 and 0.10 mg/mL in non-simplified adhesives can affect the physical-mechanical properties of functional monomer-based systems rather more than those of BisGMA systems.

How proteolytic inhibitors interact with dentin on glass-fiber post luting over 6 months

OBJECTIVES Enzyme inhibitors minimize the degradation of unprotected collagen of dentin promoted by matrix metalloproteinases (MMPs) and cysteine cathepsins (CCs). As the evidence of their effect on the root canal is limited, this study aimed to evaluate the role of EDTA, chlorhexidine and E-64 as antiproteolytic agents on the bond strength (BS) of glass-fiber posts in root canals. MATERIALS AND METHODS Ninety-six bovine roots were distributed in groups for each time point (n = 8). Adper Scotchbond Multipurpose (MP)/ RelyX ARC system was used to lute the post according to the treatment: negative control (NC)- water, EDTA- 17% ethylenediaminetetraacetic acid, CHX- 2% digluconate chlorhexidine, E-64-5- 5μM E-64, E-64-10- 10μM E-64 and positive control (PC)- MP associated with activator/ catalyst. Then, slices were subjected to push-out test (0.5mm/min) after 24h/6 mons. Data were analyzed by three-way ANOVA/Tukey tests. Failure modes were analyzed (40×). RESULTS The factors treatment, time, root canal third and the interaction between treatment and time were statistically significant. At 24h, no negative interactions were observed among the root dentin, bonding system and post. At 6 mons, CHX improved the BS for middle and apical root thirds. CONCLUSIONS CHX was able to promote beneficial BS after 6 mons, which was not noted for any other tested enzyme inhibitors.

Use of sodium trimetaphosphate in the inhibition of dentin matrix metalloproteinases and as a remineralizing agent

OBJECTIVES Because of its ability to act as an antiproteolytic agent, the effect of sodium trimetaphosphate (STMP) against specific enzymes extracted from sound dentin and its performance under acidic challenge on demineralized dentin were investigated. METHODS The antiproteolytic potential of STMP (0.5%, 1.0%, and 1.5%) was assessed in triplicate by zymography. For the evaluation of remineralization activity, 50 bovine-root dentin specimens were selected and randomly divided into 5 groups (n=10). Three areas were determined for each specimen: 1) control (no treatment); 2) demineralized (artificial caries-like challenge); 3) treated (demineralized and subjected to pH-cycling for 7days, and treated for 10min with 1.5% STMP, 1.5% STMP+calcium hydroxide (Ca[OH]2), 1.5% STMP+sodium fluoride (NaF), NaF, or deionized H2O). The dentin specimens were analyzed for superficial hardness (SH) and cross-sectional hardness (CSH) at different depths (10, 30, 50, 70, 90, 110, and 220μm) using a Knoop penetrator (10g/10s). Statistical analyses were performed with analysis of variance (ANOVA) and Tukey tests (p<0.05). RESULTS The zymographic analysis showed that 1.5% STMP promoted complete inhibition of gelatinolytic activity. Therefore, 1.5% STMP was investigated in association with supplemented calcium or fluoride; a combination of 1.5% STMP and Ca(OH)2 significantly increased the mechanical properties of the treated dentin. CONCLUSION 1.5% STMP serves as an antiproteolytic agent against matrix metalloproteinases extracted from human dentin. Furthermore, when supplemented with Ca(OH)2, 1.5% STMP may potentially induce remineralization. CLINICAL SIGNIFICANCE STMP can be introduced as a novel strategy that combines enzymatic inhibition and remineralizing potential, which can serve to strengthen dentin and improve stability. STMP may have potential in the treatment of demineralized dentin lesions, especially when supplemented with calcium.

Sodium trimetaphosphate as a novel strategy for matrix metalloproteinase inhibition and dentin remineralization

The effect of sodium trimetaphosphate (STMP) as an antiproteolytic and remineralizing agent on demineralized dentin was evaluated in vitro. The inhibitory potential of STMP at 0.5, 1.5, 3.5, and 5% against recombinant matrix metalloproteinases (MMPs) MMPs-2 and -9 was assessed by zymography. To investigate its remineralization potential, 40 bovine root specimens were obtained and subjected to a demineralization protocol to produce caries-like dentin lesions. After that, dentin surfaces were divided into 3 areas: (1) mineralized (no treatment); (2) demineralized; and (3) demineralized/treated with STMP and submitted to a pH-cycling associated or not with STMP (1.5, 3.5, or 5% STMP, 10 min of treatment). After that, superficial hardness (SH) and cross-sectional hardness (CSH) were determined. Polarized light microscopy (PLM) was used to qualitatively evaluate mineralization within the caries-like lesions. The zymographic analysis showed that STMP solution is a potent inhibitor of the gelatinolytic activity of MMPs-2 and -9 depending on the dose, since the lowest concentration (0.5%) partially inhibited the enzyme activity, while the higher concentrations completely inhibited enzyme activity. Regarding remineralization effect, only 1.5% STMP solution enhanced both the SH and CSH. PLM showed that the area treated with 1.5% STMP presented similar birefringence as mineralized sound dentin. In conclusion, 1.5% STMP solution is effective as an antiproteolytic agent against MMPs and promotes dentin remineralization.

Influence of Erosive and Abrasive Cycling on Bonding of Different Adhesive Systems to Enamel: An In situ Study

This study evaluated the impact of orange juice on the bond strength (BS) of dentin bonding systems (DBSs) to enamel surface after simulation with an in situ/ ex vivo erosive cycling. One hundred and ninety two bovine enamel fragments (4x4x2mm) were obtained and randomized regarding superficial microhardness and distributed to palatal devices for 8 volunteers, in three phases (one for each DBS), containing 8 blocks, which were, allocated in 4 pairs. Daily, these pairs were subjected extraorally to the following conditions: CONT- neither erosive nor abrasive challenge; ERO- erosive challenge only; ABR- abrasive challenge only and ERO + ABR- with erosive and abrasive challenges. Erosive cycles (immersion in orange juice, 3 times/day/5 min/5 days) or/and abrasive challenges (electric toothbrush, 3 times/day/1 min/5 days) were performed. After these cycles, all specimens were restored with the adhesive systems Adper Scotchbond Multi Purpose (MP), Adper Single Bond 2 (SB) or Clearfil SE Bond (SE), and the composite resin Filtek Z250. After 7 days, sticks (area ≅1 mm2) were obtained and subjected to the microtensile bond strength test (μTBS) at 0.5 mm/min. Data was statistically analyzed by ANOVA and Tukey tests (a=0.05). Failure modes were determined using a digital microscope (40´). DBS was the only statistical significant factor. SE was the unique DBS not affected in any challenge, whereas MP and SB performed according to the scenario. The adhesive and mixed failures were predominant in all groups. Overall performance suggested that BS to enamel after erosive /abrasive challenged by orange juice was not affected and it was material-dependent.