Gabriel Flores Abuna

Bonding performance of experimental bioactive/biomimetic self-etch adhesives doped with calcium-phosphate fillers and biomimetic analogs of phosphoproteins

OBJECTIVES This study examined the bonding performance and dentin remineralization potential of an experimental adhesive containing calcium-phosphate (Ca/P) micro-fillers, and self-etching primers doped with phosphoprotein biomimetic analogs (polyacrylic acid-(PAA) and/or sodium trimetaphosphate-(TMP)). METHODS Experimental self-etching primers doped with biomimetic analogs (PAA and/or TMP), and an adhesive containing Ca(2+), PO4(-3)-releasing micro-fillers (Ca/P) were formulated. Sound human dentin specimens were bonded and cut into sticks after aging (24h or 6 months) under simulated pulpal pressure (20cm H2O), and tested for microtensile bond strength (μTBS). Results were analyzed using two-way ANOVA and Tukeys test (p<0.05). Interfacial silver nanoleakage was assessed using SEM. Remineralization of EDTA-demineralized dentin was assessed through FTIR and TEM ultrastructural analysis. RESULTS Application of the Ca/P-doped adhesive with or without dentin pre-treatments with the primer containing both biomimetic analogs (PAA and TMP) promoted stable μTBS over 6 months. Conversely, μTBS of the control primer and filler-free adhesive significantly decreased after 6 months. Nanoleakage decreased within the resin-dentin interfaces created using the Ca/P-doped adhesives. EDTA-demineralized dentin specimens treated the Ca/P-doped adhesive and the primer containing PAA and TMP showed phosphate uptake (FTIR analysis), as well as deposition of needle-like crystallites at intrafibrillar level (TEM analysis). SIGNIFICANCE The use of Ca/P-doped self-etching adhesives applied in combination with analogs of phosphoproteins provides durable resin-dentin bonds. This approach may represent a suitable bonding strategy for remineralization of intrafibrillar dentin collagen within the resin-dentin interface.

The effect of proanthocyanidin-containing 10% phosphoric acid on bonding properties and MMP inhibition

OBJECTIVES This study evaluated the effect of etching using 2% proanthocynidin-containing 10% phosphoric acid 2% PA/10% PhA vs. 35% phosphoric acid 35% PhA on immediate (IM) and 6-months (6M) resin-enamel microshear bond strength (μSBS), resin-dentin microtensile bond strength (μTBS), nanoleakage (NL) and as well as in situ MMP inhibition potential. METHODS The dentin surface of human were exposed and then etched using 35% phosphoric acid for 15s or 2% PA/10% phosphoric acid for 30s. After rinsing with water, the dentin was bonded with Single Bond Plus (3M ESPE) and composite build-ups were constructed, followed by polymerization. The teeth were sectioned and the bonds were testing for microtensile bond strength (μTBS) and by SEM for NL analysis at IM and 6M. For MMP activity, resin-dentin slices were prepared for in situ zymography, and analyzed under confocal microscopy. For μSBS, others teeth had flattened enamel surfaces etched according the experimental groups and prepared to microshear procedure. The specimens were tested IM and after 6M by microshear bond strength. The data were submitted to two-way repeated measures ANOVA and Tukeys test (α=0.05). RESULTS Acid-etching using the 2% PA/10% phosphoric acid did not lower the μTBS in IM (p>0.05) compared to the control 35% phosphoric acid group. However, after 6M, only the 2% PA/10% PhA etched dentin had remained stable the resin-dentin bond strength (p<0.05). Bonds made with 35% PhA showed significant increase in NL% after 6M (p<0.05). Dentin bonds made with 2% PA/10% phosphoric acid showed no increase in NL% after 6 months. The MMP activity within the resin-dentin interface was almost completely reduced after 2% PA/10% PhA etching, while the 35% PhA exhibited intense MMP activity. For μSBS, the type of etchant and the storage period did not affect the resin-enamel bond strengths (p>0.05). SIGNIFICANCE Ten percent phosphoric acid containing 2% PA can produce stable resin-dentin and enamel-resin interfaces, without requiring additional steps in the bonding procedure. Future studies for longer evaluation time are required.

Cytotoxicity and the effect of cationic peptide fragments against cariogenic bacteria under planktonic and biofilm conditions

Abstract This study evaluated the cytotoxicity and effect of fragments derived from three oral cationic peptides (CP): LL-37, D6-17 and D1-23 against cariogenic bacteria under planktonic and biofilm conditions. For cytotoxicity analysis, two epithelial cell lines were used. The minimum inhibitory concentration and the minimal bactericidal concentration were determined for the CP fragments and the control (chlorhexidine-CHX) against cariogenic bacteria. The fractional inhibitory concentration was obtained for the combinations of CP fragments on Streptococcus mutans. Biofilm assays were conducted with the best antimicrobial CP fragment against S. mutans. The results indicated that D6-17 was not cytotoxic. D1-23, LL-37 and CHX were not cytotoxic in low concentrations. D1-23 presented the best bactericidal activity against S. mutans, S. mitis and S. salivarius. Combinations of CP fragments did not show a synergic effect. D1-23 presented a higher activity against S. mutans biofilm than CHX. It was concluded that D1-23 showed a substantial effect against cariogenic bacteria and low cytotoxicity.

Morphology of sealant/enamel interface after surface treatment with bioactive glass

The purpose of this study was to analyze, by confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM), the morphology of sealant/enamel interface after surface treatment with Biosilicate. Before pits and fissures sealing, the occlusal surfaces of 10 sound human molars were sectioned perpendicularly at the fissures in order to obtain three slices for each tooth. Slices were randomly assigned into three groups (n = 10) according to sealing protocol: Group 1‐ Acid etching + Biosilicate + glass ionomer‐based sealant (Clinpro XT Varnish, 3M ESPE); Group 2‐ Acid etching + glass ionomer‐based sealant (Clinpro XT Varnish, 3M ESPE); Group 3‐ No sealing. All slices were subjected to thermal cycling (5,000 cycles; 5–55°C; dwell time: 30s). Half of the slices from each group (n = 5) were analyzed by CLSM and the other half by SEM. Groups 1 and 2 were also submitted to EDS analysis and their data were evaluated by Two‐Way ANOVA e Tukeys test (α=5%). EDS data analysis showed higher amounts of silicon (Si) ions than calcium (Ca) ions in Group 1 (P < 0.05); Group 2 presented higher amounts (P < 0.05) of Ca ions than Si ions. It may be concluded that the use of Biosilicate for surface treatment did not affect the morphology of glass ionomer‐based sealant/enamel interfaces. Microsc. Res. Tech. 78:1062–1068, 2015.

Two methods to simulate intrapulpal pressure: effects upon bonding performance of self-etch adhesives

AIM To evaluate the effects of two methods to simulate physiological pulpal pressure on the dentine bonding performance of two all-in-one adhesives and a two-step self-etch silorane-based adhesive by means of microtensile bond strength (μTBS) and nanoleakage surveys. METHODOLOGY The self-etch adhesives [G-Bond Plus (GB), Adper Easy Bond (EB) and silorane adhesive (SIL)] were applied to flat deep dentine surfaces from extracted human molars. The restorations were constructed using resin composites Filtek Silorane or Filtek Z350 (3M ESPE). After 24 h using the two methods of simulated pulpal pressure or no pulpal pressure (control groups), the bonded teeth were cut into specimens and submitted to μTBS and silver uptake examination. Results were analysed with two-way anova and Tukeys test (P < 0.05). RESULTS Both methods of simulated pulpal pressure led statistically similar μTBS for all adhesives. No difference between control and pulpal pressure groups was found for SIL and GB. EB led significant drop (P = 0.002) in bond strength under pulpal pressure. Silver impregnation was increased after both methods of simulated pulpal pressure for all adhesives, and it was similar between the simulated pulpal pressure methods. CONCLUSIONS The innovative method to simulate pulpal pressure behaved similarly to the classic one and could be used as an alternative. The HEMA-free one-step and the two-step self-etch adhesives had acceptable resistance against pulpal pressure, unlike the HEMA-rich adhesive.

Optimization of the etch-and-rinse technique: New perspectives to improve resin–dentin bonding and hybrid layer integrity by reducing residual water using dimethyl sulfoxide pretreatments

OBJECTIVE To determine whether bonding effectiveness and hybrid layer integrity on acid-etched dehydrated dentin would be comparable to the conventional wet-bonding technique through new dentin biomodification approaches using dimethyl sulfoxide (DMSO). METHODS Etched dentin surfaces from extracted sound molars were randomly bonded in wet or dry conditions (30s air drying) with DMSO/ethanol or DMSO/H2O as pretreatments using a simplified (Scotchbond Universal Adhesive, 3M ESPE: SU) and a multi-step (Adper Scotchbond Multi-Purpose, 3M ESPE: SBMP) etch-and-rinse adhesives. Untreated dentin surfaces served as control. Bonded teeth (n=8) were stored in distilled water for 24h and sectioned into resin-dentin beams (0.8mm2) for microtensile bond strength test and quantitative interfacial nanoleakage analysis (n=8) under SEM. Additional teeth (n=2) were prepared for micropermeability assessment by CFLSM under simulated pulpar pressure (20cm H2O) using 5mM fluorescein as a tracer. Microtensile data was analyzed by 3-way ANOVA followed by Tukey Test and nanoleakage by Kruskal-Wallis and Dunn-Bonferroni multiple comparison test (α=0.05). RESULTS While dry-bonding of SBMP produced significantly lower bond strengths than wet-bonding (p<0.05), DMSO/H2O and DMSO/ethanol produced significantly higher bond strengths for SBMP irrespective of dentin condition (p<0.05). SU presented significantly higher nanoleakage levels (p<0.05) and micropermeability than SBMP. Improvement in hybrid layer integrity occurred for SBMP and SU for both pretreatments, albeit most pronouncedly for DMSO/ethanol regardless of dentin moisture. CONCLUSION DMSO pretreatments may be used as a new suitable strategy to improve bonding of water-based adhesives to demineralized air-dried dentin beyond conventional wet-bonding. Less porous resin-dentin interfaces with higher bond strengths on air-dried etched dentin were achieved; nonetheless, overall efficiency varied according to DMSOs co-solvent and adhesive type. CLINICAL SIGNIFICANCE DMSO pretreatments permit etched dentin to be air-dried before hybridization facilitating residual water removal and thus improving bonding effectiveness. This challenges the current paradigm of wet-bonding requirement for the etch-and-rinse approach creating new possibilities to enhance the clinical longevity of resin-dentin interfaces.

Influence of Photoinitiator on Accelerated Artificial Aging and Bond Strength of Experimental Resin Cements

The goal of this study was to evaluate in vitro the effect of the photoinitiator phenylpropanedione (PPD), alone or combined with camphorquinone (CQ), on color stability of photoactivated resin cements and their bond strength to ceramics using a micro-shear test. Four resin cements were used: a commercial brand cement (RelyX Veneer®) and 3 experimental cements with different types and concentration of photoinitiators. For color analysis, ceramic discs were cemented on bovine dentin specimens to simulate indirect restorations (n=8) and were exposed to UV for 120 h and tested for color alteration using a reflectance spectrophotometer and the CIEL*a*b* system. Data were analyzed by Anova and Tukeys test at 5% significance level. The color test results did not present statistically significant difference for the ∆E for all the studied cements, neither for ∆L, ∆a and ∆b. For the bond strength, all the studied cements showed statistically significant differences to each other, with the highest result for the RelyX Veneer® (29.07 MPa) cement, followed by the cement with CQ (21.74 MPa) and CQ+PPD (19.09 MPa) cement; the lowest result was obtained by the cement using only PPD as a photoinitiator (13.99 MPa). So, based on the studied parameters, PPD was not advantageous as photoinitiator of resin cements, because it showed a low value of bond strength to the ceramics and no superior color stability.

Microtensile Bond Strength of Adhesive Systems in Different Dentin Regions on a Class II Cavity Configuration

The aim of this study was to evaluate microtensile bond strength (µTBS) of self-etch and etch-and-rinse adhesives systems compared in different dentin regions (central-CD or proximal-PD) in a class II cavity configuration. A class II (mesial-oclusal-distal) cavity configuration was simulated on 20 extracted human third-molars (4 mm wide/3 mm deep). Etch-and-rinse adhesive (Scotchbond Multi Purpose, n=5, SBMP and Optibond FL, n=5, OPFL) and self-etch adhesives (Clearfil SE Bond, n=5, CSE and Optibond XTR, n=5, OPXTR) were applied. Class II restorations were performed by incremental technique and photo-activated (Bluephase/G2). Samples were sectioned to beam shape (1 mm² cross-section), placed on Geraldelis device for µTBS test (0.5 mm/min cross-head speed). Fracture patterns were analyzed on stereomicroscope and classified as cohesive-resin, adhesive, mixed/resin or mixed/dentin. Samples (n=4) were prepared for scanning electron microscope observation. Data were submitted to one-way ANOVA with Split-Plot arrangement and Tukeys test (α=0.05). There were no statistically significant differences among SBMP, OPFL, CSE and OPXTR on CD (p>0.05). However, on PD for SBMP and OPFL, µTBS values were significantly lower compared to CSE and OPXTR (p<0.05). In all groups, mixed failure pattern was more frequently observed, except for SBMP/CD (adhesive). In class II type cavity configuration, PD location negatively influenced bond strength of etch-and-rinse adhesive systems. Opposite to self-etching adhesives, which presented higher bond strength values compared to etch-and-rinse adhesives in PD.

KR-12-a5 is a non-cytotoxic agent with potent antimicrobial effects against oral pathogens

Abstract This study evaluated the cytotoxicity and antimicrobial activity of analogs of cationic peptides against microorganisms associated with endodontic infections. L-929 fibroblasts were exposed to LL-37, KR-12-a5 and hBD-3–1CV and chlorhexidine (CHX, control), and cell metabolism was evaluated with MTT. The minimal inhibitory concentration (MIC) and the minimal bactericidal/fungicidal concentration (MBC/MFC) of the peptides and CHX were determined against oral pathogens associated with endodontic infections. Enterococcus faecalis and Streptococcus mutans biofilms were cultivated in bovine dentin blocks, exposed to different concentrations of the most efficient antimicrobial peptide and analyzed by confocal laser scanning microscopy. CHX and peptides affected the metabolism of L-929 at concentrations > 31.25 and 500 μg ml−1, respectively. Among the peptides, KR-12-a5 inhibited growth of both the microorganisms tested with the lowest MIC/MBC/MFC values. In addition, KR-12-a5 significantly reduced E. faecalis and S. mutans biofilms inside dentin tubules. In conclusion, KR-12-a5 is a non-cytotoxic agent with potent antimicrobial and anti-biofilm activity against oral pathogens associated with endodontic infections.