Assessing the Antimicrobial Properties of Copper-Iodide Doped Adhesives in an In vitro Caries Model: A Pilot Study.

Context: Recurrent caries are the leading cause of composite resin failure. Aims: The purpose of this pilot study was to test the efficacy of a novel copper iodide (CuI) containing dental adhesive in an in vitro caries model. Subjects and Methods: Streptococcus mutans and Lactobacillus acidophilus were grown individually on the complex medium for 48 h at 37°C. The pH of the mixed medium was 7.0 initially and tested every 24 h. 40 extracted teeth were prepared with standardized cavity preparations and coated with control or experimental CuI adhesives and imaged using a micro-computed tomography (microCT). Four study groups were evaluated: (1) control (2) 0.5 µg/ml CuI (3) 1.0 µg/ml CuI, 4) 5.0 µg/ml CuI. After incubation, the teeth were re-imaged using the microCT. Utilizing AnalyzePro software the three-dimensional data sets were overlaid and demineralization was measured and statistics were run. Statistics: Stratified ANOVA models were run to determine if there were differences between the control and experimental adhesive groups. Similarly, pH and bacterial concentrations were evaluated to ensure the viability of polymicrobial specimen. Results and Conclusions: Significant differences were found between the control group and the 1.0 and 5.0 CuI adhesive groups. No differences in pH were noted between the groups. Overlaid changes in demineralization were recorded as volume loss. CuI adhesives with 5 mg/ml or higher have the potential to limit tooth demineralization after bacterial penetration of a dental restoration in an in vitro caries model. Further testing is needed.

Dental Pulp Cell Conditioning through Polyinosinic-Polycytidylic Acid Activation of Toll-like Receptor 3 for Amplification of Trophic Factors.

INTRODUCTION: Regeneration of the pulp-dentin complex hinges on functionally diverse growth factors, cytokines, chemokines, signaling molecules, and other secreted factors collectively referred to as trophic factors. The delivery of exogenous factors and the induced release of endogenous dentin-bound factors by conditioning agents have been explored toward these goals. The aim of this study was to investigate a promising regeneration strategy based on the conditioning of dental pulp cells (DPCs) with polyinosinic-polycytidylic acid (poly[I:C]) for the amplification of endogenous trophic factors. METHODS: DPCs were isolated from human dental pulps, propagated in culture, and treated with an optimized dose of poly(I:C). The 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay and metabolite analysis were conducted to monitor the cytotoxicity of poly(I:C). Enzyme-linked immunosorbent assays and quantitative polymerase chain reaction assays were performed to quantify the induction of trophic factors in response to DPC conditioning. Statistical significance was P < .05. RESULTS: The analysis of 32 trophic factors involved in Wnt signaling, cell migration and chemotaxis, cell proliferation and differentiation, extracellular matrix remodeling and angiogenesis, and immunoregulation revealed that DPCs abundantly express many trophic factors including AMF, BDNF, BMP2, FGF1, FGF2, FGF5, HGF, MCP1, NGF, SDF1, TGFß1, TIMP1, TIMP2, TIMP3, and VEGFA, many of which were further induced by DPC conditioning; induction was significant for BDNF, EGF, HGF, LIF, MCP1, SDF1, IL6, IL11, MMP9, and TIMP1. Both DPC proliferation and lactate production (P < .05) were inhibited by 8 µg/mL poly(I:C) relative to the control. CONCLUSIONS: In vitro DPC conditioning through poly(I:C) activation of toll-like receptor 3 led to the amplification of trophic factors involved in tissue repair. The strategy offers promise for endodontic regeneration and tooth repair and warrants further investigation.

Mechanical characterization and adhesive properties of a dental adhesive modified with a polymer antibiotic conjugate.

This study is a follow up investigation on recent work by our group demonstrating synthesis, release and strong antibacterial character of resins modified with penicillin V (PV)-based polymer-antibiotic conjugates (PACs). Here, we aimed to evaluate the mechanical, bonding, and other relevant biomedical properties of a commercial adhesive resin modified with PV-PAC. Single Bond Plus (SB+) was modified with PAC containing 1.8 wt% conjugated PV. Adhesive resins were bonded to dentin from extracted human molars and restorative resin added. Beams of cross-sectional area of 0.9 ± 0.1 mm (Kutsch and Young, 2011) (n = 20) were obtained from the molars and tested for micro-tensile bond strength (µTBS) at 24 h and 4 months. For cohesive strength, hourglass beams (10 × 2 × 1 mm; n = 10) were assessed for ultimate tensile strength (UTS), beam-shaped specimens (25x2x2 mm; n = 10) evaluated for flexural strength and modulus (FS/FM) via three-point bending, and cylindrical specimens (3 × 2 mm; n = 10) assessed for ultimate compressive strength (UCS). For surface micro-hardness (MH), cylindrical specimens (3 × 2 mm; n = 6) were assessed before and after an EtOH challenge. The degree of conversion (DC) (5 × 1 mm; n = 6) was determined based on changes in absorbance ratio between peaks at ∼1637 cm-1 and ∼1608 cm-1 before and after curing of adhesive resins using attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. To monitor water uptake and diffusion kinetics over a 28-day period, specimens (5 × 1mm) were desiccated, weighed and stored in deionized water. Control and PV-PAC modified adhesive resins demonstrated similar µTBS at 24 h and 4 months; both showing decrease in values after 4 months (p = 0.001 and 0.004). No significant differences between adhesive resins were shown in UTS, FS/FM or UCS (p<0.05). MH of PV-PAC adhesive resin was significantly reduced relative to the control (p<0.001). The DC values of the adhesive resins were not significantly different. While sorption and solubility were no different between materials, the diffusion coefficient of PV-PAC modified adhesive resin was higher than the control (p<0.001). We conclude that incorporation of PV-PAC with 1.8 wt% PV into an adhesive resin does not adversely affect its mechanical, bonding, and physical properties, thus providing a promising option for materials with long-term antibacterial character and on-demand release.

Synthesis and antibacterial activity of polymer-antibiotic conjugates incorporated into a resin-based dental adhesive.

This work reports on polymer-antibiotic conjugates (PACs) as additives to resin-based restorative dental materials as a new strategy to convey sustained antibacterial character to these materials. Such antibacterial performance is expected to improve their longevity in the oral cavity. Using the previously reported ciprofloxacin (Cip)-based PAC as a control, a penicillin V (PV)-based PAC was investigated. The monomer-antibiotic conjugate (MAC) containing a methacrylate monomer group and a PV moiety was prepared via nucleophilic substitution between 2-chloroethyl methacrylate (CEMA) and penicillin V potassium (PVK). The PV-based PAC was synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization of the MAC with hydroxyethyl methacrylate (HEMA), and further characterized by 1H NMR and gel permeation chromatography (GPC) analysis. Antibiotic resistance was investigated by passaging bacteria in low concentrations of the antibiotic for 19 days, followed by a 48 h challenge at higher concentrations. Our results suggest that the development of antibiotic resistance is unlikely. Zone of inhibition (ZOI) assays revealed no clearing zones around PV-containing resins indicating minimal antibiotic leakage from the material. Similarly, MTT assay demonstrated that the antibiotic-containing specimens did not release cytotoxic byproducts that may inhibit human gingival fibroblast growth. Counting of colony-forming units in an S. mutans biofilm model was used to assess bacterial survival at baseline and after subjecting the antibiotic-containing resin specimens to an enzymatic challenge for 30 days. Significantly reduced bacterial counts were observed as the biofilm aged from 24 to 72 h, and salivary enzymatic exposure did not reduce the antibacterial efficacy of the discs, suggesting that PV-resin will be effective in reducing the re-incidence of dental caries.

Biocompatibility, mechanical, and bonding properties of a dental adhesive modified with antibacterial monomer and cross-linker.

OBJECTIVES: This study investigated the antibacterial, cytotoxicity, and mechanical properties of a dental adhesive modified with quaternary ammonium monomer ((2-acryloyloxyethyl)dimethyldodecylammonium bromide) and cross-linker (bis(2-acryloyloxyethyl)methyldodecylammonium bromide). MATERIALS AND METHODS: Monomer (M), cross-linker (C), or a combination of these (M + C) were incorporated into adhesive Adper Single Bond Plus (SB) in 5, 10, or 25% (as wt%). A colony-forming unit and MTT assays were used to evaluate antibacterial properties against Streptococcus mutans and cell viability. Resin-dentin beams (0.9 ± 0.1 mm2) were evaluated for micro-tensile bond strength (µTBS) after 24 h, 6 months, and 3 years. Hourglass specimens were evaluated for ultimate tensile strength (UTS) after 24 h, 1 week, and 6 months. Micro-hardness measurements after softening in ethanol were taken as an indirect assessment of the polymer cross-linking density. Kruskal-Wallis, one-way ANOVA, two-way ANOVA, and Student's t test were used for analysis of the antibacterial, cytotoxicity, µTBS, UTS, and hardness data, all with a significance level of p < 0.05. RESULTS: 10%M and 25%M demonstrated a significant reduction in S. mutans relative to SB (p < 0.001). No differences in cytotoxicity were detected for any of the groups. After 6 months, no changes in µTBS were shown for any of the groups. After 3 years, all groups evidenced a significant decrease in µTBS (p < 0.05) except 5%M, 5%C, and 5%M + 5%C. All groups demonstrated either stable or significantly increased UTS after 6 months. Except for the cross-linker groups, a significant decrease in micro-hardness was shown for all groups after softening in ethanol (p < 0.05). CONCLUSIONS: A 5-10% of monomer may render the resin antibacterial without a compromise to its mechanical and bonding properties. CLINICAL RELEVANCE: Biomodification of a resin adhesive with an antibacterial monomer and cross-linker may help improve the life span of adhesive restorations.

Esterase Inhibition and Copper Release from Copper Iodide Dental Adhesives - An In Vitro Study.

PURPOSE: To investigate whether dental adhesives modified with polyacrylic acid copper iodide particles could inhibit esterase activity in vitro and the copper release rate from resin matrices, as well as the correlation between the two variables. MATERIALS AND METHODS: Different concentrations of copper iodide (0.1, 0.5 and 1.0 mg/ml) were incorporated into three commercially available adhesives representative of each type. Disk specimens (n = 3) were fabricated and incubated in cholesterol esterase and pseudo-cholinesterase solutions for 16 days (37°C, pH 7.0). The enzymatic activity and rate of copper release from resin matrices were evaluated at different 4, 8, 12, and 16 days with a UV/visible-light spectrophotometer. RESULTS: Increased copper release and reduced enzymatic activity were observed with higher concentrations of copper iodide (p < 0.001). Greater copper release with reduced enzymatic activity was also demonstrated at the earlier time periods with this relationship reversing over time (p < 0.001). A moderate negative correlation between the variables was evident (-0.441; p = 0.01). CONCLUSIONS: Adhesives containing copper iodide can inhibit esterase activity in a dose- and time-dependent manner. The correlation between the variables suggests that enzymatic activity may depend on the availability of copper.

Synthesis and characterization of silver nanoparticle-loaded amorphous calcium phosphate microspheres for dental applications.

The goals of this work were (1) to synthesize composite nanostructures comprised of amorphous calcium phosphate (ACP) loaded with silver nanoparticles using a spray pyrolysis method and (2) to demonstrate their potential for use in dental adhesives. Release of silver ions from these nanostructures could provide antibacterial activity, while release of calcium and phosphate ions could promote tooth remineralization. Precursor solutions were prepared with varying silver concentrations corresponding to 5, 10, and 15 mol% of the calcium content, then sprayed into a furnace (550 °C) as droplets with a mean diameter near 2 µm. In this process, each droplet is converted into a single solid microsphere via rapid heating. The synthesized particles were collected using a polymeric filter installed at the end of the reaction zone. Different quantities (2, 5, and 10 wt%) of the nanocomposite material were mixed with a commercially available dental adhesive (Single Bond, 3M ESPE) which was then polymerized into discs for incubation in a solution simulating cariogenic conditions. Release of silver, calcium and phosphorus ions into the solution was measured for 1 month. The nanostructures of ∼10 nm silver nanoparticles embedded into 100 nm to 2 µm ACP particles demonstrated good dispersion in the adhesive resin blend, which in application would shield surrounding tissues from direct contact with silver. The composite nanoparticles provided a quick initial release of ions after which the concentration of calcium, phosphorous, and silver in the incubation solution remained constant or increased slightly. The dispersibility and ion release of the new nanostructures may offer potential for use in dental materials to achieve anti-bacterial and remineralization effects.

Polymer-antibiotic conjugates as antibacterial additives in dental resins.

Affecting the vast majority of human beings, dental caries is a premier concern of worldwide dental health. As the most commonly used restorative material to treat dental caries, resin-based composites (RBCs) lack antibacterial properties leading to quite limited restoration lifetimes. The objective of this study is to develop a polymer-antibiotic conjugate (PAC) as an effective antibacterial additive for RBCs. A monomer-antibiotic conjugate (MAC) with significant solubility was prepared by an esterification reaction of tert-butyloxycarbonyl (Boc)-protected ciprofloxacin (Cip) and 2-hydroxyethyl methacrylate (HEMA). The Cip-containing PAC with well-controlled molecular weight and composition was synthesized by reversible addition-fragmentation chain transfer (RAFT) copolymerization of the MAC with HEMA (1 : 3 molar ratio), followed by the removal of Boc from the resulting copolymer. The antibacterial dental resin was then prepared by incorporating the PAC into a commercial resin, and their properties and antibacterial performance against Streptococcus mutans were tested. In vitro experiments revealed a very slow release of Cip, which resulted in significant killing effectiveness against Streptococcus mutans nonetheless, as observed through zone of inhibition assessment and SEM imaging. The promising antibacterial properties of these resins indicate that incorporating a PAC as an additive is a valid strategy to generate antibacterial materials for dental applications.

Biocompatibility and bond degradation of poly-acrylic acid coated copper iodide-adhesives.

OBJECTIVE: To investigate the effect of poly-acrylic acid (PAA) copper iodide (CuI) adhesives on bond degradation, tensile strength, and biocompatibility. METHODS: PAA-CuI particles were incorporated into Optibond XTR, Optibond Solo and XP Bond in 0.1 and 0.5mg/ml. Clearfil SE Protect, an MDPB-containing adhesive, was used as control. The adhesives were applied to human dentin, polymerized and restored with composite in 2mm-increments. Resin-dentin beams (0.9±0.1mm2) were evaluated for micro-tensile bond strength after 24h, 6 months and 1year. Hourglass specimens (10×2×1mm) were evaluated for ultimate tensile strength (UTS). Cell metabolic function of human gingival fibroblast cells exposed to adhesive discs (8×1mm) was assessed with MTT assay. Copper release from adhesive discs (5×1mm) was evaluated with UV-vis spectrophotometer after immersion in 0.9% NaCl for 1, 3, 5, 7, 10, 14, 21 and 30 days. SEM, EDX and XRF were conducted for microstructure characterization. RESULTS: XTR and Solo did not show degradation when modified with PAA-CuI regardless of the concentration. The UTS for adhesives containing PAA-CuI remained unaltered relative to the controls. The percent viable cells were reduced for Solo 0.5mg/ml and XP 0.1 or 0.5mg/ml PAA-CuI. XP demonstrated the highest ion release. For all groups, the highest release was observed at days 1 and 14. SIGNIFICANCE: PAA-CuI particles prevented the bond degradation of XTR and Solo after 1year without an effect on the UTS for any adhesive. Cell viability was affected for some adhesives. A similar pattern of copper release was demonstrated for all adhesives.

Antibacterial properties of copper iodide-doped glass ionomer-based materials and effect of copper iodide nanoparticles on collagen degradation.

OBJECTIVES: This study investigated the antibacterial properties and micro-hardness of polyacrylic acid (PAA)-coated copper iodide (CuI) nanoparticles incorporated into glass ionomer-based materials, and the effect of PAA-CuI on collagen degradation. MATERIALS AND METHODS: PAA-CuI nanoparticles were incorporated into glass ionomer (GI), Ionofil Molar AC, and resin-modified glass ionomer (RMGI), Vitrebond, at 0.263 wt%. The antibacterial properties against Streptococcus mutans (n = 6/group) and surface micro-hardness (n = 5/group) were evaluated. Twenty dentin beams were completely demineralized in 10 wt% phosphoric acid and equally divided in two groups (n = 10/group) for incubation in simulated body fluid (SBF) or SBF containing 1 mg/ml PAA-CuI. The amount of dry mass loss and hydroxyproline (HYP) released were quantified. Kruskal-Wallis, Student's t test, two-way ANOVA, and Mann-Whitney were used to analyze the antibacterial, micro-hardness, dry mass, and HYP release data, respectively (p < 0.05). RESULTS: Addition of PAA-CuI nanoparticles into the glass ionomer matrix yielded significant reduction (99.999 %) in the concentration of bacteria relative to the control groups. While micro-hardness values of PAA-CuI-doped GI were no different from its control, PAA-CuI-doped RMGI demonstrated significantly higher values than its control. A significant decrease in dry mass weight was shown only for the control beams (10.53 %, p = 0.04). Significantly less HYP was released from beams incubated in PAA-CuI relative to the control beams (p < 0.001). CONCLUSIONS: PAA-CuI nanoparticles are an effective additive to glass ionomer-based materials as they greatly enhance their antibacterial properties and reduce collagen degradation without an adverse effect on their mechanical properties. CLINICAL RELEVANCE: The use of copper-doped glass ionomer-based materials under composite restorations may contribute to an increased longevity of adhesive restorations, because of their enhanced antibacterial properties and reduced collagen degradation.

Glutaraldehyde collagen cross-linking stabilizes resin-dentin interfaces and reduces bond degradation.

This study evaluated the stability of resin-dentin interfaces treated with glutaraldehyde-containing agents, and assessed collagen degradation in dentin matrices treated with Gluma. Microtensile bond strength (µTBS) was evaluated 24 h and 6 months after treatment with three desensitizers (Gluma Desensitizer, Gluma Desensitizer Power Gel, and MicroPrime G) and two etch-and-rinse adhesives (Comfort Bond & Desensitizer and iBond TE). Demineralized beams of human dentin were treated with water or Gluma, and the degradation of collagen in these beams was assessed by quantification of the dry mass loss and of the amount of hydroxyproline released from hydrolyzed specimens after 1 or 4 wk. All groups demonstrated significant reduction in µTBS after 6 months, except for Gluma Desensitizer and iBond TE groups, which showed decreases of 7.2% and 10.8%, respectively. The most prevalent failure mode was 'mixed'. Significantly less hydroxyproline was released from Gluma-treated beams than from control beams after 4 wk. Beams treated with Gluma yielded significantly less dry mass loss than did beams in the control group. Collagen cross-linking with glutaraldehyde-containing agents may assist in the stabilization of resin-dentin bonds by reducing the amount of collagen solubilized from dental matrices in the hybrid layer. In turn, this may contribute to the preservation of adhesive interfaces.

Effect of Sodium Fluoride on the endogenous MMP Activity of Dentin Matrices.

OBJECTIVES: This study evaluated the effect of incorporating increasing concentrations of sodium fluoride in incubation media, on the loss of dry mass and solubilization of collagen from demineralized dentin beams incubated for up to 7 days. The effect of fluoride on the inhibition of matrix-bound metalloproteinases (MMPs) was also measured. METHODS: Dentin beams were completely demineralized in 10% phosphoric acid. After baseline measurements of dry mass, the beams were divided into six groups (n=10) and incubated at 37°C either in buffered media containing sodium fluoride (NaF) at 75, 150, 300, 450, 600 ppm or in fluoride-free media (control) for seven days. Following incubation, dry mass was re-measured. The incubation media was hydrolyzed with HCl for the quantitation of hydroxyproline (HYP) as an index of solubilization of collagen by endogenous dentin proteases. Increasing concentrations of fluoride were also evaluated for their ability to inhibit rhMMP-9. RESULTS: Addition of NaF to the incubation media produced a progressive significant reduction (p<0.05) in the loss of mass of dentin matrices, with all concentrations demonstrating significantly less mass loss than the control group. Significantly less HYP release from the dentin beams was found in the higher fluoride concentration groups, while fluoride concentrations of 75 and 150 ppm significantly reduced rhMMP-9 activity by 6.5% and 79.2%, respectively. CONCLUSIONS: The results of this study indicate that NaF inhibits matrix-bound MMPs and therefore may slow the degradation of dentin matrix by endogenous dentin MMPs.

Incorporation of bactericidal poly-acrylic acid modified copper iodide particles into adhesive resins.

OBJECTIVES: This study aimed to investigate incorporation of polyacrylic acid (PAA) coated copper iodide (CuI) nanoparticles into dental adhesives, and to evaluate for the first time, their antibacterial properties, bond strength and cytotoxicity. METHODS: PAA-CuI nanoparticles were synthesized and incorporated into commercially available adhesives Optibond XTR (1.0mg/ml) and XP Bond (0.5 and 1.0mg/ml). The antibacterial properties of experimental and control specimens were evaluated (n=8), after ageing for 18h or 1 year, against Streptococcus mutans (1×10(8)cells/ml). Bond strength to human dentine of the control and experimental adhesives was evaluated by shear bond strength (n=10). For cytotoxicity evaluation, HGF cells were cultured with gingival fibroblast media and exposed to control and experimental adhesive blends (n=3). An MTT cell viability assay was used to assess cell metabolic function. A one-way analysis of variance followed by Tukey's test was used for data analysis. RESULTS: Significantly greater antibacterial properties were demonstrated for PAA-CuI containing adhesives after ageing for 18h or 1 year relative to all control groups. A reduction in Streptococcus mutans viable cell count of 99.99%, 99.99% and 79.65% was shown for XP Bond - 0.5mg/ml, XP Bond - 1.0mg/ml and Optibond XTR - 1.0mg/ml PAA-CuI after ageing for 18h, and 99.99% for both XP Bond - 0.5mg/ml and XP Bond - 1.0mg/ml PAA-CuI after ageing for 1 year. No significant variations in shear bond strength or cytotoxicity were detected between the experimental resins and their corresponding controls. CONCLUSIONS: PAA-CuI nanoparticles are an effective additive to adhesive blends as it renders them antibacterial without adversely affecting their bond strength or cytotoxicity. CLINICAL SIGNIFICANCE: The incorporation of PAA-coated copper iodide particles into adhesive resins renders the adhesive antibacterial to S. mutans for at least 1 year in vitro. This may prevent or delay bacterial invasion and the consequent development of caries lesions if the adhesive interface becomes defective.

Aging of adhesive interfaces treated with benzalkonium chloride and benzalkonium methacrylate.

Inhibition of endogenous dentin matrix metalloproteinases (MMPs) within incompletely infiltrated hybrid layers can contribute to the preservation of resin-dentin bonds. This study evaluated the bond stability of interfaces treated with benzalkonium chloride (BAC) and benzalkonium methacrylate (MBAC), and the inhibitory properties of these compounds on dentin MMP activity. Single-component adhesive ALL-BOND UNIVERSAL, modified with BAC or MBAC at concentrations of 0, 0.5, 1.0, and 2.0%, was used for microtensile bond strength (µTBS) evaluation after 24 h, 6 months, and 1 yr. Beams produced from human dentin were treated with 37% phosphoric acid, dipped in 0.5% BAC, 1.0% BAC, or water (control) for 60 s, and then incubated in SensoLyte generic MMP substrate to determine MMP activity. A significant decrease in the µTBS after 6 months and 1 yr was observed for the control group only. No significant differences among groups were shown at 24 h. After 6 months and 1 yr, the control group demonstrated significantly lower µTBS than all treatment groups. When applied for 60 s, 0.5% BAC inhibited total MMP activity by 31%, and 1.0% BAC inhibited total MMP activity by 54%. Both BAC and MBAC contributed to the preservation of resin-dentin bonds, probably because of their inhibitory properties of endogenous dentin proteinases.

Preservation of resin-dentin interfaces treated with benzalkonium chloride adhesive blends.

Reducing collagen degradation within hybrid layers may contribute to the preservation of adhesive interfaces. This study evaluated the stability of resin-dentin interfaces treated with benzalkonium chloride (BAC)-modified adhesive blends and assessed collagen degradation in dentin matrices treated with BAC. The etch-and-rinse adhesive, Adper Single Bond Plus, modified with 0.5% and 1.0% BAC, was evaluated for microtensile bond strength (µTBS) and nanoleakage (NL) after 24 h and 1 yr. Thirty completely demineralized dentin beams from human molars were dipped for 60 s in deionized water (DW; control), or in 0.5% or 1.0% BAC, and then incubated in simulated body fluid (SBF). Collagen degradation was assessed by quantification of the dry mass loss and the amount of hydroxyproline (HYP) released from hydrolyzed specimens after 1 or 4 wk. Although all groups demonstrated a significant increase in NL after 1 yr, adhesive modified with 0.5% BAC showed stable bond strength after 1 yr (9% decrease) relative to the control (44% decrease). Significantly less HYP release and dry mass loss were observed for both 0.5% and 1.0% BAC relative to the control. This in vitro study demonstrates that BAC contributes to the preservation of resin-dentin bonds for up to 1 yr by reducing collagen degradation.

Inhibition of endogenous human dentin MMPs by Gluma.

OBJECTIVE: The objective of this study was to determine if Gluma dentin desensitizer (5.0% glutaraldehyde and 35% HEMA in water) can inhibit the endogenous MMPs of dentin matrices in 60 s and to evaluate its effect on dentin matrix stiffness and dry mass weight. METHODS: Dentin beams of 2 mm×1 mm×6 mm were obtained from extracted human third molars coronal dentin. To measure the influence of Gluma treatment time on total MMP activity of dentin, beams were dipped in 37% phosphoric acid (PA) for 15 s and rinsed in water. The acid-etched beams were then dipped in Gluma for 5, 15, 30 or 60 s, rinsed in water and incubated into SensoLyte generic MMP substrate (AnaSpec, Inc.) for 60 min. Controls were dipped in water for 60 s. Additional beams of 1 mm×1 mm×6 mm were completely demineralized in 37% PA for 18 h, rinsed and used to evaluate changes on the dry weight and modulus of elasticity (E) after 60 s of Gluma treatment followed by incubation in simulated body fluid buffer for 0, 1 or 4 weeks. E was measured by 3-pt flexure. RESULTS: Gluma treatment inhibited total MMP activity of acid-etched dentin by 44, 50, 84, 86% after 5, 15, 30 or 60 s of exposure, respectively. All completely demineralized dentin beams lost stiffness after 1 and 4 weeks, with no significant differences between the control and Gluma-treated dentin. Gluma treatment for 60 s yielded significantly less dry mass loss than the control after 4 weeks. SIGNIFICANCE: The use of Gluma may contribute to the preservation of adhesive interfaces by its cross-linking and inhibitory properties of endogenous dentin MMPs.

Mechanisms regulating the degradation of dentin matrices by endogenous dentin proteases and their role in dental adhesion. A review.

PURPOSE: This systematic review provides an overview of the different mechanisms proposed to regulate the degradation of dentin matrices by host-derived dentin proteases, particularly as it relates to their role in dental adhesion. Significant developments have taken place over the last few years that have contributed to a better understanding of all the factors affecting the durability of adhesive resin restorations. The complexity of dentin-resin interfaces mandates a thorough understanding of all the mechanical, physical and biochemical aspects that play a role in the formation of hybrid layers. The ionic and hydrophilic nature of current dental adhesives yields permeable, unstable hybrid layers susceptible to water sorption, hydrolytic degradation and resin leaching. The hydrolytic activity of host-derived proteases also contributes to the degradation of the resin-dentin bonds. Preservation of the collagen matrix is critical to the improvement of resin-dentin bond durability. Approaches to regulate collagenolytic activity of dentin proteases have been the subject of extensive research in the last few years. A shift has occurred from the use of proteases inhibitors to the use of collagen cross-linking agents. Data provided by 51 studies published in peer-reviewed journals between January 1999 and December 2013 were compiled in this systematic review. RESULTS: Appraisal of the data provided by the studies included in the present review yielded a summary of the mechanisms which have already proven to be clinically successful and those which need further investigation before new clinical protocols can be adopted.

Matrix metalloproteinase inhibitory properties of benzalkonium chloride stabilizes adhesive interfaces.

This study evaluated the effects of different concentrations of benzalkonium chloride (BAC) on the preservation of adhesive interfaces created with two etch-and-rinse adhesives and its inhibitory properties on dentin matrix metalloproteinase (MMP) activity. The following groups were tested with the adhesive systems Optibond Solo Plus and All-Bond 3: Group 1, adhesive without inhibitor (control); Group 2, topical 2.0% chlorhexidine (2.0% CHX); Group 3, phosphoric acid with 1.0%wt BAC (BAC-PA); Group 4, 0.25% BAC-adhesive (0.25% BAC); Group 5, 0.5% BAC-adhesive (0.5% BAC); Group 6, 1.0% BAC-adhesive (1.0% BAC); and Group 7, 2.0% BAC-adhesive (2.0% BAC). Composite cylinders were fabricated, and shear bond strength (SBS) was evaluated after 24 h, 6 months, and 18 months of storage. Extracts from concentrated demineralized human dentin powder were subjected to SDS-PAGE and incubated in the presence of 0.25, 0.5, 1.0, and 2.0% BAC. Overall, stable bonds were maintained for 18 months. Improved bond strengths were seen for 0.5% BAC and 1.0% BAC when bonding with Optibond Solo Plus, and for 0.25% BAC and 0.5% BAC when bonding with All-Bond 3. Zymographic analysis revealed complete inhibition of gelatinolytic activity with BAC. Benzalkonium chloride, at all concentrations, inhibited dentin proteolytic activity, which seems to have contributed to the improved bond stability after 18 months for specific combinations of BAC concentration and adhesive.

Effect of phosphoric acid etching on the shear bond strength of two self-etch adhesives.

OBJECTIVE: To evaluate the effect of optional phosphoric acid etching on the shear bond strength (SBS) of two self-etch adhesives to enamel and dentin. MATERIAL AND METHODS: Ninety-six bovine mandibular incisors were ground flat to obtain enamel and dentin substrates. A two-step self-etch adhesive (FL-Bond II) and a one-step self-etch adhesive (BeautiBond) were applied with and without a preliminary acid etching to both the enamel and dentin. The specimens were equally and randomly assigned to 4 groups per substrate (n=12) as follows: FL-Bond II etched; FL-Bond II un-etched; BeautiBond etched; BeautiBond un-etched. Composite cylinders (Filtek Z100) were bonded onto the treated tooth structure. The shear bond strength was evaluated after 24 hours of storage (37°C, 100% humidity) with a testing machine (Ultra-tester) at a speed of 1 mm/min. The data was analyzed using a two-way ANOVA and post-hoc Tukey's test with a significance level of p<0.05. A field emission scanning electron microscope was used for the failure mode analysis. RESULTS: Both adhesives evidenced a significant decrease in the dentin SBS with the use of an optional phosphoric acid-etching step (p<0.05). Preliminary phosphoric acid etching yielded significantly higher enamel SBS for FL-Bond II (p<0.05) only, but not for BeautiBond. FL-Bond II applied to un-etched dentin demonstrated the highest mean bond strength (37.7±3.2 MPa) and BeautiBond applied to etched dentin showed the lowest mean bond strength (18.3±6.7 MPa) among all tested groups (p<0.05). CONCLUSION: The use of a preliminary acid-etching step with 37.5% phosphoric acid had a significant adverse effect on the dentin bond strength of the self-etch adhesives evaluated while providing improvement on the enamel bond strength only for FL-Bond II. This suggests that the potential benefit that may be derived from an additional etching step with phosphoric acid does not justify the risk of adversely affecting the bond strength to dentin.

In vitro shear bond strength of three self-adhesive resin cements and a resin-modified glass ionomer cement to various prosthodontic substrates.

OBJECTIVE: To evaluate the shear bond strength (SBS) of three self-adhesive resin cements and a resin-modified glass ionomer cement (RMGIC) to different prosthodontic substrates. MATERIALS AND METHODS: The substrates base metal, noble metal, zirconia, ceramic, and resin composite were used for bonding with different cements (n=12). Specimens were placed in a bonding jig, which was filled with one of four cements (RelyX Unicem, Multilink Automix, Maxcem Elite, and FujiCEM Automix). Both light-polymerizing (LP) and self-polymerizing (SP) setting reactions were tested. Shear bond strength was measured at 15 minutes and 24 hours in a testing device at a test speed of 1 mm/min and expressed in MPa. A Student t-test and a one-way analysis of variance (ANOVA) were used to evaluate differences between setting reactions, between testing times, and among cements irrespective of other factors. Generalized linear regression model and Tukey tests were used for multifactorial analysis. RESULTS: Significantly higher mean SBS were demonstrated for LP mode relative to SP mode (p<0.001) and for 24 hours relative to 15 minutes (p<0.001). Multifactorial analysis revealed that all factors (cement, substrate, and setting reaction) and all their interactions had a significant effect on the bond strength (p<0.001). Resin showed significantly higher SBS than other substrates when bonded to RelyX Unicem and Multilink Automix in LP mode (p<0.05). Overall, FujiCEM demonstrated significantly lower SBS than the three self-adhesive resin cements (p<0.05). CONCLUSIONS: Overall, higher bond strengths were demonstrated for LP relative to SP mode, 24 hours relative to 15 minutes and self-adhesive resin cements compared to the RMGICs. Bond strengths also varied depending on the substrate, indicating that selection of luting cement should be partially dictated by the substrate and the setting reaction.