The Role of Bacterial, Dentinal, Salivary, and Neutrophil Degradative Activity in Caries Pathogenesis.

Until recently, it was widely accepted that bacteria participate in caries pathogenesis mainly through carbohydrate fermentation and acid production, which promote the dissolution of tooth components. Neutrophils, on the other hand, were considered white blood cells with no role in caries pathogenesis. Nevertheless, current literature suggests that both bacteria and neutrophils, among other factors, possess direct degradative activity towards both dentinal collagen type-1 and/or methacrylate resin-based restoratives and adhesives, the most common dental restoratives. Neutrophils are abundant leukocytes in the gingival sulcus, where they can readily reach adjacent tooth roots or gingival and cervical restorations and execute their degradative activity. In this review, we present the latest literature evidence for bacterial, dentinal, salivary, and neutrophil degradative action that may induce primary caries, secondary caries, and restoration failure.

Detection of Bacteria-Induced Early-Stage Dental Caries Using Three-Dimensional Mid-Infrared Thermophotonic Imaging.

Tooth decay, or dental caries, is a widespread and costly disease that is reversible when detected early in its formation. Current dental caries diagnostic methods including X-ray imaging and intraoral examination lack the sensitivity and specificity required to routinely detect caries early in its formation. Thermophotonic imaging presents itself as a highly sensitive and non-ionizing solution, making it suitable for the frequent monitoring of caries progression. Here, we utilized a treatment protocol to produce bacteria-induced caries lesions. The lesions were imaged using two related three-dimensional photothermal imaging modalities: truncated correlation photothermal coherence tomography (TC-PCT) and its enhanced modification eTC-PCT. In addition, micro-computed tomography (µ-CT) and visual inspection by a clinical dentist were used to validate and quantify the severities of the lesions. The observational findings demonstrate the high sensitivity and depth profiling capabilities of the thermophotonic modalities, showcasing their potential use as a non-ionizing clinical tool for the early detection of dental caries.

Streptococcus mutans Proteases Degrade Dentinal Collagen.

Here, we explored the role of S. mutans's whole cell and discrete fractions in the degradation of type I collagen and dentinal collagen. Type I collagen gels and human demineralized dentin slabs (DS) were incubated in media alone or with one of the following: overnight (O/N) or newly inoculated (NEW) cultures of S. mutans UA159; intracellular proteins, supernatant or bacterial membranes of O/N cultures. Media from all groups were analyzed for protease-mediated release of the collagen-specific imino acid hydroxyproline. Images of type I collagen and DS were analyzed, respectively. Type I collagen degradation was highest for the supernatant (p < 0.05) fractions, followed by intracellular components and O/N cultures. Collagen degradation for DS samples was highest for O/N samples, followed by supernatant, and intracellular components (p < 0.05). There was lower detectable degradation for both type I collagen and DS from NEW culture samples (p < 0.05), and there was no type I collagen or DS degradation detected for bacterial membrane samples. Structural changes to type I collagen gel and dentinal collagen were observed, respectively, following incubation with S. mutans cultures (O/N and NEW), intracellular components, and supernatant. This study demonstrates that intracellular and extracellular proteolytic activities from S. mutans enable this cariogenic bacterium to degrade type I and dentinal collagen in a growth-phase dependent manner, potentially contributing to the progression of dental caries.

Towards the development of biostable dental resin systems - design criteria and constraints beyond ester-free chemistries.

OBJECTIVE: The objective of this review article is to summarize the current literature on dental resin-based restorative (RBR) materials specifically from the perspective of emerging resin technologies, and to provide researchers with structured design criteria enabling the effective screening of new RBR developments. METHODS: The continued failure of newly introduced RBRs to address biostability without compromising function, over the last decade, are presented as a rationale to support different resin-based concepts. Several developments in the field, aimed at addressing the issues facing modern resin-based systems are summarized and their limitations discussed. A design workflow is proposed for evaluating new RBR, considering resource needs. RESULTS: While several alternative resin chemistries have been suggested over the past decade, all have shown serious limitations in replacing MA-based materials, including their limited physical and mechanical properties, and curing kinetics. Additionally, a broad and inconsistent range of laboratory methods have been used to validate these developments, leading to results that are difficult to compare across studies. A design workflow was conceptualized to facilitate the screening of novel RBRs from both a clinical and research perspective. SIGNIFICANCE: While several alternative chemistries have shown some degree of potential in emulating material property aspects of MA-based resins, a complete restorative system that is resistant to biochemical reactions in saliva has yet to achieve broad clinical adoption. To further spur development, it would be useful to have a more systematic design workflow, that may be used to easily screen new resin technologies effectively early in the design phase, so as to mitigate potential performance failures in the clinic.

Interfacial Biomaterial-Dentin Bacterial Biofilm Proliferation and Viability Is Affected by the Material, Aging Media and Period.

Biomaterial−dentin interfaces undergo degradation over time, allowing salivary, tissue fluid, and bacterial movement between the root filling or restoration and dentin. This study aims to investigate the effect of aging in simulated human salivary/bacterial/blood esterases (SHSE) on proliferation and viability of Enterococcus faecalis biofilm within the dentin interface with four materials used to fill/restore the endodontic space. Root canals of human anterior teeth were prepared and filled with gutta-percha and one of the following: self-cured resin composite (BisfilTM 2B, Bisco, Schaumburg, IL, USA) with either self-etch (SE) (EasyBond) or total-etch (TE) (ScotchbondTM, 3M, Saint Paul, MN, USA) methacrylate-based adhesives, epoxy-resin sealer (AH Plus®, Dentsply Sirona, York, PA, USA), or bioceramic sealer (EndoSequence® BC Sealer™, Brasseler USA, Savannah, GA, USA). Specimens were aged in SHSE or phosphate-buffered saline (PBS) for up to 360 days, followed by cultivation of steady-state E. faecalis biofilm. Depth and viability of interfacial bacterial biofilm proliferation were assessed by confocal laser scanning microscopy and live/dead staining. Data were analyzed using three-way ANOVA and Scheffe's post hoc analyses. Initial depths of biofilm proliferation were similar among material groups (p > 0.05). All groups showed significantly deeper biofilm proliferation with increased aging period (p < 0.05). SHSE aging increased interfacial biofilm depth for TE, SE and BC (p < 0.05) but not AH. For unaged interfaces, BC exhibited the lowest ratio of live bacteria, followed by AH, TE, and SE (p < 0.05). Interfacial bacterial biofilm proliferation and viability were dependent on the biomaterial, aging media, and period.

Comparison of Long-Wave and Mid-Wave Infrared Imaging Modalities for Photothermal Coherence Tomography of Human Teeth.

The ability to detect dental caries at early stages lies at the heart of minimal intervention dentistry, enabling the curing or arresting of carious lesions before they advance to the cavity stage. Enhanced truncated-correlation photothermal coherence tomography (eTC-PCT) using mid-wave infrared (MWIR) cameras has recently been shown to offer tomographic visualization of early caries. The tomographic slicing ability of such systems, however, is believed to be limited by direct radiative thermal emission through the translucent dental enamel in the 3-5 µm MWIR spectral range. Such radiative emissions can dominate the delayed conductive thermal contributions needed for tomographic reconstruction of internal dental defects. It has been hypothesized that long-wave infrared (LWIR) eTC-PCT systems may offer better tomographic performance by taking advantage of the intrinsic attenuation of direct radiative emission by dental enamel in the LWIR spectral range, enabling more effective delayed conductive thermal contributions from subsurface caries. More than an order of magnitude lower cost of the system is another key attribute of LWIR eTC-PCT which can open the door for downstream translation of the technology to clinics. In this report, we offer a systematic comparison of the performance/effectiveness of caries detection with LWIR and MWIR eTC-PCT systems for detecting natural caries, bacterial caries, and artificially demineralized enamel surfaces. Our results suggest that the low-cost LWIR based eTC-PCT system provides 3D visualization and 2D slice-by-slice images of early caries and internal micro-cracks similar to those obtained from the more expensive MWIR-based eTC-PCT system, albeit with ∼1.3dB lower signal-to-noise ratio.

Simulating the Intraoral Aging of Dental Bonding Agents: A Narrative Review.

Despite their popularity, resin composite restorations fail earlier and at higher rates than comparable amalgam restorations. One of the reasons for these rates of failure are the properties of current dental bonding agents. Modern bonding agents are vulnerable to gradual chemical and mechanical degradation from a number of avenues such as daily use in chewing, catalytic hydrolysis facilitated by salivary or bacterial enzymes, and thermal fluctuations. These stressors have been found to work synergistically, all contributing to the deterioration and eventual failure of the hybrid layer. Due to the expense and difficulty in conducting in vivo experiments, in vitro protocols meant to accurately simulate the oral environment's stressors are important in the development of bonding agents and materials that are more resistant to these processes of degradation. This narrative review serves to summarize the currently employed methods of aging dental materials and critically appraise them in the context of our knowledge of the oral environment's parameters.

Minimally Invasive Therapies for the Management of Dental Caries-A Literature Review.

In recent years, due to a better understanding of the caries pathology and advances in dental materials, the utilization of non-invasive and minimally invasive techniques that delay/obviate the need for traditional restorations has started gaining momentum. This literature review focuses on some of these approaches, including fluoride varnish, silver diamine fluoride, resin sealants, resin infiltration, chemomechanical caries removal and atraumatic restorative treatment, in the context of their chemistries, indications for use, clinical efficacy, factors determining efficacy and limitations. Additionally, we discuss strategies currently being explored to enhance the antimicrobial properties of these treatment modalities to expand the scope of their application.

Assessment of Root Canal Sealers Loaded with Drug-Silica Coassembled Particles Using an In Vitro Tooth Model.

INTRODUCTION: The purpose of this study was to assess the antimicrobial activity of root canal sealers modified with novel highly loaded antimicrobial drug-silica coassembled particles (DSPs) on Enterococcus faecalis-infected root canal dentin. METHODS: DSPs were synthesized through coassembly of silica and octenidine dihydrochloride (OCT) surfactant drug (35% w/w OCT). DSPs (1% wt of the total mass of the sealer) were mixed homogenously with either epoxy resin sealer (AH Plus [AH]; Dentsply Sirona, Tulsa, OK) or calcium silicate-based sealer (EndoSequence BC Sealer [BC]; Brasseler, Savannah, GA). To assess the antimicrobial activity of DSP-loaded sealers, the apical third of single-rooted teeth was obtained and infected with E. faecalis for 3 weeks followed by the application of experimental (DSP-loaded) sealers or corresponding controls for up to 28 days. Microbiological analysis and laser scanning confocal and scanning electron microscopy were used to determine the colony-forming unit (CFU)/mL, the percentage of live bacteria, and the intratubular bacterial and sealer penetrations. Factorial analysis of variance and Tukey post hoc tests were used to assess the antimicrobial effect of DSPs on different sealers. RESULTS: All experimental groups showed significant reductions in CFUs at all-time points compared with positive controls (P < .05). The addition of DSPs to BC significantly reduced the CFUs (2.11 ± 0.13, 2.22 ± 0.19, and 2.25 ± 0.17 at 1, 7, and 28 days, respectively) compared with the unmodified sealer (3.21 ± 0.11, 4.3 ± 0.15, and 4.2 ± 0.2 at 0, 7, and 28 days). DSPs enhanced the antimicrobial performance of AH only at 1 day (4.21 ± 0.17 vs 5.19 ± 0.12, P < .05). AH and AH + DSPs showed higher bacterial viability compared with BC and BC + DSPs at all incubation periods (P < .05). CONCLUSIONS: Loading endodontic sealers with DSPs had a material-dependent effect on the antimicrobial properties and could reduce the incidence of secondary infections.

Effect of processing methods on the cytotoxicity of methyl methacrylate-based ocular prostheses: An in vitro study.

The study evaluated the influence of cycles and methods of an ocular prosthesis resin on cytotoxicity toward human conjunctival cells. Resins were polymerized by water bath (WB, 74 °C or 100 °C for 30 min to 9 h), microwave (MW, 1200 W, 3 to 14 min and 30 s at 0 to 720 W), or autopolymerization (AP, room temperature for 20 min ± 60 °C for 30 min). Degree of conversion (DC), cytotoxicity, level of inflammatory mediators, gene expression of different markers, and apoptosis were evaluated. Data were submitted to ANOVA and Tukey test (p < 0.05). WB with longer processing time at higher temperature had highest DC (85.6%) and higher TGF ß1-gene expression (1.39); long cycle low power MW showed lowest DC (69.6%), lower cell proliferation (85.4%, MTT), and large IL-2 release (39,297 ng/mL). AP with additional processing time showed lower cell proliferation (75.3%, Alamar Blue), and AP polymerized at room temperature showed higher CASP 9-gene expression (1.21). AP methods showed higher IL-6 release (>277 pg/mL). Short cycle medium power MW had higher IL-23 release (534.2 pg/mL). MW (long and short cycles) and AP polymerizations have triggered a more intense inflammatory response. Among methods recommended by the manufacturer, WB showed high DC and less cytotoxicity.

Drug-Silica Coassembled Particles Improve Antimicrobial Properties of Endodontic Sealers.

INTRODUCTION: The purpose of this study was to assess the antimicrobial activity and flow of root canal sealers after incorporating novel highly loaded antimicrobial drug-silica coassembled particles (DSPs). METHODS: DSPs were synthesized through coassembly of silica and octenidine dihydrochloride (OCT) antimicrobial surfactant. DSPs were loaded (1% and 2% wt) into epoxy resin sealer (AH Plus [AH]; Dentsply DeTrey GmbH, Konstanz, Germany) or calcium silicate-based sealer (EndoSequence Bioceramic Sealer (BC); Brasseler, Savannah, GA). OCT release from DSP-modified sealers was determined using liquid chromatography. Antimicrobial activity of sealers against planktonic or biofilm form Enterococcus faecalis was assessed using direct contact and membrane restricted tests. Sealer flow was tested according to ISO6876:2012. RESULTS: OCT release from BC + 1% or 2% DSPs was above the minimum inhibitory concentration following 2 days throughout the 30-day experiment, whereas OCT release from AH + 1% or 2% DSP was significantly below the minimum inhibitory concentration against E. faecalis (4 µg/mL) over the whole 30-day experimental period. All materials (with or without DSPs) killed planktonic bacteria initially. AH ± 1% or 2% DSPs had no antimicrobial activity after 7 days. BC + 1% or 2% DSPs maintained antibacterial activity over the 30-day period. Both modified and unmodified sealers completely inhibited the growth of E. faecalis biofilms after 24 hours of contact. DSPs decreased the flow of AH and BC sealers; for AH, the reduction was proportional to the amount of DSPs added. All modified and unmodified sealers, except for AH + 2% DSPs, were within the acceptable limits of ISO 6876 flow tests. CONCLUSIONS: DSPs enhanced the antimicrobial performance of BC but not AH, whereas the material's flow remained compliant with ISO 6876 standards. Depending on the sealer, DSPs may enhance antimicrobial efficacy in root canal treatment and potentially improve treatment outcome.

Antimicrobial antidegradative dental adhesive preserves restoration-tooth bond.

OBJECTIVE: Assess the ability of an antimicrobial drug-releasing resin adhesive, containing octenidine dihydrochloride (OCT)-silica co-assembled particles (DSPs), to enhance the biostability and preserve the interfacial fracture toughness (FT) of composite restorations bonded to dentin. Enzyme-catalyzed degradation compromises the dental restoration-tooth interface, increasing cariogenic bacterial infiltration. In addition to bacterial ingress inhibition, antimicrobial-releasing adhesives may exhibit direct interfacial biodegradation inhibition as an additional benefit. METHODS: Mini short-rod restoration bonding specimens with total-etch adhesive with/without 10% wt. DSPs were made. Interfacial fracture toughness (FT) was measured as-manufactured or post-incubation in simulated human salivary esterase (SHSE) for up to 6-months. Effect of OCT on SHSE and whole saliva/bacterial enzyme activity was assessed. Release of OCT outside the restoration interface was assessed. RESULTS: No deleterious effect of DSPs on initial bonding capacity was observed. Aging specimens in SHSE reduced FT of control but not DSP-adhesive-bonded specimens. OCT inhibited SHSE degradation of adhesive monomer and may inhibit endogenous proteases. OCT inhibited bacterial esterase and collagenase. No endogenous collagen breakdown was detected in the present study. OCT increased human saliva degradative esterase activity below its minimum inhibitory concentration towards S. mutans (MIC), but inhibited degradation above MIC. OCT release outside restoration margins was below detection. SIGNIFICANCE: DSP-adhesive preserves the restoration bond through a secondary enzyme-inhibitory effect of released OCT, which is virtually confined to the restoration interface microgap. Enzyme activity modulation may produce a positive-to-negative feedback switch, by increasing OCT concentration via biodegradation-triggered release to an effective dose, then subsequently slowing degradation and degradation-triggered release.

Human neutrophils compromise the restoration-tooth interface.

Neutrophils, cells of the innate immune system, enter the mouth and release factors that are hypothesized to contribute to the degradation of tooth dentin, methacrylate resin composites, and adhesives at the restoration-tooth-dentin interface. The objectives were to characterize neutrophils' degradation towards resin composite, self-etch (SE) and total-etch (TE) adhesives, SE and TE resin-dentin interfaces and to identify proteins that could contribute to the degradation process. Neutrophils' degradation of cured resin composite, and SE and TE adhesives, was quantified by measuring the specific resin degradation by-product, bishydroxy-propoxy-phenyl-propane (bisHPPP), released after 30 days incubation of the materials with the cells. Neutrophils' degradative effect on resin-dentin interfaces was examined by recording the interfacial fracture toughness (FT), and surface analysis of the fracture mode following incubation of SE and TE miniature short-rod (mini-SR) specimens with the cells. Neutrophils increased degradation of polymerized resin composite, and TE adhesive, but not SE adhesive over 30 days (p < 0.05). Incubation of SE and TE resin-dentin interfaces with neutrophils led to a reduction in FT over time (p < 0.05). The effect was more pronounced for TE interfaces. Neutrophils also affected the fracture mode of SE and TE resin-dentin interfaces. Several proteins that could contribute to the degradative activity of neutrophils, including Neutrophil collagenase (MMP-8), Matrix metalloproteinase- 9 (MMP-9), Cathepsin G, Neutrophil- gelatinase associated lipocalin (NGAL) and Myeloperoxidase, were isolated. The ability of neutrophils to degrade resin, tooth dentin, and reduce the bond strength of resin-dentin interfaces suggest neutrophils' potential role in primary and recurrent caries and dental restoration failure.

Genetic Analysis of Mutacin B-Ny266, a Lantibiotic Active against Caries Pathogens.

Bacteriocins are ribosomally synthesized proteinaceous antibacterial peptides. They selectively interfere with the growth of other bacteria. The production and secretion of bacteriocins confer a distinct ecological advantage to the producer in competing against other bacteria that are present in the same ecological niche. Streptococcus mutans, a significant contributor to the development of dental caries, is one of the most prolific producers of bacteriocins, known as mutacins in S. mutans In this study, we characterized the locus encoding mutacin B-Ny266, a lantibiotic with a broad spectrum of activity. The chromosomal locus is composed of six predicted operon structures encoding proteins involved in regulation, antimicrobial activity, biosynthesis, modification, transport, and immunity. Mutacin B-Ny266 was purified from semisolid cultures, and two inhibitory peptides, LanA and LanA', were detected. Both peptides were highly modified. Such modifications include dehydration of serine and threonine and the formation of a C-terminal aminovinyl-cysteine (AviCys) ring. While LanA peptide alone is absolutely required for antimicrobial activity, the presence of LanA' enhanced the activity of LanA, suggesting that B-Ny266 may function as a two-peptide lantibiotic. The activation of lanAA' expression is most likely controlled by the conserved two-component system NsrRS, which is activated by LanA peptide but not by LanA'. The chromosomal locus encoding mutacin B-Ny266 was not universally conserved in all sequenced S. mutans genomes. Intriguingly, the genes encoding LanAA' peptides were restricted to the most invasive serotypes of S. mutansIMPORTANCE Although dental caries is largely preventable, it remains the most common and costly infectious disease worldwide. Caries is initiated by the presence of dental plaque biofilm that contains Streptococcus mutans, a species extensively characterized by its role in caries development and formation. S. mutans deploys an arsenal of strategies to establish itself within the oral cavity. One of them is the production of bacteriocins that confer a competitive advantage by targeting and killing closely related competitors. In this work, we found that mutacin B-Ny266 is a potent lantibiotic that is effective at killing a wide array of oral streptococci, including nearly all S. mutans strains tested. Lantibiotics produced by oral bacteria could represent a promising strategy to target caries pathogens embedded in dental plaque biofilm.

Immediate versus conventional loading of mandibular implant-supported fixed prostheses in edentulous patients: 10-year report of a randomised controlled trial.

PURPOSE: To compare the clinical outcomes of immediate versus conventional delayed loading of four dental implants in edentulous mandibles with fixed prostheses. MATERIALS AND METHODS: A blinded, two-arm, parallel group, randomised controlled trial was conducted. A total of 42 patients were included, and each received four Brånemark System implants with a TiUnite surface. The patients were randomly assigned to two study arms: 1) immediate-loading arm (IL), in which the mandibular denture was converted into an interim implant-supported fixed prosthesis (ISFP) on the day of surgery, with a permanent ISFP being inserted at least 3 months postsurgery; 2) conventional-loading arm (CL), in which the mandibular removable prosthesis was relieved at the implant site and relined with a soft tissue conditioner. Only implants with a minimum insertion torque of ≥ 35 Ncm were included in the IL group. Implants were loaded 4 to 6 months postsurgery. Independent, blind investigators assessed the patients at 2, 6 and 12 months and at 10 years. The outcome measures were prosthesis and implant success rates, type and frequency of complications and changes in peri-implant marginal bone levels. RESULTS: A total of 20 patients were allocated to the IL group and 22 to the CL group. However, one patient from the IL arm was excluded and three patients were reallocated to the CL arm. Two implants in one patient and one in another patient could not be placed with a ≥ 35 Ncm insertion torque, and a third patient developed severe sudden gag reflex and thus it was not possible to load the implants immediately. At a later stage, one of the patients who failed the initial stability test dropped out of the study. Therefore, initially, 24 patients were conventionally loaded and 16 patients were immediately loaded. At the 10-year follow-up, six patients dropped out from the IL arm and two from the CL arm. Also, at the 10-year-follow-up, the CL and IL study arms consisted of 22 and 10 participants, respectively, using the per-protocol (PP) analysis. Six implants failed in two patients of the CL arm (two implants in one patient and four implants in another patient), and three implants failed in three patients in the IL arm (PP analysis), respectively. The patient-level implant failure rate was 10% (intention-to-treat [ITT] analysis) and 14% (PP analysis) in the CL arm, and 25% (ITT) and 20% (PP) in the IL arm. The difference was not statistically significant (95% CI from -0.18 to 0.39, P = 0.65). The failure rate at the implant level was 8% (ITT) and 8% (PP) in the CL arm, and 6% (ITT) and 5% (PP) in the IL arm. The difference was not statistically significant (95% CI from -0.06 to 0.14, P = 0.44). Ten years after loading, patients in the IL arm lost an average of 0.55 ± 0.64 mm of peri-implant bone versus 0.41 ± 0.40 mm of peri-implant bone loss observed in the CL arm. The 10-year bone loss in both arms was statistically significant compared with the baseline (P < 0.001). However, there was no statistically significant difference between the two arms for peri-implant bone level changes (the difference between the arms was 0.14 mm ± 0.50 mm; 95% CI -0.23 to 0.52; P = 0.43). One prosthesis failed due to the loss of all four implants in one patient of the CL arm. Eight patients from the IL arm were affected by 13 complications (such as pain from fractures and inflammation) versus seven patients (10 complications) from the CL arm. The complication rate was 67% in the IL arm and 35% in the CL arm. The difference in complication proportions between the two arms was not statistically significant (difference in proportions = 0.32; 95% CI = -0.08 to 0.61; P = 0.14). All complications were managed successfully. CONCLUSIONS: Long-term data of immediate loading of four dental implants with a mandibular fixed prosthesis revealed comparable clinical outcomes to conventional loading. Therefore, immediate loading should be considered in the treatment of edentulous patients.

Ultrashort-pulse laser as a surface treatment for bonding between zirconia and resin cement.

OBJECTIVES: To evaluate ultrashort-pulse laser (UPL) as a surface treatment for improved bond strength to Yttria-tetragonal zirconia polycrystalline (Y-TZP). METHODS: Fully-sintered Y-TZP samples received either no treatment (CTL), or were treated by alumina blasting (ALB), tribochemical silica coating (SIL), or one of two UPL patterns: multiple pulses laser surface dots with 2.5µm spacing (8mJ, 10kHz)(LSD); or single pulse laser surface lines with 2.5µm spacing (4mJ, 6.7kHz)(LSL). Surface roughness, wettability (contact angle), and quantification of crystalline phases were evaluated for each group (n=3/group). Y-TZP treated slabs were cemented to resin composite slabs using silane and 10-methacryloyloxydecyl dihydrogen phosphate (MDP)-containing adhesive. Beams from the Y-TZP/resin blocks were microtensile tested (n=5/group) after 48h water incubation (37°C) with or without subsequent thermocycling (5-55°C, 5000 cycles). RESULTS: All surface treatments increased surface roughness values versus control (P<0.001). Contact angles were lowest for SIL (6.57±2.37°) and highest for control (50.97±6.30°). LSL and LSD were the only treatments that did not increase the relative monoclinic phase. All surface treatments significantly increased microtensile bond strengths (µTBS) compared with the control group (P<0.001), with highest values for UPL (LSD: 35.40±4.53MPa>LSL: 31.84±8.46MPa>SIL: 19.95±3.99MPa=ALB: 19.51±2.55MPa>CTL: 14.51±2.23MPa). Thermocycling significantly reduced bond strength for all treatments in a surface treatment-dependent manner. SIGNIFICANCE: The ability of UPL to alter Y-TZP surface morphology, increase wettability and µTBS without increasing the monoclinic content suggests its potential to improve bonding to the underlying resin cement and tooth without compromising the strength of the restoration.

Human neutrophils degrade methacrylate resin composites and tooth dentin.

Cholesterol esterase-like (CE) activity from saliva and esterase from cariogenic bacteria hydrolyze ester linkages of dental methacrylate resins. Collagenolytic, matrix metalloproteinase-like (MMP) activities from dentin and bacteria degrade collagen in demineralized tooth dentin. Human neutrophils in the oral cavity contain factors that are hypothesized to have CE and MMP activities that could contribute to the degradation of methacrylate resins and dentinal collagen. OBJECTIVES: To measure the CE and MMP activities from human neutrophils and their ability to degrade dental methacrylate resin composite and dentinal collagen. Neutrophils' CE and MMP activities were measured using nitrophenyl-esters or fluorimetric MMP substrates, respectively. Neutrophils' degradation of resin composite and dentinal collagen was quantified by measuring release of a universal 2,2-Bis[4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane (bisGMA)-derived resin composite degradation byproduct, bishydroxy-propoxy-phenyl-propane (bisHPPP), or a collagen degradation by-product, hydroxyproline, respectively using ultra performance liquid chromatography/mass spectrometry. Neutrophils' CE activity increased the release of bisHPPP from bisGMA monomer compared to control after 24 and 48 h (p < 0.05). Neutrophils degraded polymerized resin composite and produced higher amounts of bisHPPP than buffer after 48 h of incubation (p < 0.05). Neutrophils show generic MMP, gelatinase, MMP-2 and MMP-9, and collagenase, MMP-1 and MMP-8 activities that were stable or increased over the first 24 h (p < 0.05). Neutrophils degraded demineralized dentin more than buffer-only groups, indicated by higher amounts of hydroxyproline (p < 0.05). The ability of neutrophils to degrade both dental resin composite and tooth dentin, suggest neutrophil's potential role in root caries, and in recurrent carries by accelerating the degradation of resin-dentin interfaces, and compromising the longevity of the restoration. STATEMENT OF SIGNIFICANCE: Neutrophils are part of the innate immune system and are constantly entering the oral cavity through the gingival sulcus, in direct contact with the tooth, restoration, restoration-tooth margins and pathogenic bacteria. The current study is the first to characterize and quantify degradative activities from neutrophils toward methacrylate resin and demineralized dentin, the two main components of the restoration-tooth interface, suggesting that this interface could be negatively influenced by neutrophils, potentially contributing to increase in caries formation and progression, and premature restoration failure. This study provides a significant finding to the biomaterials and oral health fields by identifying a potential weakness in current restorative procedures and materials used to manage gingival proximal and cervical gingival or sub-gingival carious lesions.

Physical properties and cytotoxicity of antimicrobial dental resin adhesives containing dimethacrylate oligomers of Ciprofloxacin and Metronidazole.

OBJECTIVE: Antimicrobial oligomers synthesized from ciprofloxacin (CF) and metronidazole (MN) were investigated for their potential use in dental adhesives. METHODS: Susceptibility of the cariogenic bacterium Streptococcus mutans UA159 to CF, MN, and CF/MN combination was evaluated. Hydrolytic stability and drug release from the oligomers was studied in buffer and simulated human salivary esterase conditions. Cytotoxicity of films with 15wt% drug oligomers co-polymerized with commercial monomers were assessed using human gingival fibroblasts (HGFs). In-house adhesives were prepared and characterized for viscosity. Polymerized films were analysed for gel content and water swelling. Interfacial fracture toughness (KIC) of composites bonded to dentin by either a 2 or 3-step etch-and-rinse approach using the in-house formulated adhesives was measured. RESULTS: The respective minimum inhibitory concentration for CF and MN against S. mutans was 0.7 and 2400µg/mL, with the combination having an additive effect (0.35µg/mL CF with 1200µg/mL MN). Antibiotics were released upon hydrolysis of the oligomers. Films containing the drug oligomers were not cytotoxic against HGFs. Replacing 2-hydroxyethyl methacrylate with the drug oligomers increased the viscosity of the experimental adhesives, reduced gel content, and decreased swelling of films in water. Antimicrobial adhesives demonstrated bonding to dentin with interfacial KIC values comparable to the in-house control in the 2-step application, and with slightly lower KIC values in the 3-step approach. SIGNIFICANCE: The antimicrobial oligomers can be incorporated into dental adhesive systems using formulations that show comparable fracture toughness to commercial materials, and may provide a means to deliver local antimicrobial drug release at the marginal interface.

Biostable, antidegradative and antimicrobial restorative systems based on host-biomaterials and microbial interactions.

OBJECTIVES: Despite decades of development and their status as the restorative material of choice for dentists, resin composite restoratives and adhesives exhibit a number of shortcomings that limit their long-term survival in the oral cavity. Herein we review past and current work to understand these challenges and approaches to improve dental materials and extend restoration service life. METHODS: Peer-reviewed work from a number of researchers as well as our own are summarized and analyzed. We also include yet-unpublished work of our own. Challenges to dental materials, methods to assess new materials, and recent material improvements and research directions are presented. RESULTS: Mechanical stress, host- and bacterial-biodegradation, and secondary caries formation all contribute to restoration failure. In particular, several host- and bacterial-derived enzymes degrade the resin and collagen components of the hybrid layer, expanding the marginal gap and increasing access to bacteria and saliva. Furthermore, the virulence of cariogenic bacteria is up-regulated by resin biodegradation by-products, creating a positive feedback loop that increases biodegradation. These factors work synergistically to degrade the restoration margin, leading to secondary caries and restoration failure. Significant progress has been made to produce hydrolytically stable resins to resist biodegradation, as well as antimicrobial materials to reduce bacterial load around the restoration. Ideally, these two approaches should be combined in a holistic approach to restoration preservation. SIGNIFICANCE: The oral cavity is a complex environment that poses an array of challenges to long-term material success; materials testing conditions should be comprehensive and closely mimic pathogenic oral conditions.