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.

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.

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.

Enterococcus faecalis Hydrolyzes Dental Resin Composites and Adhesives.

INTRODUCTION: After root canal treatment, the dentin-sealer interface undergoes degradation, allowing for interfacial microbial biofilm proliferation and treatment failure. Saliva and cariogenic bacteria showed esterase-like activities (ie, cholesterol esterase [CE]-like and/or pseudocholinesterase [PCE]-like) that degrade methacrylate-based resin materials and/or the restoration-tooth interface, increasing microbial interfacial proliferation. Enterococcus faecalis is a gram-positive bacterium that is commonly detected in persistent endodontic infections. The aim of this study was to measure E. faecalis esterase-like, CE-like, and PCE-like activities and to assess the ability of the bacterium to degrade methacrylate-based resin composite (RC) and total-etch (TE) and self-etch (SE) adhesives. METHODS: CE-like and PCE-like activities from E. faecalis were measured using nitrophenyl and butyrylthiocholine substrates, respectively. The ability of E. faecalis to degrade resin composite, total-etch and self-etch adhesives was examined by quantifying the release of a universal resin degradation by-product (ie, Bis[hydroxypropoxy]-phenyl propane [BisHPPP]) using high-performance liquid chromatography. RESULTS: E. faecalis showed CE-like (1.23 ± 0.13 U/µg dry bacteria) but no PCE-like activity. After 30 days and/or 14 days of incubation, the amount of BisHPPP released was significantly higher in the presence of bacteria versus media for TE and RC but not SE (P < .05). The amount of BisHPPP released after 30 days of incubation with bacteria was highest for TE (23.69 ± 1.72 µg/cm2) followed by RC (3.43 ± 1.20 µg/cm2) and lowest for SE (0.86 ± 0.44 µg/cm2) (P < .05). CONCLUSIONS: E. faecalis possesses esterase-like degradative activity toward dental methacrylate resin restoration materials, which could accelerate the degradation of the dentin-methacrylate resin interface, increasing bacterial biofilm proliferation and penetration into the root canal system.