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.

A bifunctional O-antigen polymerase structure reveals a new glycosyltransferase family.

Lipopolysaccharide O-antigen is an attractive candidate for immunotherapeutic strategies targeting antibiotic-resistant Klebsiella pneumoniae. Several K. pneumoniae O-serotypes are based on a shared O2a-antigen backbone repeating unit: (→ 3)-α-Galp-(1 → 3)-ß-Galf-(1 →). O2a antigen is synthesized on undecaprenol diphosphate in a pathway involving the O2a polymerase, WbbM, before its export by an ATP-binding cassette transporter. This dual domain polymerase possesses a C-terminal galactopyranosyltransferase resembling known GT8 family enzymes, and an N-terminal DUF4422 domain identified here as a galactofuranosyltransferase defining a previously unrecognized family (GT111). Functional assignment of DUF4422 explains how galactofuranose is incorporated into various polysaccharides of importance in vaccine production and the food industry. In the 2.1-Å resolution structure, three WbbM protomers associate to form a flattened triangular prism connected to a central stalk that orients the active sites toward the membrane. The biochemical, structural and topological properties of WbbM offer broader insight into the mechanisms of assembly of bacterial cell-surface glycans.

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.