Distribution of alpha-synuclein in rat salivary glands.

Expression of alpha-synuclein (Syn), a presynaptic neuronal protein, was immunohistochemically examined in intact rat submandibular, sublingual, and lingual glands. The submandibular gland contained abundant periductal Syn-immunoreactive (-ir) nerve fibers. Abundant Syn-ir varicosities were present in acini of the sublingual and serous lingual glands. By confocal laser scanning microscopy, Syn-ir nerve fibers around smooth muscle actin (SMA)-ir cells alone were infrequent; however, those around aquaporin-5 (AQP5)-ir cells alone and both SMA- and AQP5-ir cells were abundant in the sublingual and serous lingual glands. SMA-ir cells were occasionally immunoreactive for toll-like receptor 4, a Syn receptor. Syn-ir nerve fibers contained tyrosine hydroxylase (TH) in the submandibular gland and choline acetyltransferase (ChAT) in all examined salivary glands. In the superior cervical (SCG), submandibular, and intralingual ganglia, sympathetic and parasympathetic neurons co-expressed Syn with TH and ChAT, respectively. SCG neurons innervating the submandibular gland contained mostly Syn. In the thoracic spinal cord, 14.7% of ChAT-ir preganglionic sympathetic neurons co-expressed Syn. In the superior salivatory nucleus, preganglionic parasympathetic neurons projecting to the lingual nerve co-expressed Syn and ChAT. The present findings indicate that released Syn acts on myoepithelial cells. Syn in pre- and post-ganglionic neurons may regulate neurotransmitter release and salivary volume and composition.

Distribution of alpha-synuclein in the rat cranial sensory ganglia, and oro-cervical regions.

BACKGROUND: Alpha-synuclein (Syn), an unfolded soluble cytosolic protein, is known as a disease-associated protein in the brain. However, little is known about distribution of this protein in the peripheral nervous system. In this study, expression of Syn was investigated in the sensory ganglia of the cranial nerves V, IX and X. METHODS: To analyze distribution of Syn and its co-expression with calcitonin gene-related peptide (CGRP) or the transient receptor potential cation channel subfamily V member 1 (TRPV1), immunohistochemical techniques were used in the rat cranial sensory ganglia and their peripheral tissues. RESULTS: Syn-immunoreactive (-ir) neurons were abundant in the sensory ganglia of the petrosal (56.7%), jugular (28.3%) and nodose ganglia (82.5%). These neurons had small to medium-sized cell bodies (petrosal, mean ± S.D. = 667.4 ± 310.8 µ m2; jugular, 625.1 ± 318.4 µ m2; nodose, 708.3 ± 248.3 µ m2), and were distributed throughout the ganglia. However, the trigeminal ganglion was mostly free of Syn-ir neurons. By double and triple immunofluorescence staining, Syn-ir neurons co-expressed CGRP and TRPV1 in the petrosal and jugular ganglia. Syn-immunoreactivity was expressed by nerve fibers in the epithelium and taste bud of oral and cervical viscerae. These nerve fibers were abundant in the naso-pharynx, epiglottis and laryngeal vestibule. Some taste bud cells were also immunoreactive for Syn. In addition, Syn-ir nerve fibers were detected in the vicinity of macrophages, dendritic cells and Langerhans cells. CONCLUSIONS: Syn was abundant in the visceral sensory neurons but not in somatic sensory neurons. This protein may play a role in nociceptive and chemosensory transduction in the glossopharyngeal and vagal sensory ganglia. It is possible that Syn has a function about the immune mechanism of the upper air way.

Effects of quaternary ammonium-methacrylates on the mechanical properties of unfilled resins.

OBJECTIVE: Adding antimicrobial/anti-MMP quaternary ammonium methacrylates (QAMs) to comonomer blends should not weaken the mechanical properties of dental resins. This work evaluated the degree conversion and mechanical properties of BisGMA/TEGDMA/HEMA (60:30:10) containing 0-15 mass% QAMs A-E (A: 2-acryloxyethyltrimethyl ammonium chloride; B: [3-(methacryloylamino)propyl]trimethylammonium chloride; C: [2-(methacryloxy)ethyl] trimethyl ammonium chloride; D: diallyldimethyl ammonium chloride; E: 2-(methacryloyloxy) ethyltrimethyl ammonium methyl sulfate. METHODS: Unfilled resins with and without QAM were placed on ATR-FTIR and light-polymerized for 20s in a thin film at 30°C. Unfilled resin beams were casted from square hollow glass tubings. Half of the beams were tested after 3 days of drying (control); the other half were tested wet after 3 days of water storage. RESULTS: Addition of QAMs in control resins significantly increased conversion 600 s after light termination, with the exception of 5% MAPTAC (p<0.05). Increase of QAM content within a formulation significantly increased conversion. Control beams gave dry Young's moduli of ∼700 MPa. Addition of 5, 10 or 15 mass% QAMs produced significant reductions in dry Young's moduli except for 5% B or C. 15 mass% A, B and C lowered the wet Young's moduli of the resin beams by more than 30%. The ultimate tensile stress (UTS) of control dry resin was 89±11 MPa. Addition of 5-10 mass% QAMs had no adverse effect on the dry UTS. After water storage, the UTS of all resin blends fell significantly (p<0.05), especially when 15 wt% QAMs was added. Control dry beams gave fracture toughness (KIC) values of 0.88±0.1 MPa m(1/2). Wet values were significantly higher at 1.02±0.06 (p<0.05). KIC of dry beams varied from 0.85±0.08 at 5% QAMs to 0.49±0.05 at 15% QAMs. Wet beams gave KIC values of 1.02±0.06 MPa m(1/2) that fell to 0.23±0.01 at 15% QAMs. SIGNIFICANCE: Addition of 10% QAMs increased the degree of conversion of unfilled resins, but lowered wet toughness and UTS; addition of 15% QAMs lowered the mechanical properties of wet resins below acceptable levels.

Effects of chlorhexidine in self-etching adhesive: 24 hours results.

It is known that chlorhexidine (CHX) does not inhibit micro-tensile bond strengths (MTBS) when it is used in etch-and-rinse adhesives. In that technique, CHX is applied to dentin as a primer after phosphoric acid-etching before bonding with Single Bond. It would be more convenient if it is possible to incorporate CHX into the adhesive. The purpose of this study was to compare the MTBS and the FT-IR percent conversion of an all-in-one self-etching adhesives contained varying concentration of CHX. Extracted human third molars were bonded with a control all-in-one adhesive or experimental versions containing 0.5, 1, 2 or 5% CHX. The MTBS and the percent conversion of experimental adhesives containing up to 1% CHX were not significantly CHX-free control adhesives. However, addition of 2 or especially 5% CHX experimental adhesives produced significant reductions in both the MTBS and the percent conversion.

Inhibition of MMPs by alcohols.

OBJECTIVES: While screening the activity of potential inhibitors of matrix metalloproteinases (MMPs), due to the limited water solubility of some of the compounds, they had to be solubilized in ethanol. When ethanol solvent controls were run, they were found to partially inhibit MMPs. Thus, the purpose of this study was to compare the MMP-inhibitory activity of a series of alcohols. METHODS: The possible inhibitory activity of a series of alcohols was measured against soluble rhMMP-9 and insoluble matrix-bound endogenous MMPs of dentin in completely demineralized dentin. Increasing concentrations (0.17, 0.86, 1.71 and 4.28 mol/L) of a homologous series of alcohols (i.e. methanol, ethanol, propanols, butanols, pentanols, hexanols, the ethanol ester of methacrylic acid, heptanols and octanol) were compared to ethanediol, and propanediol by regression analysis to calculate the molar concentration required to inhibit MMPs by 50% (i.e. the IC(50)). RESULTS: Using two different MMP models, alcohols were shown to inhibit rhMMP-9 and the endogenous proteases of dentin matrix in a dose-dependent manner. The degree of MMP inhibition by alcohols increased with chain length up to 4 methylene groups. Based on the molar concentration required to inhibit rhMMP-9 fifty percent, 2-hydroxyethylmethacrylate (HEMA), 3-hexanol, 3-heptanol and 1-octanol gave the strongest inhibition. SIGNIFICANCE: The results indicate that alcohols with 4 methylene groups inhibit MMPs more effectively than methanol or ethanol. MMP inhibition was inversely related to the Hoy's solubility parameter for hydrogen bonding forces of the alcohols (i.e. to their hydrophilicity).

The requirement of zinc and calcium ions for functional MMP activity in demineralized dentin matrices.

UNLABELLED: The progressive degradation of resin-dentin bonds is due, in part, to the slow degradation of collagen fibrils in the hybrid layer by endogenous matrix metalloproteinases (MMPs) of the dentin matrix. In in vitro durability studies, the storage medium composition might be important because the optimum activity of MMPs requires both zinc and calcium. OBJECTIVE: This study evaluated the effect of different storage media on changes in matrix stiffness, loss of dry weight or solubilization of collagen from demineralized dentin beams incubated in vitro for up to 60 days. METHODS: Dentin beams (1mm×2mm×6mm) were completely demineralized in 10% phosphoric acid. After baseline measurements of dry mass and elastic modulus (E) (3-point bending, 15% strain) the beams were divided into 5 groups (n=11/group) and incubated at 37°C in either media containing both zinc and calcium designated as complete medium (CM), calcium-free medium, zinc-free medium, a doubled-zinc medium or water. Beams were retested at 3, 7, 14, 30, and 60 days of incubation. The incubation media was hydrolyzed with HCl for the quantitation of hydroxyproline (HOP) as an index of solubilization of collagen by MMPs. Data were analyzed using repeated measures of ANOVA. RESULTS: Both the storage medium and the storage time showed significant effects on E, mass loss and HOP release (p<0.05). The incubation in CM resulted in relatively rapid and significant (p<0.05) decreases in stiffness, and increasing amounts of mass loss. The HOP content of the experimental media also increased with incubation time but was significantly lower (p<0.05) than in the control CM medium, the recommended storage medium. CONCLUSIONS: The storage solutions used to age resin-dentin bonds should be buffered solutions that contain both calcium and zinc. The common use of water as an aging medium may underestimate the hydrolytic activity of endogenous dentin MMPs.

The inhibitory effect of polyvinylphosphonic acid on functional matrix metalloproteinase activities in human demineralized dentin.

This study has examined the use of polyvinylphosphonic acid (PVPA) as a potential matrix metalloproteinase (MMP) inhibitor and how brief cross-linking of demineralized dentin matrix that did not affect its mechanical properties enhanced the anti-MMP activity of PVPA. The anti-MMP potential of five PVPA concentrations (100-3000 microgml(-1)) was initially screened using a rhMMP-9 colorimetic assay. Demineralized dentin beams were treated with the same five concentrations of PVPA to collagen and then aged for 30 days in a calcium- and zinc-containing medium. The changes in modulus of elasticity, loss of dry mass and dissolution of collagen peptides were measured via three-point bending, precision weighing and hydroxyproline assay, respectively. All tested PVPA concentrations were highly effective (P<0.05) in inhibiting MMP-9. Ageing in the incubation medium did not significantly alter the modulus of elasticity of the five PVPA treatment groups. Conversely, aged dentin beams from the control group exhibited a significant decline in their modulus of elasticity (P<0.05) over time. Mass loss from the dentin beams and the corresponding increase in hydroxyproline in the medium in the five PVPA treatment groups were significantly lower than for the control (P<0.05). PVPA is a potent inhibitor of endogenous MMP activities in demineralized dentin. It may be used as an alternative to chlorhexidine to prevent collagen degradation within hybrid layers to extend the longevity of resin-dentin bonds.