Bacteria in the cavity-restoration interface after varying periods of clinical service - SEM description of distribution and 16S rRNA gene sequence identification of isolates.

OBJECTIVES: To use extracted human teeth with amalgam (n = 26) or GIC (n = 3) restorations in service up to 20 years to evaluate microbiota at the cavity/restoration interface by SEM or culture. MATERIALS AND METHODS: Extracted teeth with intracoronal restorations (n = 20) of known history (2-20 years) were fixed, split, and prepared for SEM to ascertain the pattern and structure of bacterial aggregates on cavity and restoration surfaces. Another 9 teeth were anaerobically decontaminated, split and sampled (cavity/restorations), and cultured (anaerobically, aerobically); recovered isolates were identified by 16S rRNA gene sequencing. RESULTS: SEM showed rods, cocci, and filaments in 11/20 teeth (55%) on cavity and corresponding restoration surfaces; 4/20 (20%) on neither surface; 1/20 (5%) on just cavity; and 4/20 (20%) on just restoration. Microbial growth extended from marginal openings into the deeper interfacial microspace to varying extents but was not always evident. Restoration size or age did not predict bacterial presence. Bacteria-free surfaces (cavity/amalgam) showed possible calcification. Cultivation yielded 160 isolates, mainly Gram-positive (86%) and facultative (81%); and morphotypes of rods (43%), cocci (36%), and cocco-bacilli (18%) belonging to Actinobacteria (45%) and Firmicutes (50%). The most frequent genera were Staphylococcus, Streptococcus, Actinomyces, and Lactobacillus. Biofilms on cavity and restoration appeared independent of each other. CONCLUSIONS: Cavity and amalgam surfaces were independently colonised and some not. The penetration of microbiota into marginal gaps varied; resembled root caries and was dominated by Gram-positive species. CLINICAL RELEVANCE: Marginal gaps around restorations are unavoidable but are not always colonised by bacteria after long-term clinical service. Calcification of biofilms in the restorative interface may prevent further colonisation. The viable microbiota in the restorative interface resembled root caries and may be subject to ecological fluxes of activity and arrest and therefore preventative management.

Structure and properties of strontium-doped phosphate-based glasses.

Owing to similarity in both ionic size and polarity, strontium (Sr2+) is known to behave in a comparable way to calcium (Ca2+), and its role in bone metabolism has been well documented as both anti-resorptive and bone forming. In this study, novel quaternary strontium-doped phosphate-based glasses, containing 1, 3 and 5 mol% SrO, were synthesized and characterized. (31)P magic angle spinning (MAS) nuclear magnetic resonance results showed that, as the Sr2+ content is increased in the glasses, there is a slight increase in disproportionation of Q2 phosphorus environments into Q(1) and Q3 environments. Moreover, shortening and strengthening of the phosphorus to bridging oxygen distance occurred as obtained from FTIR. The general broadening of the spectral features with Sr2+ content is most probably due to the increased variation of the phosphate-cation bonding interactions caused by the introduction of the third cation. This increased disorder may be the cause of the increased degradation of the Sr-containing glasses relative to the Sr-free glass. As confirmed from elemental analysis, all Sr-containing glasses showed higher Na2O than expected and this also could be accounted for by the higher degradation of these glasses compared with Sr-free glasses. Measurements of surface free energy (SFE) showed that incorporation of strontium had no effect on SFE, and samples had relatively higher fractional polarity, which is not expected to promote high cell activity. From viability studies, however, the incorporation of Sr2+ showed better cellular response than Sr(2+)-free glasses, but still lower than the positive control. This unfavourable cellular response could be due to the high degradation nature of these glasses and not due to the presence of Sr2+.

Chlorhexidine-releasing methacrylate dental composite materials.

Light curable antibacterial, dental composite restoration materials, consisting of 80 wt% of a strontium fluoroaluminosilicate glass dispersed in methacrylate monomers have been produced. The monomers contained 40-100 wt% of a 10 wt% chlorhexidine diacetate (CHXA) in hydroxyethylmethacrylate (HEMA) solution and 60-0 wt% of a 50/50 mix of urethane dimethacrylate (UDMA) and triethyleneglycol dimethacrylate (TEGDMA). On raising HEMA content, light cure polymerisation rates decreased. Conversely, water sorption induced swelling and rates of diffusion controlled CHXA release from the set materials increased. Experimental composites with 50 and 90 wt% of the CHXA in HEMA solution in the monomer were shown, within a constant depth film fermentor (CDFF), to have slower rates of biofilm growth on their surfaces between 1 and 7 days than the commercial dental composite Z250 or fluoride-releasing dental cements, Fuji II LC and Fuji IX. When an excavated bovine dentine cylinder re-filled with Z250 was placed for 10 weeks in the CDFF, both bacteria and polymers from the artificial saliva penetrated between the material and dentine. With the 50 wt% experimental HEMA/CHXA formulation, this bacterial microleakage was substantially reduced. Polymer leakage, however, still occurred. Both polymer and bacterial microleakage were prevented with a 90 wt% HEMA/CHXA restoration in the bovine dentine due to swelling compensation for polymerisation shrinkage in combination with antibacterial release.

Sol-gel synthesis of quaternary (P2O5)55-(CaO)25-(Na2O)(20-x)-(TiO2) x bioresorbable glasses for bone tissue engineering applications (x = 0, 5, 10, or 15).

In the present study, we report a new and facile sol-gel synthesis of phosphate-based glasses with the general formula of (P2O5)55-(CaO)25-(Na2O)(20-x)-(TiO2) x , where x = 0, 5, 10 or 15, for bone tissue engineering applications. The sol-gel synthesis method allows greater control over glass morphology at relatively low processing temperature (200 °C) in comparison with phosphate-based melt-derived glasses (~1000 °C). The glasses were analyzed using several characterization techniques, including x-ray diffraction (XRD), (31)P magic angle spinning nuclear magnetic resonance ((31)P MAS-NMR), Fourier transform infrared (FTIR) spectroscopy and energy-dispersive x-ray (EDX) spectroscopy, which confirmed the amorphous and glassy nature of the prepared samples. Degradation was assessed by measuring the ion release and pH change of the storage medium. Cytocompatibility was also confirmed by culturing osteoblast-like osteosarcoma cell line MG-63 on the glass microparticles over a seven-day period. Cell attachment to the particles was imaged using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The results revealed the potential of phosphate-based sol-gel derived glasses containing 5 or 10 mol% TiO2, with high surface area, ideal dissolution rate for cell attachment and easily metabolized dissolution products, for bone tissue engineering applications.

Study of bacterial viability within human supragingival dental calculus.

BACKGROUND: There is evidence that supragingival calculus contains unmineralized channels and lacunae. The purpose of this study was to investigate the viability of bacteria within these areas. METHODS: Supragingival calculus harvested from patients with moderate to severe chronic periodontitis was immediately frozen to -70 degrees C. Six samples were cryosectioned, stained with a bacterial viability kit, and examined with fluorescence microscopy. Controls comprised heat treatment of cryosections prior to staining. Four additional samples were stained and examined whole in a confocal laser scanning microscope (CLSM). Nine additional samples were prepared for bacterial culture, after initial irradiation with ultraviolet light to kill viable organisms on the covering plaque layer. Test samples were crushed to expose internal bacteria, while two controls were used without crushing. RESULTS: Viable bacteria, as identified using the bacterial viability stain, were found within cavities/lacunae in supragingival calculus cryosections. Similar results were obtained from whole calculus samples using CLSM. Of the nine experimental samples where bacterial culture was attempted, five provided positive bacterial culture under both aerobic and anaerobic conditions; one showed positive growth under aerobic conditions only; while one showed no bacterial growth. The controls showed no bacterial growth. CONCLUSIONS: From this study, it appears that viable aerobic and anaerobic bacteria may be present within supragingival calculus, specifically within the internal channels and lacunae. Clinically, this may be important, since incomplete removal of supragingival calculus may expose these reservoirs of possible pathogenic bacteria and be a factor in the recurrence of periodontal diseases after treatment.