Comprehensive analysis of bacterial flora of a biofilm model in initial caries-inducing environment.

PURPOSE: To analyze changes in pH and bacterial flora with duration of culture and timing of sugar supply using a polymicrobial biofilm model. METHODS: The biofilm was prepared using the method of Exterkate et al. Stimulated saliva from an adult was collected on a glass slide and added to unbuffered McBain medium containing 0.2% sucrose and cultivated under anaerobic conditions for 10 hours. Cultivation continued anaerobically in saliva-free medium refreshed twice daily, with or without sucrose, in five groups: in the Control and Groups A and C, with 0.2% sucrose for 96, 192 and 288 hours, respectively; in Groups B and E, with 0.2% sucrose for 96 hours then, respectively, without for 96 and 192 hours; in Group D, with 0.2% sucrose for 96 hours, without for 96 hours, then with for 96 hours. The pH of all spent medium was measured. Total bacteria counts were determined by Q-PCR. The bacterial composition was determined by next-generation sequencing of 16S rDNA. RESULTS: The pH of spent medium depended on the presence or absence of sucrose. Total bacteria counts were higher in A, C and D than the other groups, and markedly lower in Group E. Principal components analysis and cluster analysis showed wider variation of bacterial flora of the biofilm in Groups B, D and E than other groups. CLINICAL SIGNIFICANCE: Inspection of bacterial flora of a biofilm model of the initial caries-inducing environment may lead to the development of materials and procedures for the prevention of dental caries.

Anti-demineralization effect of desensitizer containing copolymer and sodium fluoride on root dentin - a transverse microradiographic study.

Objective: To evaluate anti-demineralization effects of dentin desensitizer containing sodium fluoride and methacrylate-co-p-styrene sulfonic acid (MS polymer) on root dentin using transverse microradiography (TMR). Material and methods: Twenty-four dentin specimens were divided into four groups: MSO (no fluoride), MSF (3000 ppm F), FJL (9000 ppm F), and Control. In MSO and MSF, each desensitizer was rubbed into the dentin surfaces for 10 s then left for 20 s. In FJL, paste containing 9000 ppm F was applied onto the surface for 30 s. All specimens, including the Controls, were rinsed with deionized water, dried and an area of their surface exposed to pH 5.0 acidic solution, refreshed every 24 h, for 4 days. Sections 300-µm-thick were assessed by TMR. Mineral profiles and integrated mineral loss (IML) of lesions were analyzed by dedicated software. IML was analyzed with one-way ANOVA and Tukey's test. Results: MSF and FJL specimens showed high mineral volume % at the surface and in lesions, and significantly lower IML than the other groups (p < .05). Conclusion: Dentin desensitizer containing 3000 ppm fluoride and MS polymer has the same anti-demineralization effect as does a fluoride paste containing 9000 ppm F.

Effects of resin-based temporary filling materials against dentin demineralization.

This study investigated the in vitro anti-demineralization effects of resin-based temporary filling materials containing surface prereacted glass-ionomer (S-PRG) filler on dentin. Bovine root dentin specimens with a 3×3 mm experimental surface were divided into four treatment groups: DuraSeal (DU) as a control, S-PRG filler-free temporary material (S0), material containing 10% (S10) and 20% (S20) S-PRG filler. Each material was applied to 3×2 mm of the experimental surface, and the specimens were immersed in 8% methylcellulose gel demineralization system for one week at 37˚C. Mineral profiles and integrated mineral loss (IML) of lesions induced on the surface (3×1 mm) adjacent to the materials were computed by transversal microradiography. S10 and S20 yielded thick surface layers and shallow lesion bodies, with significantly lower IML than DU and S0 (p<0.05, Tukey's test). These findings indicate that temporary filling resin-based materials containing over 10% of S-PRG filler content have anti-demineralization effects on adjacent dentin.

Antimicrobial effects of carbamide peroxide against a polymicrobial biofilm model.

PURPOSE: To investigate the in vitro antimicrobial effects of carbamide peroxide (CP) and CP-based home bleaching agents against polymicrobial (PM) biofilms. METHODS: Using a high-throughput active attachment model, PM biofilms were cultured on glass coverslips by diluting the stimulated saliva of one healthy adult. All experiments were performed anaerobically in McBain medium, which was refreshed twice daily. After biofilm formation for 24 or 72 hours, the biofilms were treated with 0.5%, 2.5%, 5%, or 10% CP, 20-fold dilutions of HiLite Shade Up (HS) or Opalescence Regular (OR), 0.2% chlorhexidine digluconate (CHX), 0.2% NaF, or deionized water (n = 10 each). Biofilms were dispersed and the number of colony forming units (CFU) was measured on tryptic soy agar blood plates. Coverslips containing 72-hour biofilms treated with 0.5% and 10% CP and deionized water were stained and scanned by confocal laser scanning microscopy (CLSM). RESULTS: Treatment of 24- and 72-hour biofilms with HS, OR and CH yielded significantly fewer colonies than treatment with water or 0.2% NaF. No growing colonies were observed after treatment with 10% CP. CLSM showed that the percentage of dead bacteria increased as the concentration of CP increased.

Effect of the coating material on root dentin remineralization in vitro.

PURPOSE: A fluoride-releasing coating material containing surface pre-reacted glass-ionomer (S-PRG) filler has become commercially available. However, there has been no detailed investigation of its remineralization effects at various tooth surface regions. The remineralization effects of S-PRG filler-containing coating material at different sites of demineralized dentin surfaces in vitro were evaluated. METHODS: Baseline lesions were prepared on bovine root dentin surfaces by immersion in demineralization buffer and divided into four groups: (B)--baseline lesion; (P)--S-PRG filler-containing material; (V)--S-PRG filler-free coating material as negative control; and (X)--resin-modified glass- ionomer as positive control. Material was applied to half the lesion surface, then P, V and X were remineralized in a gel system. Mineral profiles, integrated mineral loss (IML) and lesion depth (LD) at four regions, i.e. 1--exposed dentin surface adjacent to the material; 2--at a distance from the material; 3--beneath the material near to the edge; and 4--at a distance from the edge, were analyzed by transversal microradiography. Data were analyzed using ANOVA and Games-Howell test with α = 0.05. RESULTS: B showed typical artificial demineralized lesion. The IMLs of V, P and X at regions 1 and 2, and P and X at region 3 were significantly lower than that of B, however, those of V at region 3 and the other three groups at region 4 were not significantly different from that of B. At region 1, P and X showed significantly lower IMLs than V. At region 2, the IML of X showed significantly lower IML than V. There was no significant difference between P and X. The LD values of V, P and X at all regions were not significantly different from that of B. Fluoride, strontium and silicate ions released from the S-PRG filler would provide a favorable environment for remineralization of the demineralized dentin in P.

Appearance of thyroid stimulating and blocking immunoglobulins after bone marrow transplantation: presentation of two contrasting cases.

Two acute leukemia cases who presented autoimmune thyroid diseases after bone marrow transplantation (BMT) are described with reference to the pathogenesis of their autoimmune clones. A 37-year old Japanese woman developed Graves' hyperthyroidism 39 months after allogeneic BMT for acute myeloid leukemia (AML) donated from her sister. Although both donor and recipient were euthyroid and negative for thyroid autoimmunity before BMT, the donor was positive for anti-nuclear and anti-single strand DNA autoantibodies. Studies on polymorphism for variable number of tandem repeat region of T-cell receptor gene suggested that the lymphocytes responsible for the hyperthyroidism were of donor origin. The second case was a 12-year-old Japanese schoolboy who presented nongoitrous hypothyroidism 2 years after autologous BMT for acute lymphoblastic leukemia (ALL). He had been clinically euthyroid before transplantation. Family history revealed that his mother and sister had a history of Graves' disease. His serum was positive for thyroid-stimulation blocking antibody. It is highly likely that the autoimmune process was activated after transient immune suppression during peri-BMT period in this patient. Pathogenesis, incidence, and observed time lag between BMT and development of autoimmune thyroid diseases were discussed.