Suppression of Streptococcus mutans Biofilm Formation and Gene Expression by PRG Barrier Coat: A Molecular and Microscopic Study for Preventing Dental Caries.

PURPOSE: This study aimed to investigate the inhibitory effect of a PRG Barrier Coat on biofilm formation and structure by Streptococcus mutans and propose an effective method for preventing dental caries. MATERIALS AND METHODS: Streptococcus mutans MT8148 biofilms were obtained from hydroxyapatite disks with and with- out a PRG Barrier Coat. Scanning electron microscopy (SEM) was used to observe the 12- and 24-h-cultured biofilms, while reverse-transcription polymerase chain reaction (qRT-PCR) was used to quantify caries-related genes. Biofilm adhe- sion assessments were performed on glass. Statistical analysis was performed using a two-sample t-test. RESULTS: A statistically significant difference in Streptococcus mutans biofilm adhesion rate was observed between the con- trol and PRG Barrier Coat-coated samples (p < 0.01). However, there was no statistically significant difference in total bacter- ial count or biofilm volume (p > 0.05). SEM revealed that the PRG Barrier Coat inhibited biofilm formation by Streptococcus mutans. Real-time RT-PCR revealed that the material restricted the expression of genes associated with caries-related bio- film formation. However, the suppression of gtfD and dexB differed from that of other genes. CONCLUSION: PRG Barrier Coat suppressed biofilm formation by Streptococcus mutans by inhibiting the expression of in- soluble glucan synthase, which is associated with primary biofilm formation. The material also affected gene expression and altered the biofilm structure. Tooth surface-coating materials, such as PRG Barrier Coat, may improve caries preven- tion in dental practice.

Effects of polymerization method on flexural and shear bond strengths of a fiber-reinforced composite resin.

This study investigated the effects of indirect composite polymerization on the postcuring mechanical properties of a fiber-reinforced composite. An indirect composite seated on glass fibers preimpregnated with polymerized monomer was polymerized by 1) photoirradiation using a halogen-fluorescent polymerizing unit for 5 min, 2) method 1 plus secondary heating at 100°C for 15 min, 3) photoirradiation using a metal halide light unit for 60 s, or 4) preliminary polymerization using a halogen light unit for 20 s followed by method 3. After polymerization, the flexural and shear bond strengths of the fiber-reinforced composite were examined, as was the flexural strength of non-fiber-reinforced composite specimens polymerized using the same methods. Among non-fiber-reinforced composite specimens, flexural strength was lower for method 1 than for the other three methods; however, among fiberreinforced composite specimens, the four methods did not significantly differ in flexural strength or shear bond strength. Composite-fiber interface separation without breakage of the fiber after flexural strength testing was not observed in specimens polymerized by methods 2 or 4. In conclusion, use of a conventional unit for high-intensity light irradiation after preliminary irradiation, or light irradiation followed by secondary heating, is recommended for polymerization of composite material seated on polymerized glass fiber.

Acute oral complications in a pediatric patient with acute lymphoid leukemia.

Pseudomonas aeruginosa is a low-virulence pathogen that can cause infection in immunocompromised patients. Among pediatric hematologists, it is known that careful attention should be paid to P. aeruginosa as the bacteria responsible for intraoral inflammation, and antibiotic therapy targeting P. aeruginosa is the first-line treatment during neutropenia in patients with hematological malignancy. Immunosuppressed patients, however, are at high risk of developing inflammation. Here, we report a case involving a 10-year-old patient with acute lymphoid leukemia (ALL), in which P. aeruginosa synergistically acted with pre-existing gingivitis to induce unusual oral necrotic lesions, leading to acute buccal cellulitis and pus discharge from external sinus tracts.

Dental manifestations of a pediatric patient with hyperimmunoglobulin e syndrome: a case report.

A 7-year-old girl with hyperimmunoglobulin E syndrome (HIES) presented to our clinic with tooth agenesis in both primary and permanent dentitions. The patient's serum immunoglobulin E level was elevated at 17,091 IU/ml, and her medical history indicated the occurrence of HIES, numerous skin abscesses, and recurrent infection by bacteria and/or fungi such as Candida from birth. She also suffered from heart disease. Dental manifestations included extensive caries, impaired root resorption of primary mandibular central incisors and absence of primary mandibular canines and permanent mandibular lateral incisors. Intraoral phenotypes in HIES patients have already been reported in detail, but no previous report has described abnormalities in the number of primary teeth in HIES patients. The purpose of this report was to describe the dental manifestations in the primary dentition of a hyperimmunoglobulin E syndrome patient and to emphasize the importance of preventive dental management from early childhood in compromised children, such as those with HIES.

Amelogenin is a negative regulator of osteoclastogenesis via downregulation of RANKL, M-CSF and fibronectin expression in osteoblasts.

Amelogenin is a novel enamel matrix protein. Knockout mice showed enhanced osteoclast formation and resorption of tooth cementum. This study investigated the effects of amelogenin on osteoclastogenesis. In co-cultures with calvaria osteoblasts and purified bone marrow cells, amelogenin inhibited osteoclastogenesis dramatically. Furthermore, amelogenin inhibited the expression of receptor activator of nuclear factor kappaB ligand (RANKL), macrophage-colony stimulating factor (M-CSF) and fibronectin in osteoblasts, while RANKL expression was induced by fibronectin and inhibited by treatment with fibronectin small interfering RNA. These results suggest that the inhibitory effects of amelogenin on osteoclastogenesis lead to downregulation of RANKL, M-CSF and fibronectin production in osteoblasts.

GD3 synthase gene found expressed in dental epithelium and shown to regulate cell proliferation.

GD3 synthase is one of the key enzymes involved with ganglioside synthesis, and its activity regulates the main profile of ganglioside expression. We analyzed the expression of the GD3 synthase gene in laser-dissected teeth germs using RT-PCR. The GD3 synthase gene was found expressed in brain, thymus, and tooth germ tissues, however, not in liver or skin specimens. Further, it was highly expressed during the early stage of tooth germ development (embryonic day 14.5), especially in dental epithelia, which gradually reduced in the molar site until postnatal day 7, whereas it was not in dental mesenchyme tissues. In addition, dental epithelial cells transiently transfected with the GD3 synthase gene showed enhanced proliferation. These results indicate that the GD3 synthase gene may be involved in early tooth development, particularly in the proliferation of dental epithelium.

Laminin alpha2 is essential for odontoblast differentiation regulating dentin sialoprotein expression.

Laminin alpha2 is subunit of laminin-2 (alpha2beta1gamma1), which is a major component of the muscle basement membrane. Although the laminin alpha2 chain is expressed in the early stage of dental mesenchyme development and localized in the tooth germ basement membrane, its expression pattern in the late stage of tooth germ development and molecular roles are not clearly understood. We analyzed the role of laminin alpha2 in tooth development by using targeted mice with a disrupted lama2 gene. Laminin alpha2 is expressed in dental mesenchymal cells, especially in odontoblasts and during the maturation stage of ameloblasts, but not in the pre-secretory or secretory stages of ameloblasts. Lama2 mutant mice have thin dentin and a widely opened dentinal tube, as compared with wild-type and heterozygote mice, which is similar to the phenotype of dentinogenesis imperfecta. During dentin formation, the expression of dentin sialoprotein, a marker of odontoblast differentiation, was found to be decreased in odontoblasts from mutant mice. Furthermore, in primary cultures of dental mesenchymal cells, dentin matrix protein, and dentin sialophosphoprotein, mRNA expression was increased in laminin-2 coated dishes but not in those coated with other matrices, fibronectin, or type I collagen. Our results suggest that laminin alpha2 is essential for odontoblast differentiation and regulates the expression of dentin matrix proteins.