Kaoru Sakurai

Oral environmental factors affecting number of microbes in saliva of complete denture wearers.

The purpose of this study was to clarify which oral environmental factors affected number of microbes in saliva in an edentulous environment. We enrolled 68 edentulous subjects in the study. Numbers of total anaerobic bacteria and Candida species in saliva were determined. Age, sex, un-stimulated salivary flow rate, pH and viscosity of saliva, histatin level in saliva, tongue coating status, tongue pressure, denture plaque status, material of denture base, duration of edentulism, frequency of self oral health care and number of cigarettes per day were also investigated as oral environmental factors. Correlation between number of total anaerobic bacteria or Candida species and each oral environmental factor was determined with the Spearman rank correlation coefficient. Stepwise logistic regression analysis was used to identify which factors were significantly associated with level of total anaerobic bacteria and Candida species. Correlation and stepwise logistic regression analyses revealed associations between un-stimulated salivary flow rate, tongue coating status, denture plaque status or frequency of self oral health care and number of total anaerobic bacteria. The correlation analysis showed a significant correlation between age and number of total anaerobic bacteria. Stepwise logistic analysis revealed associations between pH of saliva or viscosity of saliva and level of anaerobic bacteria; it also revealed associations between histatin level in saliva or un-stimulated salivary flow rate and level of Candida species. We conclude that salivary flow rate, in particular, affects number of salivary microbes in an edentulous environment.

Fatigue Strength of Ce-TZP/Al2O3 Nanocomposite with Different Surfaces

Ce-TZP/Al2O3 nanocomposite (NANOZR) has not only higher strength, but also higher fracture toughness than conventional Y-TZP, indicating its potential for use in dental implants. Surface treatment to obtain osseointegration, however, may alter its surface topography, thus affecting the cyclic fatigue strength that plays such an important role in the durability of this material. The aim of this study was to evaluate the influence of surface treatment on cyclic fatigue strength in NANOZR as compared with grit-blasted and acid-etched Y-TZP (125BE Y-TZP). Bi-axial flexure strength was measured in both static and cyclic fatigue tests, as recommended by ISO 6872. The cyclic fatigue test was performed by the staircase method in distilled water at 37°C, with a load of 106 cycles and 10 Hz. Bi-axial flexure strength of NANOZR was 1111-1237 MPa and 667-881 MPa in the static and cyclic fatigue tests, respectively. The bi-axial flexure strength of NANOZR under all conditions was greater than that of 125BE Y-TZP in the static and cyclic fatigue tests. The cyclic fatigue strength of NANOZR was more than twice that of Y-TZP as specified in ISO 13356 for surgical implants (320 MPa), indicating the promise of this material for use in dental implants.

Influence of number and inclination angle of implants on stress distribution in mandibular cortical bone with All-on-4 Concept

PURPOSE The All-on-4 is used in the therapy of edentulous mandibles. However, few studies have investigated the effect of such implants on supporting bone. To clarify differences in stress in peri-implant cortical bone between 2-patterns of 6 implants and 8-patterns of 4 implants with change in inclination angle based on the All-on-4. METHODS Three-dimensional finite element analysis models based on the mean value of the Japanese edentulous mandible were constructed. Implants 13 or 15 mm in length were inserted between the mental foramina. In the 6-implant model, implants were inserted in parallel. In the 4-implant model, the 2 anterior implants were inserted in parallel and the 2 posterior implants in parallel or on a slant. Implants were splinted with a superstructure. A 50 N load was applied to the occlusal surface under condition A, at 2mm distal from the distal implant, or condition B, at the distal end of the superstructure. Maximum von Mises stress on cortical bone was measured. RESULTS Stress was concentrated around the posterior-most implant on the right side. Under condition A, stress increased with 4 implants and increase in angulation. At 45°, stress increased by 23% of that in the 6-implant model. Under condition B, stress increased with 4 implants, although stress decreased with increase in angulation. At 45°, stress decreased by 45% of that in the 6 implants. CONCLUSIONS The use of 4 implants or inclined implants increased stress on peri-implant cortical bone. However, when used in conjunction with a short cantilever, inclined implants decreased stress on peri-implant cortical bone.

Inhibiting microbial adhesion to denture base acrylic resin by titanium dioxide coating.

Mechanical cleaning of dentures is effective in preventing infections such as aspiration pneumonia and denture stomatitis. For denture wearers with a physical handicap and the elderly, however, mechanical cleaning can present problems. The aim of this study was to investigate the effect of coating denture base acrylic resin with titanium dioxide (TiO(2)) in the inhibition of oral microbial adhesion. We prepared uniformly sized acrylic resin plates (10 mm x 10 mm x 0.5 mm), which were divided into two groups (a non-coated group and a TiO(2)-coated group). The plates were immersed in cultured Streptococcus sanguinis or Candida albicans and incubated for 24 h. After incubation, each plate was washed to remove loosely adherent microorganisms, and then incubated for a further 24 h. Adenosine triphosphate (ATP) content of the microorganisms was evaluated using a reagent containing benzalkonium, which extracts intra-cellular ATP. In addition, to determine biofilm formation, we also observed each plate by scanning electron microscopy (SEM). We found that the ATP content of both S. sanguinis and C. albicans was reduced by the TiO(2) coating (P = 0.000). Observation by SEM confirmed that the TiO(2) coating inhibited biofilm formation. The results indicate that a TiO(2) coating on a denture base acrylic resin inhibits adhesion of S. sanguinis and C. albicans.

Enhancement of adhesion strength and cellular stiffness of osteoblasts on mirror-polished titanium surface by UV-photofunctionalization

Ultraviolet (UV)-photofunctionalization of titanium substantially enhances the strength and quality of osseointegration by promoting osteogenic cellular attachment and proliferation. However, the mechanism underlying the initial interaction between the cells and the surface of the material remains to be elucidated, especially where the influence of surface roughness is excluded as a factor. The effect of UV-photofunctionalization on the adhesive strength and cellular stiffness of a single osteoblast and its association with the extent of cell spread, cytoskeletal development and focal adhesion assembly on a very smooth titanium surface was evaluated. Rat bone marrow-derived osteoblasts were cultured on UV-treated or untreated mirror-polished titanium disks. The mean critical shear force required to initiate detachment of a single osteoblast (n=10) was >2000nN on a UV-treated surface at 3h incubation, which was 17 times greater than that on an untreated surface. The mean total energy required to complete the detachment of osteoblasts (n=10) was consistently >60pJ on a UV-treated titanium surface after 24h culture, which was up to 42 times greater than that on an untreated surface. Cellular shear modulus, which represents cellular stiffness, was consistently greater on a UV-treated surface than on an untreated surface after 24h incubation (n=10). This strengthening of cell adhesion and cellular mechanical properties on UV-treated titanium was accompanied by enhanced cell spread and actin fiber development and increased levels of vinculin expression. These results indicate that UV-photofunctionalization substantially strengthens osteoblast retention on titanium bulk material with no topographical features, and that this is associated with enhancement of intracellular structural development during the cell adhesion process.

Colonisation of the oral cavity by periodontopathic bacteria in complete denture wearers

OBJECTIVE The purpose of this study was to investigate colonisation by periodontopathic bacteria and the sites of colonisation in elderly upper and lower complete denture wearers. We also investigated the relationship between level of oral hygiene and colonisation by periodontopathic bacteria. MATERIALS AND METHODS Forty edentulous and 37 dentate volunteers participated in this study. Samples were collected from whole saliva, and levels of Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia and Fusobacterium nucleatum were determined by PCR Invader technology. Detection of these species on oral mucosal and denture surfaces was performed by PCR. Fishers exact test was used for the statistical analysis. Cluster analysis was employed to investigate trends in the periodontopathic bacteria flora in each sampling area. RESULTS Detection rates of periodontopathic bacteria in whole saliva were lower under edentulous conditions than under dentulous conditions, except for A. actinomycetemcomitans and F. nucleatum (p < 0.01). Detection rate of F. nucleatum was the highest in all areas. A positive correlation was observed between DNA quantification of P. gingivalis and number of Candida species in saliva. Cluster analysis of the test species identified two clusters. Tongue-coating status was associated with the detection rate of all periodontopathic bacteria investigated, and denture plaque status was associated with the detection rate of T. denticola and F. nucleatum. CONCLUSION Results indicate the presence of periodontopathic bacteria under edentulous conditions and that the status of oral hygiene of the mucosal or denture surfaces affects colonisation by T. denticola and F. nucleatum.

Biodegradation property of beta-tricalcium phosphate-collagen composite in accordance with bone formation: a comparative study with Bio-Oss Collagen® in a rat critical-size defect model

PURPOSE The objective of this study was to compare osteoconductivity and biodegradation properties of an in-house fabricated beta-tricalcium phosphate (b-TCP)-collagen composite with those of Bio-Oss Collagen® (Osteohealth, Shirley, NY, USA) using a rat calvarial critical-size defect model. MATERIALS AND METHODS b-TCP-collagen composite material was fabricated by mixing b-TCP granules having a particle size of 0.15 to 0.8 mm and 75% porosity, with bovine dermis-derived soluble collagen sponge. The dry weight ratio of b-TCP granules-to-collagen ratios was 4:1. Bio-Oss Collagen or the b-TCP-collagen composite was used to fill a 5.0 mm-diameter calvarial defect in rats. The defects were evaluated by histological and histomorphological analyses of decalcified histological sections with hematoxylin and eosin staining 6 and 10 weeks, respectively, after surgery. RESULTS The defect implanted with the b-TCP composite contained immature bone structures with dense connective tissue in contrast to the abundant fibrous tissue, but no trabecular structure was observed within the defect implanted with Bio-Oss Collagen at 6 weeks postoperatively. Eventually, the defect filled with the b-TCP composite was covered with dense, continuous, mature bone tissue with complete replacement of the graft material. However, in defects filled with Bio-Oss Collagen, only dense connective tissue, containing limited amounts of immature trabecular bone and abundant remnant Bio-Oss particles, was observed. Histomorphological analysis revealed that the b-TCP composite caused greater tissue augmentation with a larger volume of bone tissue observed in the defect and greater bioabsorption of remnant material than Bio-Oss Collagen. CONCLUSION These results indicated that the b-TCP composite has greater osteoconductivity and better biodegradation properties than Bio-Oss Collagen; these properties of the b-TCP-collagen composite complimented bone formation and remodeling.

N-acetyl cysteine as an osteogenesis-enhancing molecule for bone regeneration

Bone regeneration often requires cues from osteogenesis-inducing factors for successful outcome. N-acetyl cysteine (NAC), an anti-oxidant small molecule, possibly modulates osteoblastic differentiation. This study investigated the potential of NAC as an osteogenesis-enhancing molecule in vitro and in vivo. Various concentrations of NAC (0, 2.5, 5.0, and 10 mM) were added to rat bone marrow stromal cell or osteoblastic cell culture in media with or without dexamethasone. The results showed marked enhancement of alkaline phosphatase activity and mineralized matrix formation together with consistent upregulation of bone-related gene markers such as collagen I, osteopontin, and osteocalcin in the osteoblastic culture with addition of 2.5 or 5.0 mM NAC regardless of the presence of dexamethasone. Micro-CT-based analysis and histological observation revealed that addition of NAC to a collagenous sponge implanted in a critical size cortical bone defect (3.0 mm × 5.0 mm) in rat femur yielded acceleration and completion of defect closure, with thick, compact, and contiguous bone after 6 weeks of healing. In contrast, with sponge alone, only sparse and incomplete bone regeneration was observed during the matching healing period. These results indicate that NAC can function as an osteogenesis-enhancing molecule to accelerate bone regeneration by activating differentiation of osteogenic lineages.

Influence of chewing force on salivary stress markers as indicator of mental stress

The aim of this study was to investigate the influence of chewing force on salivary stress markers (alpha-amylase activity, salivary cortisol level and secretory immunoglobulin A secretion rate) as indicators of mental stress. Participants comprised 20 healthy men. The first set of saliva specimens (S1) was collected at immediately after a 20-min rest to evaluate stress markers. As stress loading, the participants were required to perform arithmetic calculations for 20 min, after which the second set of saliva specimens (S2) was collected. Each participant was then required to chew a piece of tasteless gum for 10 min, after which the third set of saliva specimens (S3) was collected. After a 20-min rest, the fourth set of saliva specimens (S4) was collected. Weak, habitual and strong chewing forces were assigned. Change rates of stress markers between S2 and S3, and S2 and S4 were calculated. A significant difference was observed in the change rate of cortisol levels between S2 and S3. Cortisol level decreased more under strong chewing than under weak chewing. No significant differences were observed in the change rate of amylase activity or s-IgA secretion rate among the three chewing forces. The results suggest that differences in chewing force influence the salivary cortisol level of the three stress markers, and that a strong chewing force induces a greater reduction in mental stress than a weak one.

Exposure of P. gingivalis to noradrenaline reduces bacterial growth and elevates ArgX protease activity

OBJECTIVE Periodontitis, an infectious disease caused by periodontopathic bacteria, including Porphyromonas gingivalis, is reported to be accelerated by stress, under which noradrenaline levels are increased in the bloodstream. The purpose of this study was to evaluate the effects of noradrenaline on P. gingivalis. DESIGN P. gingivalis was incubated in the presence of 25μM, 50μM, or 100μM adrenaline or noradrenaline at 37°C for 12, 24 or 36h and growth was evaluated by OD(660). Auto-inducer-2 (AI-2) was measured by luminescence of Vibrio harveyi BB 170. Expression of P. gingivalis genes was evaluated using a microarray and RT-PCR. Rgp activity of arg-gingipainA and B (Rgp) was measured with a synthetic substrate. RESULTS Growth of P. gingivalis FDC381 was inhibited by noradrenaline at 24 and 36h. Growth inhibition by noradrenaline increased dose-dependently. Inhibition of growth partially recovered with addition of propranolol. AI-2 production from P. gingivalis showed a marked decrease with addition of noradrenaline compared with peak production levels in the control group. Microarray analysis revealed an increase in expression in 18 genes and a decrease in expression in 2 genes. Amongst these genes, expression of the protease arg-gingipainB (RgpB) gene, a major virulence factor of P. gingivalis, was further analysed. Expression of rgpB showed a significant increase with addition of noradrenaline, which was partially reduced by addition of propranolol. Cell-associated Rgp activity also increased with addition of noradrenaline. CONCLUSIONS These results suggest that stressors influence the expression of the virulence factors of P. gingivalis via noradrenaline.