Adherence of <Emphasis Type="Italic">Candida albicans</Emphasis> to denture base acrylics and silicone-based resilient liner materials with different surface finishes

This study evaluated the surface roughness and Candida albicans adherence on denture base acrylic resins and silicone-based resilient liners with different surface finishes. Four commercial denture base acrylic resins (three heat polymerized and one room temperature polymerized) and five silicone-based liner materials (two heat polymerized and three room temperature polymerized) (10 × 10 × 2 mm) were tested in this study. The materials were processed against glass or plaster or finished with a tungsten carbide bur. Surface roughness measurements were made using a profilometer with an optical scanner probe. All specimens were ultrasonically cleaned in water for 15 s, autoclave sterilized, and contaminated with C. albicans solution for adherence assay evaluation. The materials processed against the glass surface showed significantly lower surface roughness values (0.11 ± 0.1–1.66 ± 1.1 μm) than those of the materials processed against the dental plaster (2.61 ± 0.2–6.12 ± 2.8 μm) or roughening with a bur (1.48 ± 0.2–7.05 ± 1.2 μm; p < 0.05, one- or two-way analysis of variance). Also, the materials processed against the glass surface showed lower C. albicans adhesion (mean ranks 120.36) than those of the materials processed against the dental plaster (mean ranks 139.77) or roughening with a bur (mean ranks 143.06), but the differences were not statistically significant (p > 0.05, Kruskal–Wallis and Mann–Whitney). In all types of surface finishes, C. albicans adhesion on denture base acrylics was significantly less (mean ranks 90.18–90.40) than those of silicone liners (mean ranks 119.38–205.18; p < 0.01, Kruskal–Wallis).     

Inhibitory effect of alpha-mangostin on <Emphasis Type="Italic">Candida</Emphasis> biofilms

The objective of this study was to determine the inhibitory effect of alpha-mangostin on Candida biofilms. Candida species including Candida albicans, Candida krusei, Candida tropicalis, and Candida glabrata were tested. Candida biofilms were formed in flat-bottomed 96-well microtiter plates. The metabolic activity of cells within biofilms was quantified using the XTT assay. The results demonstrated that alpha-mangostin showed a significant anti-biofilm effect on both developing biofilms and preformed biofilms of Candida species. It may be concluded that alpha-mangostin could be an anti-biofilm agent against Candida species. Further in vivo investigations are needed to uncover the therapeutic values of this medicinal plant.     

Comparison of different methods to detect <Emphasis Type="Italic">Helicobacter pylori</Emphasis> in the dental plaque of dyspeptic patients

The aim of this study was to compare different methods of detection of Helicobacter pylori (H. pylori) in the dental plaque of dyspeptic patients. After recording the clinical indices, culture and polymerase chain reaction (PCR) methods were performed on plaque samples, while rapid urease test in addition to these tests was carried on gastric samples from 67 dyspeptic patients who attended for an upper gastrointestinal endoscopy. Forty-seven of 67 patients were H. pylori-positive in gastric biopsy material whereas the microbial dental plaque from 19 patients demonstrated H. pylori positivity detected by PCR. Among the patients, 25.4% harbored H. pylori both in the stomach and in microbial dental plaque. No significant correlations were found among the presence of H. pylori in the stomach, in plaque, and clinical variables (P > 0.05). Although oral hygiene was observed optimal and the mean of pocket depth was not found to be higher, the prevalence of H. pylori was observed to be higher in dental plaque. According to our results, PCR technique gave the highest detection rate both in gastric biopsy and in dental plaque compared to the other methods used.     

<Emphasis Type="Italic">Tannerella forsythia</Emphasis> and the <Emphasis Type="Italic">HLA-DQB1</Emphasis> allele are associated with susceptibility to periodontal disease in Japanese adolescents

Periodontal disease is a multiple factor disease caused by genetic factors, environmental factors, and periodontal bacteria (periodontal pathogens). The present study aimed to elucidate the risk factors for periodontal disease in Japanese adolescents. Subjects (11–16 years old) were classified into three groups: localized aggressive periodontitis (LAP), periodontal attachment loss (PAL), and periodontally healthy (PH) groups. Genomic DNA isolated from the buccal mucosa was used for single-nucleotide polymorphism analyses of the candidate genes (interleukin-1α-889; interleukin-1α +4845; interleukin-1β +3954; an immunoglobulin G Fc gamma receptor, FcγRIIa-R/H131; and a human leukocyte antigen class II allele, HLA-DQB1) of aggressive periodontitis. Subgingival plaque samples obtained from the same subjects were used for 16S rRNAbased polymerase chain reaction analysis of five important periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola, and Tannerella forsythia). Tannerella forsythia was detected in the deepest periodontal pockets in all subjects in the LAP and PAL groups. The prevalence of an atypical BamHI restriction site in HLA-DQB1 of the LAP group was significantly higher than that in the PH and PAL groups. Furthermore, all subjects who had the atypical BamHI restriction site in HLA-DQB1 had T. forsythia infection. These results suggested that T. forsythia is associated with periodontal disease in Japanese adolescents and also suggested that HLA-DQB1 is related to LAP and is associated with T. forsythia infection.     

Adhesion of<Emphasis Type="Italic"> Porphyromonas gingivalis</Emphasis> to cultured pocket epithelium: mono- and multi-layered

Adhesion of bacteria to epithelial cells might be influenced by the degree of cell differentiation, as observed in the multi-layering process of epithelial cells. In the present study, the adhesion of a large group of clinical Porphyromonas gingivalis strains (n=11) to in vitro cultured mono- and multi-layers of epithelial cells was examined and compared. The tissue samples originated from 6 patients with chronic adult periodontitis. Porphyromonas gingivalis bacteria adhered more to mono-layers as opposed to the more differentiated multi-layers. Differences between the clinical P. gingivalis strains, however, became obvious only on multi-layers. These partially differentiated cells may also better represent the individual subject variations. Mono-layer cultures, which are simpler to obtain, seem to be less precise. The importance of cell differentiation on bacterial adhesion needs more attention.     

Effects of low-concentrated chlorhexidine on growth of<Emphasis Type="Italic"> Streptococcus sobrinus</Emphasis> and primary human gingival fibroblasts

The aim of this study was to investigate the in vitro effects of low concentrations of chlorhexidine (CHX) on the proliferation of Streptococcus sobrinus (ATCC 33478) and primary human gingival fibroblasts (HGF). Liquid cultures of bacteria or human gingival fibroblasts were exposed to CHX concentrations ranging from 0.07 to 40 M in microtiter plates at 37°C for 24 h. Bacteria or cells grown without CHX served as controls. The effects of CHX were determined either by measurements of the optical density (OD) of bacterial cultures or by evaluation of cell growth with the DNA-intercalating fluorescent stain H33342 in comparison to untreated controls. Results were evaluated calculating means and standard deviations. Data were statistically analyzed by an ANOVA using Post Hoc tests (p<0.005). No growth inhibition of S. sobrinus was found at concentrations between 0.07 and 0.15 M CHX, whereas 0.3 M CHX led to an elongated (2 h more) lag phase and 0.6 M CHX induced a lag phase of 4 h more and a minor inclination of the curve in the log phase. Concentrations of CHX1.25 M completely inhibited growth of S. sobrinus. On the contrary, CHX showed no significant effect on growth of HGF at concentrations 5 M. A slight growth inhibition was only observed at a concentration of 5 M. CHX-concentrations of 10 and 20 M reduced cell growth to 88 or 75% of control assays. Data analysis showed that overall treatment effects were highly significant (p<0.005). Our data reveal that chlorhexidine inhibits proliferation of S. sobrinus even at very low concentrations while concentrations of CHX5 M are not cytotoxic to human gingival fibroblasts.     

Effect of mouthrinses on <Emphasis Type="Italic">Aggregatibacter actinomycetemcomitans</Emphasis> biofilms in a hydrodynamic model

The aim of the study was to evaluate the effects of Listerine®, Meridol®, and Perioaid® on the viability and total number of bacteria in established biofilms using an in vitro model under hydrodynamic conditions. Biofilms of Aggregatibacter actinomycetemcomitans were placed in a modified Robbins device and rinsed twice daily during 4 days. Bacteria were quantified by culture and quantitative polymerase chain reaction. Visualization of the samples was performed by scanning electron and confocal laser scanning microscopy, combined with a fluorescent vital staining. All three mouthrinses caused a significant reduction in the number of cultivable A. actinomycetemcomitans in a biofilm. Perioaid® was significantly the most powerful in killing the biofilm-protected bacteria and also in counteracting the development of thick dense microbial communities. The total amount of bacteria was not significantly affected by Listerine® and Meridol®.     

表面不同形貌对钛耐腐蚀性能影响的实验研究

目的：评价不同方法表面处理后纯钛表面粗糙程度对其耐腐蚀性能的影响。方法：制备33个纯钛试件并随机分为3组,分别进行喷砂、阳极氧化和机械抛光处理,应用扫描电镜观察不同处理后钛件表面微形貌,在酸性含氟人工唾液中测试开路电位和阳极极化曲线。结果：扫描电镜结果显示喷砂后钛表面有大量的弹坑和划痕,深度较深,且形状不规则,大小不一,边缘锐利,为微米级粗糙表面形态;而阳极氧化表面为大小均一的直径80Jim的纳米管状结构,为纳米级粗糙表面形态;机械抛光表面划痕明显,深度较浅,为微米级粗糙表面形态。在酸性含氟人工唾液中,3组之间自腐蚀电位值的比较为：阳极氧化组〉机械抛光组〉喷砂组;腐蚀电流密度值的比较为：喷砂组〉机械抛光组〉阳极氧化组,具有显著性差异（P〈0．05）。结论：不同的表面处理技术改变了钛材料表面的微形貌并影响了钛的耐腐蚀性能,经阳极氧化处理后的纳米级表面具有更好的耐腐蚀性能,喷砂会使钛的耐腐蚀性降低。     

PCR detection of <Emphasis Type="Italic">Streptococcus mutans</Emphasis> and <Emphasis Type="Italic">Aggregatibacter actinomycetemcomitans</Emphasis> in dental plaque samples from Haitian adolescents

Streptococcus mutans and Aggregatibacter actinomycetemcomitans are oral pathogens associated with dental caries and periodontitis, respectively. The aim of this study was to determine the colonization of these two microorganisms in the dental plaque of a group of Haitian adolescents using two different polymerase chain reaction (PCR) methods, standard PCR, and quantitative real-time PCR (qPCR) assays. Fifty-four pooled supra-gingival plaque samples and 98 pooled sub-gingival plaque samples were obtained from 104 12- to19-year-old rural-dwelling Haitians. The total genomic DNA of bacteria was isolated from these samples, and all participants also received caries and periodontal examinations. Caries prevalence was 42.2%, and the mean decayed, missing, and filled surface (DMFS) was 2.67 ± 5.3. More than half of the adolescents (53.3%) experienced periodontal pockets (Community Periodontal Index score ≥3). S. mutans was detected in 67.3% by qPCR and 38.8% by PCR of the supra-gingival plaque samples (p < 0.01), and 36.6% by qPCR and 8.1% by PCR of the sub-gingival samples (p < 0.01). A. actinomycetemcomitans was detected in 85.1% by qPCR and 44.0% by PCR of the sub-gingival samples (p < 0.01), but the prevalence was similar, 67.3% by qPCR and 59.2% by PCR, in the supra-gingival plaque samples. Neither age nor gender was significantly correlated to the bacterial colonization. The results demonstrated a moderate-to-high prevalence of S. mutans and A. actinomycetemcomitans in the Haitian adolescent population, and qPCR is more sensitive than standard PCR in field conditions. These findings suggest that qPCR should be considered for field oral epidemiologic studies and may be necessary in investigations having major logistic challenges.     

Clinical effects of scaling and root planing in adults infected with<Emphasis Type="Italic"> Actinobacillus actinomycetemcomitans</Emphasis>

The periodontal pathogen Actinobacillus actinomycetemcomitans can frequently be isolated from subgingival plaque of adults with chronic inflammatory periodontal disease and individuals with plaque-induced gingivitis. Problems with the persistence of the organism after thorough debridement of root surfaces have been reported. In the present study clinical effects of the hygienic phase of periodontal therapy in ten adult patients with moderate or advanced periodontitis harbouring A. actinomycetemcomitans were analysed. Since proper analysis of highly correlated data within a given patient is crucial for appropriate interpretation, a major objective of this study was to compare the results of different models derived from logistic regression of clinical and microbiological factors on gain or loss of clinical attachment under different assumptions. Subgingival samples from every tooth present were obtained before and 6 weeks after thorough subgingival scaling, and selectively cultivated for the organism. A relevant gain of clinical attachment of 2 mm or more was observed at a total of 36% of periodontitis sites after scaling. Overall, loss of attachment of 2 mm or more was observed at 8% sites. Most loss occurred at sites with gingival enlargement (15%), whereas 3% periodontitis sites lost 2 mm or more. In multivariate analyses erroneously assuming either independence of data or correctly considering the correlated structure of observations attachment gain was mainly associated with deep probing depths at the outset. Presence or absence of A. actinomycetemcomitans before or after therapy was not included into the periodontitis models. Also, loss of attachment of 2 mm or more after subgingival scaling was not influenced by the organism. A direct comparison of the results obtained with both approaches of logistic regression may be helpful in the assessment of the influence of the magnitude of correlation of the data on the regression coefficients.     

Impact of <Emphasis Type="Italic">Streptococcus mutans</Emphasis> on the generation of fluorescence from artificially induced enamel and dentin carious lesions in vitro

The purpose of this study was to examine whether Streptococcus mutans is implicated in the generation of fluorescence detected in carious lesions. Enamel surfaces and dentin cavities of extracted human teeth were subjected to artificial caries generation by exposing them either to a culture medium containing S. mutans or to a lactic acid buffer for 2 weeks. Fluorescence from the lesions was detected with confocal laser scanning microscopy or fluorescence microscopy at various excitation wavelengths, and maximum fluorescence radiance was computed using imageanalyzing software. Culture media of S. mutans were also examined for fluorescence generation. The results demonstrated that S. mutans-induced enamel and dentin lesions exhibited increased fluorescence in the red and green spectral regions, with the signal stronger in the red region. In the blue region, however, fluorescence signals in the corresponding area were below the background level. Significantly weaker or virtually no fluorescence was detected in lactic acid-demineralized lesions at all excitation wavelengths. Neither bacterial cells nor culture media generated any fluorescence. These results indicate that, although the presence of S. mutans may be a prerequisite for the emission of fluorescence from carious lesions, some interaction of S. mutans with exposed tooth matrix elements may also be required for the generation or unmasking of fluorophores.     

The effect of Streptococcus mutans and Candida glabrata on Candida albicans biofilms formed on different surfaces

Although Candida containing biofilms contribute to the development of oral candidosis, the characteristics of multi-species Candida biofilms and how oral bacteria modulate these biofilms is poorly understood. The aim of this study was to investigate interactions between Candida albicans and either Candida glabrata or Streptococcus mutans in biofilms grown on various surfaces, with or without saliva. Hydroxyapatite (HA), polymethylmetacrylate (PMMA) and soft denture liner (SL) discs were used as substratum. Counts of viable micro-organisms in the accumulating biofilm layer were determined and converted to colony forming units per unit surface area. Confocal laser scanning microscopy was used to characterize biofilms and to quantitate the number of hyphae in each condition tested. Viable counts of C. albicans and C. glabrata per mm(2) decreased in the order HA>PMMA>SL (p<0.05). Biofilms grown on saliva-coated specimens harboured fewer C. glabrata than uncoated specimens (p<0.05). Glucose and the presence of S. mutans suppressed C. albicans hyphal formation. Dual Candida species biofilms did not show competitive interaction between the two species. We conclude that Candida biofilms are significantly affected by saliva, substratum type and by the presence of other micro-organisms.     

Probiotic <Emphasis Type="Italic">Lactobacillus paracasei</Emphasis> effect on cariogenic bacterial flora

Lactobacillus paracasei has been demonstrated to inhibit the growth of many pathogenic microbes such as Streptococcus mutans, in vitro. However, its clinical application remains unclear. Here, we examined whether a novel probiotic L. paracasei GMNL-33 may reduce the caries-associated salivary microbial counts in healthy adults. Seventy-eight subjects (aged 20 to 26) had completed this double-blinded, randomized, placebo-controlled study. A probiotic/test (n = 42) and a control group (n = 36) took a L. paracasei GMNL-33 and a placebo oral tablet three times per day for 2 weeks, respectively. Bacterial counts of salivary S. mutans, lactobacilli, and salivary buffer capacity were measured with chair-side kits at the beginning (T1), the completion (T2) of medication, and 2 weeks after medication (T3). The results did not show differences in the counts of S. mutans and lactobacilli between probiotic and control groups at T1, T2, and T3. Nevertheless, within the probiotic group, an interesting probiotic effect was noticed. Between T1 and T2, no inhibitory effect against S. mutans was observed. However, a significant count reduction in the salivary S. mutans was detected between T2 and T3 (p = 0.016). Thus, a 2-week period of medication via oral administration route may be needed for L. paracasei GMNL-33 to be effective in the probiotic action.     

Dental caries experience in relation to salivary findings and molecular identification of <Emphasis Type="Italic">S. mutans</Emphasis> and <Emphasis Type="Italic">S. sobrinus</Emphasis> in subjects with Down syndrome

This study investigated the association between clinical and salivary or molecular parameters in Down syndrome subjects. Sixty individuals (1- to 48-year old) were clinically examined using DMFT/DMFS. Stimulated saliva was collected; salivary flow was calculated (mL/min), buffering capacity was measured using a standard pH tape. In addition, 25 μL of saliva was diluted using 10-fold-dilution method and then placed on Mitis-Salivarius-Bacitracin agar to count colony forming units (CFU/mL) of mutans streptococci. Polymerase chain reaction analysis identified species. Caries indexes were 0.65–13.5 (DMFT) and 0.65–26.0 (DMFS) according to groups. Ninety-four percent of subjects had low flow rate (0.7–1.0 mL/min) and 44% had low buffering capacity (pH < 4). Besides, 60% had more than 1 × 10⁶ CFU/mL, 60% had S. mutans, and 41.4% had S. sobrinus. Caries indexes did not significantly correlate with flow rate, buffering capacity, CFU/mL by Pearson’s correlation (p > 0.05), and showed no significant association with prevalence of species by Chi-square (p > 0.05). There is no association between clinical picture and salivary or molecular parameters in Down syndrome subjects.     

Three-dimensional analysis of tooth dimensions in the <Emphasis Type="Italic">MSX1</Emphasis>-missense mutation

Objectives

A novel, 3D technique to measure the differences in tooth crown morphology between the MSX1 cases and non-affected controls was designed to get a better understanding of dental phenotype-genotype associations.

Materials and methods

Eight Dutch subjects from a single family with tooth agenesis, all with an established nonsense mutation c.332 C > A, p. Ser 111 Stop in exon 1 of MSX1, were compared with unaffected controls regarding several aspects of tooth crown morphology of incisor and molar teeth. A novel method of quantitative three-dimensional analysis was used to detect differences.

Results

Statistically significant shape differences were observed for the maxillary incisor in the MSX1 family compared with the controls on the following parameters: surface area, buccolingual dimension, squareness, and crown volume (P?≤?0.002). Molar crown shape was unaffected.

Conclusions

A better understanding of dental phenotype-genotype associations may contribute to earlier diagnosis of some multiple-anomaly congenital syndromes involving dental anomalies.

Clinical relevance

A “shape database” that includes associated gene mutations resulting from developmental syndromes may facilitate the genetic identification of hypodontia cases.

     

The effect of different titanium nitride coatings on the adhesion of Candida albicans to titanium

Objectives

The aim of the present study was an in vitro evaluation of the effects of different titanium nitride (TiN_x) coatings on Candida albicans (C. albicans) adhesion to titanium and to correlate these findings to differences in specific surface characteristics (surface topography, roughness, chemical component, and surface free energy).

Methods

TiN_x coatings were prepared by physical vapour deposition (PVD), a plasma nitriding process or a dual nitriding process. Surface properties were analysed by the optical stereoscopic microscopy, scanning electron microscopy, roughmeter, and drop shape methods. Quantity comparisons of C. albicans on the four surfaces were assessed by cell count and XTT reduction assays. Types of adhesive C. albicans were explored by SEM and confocal laser scanning microscope.

Results

The nitrided modifications were found to influence the surface properties and fungal susceptivity of flat titanium. Compared to flat titanium, fewer adhered C. albicans in yeast form were observed on the TiN-coated surface, whereas the plasma nitrided surface did not show any reduced potential to adhere C. albicans in hyphal or yeast form. The dual nitrided coating showed anti-fungal characteristics, although a small quantity of hyphae were identified. Our findings indicate that the Ti₂N phase is prone to C. albicans hyphae, while the TiN phase inhibits their adhesion.

Conclusions

Different TiN_x phases could influence the characteristics of C. albicans adhesion. TiN coating by PVD could be a potential modification to inhibit C. albicans.     

Oral lichen planus treated with 13-<Emphasis Type="Italic">cis</Emphasis>-retinoic acid (isotretinoin): effects on the apoptotic process

The aim of the present study is to verify the efficacy of isotretinoin in oral lichen planus (OLP). In a double-blind study, ten patients with biopsy-proven OLP were treated for 4 months with 0.1% isotretinoin gel and another ten patients with placebo. At the end of the first period of observation, the patients who had been given the placebo were given isotretinoin for a further 4 months. A complete response was defined as the disappearance of the lesions as assessed by inspection, whereas a partial response was defined as a 50% or more reduction in the size of the lesions. All patients treated with isotretinoin showed a significant improvement of the oral lesions, whereas in the patients who were given the placebo, the size of the lesions remained the same. The patients who were given isotretinoin after the placebo showed a reduction in lesions. In total, there were ten complete and ten partial responses. Lesions were analysed histologically and immunohistochemically with antibodies against bcl-2 and Ki-67. Ki-67 and bcl-2 have statistical significant increased values from before to after treatment, whereas apoptotic bodies decreased one. All these facts could have contributed to the partial or complete regression of OLP lesions. The increase in Ki-67 positive cells show that the epithelium requires for enhanced proliferation and healing. The present results revealed a disturbed cell death programme in OLP that could underline an abnormal epithelial differentiation. The results of this pilot study show that the topical use of isotretinoin is effective in treating OLP.