An in vitro study on the anti-adherence effect of Brucea javanica and Piper betle extracts towards oral Candida

ObjectiveThe adherence of Candida to mucosal surfaces is the initial step for successful invasive process of the oral cavity. The study aimed to investigate the effect of two plant extracts on the non-specific and specific bindings of oral candida.MethodsIn the former, adsorption to hexadecane was used to measure the hydrophobic interaction of the candida cells. In the later, glass beads coated with saliva represented the experimental pellicles in specific adhesion of oral candida to hard tissue surface.ResultsCandida krusei, Candida dubliniensis and Candida tropicalis showed the highest adsorption to hexadecane at 30.23%, 26.19% and 19.70%, respectively, while the others within the range of 7–10%. All candidal species were significantly affected by the extracts (P < 0.05) with Brucea javanica exhibited more than 60% reduction of CSH than Piper betle. Candida parapsilosis showed the highest affinity in specific-bindings to pellicle with 18.72 ± 0.71 × 10⁵ CFU/ml. Exposing to P. betle-treated pellicle has drastically reduced the adherence of C. tropicalis, Candida albicans and C. krusei by 86.01%, 61.41% and 56.34%, respectively. B. javanica exhibited similar effect on C. tropicalis (89.86%), Candida lusitaniae (88.95%), C. albicans (79.74%), Candida glabrata (76.85%) and C. krusei (67.61%).ConclusionThe extracts demonstrated anti-adherence activities by modifying the CSH and the characteristics of the experimental pellicle.     

Molecular analysis of microbiota associated with peri-implant diseases

ObjectivesThe aim of this study was to identify bacteria associated with peri-implant diseases using Denaturing Gradient Gel Electrophoresis (DGGE) as a method for microbiological assessment.MethodsSubgingival plaque samples along with essential patient information and clinical indices were taken from 22 subjects showing signs of peri-implant diseases. Bacteria were detected from extracted DNA either by species specific PCR, or by using PCR coupled with DGGE and subsequent sequencing of resolved and excised bands.ResultsAltogether, approximately 26 different species were identified as components of peri-implant plaque, including non-culturable bacteria. Percentages of periodonto pathogenic bacterial species in plaque samples were: 82% of shallow pocket (<3 mm), 51% of moderate pocket depth, and 63% of deep pocket (>7.5 mm). A positive correlation was found between highly pathogenic bacteria and both Gingival Index score and pocket depth.ConclusionDGGE in combination with sequence analysis is a comprehensive and sensitive microbiological method for detection and identification of multiple bacterial species in peri-implant disease conditions. This makes it a valuable microbiological diagnostic method to help the clinician to conduct a more accurate clinical diagnosis and to plan appropriate treatment for peri-implant diseases.Based on results, Fusobacterium and Prevotella species were most prevalent in early stages of disease whilst an increased diversity of species was present during more advanced stages of disease.     

Changes of saliva microbiota in nasopharyngeal carcinoma patients under chemoradiation therapy

ObjectiveA growing body of evidence has implicated human oral microbiota in the aetiology of oral and systemic diseases. Nasopharyngeal carcinoma (NPC), an epithelial-originated malignancy, has a complex aetiology not yet fully understood. Chemoradiation therapy of NPC can affect oral microbiota and is usually accompanied by plaque accumulation. Thus, the study aimed to understand the diversity, divergence and development of the oral microbiota in NPC patients and their associated treatment, which might provide useful insights into disease aetiology and treatment side effects.DesignA longitudinal study was designed that included three Chinese adults with NPC. Saliva samples were collected at three time points: prior to the chemoradiation treatment (carcinoma baseline, or CB), 7 months post-treatment (carcinoma-after-therapy phase 1 or CA1) and 12 months post-treatment (carcinoma-after-therapy phase 2 or CA2). Pyrosequencing of the bacterial 16S ribosomal DNA (rDNA) V1–V3 hypervariable region was employed to characterise the microbiota. Saliva samples of three healthy subjects from our former study were employed as healthy controls. Principal coordinates analysis (PCoA), Metastats and random forest prediction models were used to reveal the key microbial members associated with NPC and its treatment programme.Results(1) In total, 412 bacterial species from at least 107 genera and 13 phyla were found in the saliva samples of the NPC patients. (2) PCoA revealed that not only were the microbiota from NPC patients distinct from those of healthy controls (p < 0.001) but also that separation was found on the saliva microbiota between pre- and post-therapy (p < 0.001) in the NPC samples. (3) At the genus level and the operational taxonomic unit (OTU) level, Streptococcus was found with lower abundance in NPC samples. (4) Chemoradiation therapy did not incur similar changes in microbiota structure among the three NPC patients; the microbiota in one of them stayed largely steady, while those in the other two showed significant alteration.ConclusionsThis is the first study employing culture-independent techniques to interrogate the phylogenetic diversity, divergence and temporal development of oral microbiota in NPC patients. Our results indicated that certain bacterial taxa might be associated with NPC and that oral microbiota of NPC patients might respond to the chemoradiation therapy in a host-specific manner. Further investigation with larger sample size should help to validate the links between oral microbiota and NPC.     

Microbial profile of dental plaque associated to white spot lesions in orthodontic patients immediately after the bracket removal

ObjectiveDenaturing Gradient Gel Electrophoresis (DGGE) is suggested to predict caries risk in young children. Such a tool would be valuable in orthodontic patients undergoing treatment with fixed appliances. In this cross-sectional study the applicability of DGGE and conventional microbiology for caries risk assessment in orthodontic patients were assessed.DesignDental plaque was obtained from orthodontic patients immediately prior to bracket removal. Presence of white spot lesions (WSL) was assessed immediately post debracketing. DGGE-patterns and band counts were assessed using varying automated band detection settings and compared to visually detected bands to determine optimum settings. Optimum settings were used to compare band patterns in subjects with or without WSL. Microbiological samples were assessed for total colony forming units (CFU’s) and percentages of aciduric flora, Streptococcus mutans, Lactobacillus spp. and Candida albicans.ResultsThirty-seven subjects were included with a mean age of 15.4 yr (SD 1.6 yr; 28 with WSL; 9 without WSL). Depending on settings, DGGE outcomes were different. Optimum minimum profiling absolute to the most intense band of 4% showed no significant difference in band numbers for subjects with or without WSL (p = 0.845). Optimum settings for minimum profiling relative to the most intense band of 15% showed significant lower band numbers for subjects with WSL than those without (p = 0.007). No differences between groups were observed for microbiological parameters.ConclusionThe analysis of DGGE-patterns is ambiguous. Software settings significantly affected outcomes. DGGE-patterns and band numbers like CFU counts were not predictive with respect to WSL formation in these orthodontic patients.     

Effect of different pre-irradiation times on curcumin-mediated photodynamic therapy against planktonic cultures and biofilms of Candida spp

ObjectivesThe aim of this study was to evaluate the effects of pre-irradiation time (PIT) on curcumin (Cur)-mediated photodynamic therapy (PDT) against planktonic and biofilm cultures of reference strains of Candida albicans, Candida glabrata and Candida dubliniensis.Materials and methodsSuspensions and biofilms of Candida species were maintained in contact with different concentrations of Cur for time intervals of 1, 5, 10 and 20 min before irradiation and LED (light emitting diode) activation. Additional samples were treated only with Cur, without illumination, or only with light, without Cur. Control samples received neither light nor Cur. After PDT, suspensions were plated on Sabouraud Dextrose Agar, while biofilm results were obtained using the XTT-salt reduction method. Confocal Laser Scanning Microscopy (CLSM) observations were performed to supply a better understanding of Cur penetration through the biofilms after 5 and 20 min of contact with the cultures.ResultsDifferent PITs showed no statistical differences in Cur-mediated PDT of Candida spp. cell suspensions. There was complete inactivation of the three Candida species with the association of 20.0 μM Cur after 5, 10 and 20 min of PIT. Biofilm cultures showed significant reduction in cell viability after PDT. In general, the three Candida species evaluated in this study suffered higher reductions in cell viability with the association of 40.0 μM Cur and 20 min of PIT. Additionally, CLSM observations showed different intensities of fluorescence emissions after 5 and 20 min of incubation.ConclusionPhotoinactivation of planktonic cultures was not PIT-dependent. PIT-dependence of the biofilm cultures differed among the species evaluated. Also, CLSM observations confirmed the need of higher time intervals for the Cur to penetrate biofilm structures.     

Overexpression of immunosuppressive cytokines is associated with poorer clinical stage of oral squamous cell carcinoma

ObjectiveThe aim of this study was to evaluate the expression of IL-10 and TGF-β2 in oral squamous cell carcinoma (OSCC) and its relationship with prognostic clinical and microscopic parameters.DesignImmunohistochemistry was used to assess the expression of IL-10 and TGF-β2 in OSCC samples from 43 patients who had undergone surgical excision and neck dissection. Metastatic lymph nodes were included in the study (n = 23). Samples of healthy oral mucosa (n = 20) were used as controls. The sections were evaluated using a semi-quantitative method in conjunction with staining intensity.ResultsOur findings showed that the expression of IL-10 and TGF-β2 by neoplastic and stromal cells was high in most of the OSCC samples (>70% of samples), especially when compared to the controls (≅10% of samples) (P < 0.05). OSCC neoplastic cells in cervical lymph nodes were also positive for IL-10 and TGF-β2. An association between high expression of IL-10 by neoplastic cells and advanced clinical stage (T3-T4) was verified (P = 0.02). Although not statistically significant, the expression of TGF-β2 was also augmented in advanced stage tumours.ConclusionsThese data suggest that the ability of OSCC neoplastic cells to secrete immunosuppressive cytokines could contribute to clinical progression by maintaining a microenvironment conducive to evasion and tumour proliferation.     

Increased activity of the antioxidants systems modulate the oxidative stress in saliva of toddlers with early childhood caries

ObjectiveThis study aimed to evaluate the oxidative stress levels and the enzymatic and non-enzymatic antioxidant systems in saliva of toddlers with severe early childhood caries (S-ECC).DesignUnstimulated saliva samples were collected at the morning from 0 to 3 year-old S-ECC (n = 30) or caries-free (CF) children (n = 30/group) for evaluation of oxidative stress (OS) and total antioxidant capacity (TAC), which were measured by the ferric reducing antioxidant power (FRAP) assay, as well as to assess the activity of enzymatic (superoxide dismutase, SOD) and non-enzymatic (uric acid, UA) antioxidant systems, respectively. Data were analyzed by Student’s t-test (p < 0.05).ResultsSignificantly higher protein levels were observed in saliva of S-ECC children (0.083 mg/mL) than in the CF group (0.070 mg/mL). Oxidative damage was significantly lower in saliva of S-ECC children (0.0019 μmol/L/mg protein) than in CF children (0.0039 μmol/L/mg protein), while salivary TAC (61.5 μmol/L), SOD activity (36.6 UE/mL) and uric acid (7.05 mg/mL) were significantly higher in saliva of S-ECC when compared to the CF group (49.1 μmol/L, 26.8 UE/mL and 5.02 mg/mL, respectively for TAC, SOD and UA).ConclusionOxidative stress levels were significantly lower in saliva of S-ECC children, what might be associated with the increased activity of salivary enzymatic (SOD) and non-enzymatic (uric acid) antioxidant systems.     

Evaluation of the gingival inflammation in pregnancy and postpartum via 25-hydroxy-vitamin D3, prostaglandin E2 and TNF-α levels in saliva

BackgroundPhysiological changes and immunological modifications occur during pregnancy. The clinical and biological features of periodontal infections are affected by pregnancy. The aim of the present study was to evaluate saliva levels of 25-hydroxy-vitamin D3 (25(OH)D3), prostaglandin E2 (PGE₂) and TNF-alpha (TNF-α) in pregnancy, postpartum and non-pregnant controls.MethodsWhole saliva samples together with full-mouth clinical periodontal recordings were obtained from 59 pregnant, 47 post partum and 70 systemically healthy non-pregnant women. Groups were also evaluated according to the periodontal health status. 25(OH)D3, PGE₂ and TNF-α levels in the saliva samples were determined by enzyme-linked immunoassays. Data were statistically tested by nonparametrical tests.ResultsSaliva TNF-α and PGE₂ levels were significantly lower and 25(OH)D3 levels were significantly higher in the pregnant group than postpartum group (p < 0.0001). Saliva TNF-α and 25(OH)D3 levels were significantly higher and PGE₂ levels were significantly lower in the control group than postpartum group (p < 0.0001). In the pregnant healthy, gingivitis and periodontitis groups saliva TNF-α levels were significantly lower than postpartum and control counterparts (p < 0.0001, p = 0.032, p = 0.003 and p = 0.013; p = 0.027; p = 0.007, respectively). In control healthy, gingivitis and periodontitis groups saliva 25(OH)D3 levels were significantly higher than the postpartum counterparts (p < 0.0001, p < 0.0001, p = 0.002, respectively). In the control healthy and gingivitis groups saliva 25(OH)D3 levels were significantly higher than pregnant healthy and gingivitis (p < 0.0001).ConclusionsIn conclusion, within the limits of the present study it seems that pregnancy have an effect on parameters in saliva in relation to the periodontal status of the women. Further studies are required for better understanding of the impact of periodontal diseases on pregnancy or otherwise.     

Probiotic intervention influences the salivary levels of Matrix Metalloproteinase (MMP)-9 and Tissue Inhibitor of metalloproteinases (TIMP)-1 in healthy adults

ObjectiveTo study the effect of orally administered Bifidobacterium animalis subsp. lactis BB-12 and Lactobacillus rhamnosus GG on the salivary levels of Matrix Metalloproteinases (MMP)-8, MMP-9 and of Tissue Inhibitor of Metalloproteinases (TIMP)-1 in healthy adults. Furthermore, the correlations between MMP-8, MMP-9 and TIMP-1 and plaque and gingival indices, salivary mutans streptococci and lactobacilli counts, and stimulated saliva secretion rate were analysed.DesignThe salivary samples originated from a randomized controlled trial where healthy student volunteers consumed probiotic or placebo lozenges twice a day for four weeks. The saliva samples were collected and clinical parameters measured at the baseline and at the end of the original study. For this study, the salivary levels of MMP-8, MMP-9 and TIMP-1 were analysed with immunofluorometric assay (IFMA) and enzyme-linked immunosorbent assay (ELISA).ResultsIn the probiotic group (n = 29), salivary MMP-9 levels increased (p < 0.01) and TIMP-1 levels decreased (p < 0.01) significantly during the intervention. Furthermore, MMP-9/TIMP-1 ratio differed significantly from the baseline level (p < 0.01). These changes were not observed in the control group (n = 31). In the whole data, salivary MMP-9 and gingival index correlated (r = 0.260, p < 0.05 at baseline and r = 0.354, p < 0.01 at the end of the study). Intergroup differences or correlations with other clinical parameters were not found. Probiotic consumption did not affect the saliva flow rate.ConclusionsIncreased MMP-9 and decreased TIMP-1 levels in saliva may indicate that probiotics have immunomodulatory effects in the oral cavity. Furthermore, increased salivary MMP-9 levels may be an indication of the defensive potential of matrix metalloproteinases.     

The effect of initial periodontal treatment on plasma,gingival crevicular fluid and salivary levels of 8-hydroxy-deoxyguanosine in obesity

ObjectiveRecent studies have shown adverse effects on the periodontium from the increased production of reactive oxygen species (ROS) in obesity. The purpose of this study was to investigate the effects of obesity on 8-hydroxy-deoxyguanosine (8-OHdG) levels in the bodily fluids of patients with and without periodontal disease and to evaluate changes after initial periodontal treatment.DesignForty-five obese individuals and 45 normal-weight individuals were included in this study. Obese and normal-weight groups were classified into three sub-groups: chronic periodontitis (CP), gingivitis (G) and periodontally healthy controls (CTRL). Gingival crevicular fluid (GCF), plasma, saliva samples and clinical measurements were obtained at baseline and a month after initial periodontal treatment. Levels of 8-OHdG were analysed by ELISA.ResultsWhile plasma 8-OHdG levels were significantly higher at baseline in the obese patients with periodontal disease than in the normal-weight individuals (P < 0.05), no significant differences in GCF and saliva 8-OHdG levels were found (P ˃ 0.05). GCF and salivary 8-OHdG levels in obese patients with G and CP were significantly higher than in CTRL groups at baseline (P < 0.05). After treatment, 8-OHdG levels were decreased in all groups with periodontal disease (P < 0.01). Statistically significant positive correlations were observed between GCF 8-OHdG levels and GI in all the groups (P < 0.001).ConclusionsThe significant increase of plasma 8-OHdG levels in obese patients did not correlate with saliva and GCF 8-OHdG levels when compared to normal-weight individuals. Periodontal treatment had a positive effect on the periodontal parameters and 8-OHdG levels of both obese and normal-weight individuals.     

Salivary pellicles equalise surfaces’ charges and modulate the virulence of Candida albicans biofilm

IntroductionNumerous environmental factors influence the pathogenesis of Candida biofilms and an understanding of these is necessary for appropriate clinical management.AimsTo investigate the role of material type, pellicle and stage of biofilm development on the viability, bioactivity, virulence and structure of C. albicans biofilms.MethodsThe surface roughness (SR) and surface free energy (SFE) of acrylic and titanium discs was measured. Pellicles of saliva, or saliva supplemented with plasma, were formed on acrylic and titanium discs. Candida albicans biofilms were then generated for 1.5 h, 24 h, 48 h and 72 h. The cell viability in biofilms was analysed by culture, whilst DNA concentration and the expression of Candida virulence genes (ALS1, ALS3 and HWP1) were evaluated using qPCR. Biofilm metabolic activity was determined using XTT reduction assay, and biofilm structure analysed by Scanning Electron Microscopy (SEM).ResultsWhilst the SR of acrylic and titanium did not significantly differ, the saliva with plasma pellicle increased significantly the total SFE of both surface. The number of viable microorganisms and DNA concentration increased with biofilm development, not differing within materials and pellicles. Biofilms developed on saliva with plasma pellicle surfaces had significantly higher activity after 24 h and this was accompanied with higher expression of virulence genes at all periods.ConclusionInduction of C. albicans virulence occurs with the presence of plasma proteins in pellicles, throughout biofilm growth. To mitigate such effects, reduction of increased plasmatic exudate, related to chronic inflammatory response, could aid the management of candidal biofilm-related infections.     

A preliminary study of the oral microbiota in Chinese patients with Sjögren’s syndrome

ObjectiveTo investigate the oral microbiota in Sjögren’s syndrome (SS) as opposed to that of healthy subjects.Study designTen patients with primary SS, [6 patients daily taking stable dosage of hydroxychloroquine (HC) and 4 patients taking hydroxychloroquine combined with Prednisone acetas (HC + PA)], along with 10 age-matched healthy controls were examined in regard of number of teeth, stimulated/unstimulated saliva secretion rate. Microflora on bilateral buccal mucosa was analyzed by high throughput sequencing. Statistical analyses were performed using the chi-square test, t test and Mann-Whitney U test. The Venn diagrams and Redundancy Analysis (RDA) were also used to evaluate effects of the disease and treatment on the bacterial community composition.ResultsThe relative abundance of Proteobacteria in SS group was lower compared to controls (P = 0.002). The total richness of genera for all groups was 339. The numbers of genera in SS group and in control group were 248 and 270, respectively. Some taxa with different prevalence and/or relative abundance were found between two groups.ConclusionsSS affects the oral microbiota and SS patients carry a different and less diverse microorganism community compared with healthy subjects. Prednisone acetas is an influence on the oral microbiome. This study provides a basic data on the oral flora in SS patients.     

Frequency of periodontal pathogens in equivalent peri-implant and periodontal clinical statuses

ObjectivesThis study tested the hypotheses that there is: (1) higher bacterial frequency in peri-implantitis/periodontitis, followed by mucositis/gingivitis and peri-implant/periodontal health; (2) similar bacterial frequency between comparable peri-implant and periodontal clinical statuses.Design of studyThe presence of Porphyromonas gingivalis, Tannerella forsythia, Campylobacter rectus, Prevotella intermedia, Treponema denticola and Aggregatibacter actinomycetemcomitans was evaluated in peri-implant (n = 53) and periodontal (n = 53) health; mucositis (n = 50), gingivitis (n = 50), peri-implantitis (n = 50) and periodontitis (n = 50).ResultsThe pattern of peri-implant bacterial frequency was not as expected (peri-implantitis > mucositis > health). Except for P. intermedia (p > 0.05), bacterial frequency was higher in peri-implantitis than health (p < 0.05). The frequency of P.gingivalis and red complex species were higher in peri-implantitis than mucositis (p < 0.05). In periodontal samples, T. forsythia and T. denticola showed the expected pattern of frequency (periodontitis > gingivitis > health). The frequencies of C. rectus and T. forsythia were higher in healthy teeth/gingivitis than healthy implants/mucositis, respectively (p < 0.05). The frequency of P. gingivalis and A. actinomycetemcomitans were similar between periodontitis and peri-implantitis (p > 0.05) while all other species occurrences were higher in periodontitis than peri-implantitis (p < 0.05).ConclusionsBacterial frequency increased from peri-implant/periodontal health to peri-implantitis/periodontitis but not from mucositis/gingivitis to peri-implantitis/periodontitis. There was a trend towards higher bacterial frequency in teeth than implants.     

Nickel and chromium levels in the saliva of a Saudi sample treated with fixed orthodontic appliances

AimThe aim of this study was to measure the amount of nickel (Ni) and chromium (Cr) released into the saliva of Saudi patients treated with fixed orthodontic appliances.Materials and methodsNinety salivary samples were collected in a cross-sectional manner. Forty samples were collected from patients (17 males, 23 females) with fixed orthodontic appliances after different periods of orthodontic treatment ranging from the first month and up to 32 months into treatment. The fixed orthodontic appliance consisted of 4 bands, 20 stainless steel brackets, and upper and lower nickel titanium or stainless-steel arch wires. The other 50 samples were collected from people without appliances (24 males, 26 females). Samples were analyzed using Inductive Coupled Plasma/Mass Spectrometry and Inductively Coupled Plasma Optical Emission Spectroscopy to measure Ni and Cr levels, respectively. Student’s t-test was used to compare Ni and Cr levels in the treated and untreated control groups.ResultsThe mean Ni level was 4.197 μg/L in the experimental group and 2.3 μg/L in the control group (p < 0.05). The mean Cr level was 2.9 μg/L in the experimental group and 3.3 μg/L in the control group (p < 0.05).ConclusionFixed orthodontic appliances resulted in a non-toxic increase in salivary levels of Ni, but no change in Cr levels. Duration of orthodontic treatment did not affect Ni and Cr levels in the saliva.     

Impact of glycemic control on oral health status in type 2 diabetes individuals and its association with salivary and plasma levels of chromogranin A

ObjectiveTo evaluate the effect of glycemic control status in type 2 diabetes mellitus (T2DM) individuals on clinical oral health indicators and to compare the concentrations of plasma and salivary chromogranin A (CHGA) among nondiabetic subjects and T2DM patients, exploring their associations.DesignIn this cross-sectional study, 32 patients with controlled T2DM, 31 with poorly controlled T2DM and 37 nondiabetic subjects underwent a clinical and periodontal examination. CHGA concentrations were determined in saliva and plasma with ELISA.ResultsPoorly controlled T2DM group exhibited significantly higher mean buffering capacity, plaque index and bleeding on probing than other groups (P < 0.05). No difference was found to DMFT (decayed, missed and filled teeth) index between groups. Sites with clinical attachment loss (CAL) of 4 and 5–6 mm were significantly higher in both diabetic groups compared to control group (P < 0.05). Poorly controlled T2DM group had significantly higher sites with CAL ≥ 7 mm than other groups (P = 0.001). Significantly higher plasma and salivary CHGA levels were found in T2DM groups (P < 0.05). In both diabetic groups, probing depths 5–6 mm and CAL 5–6 mm were associated with higher salivary CHGA concentration (P < 0.05).ConclusionsThe findings revealed that T2DM patients were more prone to periodontal tissue damage than to caries risk. The results also provide some evidence that the degree of attachment loss deteriorates significantly with poor glycemic control in T2DM (CAL ≥ 7 mm). Moreover, the results suggest that high concentrations of salivary CHGA are associated with worse periodontal parameters and T2DM, and this could be related to the pathogenesis of both diseases.     

DEFB1 polymorphisms and salivary hBD-1 concentration in Oral Lichen Planus patients and healthy subjects

ObjectivesThe aetiology of Oral Lichen Planus (OLP), a chronic inflammatory disease of oral mucosa, is not yet well understood. Since innate immunity may be hypothesized as involved in the susceptibility to OLP, we studied human beta defensin 1 (hBD-1) an antimicrobial peptide constitutively expressed in the saliva, looking at functional genetic variants possibly able to diminish hBD-1 production an consequently conferring major susceptibility to OLP.DesignWe analysed three DEFB1 polymorphisms at 5′ UTR, −52G > A (rs1799946), −44C > G (rs1800972), −20G > A (rs11362) and two DEFB1 polymorphisms at 3′UTR, c*5G > A (rs1047031), c*87A > G (rs1800971), with the aim of correlating these genetic variants and hBD-1 salivary level in a group of OLP patients and in healthy subjects. We also evaluated hBD-1 salivary concentrations, using ELISA, in OLP and healthy controls.ResultsWe compared hBD-1 concentrations in OLP and healthy subjects: hBD-1 concentration was significantly higher in OLP patients respect to control.When considering the correlation between DEFB1 polymorphisms genotypes and hBD-1 expression levels, significant results were obtained for SNPs −52G > A (p = 0.03 both in OLP patients and healthy individuals) and −44C > G (p = 0.02 in OLP patients).ConclusionshBD-1 production was different between OLP and healthy subjects (not age-matched with OLP). DEFB1 gene polymorphisms, −52G > A and −44C > G, correlated with hBD-1 salivary concentrations.     

Macrophage activating factor: A potential biomarker of periodontal health status

ObjectiveIn periodontitis, activated macrophages not only initiate immune responses to periodontal-pathogen infections, but also damage the periodontal tissues by releasing a series of inflammatory cytokines. Macrophage-activating factor (MAF) and macrophage-chemotactic factor (MCF) are two important mediators involved in macrophage accumulation, activation and function. This study analyzed the levels of salivary MAF and MCF in healthy individuals and those with different periodontal diseases, and assessed the usefulness of salivary MAF and MCF as diagnostic biomarkers in periodontal tissue health status.DesignNinety-five saliva specimens were collected from healthy individuals (n = 19), and patients with gingivitis (n = 19), mild periodontitis (n = 17), moderate periodontitis (n = 20), and severe periodontitis (n = 20). Pocket probing depth (PPD) and alveolar bone loss (ABL) were recorded via periodontal probing and dental radiography, respectively. Salivary MAF and MCF concentrations were assayed using enzyme-linked immunosorbent assays.ResultsMAF level tended to increase in saliva as periodontal diseases progressed (healthy periodontium < gingivitis < mild periodontitis < moderate periodontitis < severe periodontitis). The concentration of salivary MAF in periodontitis correlated positively with ABL (r = 0.758) and PPD (r = 0.779). In contrast, salivary MCF levels increased significantly only in periodontitis.ConclusionsSalivary MAF levels correlate positively with tissue destruction in periodontal diseases. It is a potential valuable biomarker that could be used to assess periodontal health status.     

Bioactive resin-based composite with surface pre-reacted glass-ionomer filler and zwitterionic material to prevent the formation of multi-species biofilm

ObjectiveThis study evaluated the synergetic effect between surface pre-reacted glass-ionomer (SPRG) filler and 2-methacryloyloxyethyl phosphorylcholine (MPC), for inhibiting multi-species biofilm formation, while maintaining or even improving the original beneficial features of SPRG-filled resin-based composite (RBC).MethodsMPC (1.5–10 wt%) was incorporated into commercial SPRG-filled RBC. Then, the inherent properties of RBC, and ion release and acid-neutralising properties associated with SPRG were investigated. Further, protein adsorptions and bacterial adhesion and viability on the SPRG-filled RBC surfaces were studied using four kinds of oral bacteria; Streptococcus mutans, Actinomyces naeslundii, Veillonella parvula, and Porphyromonas gingivalis. Finally, the thickness and biomass of the human saliva-derived biofilm model cultured on test and control samples were analysed.ResultsAddition of MPC content resulted in decreased flexural strength and wettability of SPRG-filled RBC. SPRG-filled RBC released significantly higher amounts of multiple ions as contents of MPC increased. Meanwhile, SPRG-filled RBC with 5-wt% MPC significantly improved acid-neutralising properties than those of other test and control samples (P < 0.001). SPRG-filled RBC with 3 wt% MPC significantly reduced the amount of adsorbed bovine serum albumin and proteins from the brain heart infusion medium as compared to the control (P < 0.01). A similar trend was observed in the attachment of four types of bacteria and multi-species biofilm (P < 0.01).SignificanceDespite limitation in terms of deteriorations of some physical properties, addition of 3% MPC to SPRG-filled RBC leads to inhibition of the attachment of multi-species bacteria on its surface, as well as inhibition of biofilm growth. Moreover, the original important bioactive features of SPRG-filled RBC such as ion release and acid neutralisations are either maintained or improved upon adding MPC.     

The membrane-associated MUC1 improves adhesion of salivary MUC5B on buccal cells. Application to development of an in vitro cellular model of oral epithelium

ObjectivesThe mucosal pellicle is a thin layer of salivary proteins, mostly MUC5B mucins, anchored to epithelial oral cells. This pellicle is involved in protection of oral mucosae against abrasion, pathogenic microorganisms or chemical xenobiotics. The present study aimed at studying the involvement of MUC1 in mucosal pellicle formation and more specifically in salivary MUC5B binding using a cell-based model of oral epithelium.DesignMUC1 mRNAs were not detected in TR146 cells, and therefore a stable cell line named TR146/MUC1 expressing this protein was developed by transfection. TR146 and TR146/MUC1 were incubated with human saliva in order to evaluate retention of MUC5B by epithelial cells.ResultsThe cell surface of both TR146 and TR146/MUC1 was typical of a squamous non-keratinized epithelium, with the presence of numerous microplicae. After incubation for 2 h with saliva diluted in culture medium (1:1) and two washes with PBS, saliva deposits on cells appeared as a loose filamentous thin network. MUC5B fluorescent immunostaining evidenced a heterogeneous lining of confluent cell cultures by this salivary mucin but with higher fluorescence on TR146/MUC1 cells. Semi-quantification of MUC5B bound to cells confirmed a better retention by TR146/MUC1, evaluated by Dot Blot (+34.1%, p < 0.05) or by immunocytochemistry (+44%, p < 0.001).ConclusionThe membrane-bound mucin MUC1 is a factor enhancing the formation of the mucosal pellicle by increasing the binding of salivary MUC5B to oral epithelial cells. An in vitro model suitable to study specifically the function and properties of the mucosal pellicle is proposed.