Gene variants for the WNT pathway are associated with severity in periodontal disease.

OBJECTIVE: Studies of Wnt variants-related to bone resorption in periodontitis are limited. The aim of this study was to establish the genotype and allele frequency of gene variants associated with the Wnt pathway in systemically healthy individuals with and without periodontitis (PD). MATERIALS AND METHODS: One hundred fifty-seven systemically healthy individuals were evaluated, 90 with PD and 67 without PD. Periodontal clinical indexes, serological and clinical indices of inflammation, and the following variants associated with the Wnt pathway: DKK, SOST, LRP5, and KREMEN were analyzed by high resolution melting and confirmed by Sanger sequencing. RESULTS: In the PD-free group, 67.2% of the individuals presented the variant for DKKrs1896367 (p = 0.008) and 82.6% had the variant for KREMEN rs132274 (p = 0.016). The heterozygous variant for the DKK rs1896367 polymorphism was associated with the absence of PD and lower severity OR: 0.33 (CI95% 0.15-0.70) and OR: 0.24 (CI95% 0.11-0.53), respectively. Similarly, KREMEN rs132274 was the homozygous variant associated with the absence of PD (OR: 0.33 (CI95% 0.13-0.88)). On the contrary, 85.6% of individuals with PD presented a variant for DKK rs1896368 (p = 0.042), all suffering severe forms of periodontitis. CONCLUSION: The presence of DKKrs1896367 and KREMENrs132274 variants in individuals without PD suggests that these single nucleotide polymorphisms could be protective factors for bone loss in PD. A very interesting finding is that the DKKrs1896368 variant was found in a high percentage of severe cases, suggesting that the presence of this variant may be related to the severe bone loss observed in PD.

A new model for the formation of an Enterococcus faecalis endodontic biofilm with nutritional restriction.

An in vitro model for the formation of an Enterococcus faecalis endodontic biofilm under nutritional restriction was established, simulating clinical conditions for the evaluation of antimicrobial substances. Biofilm formation in dentin was standardized using root quarters incubated with E. faecalis ATCC 29212 at 37°C without nutritional changes. Biofilms were evaluated at 7, 14, and 30 days, counting bacterial colony-forming units using conventional culture and verified scanning electron microscopy. Bacterial viability and biovolume were determined with confocal laser microscopy. Colonization of E. faecalis and biofilm formation on the dentinal surface was confirmed after 7 and 14 days, respectively. Microorganism colonization was homogeneous over the entire root surface at each time point, without significant differences in the viability percentage and biovolume. On the contrary, a decrease in viability and an increase in biovolume were observed when the time was increased. Compared with other incubation times, 14 days was found to be the best time for the establishment of the biofilm in terms of biovolume and bacterial viability. This in vitro model for the formation of endodontic biofilm will allow future evaluation of the efficacy of antimicrobial substances with a more adequate clinical approach.

Characterization of the Subgingival Cultivable Microbiota in Patients with Different Stages of Periodontitis in Spain and Colombia. A Cross-Sectional Study.

The objective was to characterize and compare the subgingival microbiota in patients diagnosed according to the World Workshop on the Classification of Periodontal and Peri-Implant Diseases and Conditions 2018. For this cross-sectional study, Spanish and Colombian subjects (characterized as health/gingivitis, periodontitis in stages I-II or stages III-IV) were clinically assessed, and subgingival samples were taken and processed by culture. The comparisons among patients with periodontal status (and between countries) was made using Mann-Whitney, Kruskal-Wallis, ANOVA and chi-square tests. The final sample consisted of 167 subjects. Eikenella corrodens and Parvimonas micra were more frequently detected in health/gingivitis and Porphyromonas gingivalis in periodontitis (p < 0.05). Higher total counts were observed in Colombia (p = 0.036). In Spain, significantly higher levels of P. gingivalis and Campylobacter rectus were observed, and of Tannerella forsythia, P. micra, Prevotella intermedia, Fusobacterium nucleatum, Actinomyces odontolyticus and Capnocytophaga spp. in Colombia (p < 0.001). P. micra was more prevalent in health/gingivitis and stage I-II periodontitis in Colombia, and P. gingivalis in all periodontitis groups in Spain (p < 0.05). As conclusions, significant differences were detected in the microbiota between health/gingivitis and periodontitis, with minor differences between stages of periodontitis. Differences were also relevant between countries, with Colombia showing larger counts and variability of bacterial species.

Comparison of Cell Viability and Chemical Composition of Six Latest Generation Orthodontic Wires.

Orthodontic wires are made of alloys containing different metals, including nickel. It is important to evaluate their biocompatibility prior to use, owing to their long-term use in patients. This in vitro study compared the cytotoxicity and chemical composition of six latest orthodontic wires: Fantasia®, Tanzo®, FLI®, NT3®, DuoForce®, and Gummetal®. The before-use group consisted of wires that were not used in the mouth, and the after-use group consisted of wires that were used in the mouth for two months. The wires were placed in contact with human gingival fibroblasts (HGF) for 72 h, and cytotoxicity was determined using the resazurin test. The chemical composition and surface characterisation were evaluated by spectrometry and scanning electron microscopy. The groups were compared using ANOVA and Kruskal-Wallis test. Only the FLI® wires produced a 36% reduction in HGF viability (p < 0.05) and presented greater irregularities and loss of polymer structure. After-use wires showed a significant reduction in the percentage of nickel and the appearance of new elements (oxygen and carbon). Therefore, it can be concluded that no toxic ion release was noticed in this study. Rhodium-coated wires were more stable than PTFE-coated wires, and only the FLI® wires showed a slight cytotoxic effect.

Prevalencia de la enfermedad periodontal en pacientes con antecedente de síndrome coronario agudo en un programa de rehabilitación cardiaca / Prevalence of periodontal disease in patients with a history of acute coronary syndrome in a cardiac rehabilitation programme

Resumen Objetivo: establecer la prevalencia y la gravedad de la enfermedad periodontal en pacientes con síndrome coronario agudo sin comorbilidades, que ingresan al programa de rehabilitación cardíaca en la Fundación Clínica Shaio. Métodos: se examinaron 83 pacientes con diagnóstico de síndrome coronario agudo sin comorbilidades y los siguientes diagnósticos: angina inestable (n = 27), infarto de miocardio con elevación del segmento ST (STEMI) (n = 34) e infarto de miocardio sin elevación del segmento ST (NSTEMI) (n = 22). La prevalencia y la gravedad de la enfermedad periodontal fueron evaluadas con el índice periodontal de los Centros para el Control y la Prevención de Enfermedades (CDC). Se compararon entre los grupos índice clínico, placa, cálculo, hemorragia gingival, profundidad de sondaje y nivel de inserción clínica. Resultados: la prevalencia de enfermedad periodontal fue del 97,6% y se observó periodontitis avanzada en el 38,3%. Los pacientes con STEMI mostraron la mayor gravedad de la periodontitis. Los índices clínicos inflamatorios de la enfermedad periodontal se elevaron en todos los grupos sin diferencias significativas. El nivel de inserción clínica no mostró diferencias significativas entre las condiciones cardíacas. Sin embargo, se observó mayor porcentaje de sitios con pérdida de inserción clínica (PIC) ≥ 6 mm, y sitios con una profundidad de bolsas > 6 mm, en pacientes con STEMI. Conclusión: los pacientes con síndrome coronario agudo sin comorbilidades, que ingresaron al programa de rehabilitación cardíaca para completar su tratamiento, tuvieron alta prevalencia y gravedad de la enfermedad periodontal. Es necesario hacer énfasis en el control de la enfermedad periodontal en pacientes con enfermedad coronaria.

Abstract Objective: To determine the prevalence and severity of periodontal disease in patients with acute coronary syndrome with no comorbidities admitted to a cardiac rehabilitation programme in the Clínica Shaio Foundation, Colombia. Methods: The study included a total of 83 patients diagnosed with acute coronary syndrome with no comorbidities and the following diagnoses: unstable angina (n = 27), ST-segment elevation myocardial infarction (STEMI) (n = 34), and non-ST-elevation myocardial infarction (NSTEMI) (n = 22). The prevalence and severity of periodontal disease were evaluated using the Periodontal Index of the Centres for the Control and Prevention of Diseases. A between-group comparison was made of the clinical index, plaque, calculi, gingival bleeding, probing depth, and level of clinical attachment. Results: The prevalence of periodontal disease was 97.6%, and advance periodontitis was observed in 38.3% of subjects. The patients with a STEMI showed more severe periodontitis. The inflammatory clinical indices of periodontal disease were increased in all groups, with no significant differences being observed. There were no significant differences between clinical attachment and heart conditions. However, it was observed that there was a higher percentage of locations with a loss of clinical attachment ≥ 6 mm, and locations with bag depths > 6 mm in patients with a STEMI. Conclusión: The patients with acute coronary syndrome with no comorbidities, and who entered the cardiac rehabilitation programme to complete their treatment, had a high prevalence and severity of periodontal disease. The control of periodontal disease should be emphasised in patients with coronary disease..

Differential responses of endothelial cells on three-dimensional scaffolds to lipopolysaccharides from periodontopathogens.

Studies have been conducted on the pathogenicity of periodontopathogens in cultures of endothelial cells on two-dimensional (2D) polystyrene surfaces, where the monolayer formed is not exposed to proteins of the subendothelial matrix. In this work, we developed a culture system by seeding human coronary artery endothelial cells (HCAECs) onto three-dimensional (3D) scaffolds of collagen type I, a subendothelial protein. The inflammatory responses of the HCAEC monolayers, formed either on 3D scaffolds or directly on a 2D polystyrene plate, to lipopolysaccharide (LPS) from Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg) were evaluated. The transcription of 3 genes, the secretion of 40 cytokines and 2 prostanoids, and the adhesion of monocytes to 2D and 3D cultures with or without exposure to lipopolysaccharides (control) were assessed. HCAECs exhibited differences in transcriptional and secretory profiles between the 3D and 2D models. In addition, the inflammatory responses of HCAEC to Aa-LPS and Pg-LPS differed between the two models. In 3D cultures treated with Aa-LPS, the levels of IL-8, RANTES, G-CSF, ICAM-1, IL-6, and TXA2 were significantly higher than those in the controls. In 2D cultures treated with Aa-LPS, IL-8, RANTES, G-CSF, ICAM-1, TNF-RI, PGI2, and TXA2 levels were significantly higher than those in their controls. In the presence of Aa-LPS, monocyte adhesion did not differ between treated and control 3D cultures but was significantly higher in treated 2D cultures than in the controls. In response to Pg-LPS, cytokine-prostaglandin secretion and monocyte adhesion did not differ between 3D and 2D cultures. These data indicate that HCAECs respond differently to these two types of LPS.

Oral and uro-vaginal intra-amniotic infection in women with preterm delivery: A case-control study.

AIM: The aim of the present study was to establish the association between the presence of oral and uro-vaginal microorganisms in the placental membrane and preterm delivery (PTD), the premature rupture of membranes (PRM), and the clinical signs of intra-amniotic infection. METHODS: Eighty-four women with PTD and 127 women with delivery at term were assessed for the PRM, clinical signs of intra-amniotic infection, and the presence of periodontitis. Twenty-seven microorganisms were identified in the placental tissue using nested polymerase chain reaction (PCR). Porphyromonas gingivalis (P. gingivalis) was quantified by droplet digital PCR. RESULTS: The prevalence of microorganisms was 9.47% (20/211). P. gingivalis was the most prevalent (12/211, 5.68%). Mycoplasma hominis, Ureaplasma urealyticum, Staphylococcus spp, and Fusobacterium nucleatum were isolated at a very low frequency in the placenta. Candida albicans was associated with PTD (P = 0.027). Periodontitis was associated with clinical signs of infection (odds ratio [OR] = 3.8, 95% confidence interval [CI]: 1.28-13.5) and with PTD (OR = 1.99; 95% CI: 1.07-3.72). CONCLUSION: The presence of P. gingivalis in the placenta was not associated with perinatal complications. Detecting microorganisms in the placenta by nested PCR is not relevant, as it has a poor association with clinical variables that establish the diagnosis of chorioamnionitis. However, periodontitis was associated with the clinical signs of intra-amniotic infection and PTD.

Eikenella corrodens lipopolysaccharide stimulates the pro-atherosclerotic response in human coronary artery endothelial cells and monocyte adhesion.

Eikenella corrodens is a gram-negative bacterium, and although primarily associated with periodontal infections or infective endocarditis, it has been identified in coronary atheromatous plaques. The effect of its lipopolysaccharide (LPS) on human coronary artery endothelial cells (HCAECs) is unknown. Our aim was to examine the mechanism underlying the inflammatory response in HCAECs stimulated with E. corrodens-LPS and to evaluate monocyte adhesion. Endothelial responses were determined by measuring the levels of chemokines and cytokines using flow cytometry. The surface expression of intercellular adhesion molecule 1 (ICAM-1) was determined using a cell-based ELISA, and the adhesion of THP-1 monocytes to HCAECs was also monitored. The involvement of toll-like receptors (TLRs) 2 and 4 was examined using TLR-neutralizing antibodies, and activation of extracellular signal-regulated kinase (ERK)1/2 and nuclear factor-kappa B (NF-κB) p65 were measured by western blotting and ELISA, respectively. Eikenella corrodens-LPS increased secretion of interleukin-8 (IL-8), monocyte chemotactic protein 1 (MCP-1), and granulocyte-macrophage colony-stimulating factor (GM-CSF), and expression of ICAM-1 on the surface of HCAECs, consistent with the increased adhesion of THP-1 cells. Moreover, E. corrodens-LPS interacted with TLR4, a key receptor able to maintain the levels of IL-8, MCP-1, and GM-CSF in HCAECs. Phosphorylation of ERK1/2 and activation of NF-κB p65 were also increased. The results indicate that E. corrodens-LPS activates HCAECs through TLR4, ERK, and NF-κB p65, triggering a pro-atherosclerotic endothelial response and enhancing monocyte adhesion.

Electrostimulation of the lingual nerve by an intraoral device may lead to salivary gland regeneration: A case series study

BACKGROUND: Salivary gland function is controlled by the salivary reflex, whose efferent arm is composed by the parasympathetic and the sympathetic divisions of the autonomic nervous system. Parenchymal injury is the main salivary gland involvement of Sjögren's syndrome and head and neck radiotherapy, but neural damage has been reported as well. Recently an intraoral device for electrostimulation of the lingual nerve in vicinity to the lower third molar has been introduced. At this point this nerve carries efferent fibers for the innervation of the submandibular, sublingual and several minor salivary glands and afferent fibers of the salivary reflex. Therefore, excitation of these fibers potentially leads to increased secretion of all salivary glands. Thus, the study objective was to assess whether comprehensive neural activation by electrostimulation of the lingual nerve carries the potential to induce the regeneration of damaged salivary glands. MATERIAL AND METHODS: The device was tested on three patients with no collectable resting and stimulated secretion of saliva during a double blind, sham controlled period of two months and nine open-label months. RESULTS: All three subjects developed the capacity to spit saliva, not only in direct response to the electrostimulation but also after free intervals without electrostimulation. In addition, their symptoms of dry mouth severity and frequency improved. CONCLUSIONS: This recovery is probably due to the combined effect of increase in secretory functional gland mass and regain of nervous control of the secretory elements and blood vessels. Both are phenomena that would contribute to gland regeneration

Human papilloma virus: An etiological and prognostic factor for oral cancer?

The increasing prevalence of human papilloma virus (HPV)-positive oral tumors can be considered an epidemic. Although the incidence of HPV cervical cancer is decreasing, the incidence of oral cavity and oropharyngeal cancers associated with HPV is increasing. The presence of certain HPV genotypes could be a predictor of future oral cancer lesions, although lesions associated with HPV could be less aggressive and exhibit a higher survival rate. In the present study, we review the most important biologic, clinic, epidemiologic, and prognostic factors associated with HPV infection and oral cancer.

Are obesity, ACPAs and periodontitis conditions that influence the risk of developing rheumatoid arthritis in first-degree relatives?

The aim of this study was to investigate the body mass index (BMI), anti-citrullinated protein antibodies (ACPAs) status and the presence of periodontitis and IgG-1/IgG-2 antibodies against Porphyromonas gingivalis (Pg) in the first-degree relatives (FDRs) of rheumatoid arthritis (RA) patients and compare these variables with a control group of healthy individuals from the general population. In total, 100 FDR individuals and 200 healthy controls matched by age and gender were included. Rheumatologic and periodontal assessment was performed, and the presence of ACPAs and anti-P. gingivalis antibodies was evaluated. Groupwise comparisons were analysed using the McNemar and Wilcoxon tests. A conditional logistic regression analysis was performed to establish the associations between BMI, ACPAs and periodontitis in both groups. In the FDR group, 70% of the subjects were female, with a mean age of 37.3 ± 13 years. Obesity was observed in 17 and 7% of the FDRs and controls, respectively. ACPAs were found in 7% of the FDRs vs. 2.5% of the controls. Periodontitis was diagnosed in 79 and 56% of the FDRs and controls, respectively. Among the FDRs, 15% had severe periodontitis. There were associations in the FDR group related to the presence of obesity (OR 2.93, 95% CI 1.03-8.28), ACPAs (OR 2.45, 95% CI 0.7-8.32) and periodontitis (OR 3.70 95% CI 1.89-7.29). Regarding anti-P. gingivalis antibodies and smoking history, no differences were found between the groups. Obesity, ACPAs and periodontitis (diagnosis and severity) can be considered as relevant conditions associated with the development of RA in FDRs.

Rosuvastatin Inhibits Interleukin (IL)-8 and IL-6 Production in Human Coronary Artery Endothelial Cells Stimulated With Aggregatibacter actinomycetemcomitans Serotype b.

BACKGROUND: Rosuvastatin exhibits anti-inflammatory effects and reduces periodontal diseases and atherosclerosis; however, its role in regulating periodontopathogen-induced endothelial proinflammatory responses remains unclear. The purpose of this study is to determine whether rosuvastatin can reduce the proinflammatory response induced by Aggregatibacter actinomycetemcomitans (Aa) in human coronary artery endothelial cells (HCAECs). METHODS: HCAECs were stimulated with purified Aa serotype b lipopolysaccharide (LPS) (Aa-LPS), heat-killed (HK) bacteria (Aa-HK), or live bacteria. Expression of Toll-like receptors and cellular adhesion molecules were evaluated by fluorometric enzyme-linked immunosorbent assay. Endothelial cell activation was evaluated by quantifying nuclear factor (NF)-kappa B-p65 and cytokine expression levels by quantitative polymerase chain reaction and flow cytometry. Effect of rosuvastatin in expression of the atheroprotective factor Krüppel-like factor 2 (KLF2) and cytokines were also studied using similar approaches. RESULTS: HCAECs showed increased interleukin (IL)-6, IL-8, intercellular adhesion molecule 1, and platelet endothelial cell adhesion molecule 1 expression when stimulated with Aa-LPS or Aa-HK. NF-κB-p65 activation was induced by all antigens. Aa-induced IL-6 and IL-8 production was inhibited by rosuvastatin, particularly at higher doses. Interestingly, reduced IL-6 and IL-8 levels were observed in HCAECs stimulated with Aa in the presence of higher concentrations of rosuvastatin. This anti-inflammatory effect correlated with a significant increase of rosuvastatin-induced KLF2. CONCLUSIONS: These results suggest Aa-induced proinflammatory endothelial responses are regulated by rosuvastatin in a mechanism that appears to involve KLF2 activation. Use of rosuvastatin to prevent cardiovascular disease may reduce risk of endothelial activation by bacterial antigens.

Is there a relationship between spondyloarthritis and periodontitis? A case-control study.

OBJECTIVE: To compare the frequency and severity of periodontitis in patients with spondyloarthritis (SpA) with healthy control individuals, through the evaluation of clinical, serological and microbiological periodontal condition. METHODS: Patients with a diagnosis of SpA (n=78) and biological disease-modifying antirheumatic drug (bDMARD) naive fulfilling the Assessment of SpondyloArthritis international Society (ASAS) classification criteria as well as 156 healthy controls matched for age/gender were included. Two trained and calibrated periodontologists performed the periodontal clinical assessment. The presence of periodontitis and its severity were determined according to the criteria established by the Centers for Disease Control and Prevention-American Academy of Periodontology. The clinical periodontal variables, IgG1/IgG2 antibodies against Porphyromonas gingivalis andperiodontopathic bacterial identification, were also established. Comparisons of periodontal characteristics between the patients with SpA and the control group were performed using univariable analyses. A logistic regression analyses was performed to calculate the OR (95% CI) for diagnosis of periodontitis in patients with SpA and matched controls. RESULTS: A diagnosis of periodontitis was established in 56% in patients with SpA versus 69% of healthy controls (P≤ 0.01). Severe periodontitis was found in 3% versus 12% in SpA versus healthy controls, respectively (P≤ 0.01). There was no significant increase of frequency of any periodontal variable, IgG1/IgG2 antibodies against P. gingivalis or the presence of periodontopathic bacteria between patients with SpA and control group. Periodontitis was not positively associated with a diagnosis of SpA (OR: 0.57, 95% CI 0.32 to 1.00, P=0.05) in the logistic regression analyses. CONCLUSIONS: We found a lower rather than a higher frequency and severity of periodontitis in patients with SpA in comparison with healthy control individuals. Our findings suggest that there is no positive association between SpA and periodontitis in Colombian patients.

Periodontal Disease in Individuals With a Genetic Risk of Developing Arthritis and Early Rheumatoid Arthritis: A Cross-Sectional Study.

BACKGROUND: Recent consensus emphasizes the importance of studying individuals at risk for rheumatoid arthritis (pre-RA) and those with early RA (eRA). Periodontal tissues have been recently evaluated, but these studies are limited. To evaluate the periodontal condition, immunoglobulin (Ig)G subclasses against Porphyromonas gingivalis in individuals with pre-RA and eRA were compared with controls to establish an association between periodontal infection markers and rheumatic activity. METHODS: Rheumatologic and periodontal condition was evaluated in 119 individuals with pre-RA, 48 patients with eRA, and matched controls. P. gingivalis IgG1 and IgG2 were analyzed. C-reactive protein, erythrocyte sedimentation rate (ESR), rheumatoid factor, anticitrullinated protein antibodies (ACPAs), and RA activity were measured. The groups were compared with McNemar test and paired t-test. Conditional logistic regression was performed for pre-RA confounders, and χ(2) test was used to evaluate periodontal variables and RA activity indices. RESULTS: Pre-RA individuals showed significantly higher levels of plaque index (P = 0.01) and bleeding on probing (P = 0.03) and higher severity of periodontal disease (P = 0.02). Periodontitis was associated with pre-RA (odds ratio, 3.39; 95% confidence interval, 1.64 to 7.01) but not with eRA. In pre-RA, P. gingivalis-specific IgG2 was associated with ACPAs (P = 0.049) and disease severity visual analog scale (P = 0.03). In eRA, IgG2 against P. gingivalis was associated with ESR (P = 0.046) and ACPAs (P = 0.04). P. gingivalis was associated with ACPAs (P = 0.04). CONCLUSIONS: This study shows that individuals with pre-RA have significant inflammatory periodontal involvement. There was a significant association between IgG against P. gingivalis and ACPAs in pre-RA and markers of RA activity in individuals with eRA.

Evaluation of two human dental pulp stem cell cryopreservation methods.

Dental pulp is a promising source of mesenchymal stem cells for use in cell therapy and regenerative medicine. Methods for storing stem cells with minimum compromise of cell viability, differentiation capacity and function should be developed for clinical and research applications. The aim of this study was to evaluate whether human dental pulp stem cells (hDPSCs) isolated and cryopreserved for 1, 7 and 30 days maintain viability and expression of specific stem cell markers. Human dental pulp stem cells were isolated from 23 healthy patients aged 18 to 31 years. Dental pulp was enzymatically dissociated, and CD105+ cells were separated using the Miltenyi™ system. The hDPSCs were cryopreserved using the Kamath and Papaccio methods. Post-cryopreservation viability was measured by flow cytometry (7AAD) and by the expression of the phenotype markers CD105+/ CD73+, CD34-/CD45-. The Papaccio method showed greater cell viability for cells that had been frozen for 30 days (59.5%) than the Kamath method (56.2%), while the Kamath method provided better results for 1 day (65.5%) and 7 days (56%). Post-cryopreservation expression of the markers CD105+/CD34- was greater after 1 and 7 days with the Kamath method and CD105+/CD45- were expressed after all 3 cryopreservation times. There was greater expression of CD73+ in the hDPSCs after 1 and 7 days with the Kamath method, and after 30 days with the Papaccio method. These results suggest that hDPSCs express mesenchymal stem cell markers after cryopreservation. However, cryopreservation time may affect marker expression, probably by altering the spatialconfiguration of cell membrane proteins or by compromising cells at a certain level of differentiation.

Intraoral electrostimulator for xerostomia relief: a long-term, multicenter, open-label, uncontrolled, clinical trial.

OBJECTIVE: A previous sham-controlled multinational study demonstrated the short-term efficacy and safety for xerostomia treatment of an intraoral device that delivers electrostimulation to the lingual nerve. The objective of this study was to test the hypothesis that those beneficial effects would be sustained over an 11-month period. STUDY DESIGN: The device was tested on a mixed sample of 94 patients with xerostomia in an open-label, uncontrolled, prospective multicenter trial. Statutory outcome assessments were done at 5th, 8th, and 11th months and analyzed by multiple comparisons. RESULTS: Improvements achieved at month 5 from baseline were sustained throughout the follow-up period for the primary outcome, xerostomia severity, and the secondary outcomes resting whole salivary flow rate, xerostomia frequency, oral discomfort, and difficulties in speech, swallowing, and sleeping. No significant side effects were detected. CONCLUSIONS: The beneficial effects of a removable intraoral electrostimulating device were sustained for an 11-month period.

Efficacy and safety of an intraoral electrostimulation device for xerostomia relief: a multicenter, randomized trial.

OBJECTIVE: To evaluate the efficacy and safety of an intraoral electrostimulation device, consisting of stimulating electrodes, an electronic circuit, and a power source, in treating xerostomia. The device delivers electrostimulation through the oral mucosa to the lingual nerve in order to enhance the salivary reflex. METHODS: The device was tested on a sample of patients with xerostomia due to Sjögren's syndrome and other sicca conditions in a 2-stage prospective, randomized, multicenter trial. Stage I was a double-blind, crossover stage designed to compare the effects of the electrically active device with the sham device, each used for 1 month, and stage II was a 3-month open-label stage designed to assess the long-term effects of the active device. Improvement in xerostomia severity from baseline was the primary outcome measure. RESULTS: A total of 114 patients were randomized. In stage I, the active device performed better than the sham device for patient-reported xerostomia severity (P<0.002), xerostomia frequency (P<0.05), quality of life impairment (P<0.01), and swallowing difficulty (P<0.02). At the end of stage II, statistically significant improvements were verified for patient-reported xerostomia severity (P<0.0001), xerostomia frequency (P<0.0001), oral discomfort (P<0.001), speech difficulty (P<0.02), sleeping difficulty (P<0.001), and resting salivary flow rate (P<0.01). CONCLUSION: Our findings indicate that daily use of the device alleviated oral dryness, discomfort, and some complications of xerostomia, such as speech and sleeping difficulties, and increased salivary output. The results show a cumulative positive effect of the device over the period of the study, from baseline to the end of the trial.

Porphyromonas gingivalis LIBRE DE POLISACÁRIDOS UTILIZANDO CROMATOGRAFÍA DE ALTA RESOLUCIÓN SEPHACRYL S-200 / Purification of Porphyromonas gingivalis polysaccharide free lipopolysaccharide using Sephacryl S-200 high resolution chromatography

El objetivo de este trabajo fue mejorar un método estándar para la purificación de lipopolisacárido (LPS) de Porphyromonas gingivalis libre de polisacáridos usando una estrategia de extracción, digestión enzimática y cromatografía de alta resolución. La bacteria P. gingivalis se cultivó en condiciones de anaerobiosis y se hizo extracción de las membranas con el método de fenol-agua. Luego de una digestión enzimática (DNAsa, RNAsa y proteasa) se separó el extracto por filtración por gel con Sephacryl S-200. La muestra purificada se caracterizó por electroforesis en gel de acrilamida con tinción de plata y por el método Purpald se detecto el ácido 2-ceto-3-desoxioctu-losónico (KDO). Se obtuvo una preparación libre de ácidos nucleicos, proteínas y polisacáridos. La separación por cromatografía fue de alta resolución al permitir la obtención de dos picos con diferentes componentes. El protocolo de purificación nos permitió obtener LPS de P. gingivalis con alto grado de pureza, el cual podría ser usado en próximos ensayos para evaluar su función en ensayos in vitro e in vivo; así como iniciar la obtención de LPS de otras bacterias periodontopáticas, con el fin de investigar la asociación de enfermedad periodontal con enfermedades cardiovasculares.

The aim of this work was to improve a standard methodology to purify Porphyromonas gingivalis lipopolysaccharide (LPS) using a protocol of extraction, enzymatic digestion and high resolution chromatography. P. gingivalis bacteria was cultured in anaerobiosis, their membranes were extracted using the phenol-water method, then subjected to DNAse, RNAse and protease digestion and finally, the extract was separated by chromatography using Sephacryl S-200. The purified extract was characterized by silver staining after polyacrylamide gel electrophoresis and 2-keto-3-deoxioctanoic acid (KDO) was detected using the Purpald’s method. A preparation free of nucleic acid-, protein-or polysaccharides was obtained. The chromatographic separation showed high resolution since there was two discrete peaks with different components. The purification protocol allowed us to obtain a highly purified P. gingivalis LPS which could be used in future tests to evaluate its behavior in vitro and in vivo and elucidate its function, as well as to obtain LPS from other periodontopatic bacteria to address the association of periodontal disease with cardiovascular diseases.

[Subgingival microbiota in chronic and aggressive periodontitis in Bogotá, Colombia: an epidemiological approach]. / Microflora subgingival en periodontitis crónica y agresiva en Bogotá, Colombia: un acercamiento epidemiológico.

INTRODUCTION: Periodontitis-associated microbiotas differ in regions around the world and vary with a variety of factors. This suggests that each country must establish its own dental microbiobiotic profile in order to formulate adequate prevention measures and more specific treatments. OBJECTIVE: The prevalence and proportions of the following microorganisms were investigated: Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Prevotella intermedia/Prevotella nigrescens, Eikenella corrodens, Campylobacter rectus, Micromonas micros, Fusobacterium spp, Dialister pneumosintes and enteric rods. The patients selected had been diagnosed with chronic periodontitis and aggressive periodontitis from clinical populations in Bogotá, Colombia. The variables of age, gender and socioeconomic level were included in the comparisons. MATERIALS AND METHODS: 183 patients were classified in two groups, chronic periodontitis (n=84) and aggressive periodontitis (n=59); 40 healthy subjects were selected for comparison. A pooled subgingival plaque sample was obtained from the 6 deepest periodontal pockets in every patient and processed by culture for the isolation and identification of bacteria. RESULTS: In chronic periodontitis and aggressive periodontitis, P. gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp and E. corrodens showed high and similar prevalence, whereas M. micros, A. actinomycetemcomitans, D. pneumosintes and enteric rods (commonly from the tribe Klebsielleae) showed less and similar prevalence. In the healthy group, P. intermedia/P. nigrescens, C. rectus and Fusobacterium spp showed high frequency whereas M. micros and E. corrodens showed moderate frequency. CONCLUSIONS: Porphyromonas gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp. and E. corrodens are important constituents of the microbiologic profile of the periodontitis in Colombian populations. Porphyromonas gingivalis was found more frequently than A. actinomycetemcomitans in aggressive periodontitis.

Microflora subgingival en periodontitis crónica y agresiva en Bogotá, Colombia: un acercamiento epidemiológico / Subgingival microbiota in chronic and aggressive periodontitis in Bogotá, Colombia: an epidemiological approach

Introducción. Los microorganismos involucrados en la periodontitis varían en cada región dependiendo de diferentes factores. Por ello cada país debe establecer su propio perfil microbiológico con el objeto de proponer esquemas adecuados de prevención y tratamiento. Objetivo. Investigar la presencia y concentración subgingival de Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Prevotella intermedia/Prevotella nigrescens, Eikenella corrodens, Campylobacter rectus, Micromonas micros, Fusobacterium spp , Dialister pneumosintes y bacilos entéricos en pacientes con periodontitis en una población colombiana considerando edad, género y nivel socioeconómico. Materiales y métodos. Participaron 183 pacientes: 84 con periodontitis crónica, 59 con periodontitis agresiva y 40 individuos sanos. De cada paciente se obtuvo un grupo de muestras de los seis sitios con mayor profundidad de bolsa que se procesaron por cultivo para el aislamiento e identificación de las bacterias. Resultados. En periodontitis crónica y agresiva se detectaron con alta y similar frecuencia P. gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp y E. corrodens y con menor y similar frecuencia, M. micros, A. actinomycetemcomitans, D. pneumosintes y bacilos entéricos, principalmente de la tribu Klebsielleae. En los sujetos sanos se encontraron con frecuencia elevada P. intermedia/P. nigrescens, C. rectus y Fusobacterium spp y con frecuencia moderada, M. micros y E. corrodens. Conclusión. Los resultados indican que P. gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp y E. corrodens constituyen parte importante del perfil microbiológico de las periodontitis en la población colombiana. En la periodontitis agresiva, P. gingivalis se encontró más frecuentemente que A. actinomycetemcomitans.

Introduction. Periodontitis-associated microbiotas differ in regions around the world and vary with a variety of factors. This suggests that each country must establish its own dental microbiobiotic profile in order to formulate adequate prevention measures and more specific treatments. Objective. The prevalence and proportions of the following microorganisms were investigated: Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Prevotella intermedia/Prevotella nigrescens, Eikenella corrodens, Campylobacter rectus, Micromonas micros, Fusobacterium spp, Dialister pneumosintes and enteric rods. The patients selected had been diagnosed with chronic periodontitis and aggressive periodontitis from clinical populations in Bogotá, Colombia. The variables of age, gender and socioeconomic level were included in the comparisons. Materials and methods. 183 patients were classified in two groups, chronic periodontitis (n=84) and aggressive periodontitis (n=59); 40 healthy subjects were selected for comparison. A pooled subgingival plaque sample was obtained from the 6 deepest periodontal pockets in every patient and processed by culture for the isolation and identification of bacteria. Results. In chronic periodontitis and aggressive periodontitis, P. gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp and E. corrodens showed high and similar prevalence, whereas M. micros, A. actinomycetemcomitans, D. pneumosintes and enteric rods (commonly from the tribe Klebsielleae) showed less and similar prevalence. In the healthy group, P. intermedia/P. nigrescens, C. rectus and Fusobacterium spp showed high frequency whereas M. micros and E. corrodens showed moderate frequency. Conclusions. Porphyromonas gingivalis, T. forsythia, P. intermedia/P. nigrescens, C. rectus, Fusobacterium spp. and E. corrodens are important constituents of the microbiologic profile of the periodontitis in Colombian populations...