Immunohistochemical Comparative Study of Aggressive and Non-aggressive Central Giant Cell Lesions of the Jaws Based on the Tenascin-C Expression Profile.

The purpose of this study was to compare the immunohistochemical expression of tenascin-C (Tn-C) regarding clinicopathological variables and its association with the clinical behavior of central giant cell lesions (CGCLs). Forty-eight paraffin-embedded samples of CGCLs were selected. Based on clinical and radiographic features, the lesions were classified as aggressive (A-CGCLs) and non-aggressive (NA-CGCLs) subtypes. Histological assessment included the microvessel count (MVC), multinucleated giant cell (MGC) count, and the proportion of tissue area involved by mononuclear stromal cells/interstitial fibrosis. Immunoreactivity, immunolocalization, and distribution patterns of Tn-C were studied immunohistochemically. The association between Tn-C expression and clinicopathological characteristics was analyzed separately and adjusted for confounders using logistic regression models. A significantly greater proportion of cases with moderate-to-intense, intracellular, and diffuse staining of Tn-C was observed in A-CGCLs. CGCLs with a size ≥3.3 cm, fast growth, cortical disruption, high MVC/MGC counts, and low interstitial fibrosis showed a significantly greater frequency of moderate-to-intense, intracellular, and diffuse staining. Logistic regression analysis indicated a strong/independent association of these three immunohistochemical parameters with the aggressiveness of lesions. These data appear to suggest a possible role for Tn-C in the etiopathogenesis of CGCLs of the jaws, where its upregulation might favor the destructive behavior of A-CGCLs.

Analysis of salivary detection of P16INK4A and RASSF1A promoter gene methylation and its association with oral squamous cell carcinoma in a Colombian population.

BACKGROUND: Epigenetic factors play a fundamental role in the etiopathogenesis of oral squamous cell carcinoma (OSCC). This study evaluated if salivary detection of P16INK4A/RASSF1A gene promoter methylation might be linked to the clinical/histological features of OSCC in a Colombian population. MATERIAL AND METHODS: Methylation-specific polymerase chain reaction (MSP-PCR) was used to detect the methylation frequency of P16INK4A/RASSF1A genes in DNA obtained from whole saliva collected of 40 healthy controls (HC) and 43 OSCC patients. Determination of the clinical performance of MSP-PCR assay was based on standard algorithms derived from two-way contingency table analysis. The association of methylation status of targeted genes with OSCC was analyzed in a multivariate binary logistic regression model. RESULTS: There were significantly higher proportions of promoter methylation of these target genes in OSCC patients when compared with HC. The analysis of single methylated genes showed high specificity, good positive and negative predictive values, but was accompanied by a low sensitivity. OSCC cases with clinical stage III/IV, poorly differentiated, and severe cellular atypia showed a significantly greater proportion of methylated than that of unmethylated targeted genes in saliva samples. Logistic regression analysis indicated an independent association of P16INK4A and RASSF1A promoter methylation with OSCC diagnosis. A significant interaction effect between ageing and P16INK4A promoter methylation was also detected. CONCLUSIONS: Salivary detection of P16INK4A and RASSF1A promoter methylation appears to be independently associated with OSCC and may be linked to the tumor activity in the present population. Consequently, the targeting of these genes in saliva samples might constitute an important tool for diagnosis and prognosis purposes. Key words:Gene methylation, oral squamous cell carcinoma, P16INK4A, RASSF1A, saliva.

Assessment of adherence to preventive treatment of plaque-induced oral diseases using sugar-free gums: a controlled clinical trial / Evaluación de la adhesión al tratamiento preventivo de enfermedades bucales inducidas por placa bacteriana mediante el uso de gomas de mascar sin azúcar: ensayo clínico controlado

Abstract Introduction: the aim of this study was to assess the adherence to a preventive treatment of plaque-induced oral diseases using two sugar-free gums, establishing their effect on cariogenic bacteria counts as a biological marker of treatment response. Methods: a double-blind, randomized, parallel-group, controlled intervention study was conducted in 96 young adult university students in an experimental period of 30 days. Participants were distributed into two groups and given a chewing gum with either pentitol or hexitol + CPP-ACP complex to be taken three times a day for 20 minutes after conventional oral hygiene. Saliva samples were collected at baseline to evaluate counts of Streptococcus mutans (SM) and Lactobacillus spp. A survey was conducted at the end of the experimental period to assess the levels of adherence to treatment, remeasuring bacterial counts. Results: SM counts showed statistically significant differences between the study groups at the end of the experimental period, with a decreasing tendency in both groups. Intragroup difference was observed in patients who took pentitol gum with a marked reduction in SM counts. Conclusion: the final estimate of adherence to treatment showed that the type of chewing gum patients were given had no influence and therefore such gums can be used, considering the therapeutic agents that most contribute to patient's particular case and preferences.

Resumen Introducción: el objetivo consistió en evaluar la adherencia a un tratamiento preventivo de enfermedades bucales inducidas por placa bacteriana, utilizando dos gomas de mascar sin azúcar y establecer su efecto sobre recuentos de bacterias cariogénicas como indicador biológico de los efectos del tratamiento. Métodos: se realizó un estudio de intervención, doble ciego, de grupos paralelos, aleatorizado controlado, en 96 adultos jóvenes universitarios, en un período experimental de 30 días. Los participantes fueron distribuidos en dos grupos con asignación de una goma de mascar con pentitol o con hexitol + complejo CPP-ACP que debían ser consumidas 20 minutos/tres veces al día después de la higiene oral convencional. Al inicio del studio fueron recolectadas muestras de saliva para evaluar recuentos de Estreptococos del grupo mutans (SM) y Lactobacillus spp. Al finalizar el período experimental, se realizó una encuesta para evaluar los niveles deadherencia al tratamiento y los recuentos bacterianos fueron medidos nuevamente. Resultados: Los recuentos de SM mostraron diferencias estadísticamente significativas entre los grupos de estudio al final del período experimental, con tendencia hacia la disminución en ambos grupos. Se observó diferencia intragrupo en los pacientes que consumieron la goma de mascar con pentitol, con marcada reducción de los recuentos de SM. Conclusión: la estimación final de la adherencia al tratamiento mostró que el tipo de goma de mascar asignada al paciente no tuvo influencia y, por lo tanto, estas pueden ser utilizadas teniendo en cuenta los agentes terapéuticos que más aporten al caso particular del paciente y las preferencias del mismo.

Salivary levels of specialized pro-resolving lipid mediators as indicators of periodontal health/disease status.

AIM: This cross-sectional case-control study aimed to determine if salivary levels of lipoxin A4 (LXA4), protectin D1 (PD1), resolvin E1 (RvE1) and maresin 1 (MaR1) might constitute a reflection of periodontal health/disease status. MATERIALS AND METHODS: One hundred and two periodontitis patients and 61 healthy controls were recruited. Periodontal clinical status was determined by criteria based on full-mouth clinico-radiographical data. Salivary concentration of the analytes was calculated by enzyme-linked immunosorbent assay. The association between the biomarkers with disease status was assessed individually and adjusted for confounding using multivariate binary logistic regression models. RESULTS: Significantly decreased LXA4 and increased PD1/MaR1 salivary levels were detected in periodontitis patients in comparison with healthy controls. However, no significant differences were observed for RvE1 levels between clinical groups. Clinical parameters such as probing depth, clinical attachment loss and extent were negatively correlated with LXA4, positively correlated with PD1/MaR1 and not correlated with RvE1 salivary levels. Logistic regression analyses revealed a strong/independent association of LXA4, PD1 and MaR1 salivary levels regarding disease status. Interaction effects between demographic predictor variables and salivary concentration of LXA4, PD1 and MaR1 were also identified. CONCLUSION: The results of this study demonstrated a strong/independent association between reduced LXA4 and increased PD1/MaR1 salivary levels with periodontitis suggesting an imbalance in the specialized pro-resolving lipid mediators (SPMs) in periodontal disease.

Decreased salivary concentration of CD9 and CD81 exosome-related tetraspanins may be associated with the periodontal clinical status.

AIM: This cross-sectional case-control study was designed to determine the association of the salivary concentration of CD9/CD81 exosome-related tetraspanins with the periodontal clinical status. MATERIALS AND METHODS: Saliva samples from 104 periodontitis patients and 45 healthy controls were collected. Periodontal status was assessed based on full-mouth clinico-radiographical data, and salivary concentration of the analytes was calculated by ELISA. The association between the biomarkers with disease status was analysed using multivariate binary logistic regression models. RESULTS: Significantly decreased salivary levels of CD9 and CD81 exosomes were detected in periodontitis patients in comparison with healthy controls. Also, negative significant correlations between salivary concentrations of CD9/CD81 exosomes regarding clinical measurements were observed. Likewise, a significant downward trend of the concentration of these two biomarkers concerning the stage and grade of disease could be identified. Logistic regression analyses revealed a strong/independent association for decreased salivary concentration of CD81 exosomes regarding disease status. Confounding and interaction effects between age and salivary concentration of CD9 exosomes were also noted. CONCLUSION: Reduced salivary concentration of CD9/CD81 exosomes might be of significance in the context of periodontal disease pathogenesis.

Association analysis between rs6184 and rs6180 polymorphisms of growth hormone receptor gene regarding skeletal-facial profile in a Colombian population.

Background/Objective: There is strong evidence that genetic factors may affect the craniofacial morphology. This study aimed to examine the association between the rs6184 and rs6180 polymorphic variants of the growth hormone receptor (GHR) gene and skeletal-facial profile in a Colombian population. Subjects/Methods: Saliva samples from 306 individuals ranging in age from 15 to 53 (mean 24.33) years were collected. Cephalometric parameters were used to categorize the participants as Class I, Class II, or Class III skeletal-facial profile. The polymerase chain reaction-restriction fragment length polymorphism method was used to identify genotypes of the rs6184 and rs6180 single nucleotide polymorphisms (SNPs). The association of polymorphisms with the skeletal-facial profile was assessed separately and adjusted for confounding using a multivariate binary logistic regression model, alongside with analysis of linkage disequilibrium and haplotype associations. Results: Although individuals carrying the CA genotype of the rs6184 SNP showed both significantly decreased values for ANB angle and increased measures concerning mandibular body length and mandibular length, no significant differences amongst genotype groups of rs6180 SNP were observed. Moreover, chi-square test and logistic regression analysis revealed that the CA genotype of rs6184 SNP and the A-A haplotype were highly associated with Class III skeletal-facial profile. Conclusions: Although these results do not support that rs6180 SNP could be identified as a predictor for skeletal-facial profile, they suggest that the allele A of rs6184 SNP alone or in combination with other SNPs in the GHR gene yields significant horizontal and longitudinal variations of the mandibular morphology and might be a strong/independent prognostic indicator for Class III skeletal-facial profile in the present population.

Association Study of Vitamin D Receptor (VDR) - Related Genetic Polymorphisms and their Haplotypes with Chronic Periodontitis in Colombian Population.

INTRODUCTION: There is strong evidence that both genetic and environmental factors may affect the periodontal clinical status. However, epidemiological evidence on the association between Vitamin D Receptor (VDR) polymorphisms and Chronic Periodontitis (CP) has been inconsistent. AIM: The focus of this study was to identify if a possible association between VDR Single-Nucleotide Polymorphisms (SNPs) may be implicated in the aetiopathogenesis of CP in Colombian population. MATERIALS AND METHODS: One hundred and ten CP patients and 50 Healthy Controls (HC) were recruited. Periodontal status was assessed based on probing depth, clinical attachment level, extent, and severity of periodontal breakdown. The polymerase chain reaction-restriction fragment length polymorphism method was used to identify the VDR rs7975232, rs1544410, rs2228570, and rs731236 SNPs from saliva samples. Odds Ratios (ORs) along with their 95% Confidence Intervals (CIs) were computed to compare the distribution of genotypes/alleles between HC and CP patients, alongside with analysis of Linkage Disequilibrium (LD) and haplotype associations between SNPs. Also, an analysis of the interaction between genetic findings and those significant demographic factors was performed for all SNPs. RESULTS: There was no association neither between the different genotypes/allele frequencies nor haplotypes and CP. Similarly, no significant differences in extent or severity amongst genotype/allele groups were observed. Even so, interaction analysis revealed significant synergistic interactions between each SNP and age associated with the disease status. CONCLUSION: Although these results do not support that VDR SNPs could be identified as independent risk predictor variables for CP in the Colombian population, synergistic biological interactive effects of all these SNPs related to age might play a significant role in the pathogenic pathways of CP.

Influence of Periodontal Clinical Status on Salivary Levels of Glutathione Reductase.

BACKGROUND: Inadequate antioxidant balance may play a role in the excessive tissue breakdown in periodontitis. Because aggressive periodontitis (AgP) not only differs from chronic periodontitis (CP) in terms of clinical manifestations, this study investigates whether the salivary levels of glutathione reductase (GR) may be linked with periodontal status. METHODS: Saliva samples from patients with CP (n = 121), patients with AgP (n = 18), and healthy controls (n = 69) were collected. Periodontal status was assessed by criteria based on probing depth, clinical attachment level, and extent and severity of periodontal breakdown. GR salivary levels were analyzed by spectrophotometry. The association among GR concentration and CP or AgP was analyzed individually and adjusted for confounding using multivariate binary logistic regression models. RESULTS: GR levels not only differed significantly between the two periodontitis groups, being significantly greater in patients with AgP, but also were significantly greater than those observed in healthy controls. Synchronously, positive significant correlations between salivary GR concentration and clinical parameters were observed. After binary logistic regression analysis, both GR salivary levels ≥15.38 and ≥24.20 mU/mL were associated independently with CP and AgP, respectively. A significant interaction effect was also detected between increased GR salivary concentration and aging in the CP group. CONCLUSIONS: Increased GR salivary concentration may be a strong/independent prognostic indicator of the amount and extent of oxidative stress-induced periodontal damage in both CP and AgP. Likewise, saliva samples might reflect an interactive effect of GR levels associated with the aging-related cumulative characteristics of periodontal damage in CP.

Immunoexpression of NF-ĸB and their inhibitory subunits IĸBα and IĸBß in giant cell lesions of the jaws: implications for their clinical behavior.

BACKGROUND: The unpredictable behavior of giant cell lesions (GCLs) of the jaws parallels its controversial histogenesis. This study evaluated a possible association between the immunohistochemical expression of NF-ĸB, the inhibitory subunits IĸBα/IĸBß, and clinicopathological variables with the behavior of central and peripheral GCLs of the jaws. MATERIALS AND METHODS: Paraffin-embedded samples of GCLs of the jaws (n = 68) were prepared for histological/immunohistochemical assessment. Demographic and clinicopathological parameters were assessed to determine the behavior of the lesions. A staining-intensity-distribution (SID) score was used to assess the immunomarkers reactivity. The association between significant candidate immunohistochemical predictor variables regarding clinical behavior was analyzed individually and adjusted for confounding using a binary logistic regression model. RESULTS: While univariate analysis revealed a positive association of NF-ĸB SID score, NF-ĸB nuclear expression, IĸBα SID score, and NF-ĸB to inhibitors average ratio with the aggressive status of GCLs, after bivariate logistic regression analysis, only NF-ĸB nuclear expression, IĸBα SID score, and NF-ĸB to inhibitors average ratio remained as robust predictors of aggressiveness. Confounding and interaction effects regarding clinicopathological candidate predictor variables were also noted. CONCLUSION: It looks that clinical behavior of GCLs of the jaws may be strong/independently linked to the increased nuclear expression of NF-ĸB, higher NF-ĸB to inhibitors average ratio, and decreased IĸBα SID score. Notwithstanding, there are simultaneously synergistic and opposing interactive effects with respect to age stratum, growth rate, multinucleated giant cells count, and mononuclear stromal cells density in the susceptible host that may increase the tissue destruction observed in aggressive GCLs.

Time-related changes in salivary levels of the osteotropic factors sRANKL and OPG through orthodontic tooth movement.

INTRODUCTION: In this study, we aimed to determine whether the variations in salivary concentrations of soluble receptor activator of nuclear factor kappa B ligand (sRANKL) and osteoprotegerin (OPG), and their ratios, might be linked with the different phases of orthodontic tooth movement. METHODS: Twenty healthy subjects who required fixed appliance therapy not involving tooth extractions or surgical procedures were selected. Unstimulated whole saliva samples were collected from each patient before fitting the orthodontic appliances, and at 24 to 48 hours, 2 weeks, 5 weeks, and 8 weeks after the activation. Salivary sRANKL and OPG concentrations were determined by enzyme-linked immunosorbent assays. The data were analyzed by using nonparametric statistics. RESULTS: Overall, median values of sRANKL showed significant increases, median OPG salivary values showed a significant downward trend, and the sRANKL/OPG ratio tended to increase significantly over time after the activation visit. However, clear fluctuations in the immunoenzymatic findings were noted at the different sampling times, indicating nonlinear trends in the levels of the biomarkers through time. Post-hoc pairwise comparisons showed significant differences between (1) all sRANKL values relative to those of the 8-week sampling time; (2) baseline/8-week OPG salivary levels; and (3) baseline, 24 to 48 hours, and 2-week sRANKL/OPG ratios compared with those of the 8-week test. CONCLUSIONS: The findings indicate that variations in salivary concentrations of sRANKL and OPG and their ratios might be linked to the different phases of orthodontic tooth movement. Hence, these analytes might serve in a panel of salivary functional biomarkers that could assist in the screening of orthodontic treatment in clinical practice.

Overexpression of matrix metalloproteinase (MMP)-1 and -9 in central giant cell lesions of the jaws: implications for clinical behavior.

OBJECTIVE: The aim of this study was to investigate the relationship between the immunohistochemical expression of MMP-1 and MMP-9 with the clinical behavior of central giant cell lesions (CGCLs) of the jaws. STUDY DESIGN: Paraffin-embedded tissue from 30 aggressive and 12 nonaggressive CGCLs was assessed for the expression of MMP-1 and MMP-9 using immunohistochemistry. RESULTS: Although cellular immunolocalization patterns of MMP-1 and MMP-9 were similar, mean values of expression estimation/SID scores of each protease were significantly higher in aggressive CGCLs in comparison with nonaggressive lesions. Moreover, linear regression analysis showed that there was a reasonably good correlation not only between the expression estimation but also among SID scores of the 2 proteolytic enzymes. CONCLUSION: The findings of this study suggest a role for MMP-1 and MMP-9 in the resorptive activity of different cellular groups in CGCLs and indicate that differences in immunoreactivity of these 2 proteolytic enzymes may underlie the distinct clinical behavior.

Expression of hMLH1 and hMSH2 proteins in pleomorphic adenoma of minor salivary glands: relationship with clinical and histologic findings.

OBJECTIVE: The aim was to examine the relationship between clinicopathological features and immunoexpression of hMLH1 and hMSH2 proteins in pleomorphic adenoma (PA) of minor salivary glands. STUDY DESIGN: Paraffin-embedded samples of typically benign PA lesions (n = 35) were prepared for histologic and immunohistochemical assessment. Based on the clinicopathologic features, the samples were categorized into low- and high-risk subtypes for their estimated potential for malignant transformation. RESULTS: Immunohistochemical analysis revealed strong correlations regarding the expression estimation and staining-intensity distribution (SID) scores between the two proteins. Although there was no relationship between marker immunoexpression and SID scores regarding clinical parameters, statistically significant variations for these parameters were evident regarding some histologic criteria or for risk stratification subtypes. CONCLUSION: The findings of this study could suggest the relationship of a DNA mismatch repair deficiency with high-risk subtypes of PA and the implication of its role in the origin and progression of these subtypes.

Detection of Treponema denticola in saliva obtained from patients with various periodontal conditions.

The aim of the study was to determine the prevalence of Treponema denticola in saliva of periodontally diseased and healthy patients and its relationship with the periodontal status. A 16S rRNA-based polymerase chain reaction detection method was used to determine the prevalence of T. denticola in whole saliva samples from patients with chronic periodontitis (CP, n = 37), aggressive periodontitis (AgP, n = 24), and healthy subjects (n = 28). The periodontal status of each subject was assessed by criteria based on probing depth, clinical attachment loss, and extent of periodontal breakdown. Risk factors were assessed individually and adjusted for confounding using a binary logistic regression model. The results showed that the prevalence of T. denticola in CP patients was significantly higher than those in healthy and AgP subjects (P < 0.05). Odds ratio analysis revealed a positive association for CP group/T. denticola-positive and smoking/T. denticola-positive subjects. Furthermore, all clinical measurements were significantly greater (P < 0.05) for T. denticola-positive subjects compared to T. denticola-negative subjects. After binary logistic regression analysis, both T. denticola and smoking were independently and strongly associated with development of CP. It was concluded that when used in conjunction with an optimized clinical examination protocol, this assay may offer a rapid, useful, and cost-effective tool for monitoring the presence of T. denticola in noninvasive clinical samples from both healthy and diseased patients and correlating it with the amount and extent of periodontal breakdown.

Efectos del virus del dengue durante el embarazo. Medellín, Colombia / Dengue virus effects during pregnancy, Medellín, Colombia

El riesgo de infección durante la gestación aumenta ante mayores y más severas epidemias de dengue y sus consecuencias sobre el embarazo y el feto han sido poco estudiadas o muestran resultados contradictorios. Por este motivo el presente estudio tiene como fin describir los efectos del virus del dengue en el embarazo. Se evaluaron, mediante estudio descriptivo retrospectivo, 24gestantes que presentaron dengue durante la epidemia ocurrida en Medellín en 1998 y se compararon con 24 embarazadas sin dengue. Se consideró caso de dengue toda paciente con presencia de anticuerpos IgM contra el virus del dengue o con clínica compatible y asociaciónepidemiológica. Los hallazgos presentados en la cohorte expuesta vs cohorte no expuesta fueron los siguientes: siete casos (29.2 por ciento) de amenaza de aborto Vs dos (8.3 por ciento); dos casos (8.3 por ciento) de abortoVs ninguno; seis casos (25 por ciento) de amenaza de parto prematuro Vs cuatro (16.6 por ciento); tres casos (12.5 por ciento) de parto prematuro Vs ninguno; cuatro casos (16.6 por ciento) de ruptura prematura de membranas Vs uno (4.1 por ciento), nueve casos (37.5 por ciento) de hemorragia vaginal Vs uno (4.1 por ciento) y en cuatro pacientes de la cohorte expuesta se desencadenó trabajo de parto concomitante con el dengue. De las anteriores observaciones, sólo hubo diferencia estadísticamente significativa con respecto a la presencia de hemorragia vaginal entre los grupos de estudio (Prueba de Fisher, p < 0.05). En conclusión, las mujeres que desarrollaron infección por dengue durante la gestación, tuvieron riesgo de hemorragia vaginal y presentan con mayor frecuencia actividad uterina desencadenada por la infección, acontecimientos con efectos negativos sobre el binomio madre - hijo