16 research outputs found

    Accuracy of cotinine serum test to detect the smoking habit and its association with periodontal disease in a multicenter study

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    The validity of the surveys on self-reported smoking status is often questioned because smokers underestimate cigarette use and deny the habit. It has been suggested that self-report should be accompanied by cotinine test. This report evaluates the usefulness of serum cotinine test to assess the association between smoking and periodontal status in a study with a large sample population to be used in studies with other serum markers in epidemiologic and periodontal medicine researches. 578 patients who were part of a multicenter study on blood biomarkers were evaluated about smoking and its relation to periodontal disease. Severity of periodontal disease was determinate using clinical attachment loss (CAL). Smoking was assessed by a questionnaire and a blood sample drawn for serum cotinine determination. The optimal cut-off point for serum cotinine was 10 ng/ml. Serum cotinine showed greater association with severity of CAL than self-report for mild-moderate CAL [OR 2.03 (CI95% 1.16-3.53) vs. OR 1.08 (CI95% 0.62-1.87) ] advanced periodontitis [OR 2.36 (CI95% 1.30- 4.31) vs. OR 2.06 (CI95% 0.97-4.38) ] and extension of CAL > 3 mm [ OR 1.78 (CI95% 1.16-1.71) vs. 1.37 (CI95% 0.89-2.11)]. When the two tests were evaluated together were not shown to be better than serum cotinine test. Self-reported smoking and serum cotinine test ? 10ng/ml are accurate, complementary and more reliable methods to assess the patient?s smoking status and could be used in studies evaluating serum samples in large population and multicenter studies. Clinical Relevance: The serum cotinine level is more reliable to make associations with the patient?s periodontal status than self-report questionnaire and could be used in multicenter and periodontal medicine studies

    Desarrollo de un modelo celular endotelial tridimensional para la evaluaci贸n del efecto de LPS de Porphyromonas gingivalis en la expresi贸n de algunos marcadores inflamatorios

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    Se desarroll贸 un cultivo endotelial tridimensional sobre soportes de col谩geno I (3DCE) para evaluar la respuesta inflamatoria y funci贸n endotelial inducida por lipopolisac谩ridos (LPS) de Porphyromonas gingivalis; y compararla con cultivos bidimensionales (2DCE). Los cultivos 3DCE y 2DCE fueron estimulados con 1 碌g/mL de LPS-P.g por 24 horas. Como control se utilizaron cultivos sin estimular. Adem谩s, se compar贸 en ambos modelos la respuesta inducida por LPS aislado de Aggregatibacter actinomycetemcomitans. Se evalu贸 la expresi贸n de genes IL-8, MCP-1, COX-2, eNOS, vWF, la secreci贸n de 40 citoquinas y prostanglandinas (PGI2 y TXA2), y la adhesi贸n de monocitos. Los experimentos se realizaron por triplicado y las diferencias fueron consideradas significativas con un p0,05. En 3DCE se observ贸 una disminuci贸n en la transcripci贸n de COX-2, eNOS y vWF inducida por ambos LPS, mientras que se increment贸 IL-8 y MCP-1 solamente con LPS-A.a. En 2DCE la transcripci贸n de COX-2, IL-8 y MCP-1 se increment贸 por LPS-A.a pero no por LPS-P.g. La secreci贸n de 12 citoquinas en 3DCE fue incrementada comparado con el cultivo 2DCE en los controles. El tratamiento con LPS-A.a en 3DCE increment贸 la secreci贸n de IL-8, RANTES, G-CSF, ICAM-1, IL-6 y TXA2; mientras que en 2DCE se incrementaron G-CSF, IL-8, RANTES, ICAM-1, TNF-RI, PGI2 y TXA2. La adhesi贸n de monocitos al cultivo 2DCE fue incrementada tras el est铆mulo con LPS-A.a pero no en el cultivo 3DCE. LPS-P.g no indujo la secreci贸n de citoquinas, prostanglandinas y adhesi贸n de monocitos. El cultivo 3DCE refleja un comportamiento influenciado por el entorno tridimensional, lo que permite exhibir diferentes respuestas celulares frente a LPS de periodontopat贸genos.Abstract: A three-dimensional endothelial culture on collagen I scaffold (3DCE) was developed to evaluate the inflammatory response and endothelial function induced by lipopolysaccharides (LPS) of Porphyromonas gingivalis; and compare it with twodimensional cultures (2DCE). The 3DCE and 2DCE cultures were stimulated with 1 碌g / mL of LPS-P.g for 24 hours. As a control, cultures without stimulation were used. In addition, the LPS-induced response isolated from Aggregatibacter actinomycetemcomitans was compared in both models. The expression of IL-8, MCP-1, COX-2, eNOS, vWF genes, secretion of cytokines and prostanglandins (PGI2 and TXA2), and monocyte adhesion were evaluated. The experiments were performed in triplicate and the differences were considered significant with a p 0.05. In 3DCE a decrease in the transcription of COX-2, eNOS and vWF induced by both LPS was observed, while IL-8 and MCP-1 was increased only with LPS-A.a. In 2DCE the transcription of COX-2, IL-8 and MCP-1 was increased by LPS-A.a but not by LPS-P.g. The secretion of 12 cytokines in 3DCE was increased compared to 2DCE culture in the controls. Treatment with LPS-A.a in 3DCE increased the secretion of IL-8, RANTES, G-CSF, ICAM-1, IL-6 and TXA2; while in 2DCE G-CSF, IL-8, RANTES, ICAM-1, TNF-RI, PGI2 and TXA2 were increased. Monocyte adhesion to 2DCE culture was increased after stimulation with LPS-A.a but not in 3DCE culture. LPS-P.g did not induce the secretion of cytokines, prostanglandins and monocyte adhesion. The 3DCE culture reflects a behavior influenced by the three-dimensional environment, which allows different cell responses to be exhibited against LPS of periodontopathogens.Doctorad

    Inducci贸n de disfunci贸n endotelial in vitro por lipopolisacarido de bacterias periodontopaticas e inhibici贸n de la inflamaci贸n por resolvina (rvd1) y estatina

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    Este estudio eval煤a los mecanismos de disfunci贸n endotelial mediado por lipopolisacaridos de periodontopat贸genos y valora el uso de resolvina D1 (RvD1) y Rosuvastatina en regular la disfunci贸n endotelial in vitro, para establecer los efectos de estos terap茅uticos y dar bases para la aplicaci贸n cl铆nica de las resolvinas en la terapia periodontal en pacientes con riesgo cardiovascular. El objetivo de esta fase de estudio fue evaluar la capacidad del lipopolisacarido de P. gingivalis y A. actinomycetemcomitans y de bacteria completa aniquilada por calor para inducir disfunci贸n endotelial en c茅lulas de arteria coronaria humana.Departamento Administrativo de Ciencia, Tecnolog铆a e Innovaci贸n [CO] Colciencias1308-519-28960Inducci贸n de disfunci贸n endotelial in vitro por lipopolisacarido de bacterias periodontopaticas e inhibici贸n de la inflamaci贸n por resolvina (rvd1) y estatina (rosuvastatina)s

    Efecto de rosuvastatina sobre la expresi贸n in vitro de pecam-1 en c茅lulas endoteliales humanas estimuladas con los de porphyromonas gingivalis

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    La presencia de bacterias periodonto-pat贸genas como Pophyromonas gingivalis en lesiones ateroscler贸ticas y procesos de endotoxemias en pacientes con enfermedad periodontal pueden ser factores que desencadenar铆an a nivel endotelial una respuesta inflamatoria pro-ateroscler贸tica. La expresi贸n de mol茅culas de adhesi贸n tras la activaci贸n endotelial como PECAM-1 toma gran importancia en la adhesi贸n y migraci贸n de monocitos durante el proceso inflamatorio pro-ateroscler贸tico. El manejo en la reducci贸n de estos marcadores inflamatorios ha motivado a buscar minuciosamente posibles efectos anti-inflamatorios en medicamentos convencionalmente formulados para prevenir el riesgo de enfermedad vascular siendo actualmente la Rosuvastatina un f谩rmaco con potencial antiinflamatorio, por lo que se evalu贸 sobre la expresi贸n in vitro de PECAM-1 en c茅lulas HCAEC, estimuladas con LPS de P. gingivalis 33277 mediante la t茅cnica Cell ELISA Fluorom茅trica, de igual modo se evalu贸 el efecto del f谩rmaco sobre la viabilidad celular utilizando la prueba espectrofotom茅trica de resazurina. Los resultados obtenidos demostraron que Rusuvastatina disminuye la expresi贸n de PECAM-1 en c茅lulas HCAEC de manera concentraci贸n dependiente (p<0.05) y no inducen citotoxicidad sobre las c茅lulas, a concentraciones de 100 uM- 6,5uM durante un periodo de exposici贸n de 24 horas. Los hallazgos de este estudio sugieren que Rosuvastatina posee un perfil farmacol贸gico promisorio dado que posee efectos moduladores en la adhesi贸n, vislumbrando propiedades anti-inflamatorias y un alto rango de seguridad toxicol贸gica.Departamento Administrativo de Ciencia, Tecnolog铆a e Innovaci贸n [CO] Colciencias1308-519-28960Inducci贸n de disfunci贸n endotelial in vitro por lipopolisacarido de bacterias periodontopaticas e inhibici贸n de la inflamaci贸n por resolvina (rvd1) y estatina (rosuvastatina)n

    Efecto de enjuagues de 谩cido hipocloroso sobre el pH de la saliva: estudio in vitro

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    聽Background: Hypochlorous acid (HOCl) has been proposed as antiplaque agent. The potential use of anti-plaque mouthwashes must be previously evaluated to determine whether it affects damping properties of saliva favoring tooth demineralization processes. Aim: To evaluate in vitro the effect of mouthwashes with HOCl at different concentrations on saliva pH. Methods: 20 whole saliva samples were collected. 1.1 ml of saliva were titrated with 0.1 or 0.4 mL of HOCl at different concentrations (125, 250 y 500 ppm) until a volume ratio 1:1 and 4:1. 0.5% NaCl was used as a titration control. HOCl volume required to induce a critical pH in saliva was assessed at 鈮 5.5. A descriptive analysis for all variables and ANOVA with post hoc Bonferroni with multiple comparisons was conducted. Results: None of the HOCl concentrations evaluated affects the ability of the saliva to neutralize acids in solution at a 1:1 ratio. However, it is reached at pH < 5.5 when the proportion of HOCl at 500 ppm was increased in relation to the volume of saliva (3:1; p = 0.016). Concentrations of 250 and 125 ppm do not affect saliva pH even at proportions in volume of 6:1 and 9:1. Conclusion: HOCl at 125 ppm and 250 ppm does not affect the ability of saliva to neutralize acids in solution and these concentrations are suitable for use as active agent of an antiplaque mouthwash.Antecedentes: Se ha propuesto el 谩cido hipocloroso (HOCl) como un agente antiplaca. El potencial uso de enjuagues con HOCl debe valorarse para establecer si afecta el pH y las propiedades amortiguadoras de la saliva que favorezcan procesos de desmineralizaci贸n dental. Objetivo: Evaluar el efecto in vitro de enjuagues con HOCl a diferentes concentraciones sobre el pH de la saliva. M茅todos: Se recolectaron 20 muestras de saliva total. 1,1 mL de saliva fueron titulados con 0,1 y 0,4 mL de HOCl a diferentes concentraciones (125, 250 y 500 ppm) hasta una proporci贸n en volumen 1:1 o 4:1. El NaCl 0,5 % se utiliz贸 como control de titulaci贸n. Se evalu贸 el volumen requerido de HOCl para inducir un pH cr铆tico de la saliva 鈮 5,5. Se efectu贸 un an谩lisis descriptivo para todas las variables y un Anova con post hoc de comparaciones m煤ltiples de Bonferroni. Resultados: Ninguna de las concentraciones evaluadas de HOCl afect贸 la capacidad de la saliva en amortiguar los 谩cidos en soluci贸n a una proporci贸n 1:1. Sin embargo, se alcanz贸 un pH < 5,5 cuando se aument贸 la proporci贸n de HOCl 500 ppm en relaci贸n con el volumen de saliva (3:1; p = 0,016). Las concentraciones 250 y 125 ppm no afectan considerablemente el pH de la saliva incluso a proporciones en volumen 6:1 y 9:1, respectivamente. Conclusi贸n: El HOCl a 125 ppm y a 250 ppm no afecta la capacidad de la saliva para neutralizar los 谩cidos en soluci贸n, por lo que estas concentraciones son 贸ptimas para su potencial uso como principio activo de enjuague bucal antiplaca

    Efecto de enjuagues de 谩cido hipocloroso sobre el pH de la saliva: estudio in vitro / Effect of Hypochlorous Acid as a Mouthwash on Salivary pH: in vitro Study

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    Antecedentes: Se ha propuesto el 谩cido hipocloroso (HOCl) como un agente antiplaca. El potencial uso de enjuagues con HOCl debe valorarse para establecer si afecta el pH y las propiedades amortiguadoras de la saliva que favorezcan procesos de desmineralizaci贸n dental. Objetivo: Evaluar el efecto in vitro de enjuagues con HOCl a diferentes concentraciones sobre el pH de la saliva. M茅todos: Se recolectaron 20 muestras de saliva total. 1,1 mL de saliva fueron titulados con 0,1 y 0,4 mL de HOCl a diferentes concentraciones (125, 250 y 500 ppm) hasta una proporci贸n en volumen 1:1 o 4:1. El NaCl 0,5 % se utiliz贸 como control de titulaci贸n. Se evalu贸 el volumen requerido de HOCl para inducir un pH cr铆tico de la saliva 鈮 5,5. Se efectu贸 un an谩lisis descriptivo para todas las variables y un Anova con post hoc de comparaciones m煤ltiples de Bonferroni. Resultados: Ninguna de las concentraciones evaluadas de HOCl afect贸 la capacidad de la saliva en amortiguar los 谩cidos en soluci贸n a una proporci贸n 1:1. Sin embargo, se alcanz贸 un pH &lt; 5,5 cuando se aument贸 la proporci贸n de HOCl 500 ppm en relaci贸n con el volumen de saliva (3:1; p = 0,016). Las concentraciones 250 y 125 ppm no afectan considerablemente el pH de la saliva incluso a proporciones en volumen 6:1 y 9:1, respectivamente. Conclusi贸n: El HOCl a 125 ppm y a 250 ppm no afecta la capacidad de la saliva para neutralizar los 谩cidos en soluci贸n, por lo que estas concentraciones son 贸ptimas para su potencial uso como principio activo de enjuague bucal antiplaca.聽Background: Hypochlorous acid (HOCl) has been proposed as antiplaque agent. The potential use of anti-plaque mouthwashes must be previously evaluated to determine whether it affects damping properties of saliva favoring tooth demineralization processes. Aim: To evaluate in vitro the effect of mouthwashes with HOCl at different concentrations on saliva pH. Methods: 20 whole saliva samples were collected. 1.1 ml of saliva were titrated with 0.1 or 0.4 mL of HOCl at different concentrations (125, 250 y 500 ppm) until a volume ratio 1:1 and 4:1. 0.5% NaCl was used as a titration control. HOCl volume required to induce a critical pH in saliva was assessed at 鈮 5.5. A descriptive analysis for all variables and ANOVA with post hoc Bonferroni with multiple comparisons was conducted. Results: None of the HOCl concentrations evaluated affects the ability of the saliva to neutralize acids in solution at a 1:1 ratio. However, it is reached at pH &lt; 5.5 when the proportion of HOCl at 500 ppm was increased in relation to the volume of saliva (3:1; p = 0.016). Concentrations of 250 and 125 ppm do not affect saliva pH even at proportions in volume of 6:1 and 9:1. Conclusion: HOCl at 125 ppm and 250 ppm does not affect the ability of saliva to neutralize acids in solution and these concentrations are suitable for use as active agent of an antiplaque mouthwash

    Human Coronary Artery Endothelial Cell Response to <i>Porphyromonas gingivalis</i> W83 in a Collagen Three-Dimensional Culture Model

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    P. gingivalis has been reported to be an endothelial cell inflammatory response inducer that can lead to endothelial dysfunction processes related to atherosclerosis; however, these studies have been carried out in vitro in cell culture models on two-dimensional (2D) plastic surfaces that do not simulate the natural environment where pathology develops. This work aimed to evaluate the pro-inflammatory response of human coronary artery endothelial cells (HCAECs) to P. gingivalis in a 3D cell culture model compared with a 2D cell culture. HCAECs were cultured for 7 days on type I collagen matrices in both cultures and were stimulated at an MOI of 1 or 100 with live P. gingivalis W83 for 24 h. The expression of the genes COX-2, eNOS, and vWF and the levels of the pro-inflammatory cytokines thromboxane A2 (TXA-2) and prostaglandin I2 (PGI2) were evaluated. P. gingivalis W83 in the 2D cell culture increased IL-8 levels at MOI 100 and decreased MCP-1 levels at both MOI 100 and MOI 1. In contrast, the 3D cell culture induced an increased gene expression of COX-2 at both MOIs and reduced MCP-1 levels at MOI 100, whereas the gene expression of eNOS, vWF, and IL-8 and the levels of TXA2 and PGI2 showed no significant changes. These data suggest that in the collagen 3D culture model, P. gingivalis W83 induces a weak endothelial inflammatory response

    Rosuvastatin inhibits IL-8 and IL-6 production in human coronary artery endothelial cells stimulated with Aggregatibacter actinomycetemcomitans

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    Aggregatibacter actinomycetemcomitans (A.a) is a Gram negative periodontopathogen that has been highly associated with endocarditis and atherosclerosis. However, the potential mechanisms by which A.a could be contributing to atherosclerosis remain unclear. The purpose of this study was to determine the effect of purified LPS from A.a (Aa-LPS) on the expression of pro-inflammatory molecules (i.e., adhesion molecules, Toll-like receptors and cytokines/chemokines) associated with the pathogenesis of atherosclerosis in human coronary artery endothelial cells (HCAECs), as well as evaluating the potential of Rosuvastatin (RSV) for inhibiting the A.a-induced endothelial responses. HCAECs were stimulated with purified A. a-LPS and cytokine expression levels determined by qPCR and flow cytometry, and TLR2 and TLR4 expression evaluated by ELISA fluorometric assay. The effect of RSV in Aa-LPS-induced pro-inflammatory responses was also studied using similar experimental approaches. A. a-LPS increased the expression of IL-6, IL-8, and TLR2 in HCAECs. No effects in the expression of adhesion molecules were observed. Aa-induced IL-6 and IL-8 production was inhibited by RSV particularly at higher doses. These results suggest that Aa-LPS plays a role in pro-inflammatory endothelial responses that could be contributing to the atherosclerotic process, and the use of statins (i.e., RSV) could be reducing the likelihood for Aa-induced pro-atherosclerotic endothelial responses.Departamento Administrativo de Ciencia, Tecnolog铆a e Innovaci贸n [CO] Colciencias1308-519-28960Inducci贸n de disfunci贸n endotelial in vitro por lipopolisacarido de bacterias periodontopaticas e inhibici贸n de la inflamaci贸n por resolvina (rvd1) y estatina (rosuvastatina)n

    Efecto de enjuagues de 谩cido hipocloroso sobre el pH de la saliva: estudio in vitro / Effect of Hypochlorous Acid as a Mouthwash on Salivary pH: in vitro Study

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    Antecedentes: Se ha propuesto el 谩cido hipocloroso (HOCl) como un agente antiplaca. El potencial uso de enjuagues con HOCl debe valorarse para establecer si afecta el pH y las propiedades amortiguadoras de la saliva que favorezcan procesos de desmineralizaci贸n dental. Objetivo: Evaluar el efecto in vitro de enjuagues con HOCl a diferentes concentraciones sobre el pH de la saliva. M茅todos: Se recolectaron 20 muestras de saliva total. 1,1 mL de saliva fueron titulados con 0,1 y 0,4 mL de HOCl a diferentes concentraciones (125, 250 y 500 ppm) hasta una proporci贸n en volumen 1:1 o 4:1. El NaCl 0,5 % se utiliz贸 como control de titulaci贸n. Se evalu贸 el volumen requerido de HOCl para inducir un pH cr铆tico de la saliva 鈮 5,5. Se efectu贸 un an谩lisis descriptivo para todas las variables y un Anova con post hoc de comparaciones m煤ltiples de Bonferroni. Resultados: Ninguna de las concentraciones evaluadas de HOCl afect贸 la capacidad de la saliva en amortiguar los 谩cidos en soluci贸n a una proporci贸n 1:1. Sin embargo, se alcanz贸 un pH &lt; 5,5 cuando se aument贸 la proporci贸n de HOCl 500 ppm en relaci贸n con el volumen de saliva (3:1; p = 0,016). Las concentraciones 250 y 125 ppm no afectan considerablemente el pH de la saliva incluso a proporciones en volumen 6:1 y 9:1, respectivamente. Conclusi贸n: El HOCl a 125 ppm y a 250 ppm no afecta la capacidad de la saliva para neutralizar los 谩cidos en soluci贸n, por lo que estas concentraciones son 贸ptimas para su potencial uso como principio activo de enjuague bucal antiplaca.聽Background: Hypochlorous acid (HOCl) has been proposed as antiplaque agent. The potential use of anti-plaque mouthwashes must be previously evaluated to determine whether it affects damping properties of saliva favoring tooth demineralization processes. Aim: To evaluate in vitro the effect of mouthwashes with HOCl at different concentrations on saliva pH. Methods: 20 whole saliva samples were collected. 1.1 ml of saliva were titrated with 0.1 or 0.4 mL of HOCl at different concentrations (125, 250 y 500 ppm) until a volume ratio 1:1 and 4:1. 0.5% NaCl was used as a titration control. HOCl volume required to induce a critical pH in saliva was assessed at 鈮 5.5. A descriptive analysis for all variables and ANOVA with post hoc Bonferroni with multiple comparisons was conducted. Results: None of the HOCl concentrations evaluated affects the ability of the saliva to neutralize acids in solution at a 1:1 ratio. However, it is reached at pH &lt; 5.5 when the proportion of HOCl at 500 ppm was increased in relation to the volume of saliva (3:1; p = 0.016). Concentrations of 250 and 125 ppm do not affect saliva pH even at proportions in volume of 6:1 and 9:1. Conclusion: HOCl at 125 ppm and 250 ppm does not affect the ability of saliva to neutralize acids in solution and these concentrations are suitable for use as active agent of an antiplaque mouthwash
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