Cytokines and MMPs levels in gingival crevicular fluid from patients with chronic periodontitis before and after non-surgical periodontal therapy

The study of host response in periodontal disease may provide a mechanism to monitor disease progression. the purpose of the present research was to determine the levels of IL-1alfa, IL-1beta, TNF-alfa, IL-6, IL-6sR, IL-8, IL-10, MMP-3 and MMP-8 in gingival crevicular fluid (GCF), before and after non-surgical periodontal treatment (NSPT) in order to evaluate therapy response. methodology: eleven patients diagnosed with chronic periodontitis and eleven healthy subjects were selected for this study. clinical measurements, including probing depth (PD) and clinical attachment loss (CAL) were carried out in patients diagnosed with chronic periodontitis and periodontal healthy controls. the clinical indexes evaluated were: gingival index (GI) and plaque index (PI). samples of GCF were taken from one tooth per quadrant before and 45 days after NSPT. the levels of inflammatory mediators were measured by ELISA. results: the values of all clinical parameters decressed significsntly after treatment. the concentration levels of all cytokines and MMP-3 and MMP-8 in the GCF sample were higher in patients diagnosed with chronic periodontitis compared to the healthy group. all inflammatory mediators decreased after therapy, but did not reach control values; IL-6, Il-6sR, IL-10 and TNF-alfa, attained the highest reduction (70 percent -54 percent); the vales of MMP3, IL-1alfa, IL-1beta and IL-8 were reduced between 50 percent ­ 34 percent; and MMP-8 showed the lowest decrease (28 percent). conclusion: all clinical parameters and cytokines levels decreased after NSPT. the mediators TNF-alfa IL-6, IL-6sR, and IL-10 showed the largest variation between before and after NSPT and could thus be used to evaluate therapy response.

Cytokine and metalloproteinases in gingival fluid from patients with chronic periodontitis / Niveles de citocinas y metaloproteinasas en fluido gingival de pacientes con periodontitis crónica

The purpose of the present research was to determine the levels of IL-1α, IL-1β, TNF-α, IL-6, IL-6sR, IL-8, IL-10, MMP-3 and MMP-8 in gingival crevicular fluid (GCF) of subjects with chronic periodontitis. Clinical measurements were carried out in 20 patients with chronic periodontitis and 11 periodontally healthy controls. The clinical indexes evaluated were: gingival index (GI), plaque index (PI), bleeding on probing (BOP), probing depth (PD) and clinical attachment loss (CAL); the measurements were taken at six sites per tooth in all teeth in each subject. GCF samples were taken from one tooth per quadrant, and the levels of mediators were measured using an ELISA test. Statistically significant differences were observed between patients and control group in relation to all clinical parameters evaluated (p<0.05). The gingival concentrations, in pg/mL, of IL-1α (patients: 239.06 ± 65.5 vs control: 97.79 ± 15.81), IL-1β (patients: 157.19 ± 36.4 vs control: 63.44 ± 19.04), TNF-α (patients: 10.87 ± 1.7 vs control: 1.15 ± 0.84), IL-6 (patients: 3.77 ±1.7 vs control: 0.43 ± 0.22), IL-6Sr (patients: 655.59 ± 185.8 vs control: 73.59 ± 23.18), IL-8 (patients: 496.3 ± 155.3 vs control: 206.13 ± 46.63), IL-10 (patients: 10.75 ± 3.6 vs control: 2.41 ± 0.57), MMP-3 (patients: 3531 ± 1558.2 vs control: 724.84 ± 289.51) and MMP-8 (patients: 8231.70± 1279.2 vs control: 1534.67± 814.90) were significantly greater in patients with periodontal disease than in the control group (p<0.001). The higher levels of the cytokines and metalloproteinases obtained in this study were significantly associated with the severity of the periodontal disease.

El propósito de la presente investigación fue determinar los niveles de IL-1α, IL-1β, TNF-α, IL-6, IL-6sR, IL-8, IL-10, MMP-3 and MMP-8 en fluido gingival crevicular (FGC) de sujetos con periodontitis crónica. Se evaluaron los parámetros clínicos en 20 pacientes con periodontitis crónica y 11 controles periodontalmente sanos. Los índices clínicos evaluados fueron: indice gingival (IG), indice de placa dental (IP), sangramiento al sondaje (SS), profundidad del saco (PS) y nivel de inserción (NI). Las muestras de FGC fueron tomadas de un diente por cada cuadrante y los niveles de los mediadores fueron medidos utilizando la prueba de ELISA. Se observaron diferencias estadísticamente significativas entre los pacientes y el grupo control en relación a todos los parámetros clínicos evaluados (p<0,05). Las concentraciones en fluido gingival en pg/mL de IL-1α(pacientes: 239,06 ± 65,5 vs control: 97,79 ± 15,81), IL-1β (pacientes: 157,19 ± 36,4 vs control: 63,44 ± 19,04), TNF-α (pacientes: 10,87 ± 1,7 vs control: 1,15 ± 0,84), IL-6 (pacientes: 3,77 ±1,7 vs control: 0,43 ± 0,22), IL-6Sr (pacientes: 655,59 ± 185,8 vs control: 73,59 ± 23,18), IL-8 (pacientes: 496,3 ± 155,3 vs control: 206,13 ± 46,63), IL-10 (pacientes: 10,75 ± 3.6 vs control: 2,41 ± 0,57), MMP-3 (pacientes: 3531 ± 1558,2 vs control: 724,84 ± 289,51) and MMP-8 (pacientes: 8231,70± 1279,2 vs control: 1534,67± 814,90), estuvieron significativamente mayores en pacientes con enfermedad periodontal que en el grupo control. (p<0,001). Los niveles elevados de citocinas y metaloproteinasas obtenidos en este estudio estuvieron significativamente asociados con la severidad de la enfermedad periodontal.

A novel PDE1A coupled to M2AChR at plasma membranes from bovine tracheal smooth muscle.

Muscarinic antagonists, via muscarinic receptors increase the cAMP/cGMP levels at bovine tracheal smooth muscle (BTSM) through the inhibition of phosphodiesterases (PDEs), displaying a similar behavior of vinpocetine (a specific-PDE1 inhibitor). The presence of PDE1 hydrolyzing both cyclic nucleotides in BTSM strips was revealed. Moreover, a vinpocetine and muscarinic antagonists inhibited PDE1 located at plasma membranes (PM) fractions from BTSM showing such inhibition, an M(2)AChR pharmacological profile. Therefore, a novel Ca(2+)/CaM dependent and vinpocetine inhibited PDE1 was purified and characterized at PM fractions from BTSM. This PDE1 activity was removed from PM fractions using a hypotonic buffer and purified some 38 fold using two columns (Q-Sepharose and CaM-agarose). This PDE1 was stimulated by CaM and inhibited by vinpocetine showing two bands in PAGE-SDS (56, 58 kDa) being the 58 kDa identified as PDE1A by Western blotts. This PDE1A activity was assayed with [(3)H]cGMP and [(3)H]cAMP exhibiting a higher affinity as Km (µM) for cGMP than cAMP but being close values with V(max) cAMP/cGMP ratio of 1.5. The co-factor Mg(2+) showed similar K(A) (mM) for both cyclic nucleotides. Vinpocetine showed similar inhibition concentration 50% (IC(50) of 4.9 and 4.6 µM) for cAMP and cGMP, respectively. CaM stimulated the cyclic nucleotides hydrolysis by PDE1A exhibiting similar activation constant as K(CaM), in nM range. The original finding was the identification and purification of a vinpocetine and muscarinic antagonist-inhibited and CaM-activated PM-bound PDE1A, linked to M(2)AChR. A model of this novel signal transducing cascade for the regulation of cyclic nucleotides levels at BTSM is proposed.

Cytokine and metalloproteinases in gingival fluid from patients with chronic periodontitis.

The purpose of the present research was to determine the levels of IL- 1a, IL- 1 P, TNF-a, IL-6, IL-6sR, IL-8, IL-10, MMP-3 and EMP-8 in gingival crevicular fluid (GCF) of subjects with chronic periodontitis. Clinical measurements were carried out in 20 patients with chronic periodontitis and 11 periodontally healthy controls. The clinical indexes evalua- ted were: gingival index (GI), plaque index (PI), bleeding on probing (BOP), probing depth (PD) and clinical attachment loss (CAL); the measurements were taken at six sites per tooth in all teeth in each subject. GCF samples were taken from one tooth per quadrant, and the levels of mediators were measured using an ELISA test. Statistically significant differences were observed between patients and control group in relation to all clinical parameters evaluated (p<0.05). The gingival concentrations, in pg/mL, of IL-la (patients: 239.06 ± 65.5 vs control: 97.79 ± 15.81), IL-10 (patients: 157.19 ± 36.4 vs control: 63.44 ± 19.04), TNF-a (patients: 10.87 ± 1.7 vs control: 1.15 ± 0.84), IL-6 (patients: 3.77 ± 1.7 vs control: 0.43 ± 0.22), IL-6Sr (patients: 655.59 ± 185.8 vs control: 73.59 ± 23.18), IL-8 (patients: 496.3 ± 155.3 vs control: 206.13 ± 46.63), IL-10 (patients: 10.75 ± 3.6 vs control: 2.41 ± 0.57), MMP-3 (patients: 3531 ± 1558.2 vs control: 724.84 ± 289.51) and MMiP-8 (patients: 8231.70 ± 1279.2 vs control: 1534.67± 814.90) were significantly greater in patients with periodontal disease than in the control group (p<0.00 1). The higher levels of the cytokines and metalloproteinases obtained in this study were significantly associated with the severity of the periodontal disease.

[MMP-3 and MMP-8 levels in patients with chronic periodontitis before and after nonsurgical periodontal therapy]. / Niveles de MMP-3 y MMP-8 en pacientes con periodontitis crónica antes y después del tratamiento periodontal no quirúrgico.

Immune-inflammatory processes are trigged in chronic periodontitis (CP), where matrix metalloproteinases (MMPs) are released and involved in the degradation of the extracellular matrix components that can be detected in the gingival crevicular fluid (GCF). The purpose of the study was to determine the levels of MMP-3 and MMP-8 in GCF, before and after nonsurgical periodontal treatment (NSPT), to evaluate disease activity and therapy response. Eleven patients with PC and eleven healthy controls were selected. Clinical measurements to evaluate gingival index (GI), plaque index (PI), probing depth (PD) and clinical attachment loss (CAL) were made in all the teeth of each individual and in six sites per tooth. GCF samples were taken from one tooth per quadrant, with a pocket depth > or =4 mm and a clinical attachment loss > or =5 mm, and the levels of MMP-3 and MMP-8 measured using an ELISA test. Statistically significant differences in clinical parameters were observed (p < 0.05) between patients with CP and control groups before the periodontal treatment, with significant decrease in all indexes after the NSPT. The initial concentrations of MMP-3 and MMP-8 were significantly higher than those obtained after the NSPT and in the control group, without observing a correlation between the clinical parameters and the levels of MMPs. Increased levels of MMP-3 and MMP-8 in the GCF of patients with PC declined significantly after NSPT, and the difference between the levels in healthy individuals and patients, suggests the important participation of these MMPs in tissue destruction in PC disease..

Niveles de MMP-3 y MMP-8 en pacientes con periodontitis crónica antes y después del tratamiento periodontal no quirúrgico / MMP-3 and MMP-8 levels in patients with chronic periodontitis before and after nonsurgical periodontal therapy

En la periodontitis crónica (PC) se desencadenan procesos inmunoinflamatorios, donde se liberan metaloproteinasas de la matriz (MMPs), enzimas involucradas en la degradación de la matriz extracelular, las cuales pueden ser detectadas en el fluido gingival crevicular (FGC). El propósito del estudio fue determinar los niveles de MMP-3 y MMP-8 en FGC, antes y después del tratamiento periodontal no quirúrgico (TPNQ) para evaluar actividad de la enfermedad y respuesta terapéutica. Once pacientes con periodontitis crónica y 11 controles sanos fueron seleccionados. Se evaluaron los parámetros clínicos: índice gingival, índice de placa, profundidad al sondaje y pérdida de inserción; en todos los dientes de cada individuo y en seis sitios por diente. Muestras de FGC fueron tomadas de un diente por cuadrante, con profundidad de saco ≥ 4mm y pérdida de inserción ≥ 5 mm, los niveles de MMP-3 y MMP-8 fueron determinados por ELISA. Diferencias estadísticamente significativas fueron observadas entre los parámetros clínicos del grupo control y los pacientes con PC antes del tratamiento, registrándose posterior al TPNQ disminución significativa de todos los índices. Las concentraciones iniciales de MMP-3 y 8 en el grupo con PC fueron significativamente mayores a las obtenidas luego del TPNQ y en el grupo control, sin observar correlación entre parámetros clínicos y niveles de MMPs. La disminución significativa de los valores de MMP-3 y 8 en FGC de los pacientes con PC, posterior al TPNQ, indican la participación importante de estas enzimas en la degradación del tejido, y la efectividad del tratamiento periodontal para su control.

Immune-inflammatory processes are trigged in chronic periodontitis (CP), where matrix metalloproteinases (MMPs) are released and involved in the degradation of the extracellular matrix components that can be detected in the gingival crevicular fluid (GCF). The purpose of the study was to determine the levels of MMP-3 and MMP-8 in GCF, before and after nonsurgical periodontal treatment (NSPT), to evaluate disease activity and therapy response. Eleven patients with PC and eleven healthy controls were selected. Clinical measurements to evaluate gingival index (GI), plaque index (PI), probing depth (PD) and clinical attachment loss (CAL) were made in all the teeth of each individual and in six sites per tooth. GCF samples were taken from one tooth per quadrant, with a pocket depth ≥ 4 mm and a clinical attachment loss ≥ 5 mm, and the levels of MMP-3 and MMP-8 measured using an ELISA test. Statistically significant differences in clinical parameters were observed (p < 0.05) between patients with CP and control groups before the periodontal treatment, with significant decrease in all indexes after the NSPT. The initial concentrations of MMP-3 and MMP-8 were significantly higher than those obtained after the NSPT and in the control group, without observing a correlation between the clinical parameters and the levels of MMPs. Increased levels of MMP-3 and MMP-8 in the GCF of patients with PC declined significantly after NSPT, and the difference between the levels in healthy individuals and patients, suggests the important participation of these MMPs in tissue destruction in PC disease.