Estimation of salivary malondialdehyde Levels in smokeless tobacco chewers with chronic periodontitis-A cross sectional clinico biochemical study / Estimación de los niveles de malondialdehído salival en masticadores de tabaco sin humo con periodontitis crónica: un estudio bioquímico clínico transversal

ABSTRACT: Despite the reported effects of smokeless tobacco (ST) on the periodontium and high prevalence of ST use in rural populations and in males studies on this specific topic are limited. The purpose of this cross-sectional investigation was to measure lipid peroxidation (as an end product of oxidative stress) end product i.e. Malondialdehyde (MDA) in saliva of patients with gingivitis, chronic periodontitis and to assess the influence of smokeless tobacco on Salivary Malondialdehyde (S-MDA). Total 30 patients with gingivitis, 30 with chronic periodontitis and 30 Smokeless Tobacco Chewers with Chronic Periodontitis and 30 periodontally healthy subjects were included in the study. Plaque Index (PI), Gingival Index (GI), Probing Pocket Depth (PD), and Clinical Attachment Loss (CAL) were recorded followed by stimulated Saliva sample collection. Salivary MDA Levels were assessed by UV Spectrophotometry. There was a statistically significant increase in the salivary MDA levels in gingivitis, chronic periodontitis and in smokeless tobacco chewers with chronic periodontitis when compared with healthy group. Higher salivary MDA levels in gingivitis group, chronic periodontitis, and smokeless tobacco chewers with chronic periodontitis reflects increasedoxygen radical activity during periodontal inflammation.

RESUMEN: A pesar de los efectos reportados del tabaco sin humo (TS) sobre el periodonto y la alta prevalencia del uso de TS en poblaciones rurales y en hombres, los estudios sobre este tema específico son limitados. El propósito de esta investigación transversal fue medir el producto final de la peroxidación lipídica (como producto final del estrés oxidativo), es decir, malondialdehído (MDA) en la saliva de pacientes con gingivitis, periodontitis crónica y evaluar la influencia del tabaco sin humo en el malondialdehído salival (S-MDA). Se incluyeron en el estudio un total de 30 pacientes con gingivitis, 30 con periodontitis crónica y 30 masticadores de tabaco sin humo con periodontitis crónica y 30 sujetos periodontalmente sanos. Se registraron el índice de placa (PI), el índice gingival (GI), la profundidad de la bolsa de sondeo (PD) y la pérdida de adherencia clínica (CAL), seguidos de la recogida de muestras de saliva estimuladas. Los niveles de MDA en saliva se evaluaron mediante espectrofotometría UV. Hubo un aumento estadísticamente significativo en los niveles de MDA en saliva en gingivitis, periodontitis crónica y en masticadores de tabaco sin humo con periodontitis crónica en comparación con el grupo sano. Los niveles más altos de MDA en saliva en el grupo de gingivitis, periodontitis crónica y masticadores de tabaco sin humo con periodontitis crónica reflejan un aumento de la actividad de los radicales de oxígeno durante la inflamación periodontal.

Physalis angulata reduces the progression of chronic experimental periodontitis by immunomodulatory mechanisms.

ETHNOPHARMACOLOGICAL RELEVANCE: Physalis angulata is an herb found in tropical and subtropical regions of the world; it is widely applied in popular medicine due to the therapeutic properties of the whole plant and its parts. Extracts and infusions of this plant have been extensively applied in folk medicine worldwide to treat inflammatory and immune-mediated diseases, including oral inflammatory conditions such as sore throat and gingivitis. AIM OF THE STUDY: The present study was designed to investigate the protective effects of the ethanolic extract of P. angulata (EEPA) in a murine model of chronic periodontitis, aiming to corroborate its traditional use as an anti-inflammatory and immunomodulatory agent, and to point out possible mechanisms involved in these effects. MATERIALS AND METHODS: EEPA was obtained from the stems of P. angulata collected in Belém (PA, Brazil). Chronic periodontitis was induced in male C57BL/6 mice by 12 administrations of lipopolysaccharide (LPS; 20 µg/1µL) into the gingival papilla in the course of 28 days. Starting from the 15th day after the first LPS injection, mice were daily treated with EEPA (50 or 100 mg/kg), nimesulide (25 mg/kg, reference drug), or vehicle by oral route for 14 days. At the end of the experimental period, alveolar bone loss was evaluated along with the gingival expression of biomarkers of periodontitis and cytokines by RT-q-PCR and ELISA. Hematological and biochemical parameters suggestive of systemic toxicity were also evaluated. The transcriptional activity of NF-κB was investigated using the luciferase assay in macrophages. RESULTS: Mice with chronic experimental periodontitis suffered alveolar bone loss that was prevented by the treatment with EEPA (50 or 100 mg/kg) or nimesulide (25 mg/kg). EEPA (50 and 100 mg/kg) and nimesulide (25 mg/kg) reduced mRNA levels of MMP-9 mRNA, but not of TIMP-1 in gingival tissue of periodontitis-induced mice. Both treatments also reduced the production of the pro-inflammatory cytokines IL-1ß and IL-6. The treatment with EEPA (100 mg/kg) increased the production of the anti-inflammatory cytokine TGF-ß. No hematological or biochemical alterations were caused by the daily treatment with EEPA. In vitro luciferase assay suggested that a putative mechanism of EEPA is reducing the transcriptional activity of NF-κB. CONCLUSIONS: EEPA exhibited a disease-modifying effect in the chronic experimental periodontitis, along with unidentifiable systemic toxicity. This work corroborates the traditional use of P. angulata in oral inflammatory conditions and provides mechanistic hypotheses to explain its therapeutic effects.

Clinical and Microbiological Effects of Weekly Supragingival Irrigation with Aerosolized 0.5% Hydrogen Peroxide and Formation of Cavitation Bubbles in Gingival Tissues after This Irrigation: A Six-Month Randomized Clinical Trial.

INTRODUCTION: The study investigated the effect of weekly supragingival irrigation with aerosolized 0.5% hydrogen peroxide (H2O2) solution as a maintenance periodontal therapy on clinical and microbiological parameters in patients with chronic periodontitis. The other purpose was to investigate whether cavitation bubbles can penetrate not only into periodontitis-damaged tissues but also into ex vivo porcine healthy periodontal tissues. MATERIALS AND METHODS: The study included 35 systemically healthy patients with chronic periodontitis (CP). After nonsurgical periodontal debridement (NSPD), all patients were randomized into two groups: the Control group (NSDP alone, n = 18) and the Test group (NSDP plus supragingival irrigation, n = 17). Clinical (Approximal Plaque Index (API), Bleeding Index (BI), and Modified Gingival Index (MGI)) and microbiological (Polymerase Chain Reaction technology (using a micro-IDent® kit)) measurements were performed at the initial time point, 3 months, and 6 months after NSPD. The impact of supragingival irrigation on diseased gingival tissues of CP patients (n = 5) and on ex vivo porcine healthy gingival tissue samples (n = 3) was evaluated to estimate morphological changes in healthy and diseased gingival tissues. RESULTS: Morphological data revealed that supragingival irrigation caused the formation of cavitation bubbles in diseased gingival tissue of CP patients and in healthy porcine gingival tissues. The decrease in API, BI, and MGI scores after 6 months in the Test group significantly (p ≤ 0.01, p ≤ 0.05, and p ≤ 0.01, respectively) exceeded that in the Control group. Test group patients demonstrated a decrease in periodontal sites showing Pocket Probing Depth > 4 mm and, after 6 months, a statistically significant decrease in the proportion of periopathogenic bacteria. CONCLUSION: The effectiveness of mechanical periodontal treatment combined with weekly supragingival irrigation with aerosolized 0.5% H2O2 solution on clinical and microbiological parameters of periodontal tissues of periodontitis patients is reliably higher than that of mechanical periodontal debridement alone. It has been found that cavitation bubbles as a result of irrigation with the aerosolized 0.5% hydrogen peroxide solution can form not only in periodontal tissues of periodontitis patients but also in ex vivo porcine healthy gingival tissues.

Effect of antihypertensive therapy with angiotensin-converting enzyme inhibitors on chronic periodontitis: a case-control study.

BACKGROUND: Angiotensin-converting enzyme (ACE) downregulates the activity of bradykinin, a potent proinflammatory and immunostimulatory peptide liberated from an internal portion of kininogens. Here, we asked whether periodontitis is worsened in patients under antihypertensive treatment with ACE inhibitors. METHODS: Periodontal parameters were recorded from 30 individuals taking ACE inhibitors (case) and 35 taking a non-ACE inhibitor medication (control). Data were analyzed by nonparametric and parametric statistical tests. RESULTS: Most sociodemographic figures were similar in both groups. However, family income was statistically higher in the control group, and the percentage of sites with visible plaque (PL) was statistically higher in the case group (P = 0.043 and P = 0.005, respectively). The prevalence of individuals with chronic periodontitis varied from 31.5% in the control group to 63.4% in the case group (P = 0.001). Patients in the case group presented a 3.2-fold higher risk of having sites with pocket depth ≥5 mm and a 2.9-fold higher risk of having sites with clinical attachment loss ≥5 mm in comparison with those in the control group (P = 0.009 and P = 0.001, respectively; adjusted for family income and visible PL). CONCLUSION: Angiotensin-converting enzyme inhibitors may increase the prevalence and extent of chronic periodontitis in Brazilian patients.

Phenytoin-induced gingival overgrowth management with periodontal treatment.

Phenytoin-induced gingival overgrowth (PIGO) is a common complication of the continuous use of medications. This paper presents a case of PIGO hindering oral function and compromising oral hygiene and aesthetics, which was treated with a combination of nonsurgical and surgical periodontal therapies. A 39-year-old male patient was referred for dental treatment with several complaints, especially upper and lower gingival overgrowth that hindered speech and swallowing. Generalized deep probing pockets and bone loss were detected. Diagnosis of gingival overgrowth associated with phenytoin and chronic periodontitis was established. The treatment plan consisted of conservative therapy with education on oral health, motivation and meticulous oral hygiene instruction in combination with scaling and root planing. During the revaluation period, a marked reduction in the clinical parameters was noted, particularly probing pocket depth reduction. Surgical therapy for removal of gingival overgrowth was also performed to achieve pocket reduction. Supportive periodontal therapy was proposed and the patient is currently under follow-up for 4 years. Management of PIGO may be obtained by the use of periodontal procedures combined with good oral hygiene and periodontal supportive care.

Phenytoin-Induced Gingival Overgrowth Management with Periodontal Treatment

Phenytoin-induced gingival overgrowth (PIGO) is a common complication of the continuous use of medications. This paper presents a case of PIGO hindering oral function and compromising oral hygiene and aesthetics, which was treated with a combination of nonsurgical and surgical periodontal therapies. A 39-year-old male patient was referred for dental treatment with several complaints, especially upper and lower gingival overgrowth that hindered speech and swallowing. Generalized deep probing pockets and bone loss were detected. Diagnosis of gingival overgrowth associated with phenytoin and chronic periodontitis was established. The treatment plan consisted of conservative therapy with education on oral health, motivation and meticulous oral hygiene instruction in combination with scaling and root planing. During the revaluation period, a marked reduction in the clinical parameters was noted, particularly probing pocket depth reduction. Surgical therapy for removal of gingival overgrowth was also performed to achieve pocket reduction. Supportive periodontal therapy was proposed and the patient is currently under follow-up for 4 years. Management of PIGO may be obtained by the use of periodontal procedures combined with good oral hygiene and periodontal supportive care.

O crescimento gengival induzido pela fenitoína é uma complicação comum do uso contínuo da medicacão. Este artigo apresenta um caso de crescimento gengival excessivo que dificultava a função oral e comprometia a higiene oral e a estética, o qual foi tratado com uma combinação de terapias periodontais não-cirúrgicas e cirúrgicas. Paciente masculino de 39 anos de idade foi encaminhado para tratamento odontológico com várias queixas, especialmente do crescimento gengival superior e inferior que prejudicava a fala e deglutição. Profundidades de sondagens severas generalizadas e perda óssea foram detectadas. Diagnóstico de crescimento gengival induzido pela fenitoína e periodontite crônica foi estabelecido. O plano de tratamento consistiu de terapia conservadora com educação, motivação e meticulosa instrução de higiene oral em associação com raspagem e alisamento corono-radicular. Durante o período de reavaliação, uma acentuada redução nos parâmentros clínicos foi observada, principalmente uma redução das profundidades de sondagem. Terapia cirúrgica para remoção do excesso de tecido gengival também foi realizada para conseguir redução das bolsas. Terapia periodontal de suporte foi proposta e o paciente está atualmente sob acompanhamento por um período de 4 anos. O manejo do crescimento gengival induzido pela fenitoína pode ser obtido pelo uso de procedimentos periodontais combinados com uma boa higiene oral e cuidados periodontais de suporte.

Oxidized low-density lipoprotein-induced periodontal inflammation is associated with the up-regulation of cyclooxygenase-2 and microsomal prostaglandin synthase 1 in human gingival epithelial cells.

Periodontitis is characterized by chronic gingival tissue inflammation, and inflammatory mediators such as IL-8 and prostaglandin E(2) (PGE(2)) are associated with disease progression. Previously we showed that oxidatively modified low-density lipoprotein (oxLDL) was present in gingival crevicular fluid. In this study, the role of oxLDL in the gingival epithelial cell inflammatory response was further investigated using Ca9-22 cells and primary human oral keratinocytes (HOK). Treatment of Ca9-22 cells and HOK with oxLDL induced an up-regulation of IL-8 and the PGE(2)-producing enzymes, cyclooxygenase-2 and microsomal PGE(2) synthase-1. These responses induced by oxLDL were significantly suppressed by a nuclear factor-kappa B (NF-κB) inhibitor. However, unlike the result in macrophages, oxLDL did not lead to an increase in CD36 expression in these two cells. These results suggest that oxLDL elicits gingival epithelial cell inflammatory responses through an activation of the NF-κB pathway. These data suggest a mechanistic link between periodontal disease and lipid metabolism-related disorders, including atherosclerosis.

Micro-computerized tomography analysis of alveolar bone loss in ligature- and nicotine-induced experimental periodontitis in rats.

BACKGROUND AND OBJECTIVE: Nicotine reportedly is a risk factor for periodontitis, but accurate data regarding nicotine-induced alveolar bone loss is lacking. The aim of this study was to quantitatively assess alveolar bone loss in ligature- and nicotine-induced periodontitis in rats using micro-computerized tomography (micro-CT). MATERIAL AND METHODS: Thirty-six adult male rats were treated by placing silk ligatures around the cervixes of the right second maxillary molar; the contralateral tooth was untreated. After ligation, the animals were randomly divided into three groups: group A received intraperitoneal injections of saline solution, group B received 0.83 mg of nicotine/kg/d, and group C received 1.67 mg of nicotine/kg/d. Six animals in each group were killed on days 14 and 28 after ligature placement, and then micro-CT examinations were conducted. RESULTS: In all groups, bone mineral density (BMD), bone volume fraction (BVF), trabecular number (Tb.N) and trabecular thickness (Tb.Th) values of the ligated sides were significantly lower than those of the unligated sides (p < 0.001), whereas alveolar bone height loss (ABHL) and trabecular separation (Tb.Sp) of the ligated sides were significantly higher than those of the unligated sides (p < 0.001). Compared with the control group, nicotine administration increased the ABHL value and decreased the BMD, BVF and Tb.Th values of both sides in a dose-dependent manner (p < 0.05). CONCLUSION: Our results confirmed that ligature could cause significant loss in the trabecula of alveolar bone, and daily administration of nicotine resulted in further bone loss and microstructure deterioration.

Isolation and characterization of gingival fibroblasts positive for alkaline phosphatase in patients with chronic periodontitis and drug-induced gingival hyperplasia / Isolamento e caracterização de fibroblastos gengivais positivos para fosfatase alcalina de pacientes portadores de periodontite crônica e hiperplasia gengival medicamentosa

Purpose: To investigate the expression of alkaline phosphatase (ALP) activity in gingival fibroblasts from individuals with chronic periodontitis (CP) and drug-induced gingival hyperplasia (DGH) induced by diphenylhydantoin. Methods: Gingival fragments were obtained from 13 patients (8 women and 5 men, from 22 to 74 years of age), with 4 fragments from clinically normal gingiva (NG), 5 from biopsy of periodontal pockets with CP, and 4 from DGH induced by diphenylhydantoin. Using an enzymatic digestion procedure, gingival cell suspensions containing ALP-positive fibroblasts were prepared without affecting ALP activity. Cytochemistry and histochemistry analyses were performed. Results: Fibroblasts from NG presented low levels of ALP when compared to CP and DGH, which showed elevated and intermediate levels of ALP, respectively. Little cell proliferation was observed for fibroblasts from CP and DGH as compared to NG. However, the quantity of cells recovered from the subcultures was similar to the quantity recovered from the initial cell culture for the three sources. Conclusion: The expression of ALP is increased in CP and DGH, and fibroblasts in CP and DGH show low proliferation. This suggests that periodontal inflammation and diphenylhydantoin may influence ALP expression and human gingival fibroblast expansion. Other studies are necessary to better assess the importance of ALP in the development and progression of CP and DGH.

Objetivo: Investigar a expressão da atividade de fosfatase alcalina (ALP) em fibroblastos gengivais de sujeitos com periodontite crônica (PC) e hiperplasia gengival medicamentosa (HGM) induzida por difenilhidantoína. Metodologia: Os fragmentos gengivais foram obtidos de 13 pacientes (8 mulheres e 5 homens, 22 a 74 anos), sendo 4 fragmentos procedentes de gengiva clinicamente normal (GN), 5 de biópsias de bolsas periodontais com PC e 4 de HGM induzida pela difenilhidantoína. Através de digestão enzimática, suspensões de células gengivais foram preparadas contendo fibroblastos ALP-positivos sem afetar a atividade da enzima ALP. Análises citoquímica e histoquímica foram realizadas. Resultados: Fibroblastos de GN apresentaram fraca positividade para ALP quando comparados com PC e HGM. PC e HGM exibiram níveis elevados e intermediários, respectivamente, de ALP. Observou-se menor proliferação celular na cultura de células em PC e HGM que em GN. Entretanto, a quantidade de células recuperadas das sub-culturas foi similar à quantidade recuperada na cultura inicial para as três linhagens. Conclusões: A expressão de ALP na PC e na HGM encontra-se aumentada e ocorre baixa proliferação de fibroblastos na PC e na HGM. Isto sugere que a inflamação periodontal e a difenilhidantoína podem influenciar a expressão de ALP e a multiplicação de fibroblastos gengivais humanos. Outros estudos são necessários para melhor avaliar a importância de ALP no desenvolvimento e na progressão de PC e HGM.

A novel murine model for chronic inflammatory alveolar bone loss.

BACKGROUND AND OBJECTIVE: Chronic inflammatory bowel disease (IBD) demonstrates some similarities to the dysregulated chronic immunoinflammatory lesion of periodontitis. Trinitrobenzene sulphonic acid (TNBS) and dextran sodium sulphate (DSS) administered to rodents have been shown to elicit inflammatory responses that undermine the integrity of the gut epithelium in a similar manner to IBD in humans. The objective of this study was to evaluate the ability of these chemicals to elicit periodontal inflammation as a novel model for alveolar bone loss. MATERIAL AND METHODS: Mice were treated by oral application of TNBS twice a week, or with DSS in the diet over a period of 18 weeks. Alveolar bone loss was assessed on the defleshed skull using morphometric measures for area of bone resorption. RESULTS: The TNBS-treated animals tolerated oral administration with no clinical symptoms and gained weight at a similar rate to normal control animals. In contrast, DSS exerted a systemic response, including shortening of colonic tissue and liver enzyme changes. Both TNBS and DSS caused a localized action on periodontal tissues, with alveolar bone loss observed in both maxilla and mandibles, with progression in a time-dependent manner. Bone loss was detected as early as week 7, with more severe periodontitis increasing over the 18 weeks (p < 0.001). Young (7-month-old) and old (12-month-old) mice with severe combined immunodeficiency were treated with TNBS for a period of 7 weeks and did not develop significant bone loss. CONCLUSION: These data show that oral administration of TNBS or DSS provokes alveolar bone loss in concert with the autochthonous oral microbiota.