Salivary Leukocytes Research Articles

Salivary leukocyte esterase activity by SillHa is a risk indicator of periodontal disease

BackgroundThere is increasing evidence that diagnostic salivary tests measuring inflammatory biomarkers are being developed to assess inflammatory status for early detection, prevention, and progression of periodontal disease. Therefore, the aim of the present study was to investigate and identify the salivary biomarker that can predict the inflammatory status of periodontal disease.MethodsA total of 36 patients (28 women and 8 men) with an average age of 57 years were investigated. Unstimulated saliva was collected from the recruited subjects and analyzed using SillHa, a saliva-testing device that measures bacteria count, saliva buffer capacity, acidity, leukocyte esterase, protein, and ammonia. Periodontal parameters were then obtained by clinical examination and initial periodontal therapy was performed. Data obtained with SillHa were compared with clinical periodontal parameters at baseline, re-examination (three months from baseline), and final examination (six months from re-examination).ResultsLeukocyte esterase activity in saliva measured by SillHa; BOP and PCR measured by clinical examination showed a significant difference between baseline and final examination and between re-examination and final examination. Patients in the lower median group (group 1) had a significant difference in leukocyte esterase activity between baseline and final examination and re-examination and final examination. In addition, patients in Group 1 had significantly lower BOP between baseline and final examination. While patients in the higher median group (group 2) showed a modest decrease in leukocyte esterase activity, which was significant only between baseline and final examination, no significant changes were observed concerning BOP. Furthermore, the associated systemic disease was observed in 30% and 81.2% of group 1 and 2 patients, respectively.ConclusionThe results suggest that leukocyte esterase activity in saliva measured by SillHa could serve as a reliable diagnostic marker for monitoring inflammatory status in periodontal disease.

The role of saliva in maintaining oral health: Aid to diagnose systemic ailments-A review

Saliva is a complex secretion from three major and various minor salivary glands present in mouth. From clinical point of view saliva is an ideal diagnostic fluid because it is non-invasive fluid and can be easily be collected from the patients for biochemical analysis. The other significant characteristic pertinent to saliva is the handling procedure which is much simpler in comparison with serum or blood. There is plethora of studies which shows a positive correlation between blood biomarkers and salivary biomarkers in different clinical conditions of oral and systemic diseases. Saliva has an important role in maintain oral health through variety of its secretions like calcium, phosphorus, Bacteriocins, salivary leukocyte protease inhibitors different enzymes like ptyalin, amylase for carbohydrate and protein metabolism which maintains tooth integrity when there is demineralization and also acts as first line defense against various bacterial and viral infections. In 21 century endodontic there is a paradigm shift in research on the change in composition of the saliva and biomarkers in presence of periodontal / pulpal and other systemic diseases. Molecular methods for identification and quantification of salivary biomarkers like IL-6, IL-8, MMPs are available at chair side for rapid diagnosis of various inflammatory or infective etiology of oral diseases.

Suitability of salivary leucocytes to assess DNA repair ability in human biomonitoring studies by the challenge-comet assay

The challenge-comet assay is a simple but effective approach that provides a quantitative and functional determination of DNA repair ability, and allows to monitor the kinetics of repair process. Peripheral blood mononuclear cells (PBMC) are the cells most frequently employed in human biomonitoring studies using the challenge-comet assay, but having a validated alternative of non-invasive biomatrix would be highly convenient for certain population groups and circumstances. The objective of this study was to validate the use of salivary leucocytes in the challenge-comet assay. Leucocytes were isolated from saliva samples and challenged (either in fresh or after cryopreservation) with three genotoxic agents acting by different action mechanisms: bleomycin, methyl methanesulfonate, and ultraviolet radiation. Comet assay was performed just after treatment and at other three additional time points, in order to study repair kinetics. The results obtained demonstrated that saliva leucocytes were as suitable as PBMC for assessing DNA damage of different nature that was efficiently repaired over the evaluated time points, even after 5 months of cryopreservation (after a 24 h stimulation with PHA). Furthermore, a new parameter to determine the efficacy of the repair process, independent of the initial amount of damage induced, is proposed, and recommendations to perform the challenge-comet assay with salivary leucocytes depending on the type of DNA repair to be assessed are suggested. Validation studies are needed to verify whether the method is reproducible and results reliable and comparable among laboratories and studies.

VARIABLES RELATED TO GINGIVITIS IN CHILDREN WITH PYELONEPHRITIS AND NEPHROTIC SYNDROME

Purpose: The purpose of the research is to be investigated and assessed the significance of variables related to gingivitis in children with diagnosed pyelonephritis and nephrotic syndrome. Materials and Methods: The subject of the study is represented by participants in the age interval from 0 to 18 years. They are divided into two main groups, namely a group of 92 children with diagnosed pyelonephritis and 24 children with nephrotic syndrome. There is a group of 41 healthy children. Clinical, para-clinical, sociological and statistical methods are implemented in association with the study’s aim. The model of the research is based on an investigation of the significance of definite variables: gingival index GI Lõe-Silness, level of salivary leucocytes, salivary blood and salivary glucose. In addition to these indicators are recorded some patterns of environmental matter. Results: Among the participants with the diagnosis of nephrotic syndrome is recorded the maximal value of GI= 2.04. A similar maximal value of GI=2.00 corresponds to the group of children with pyelonephritis. In both of the subgroups of children suffering from renal disorders is registered the considerable maximal value of salivary blood equal to 250.00 RBC/µl, compared to the maximal value of 50.00 RBC /µl for healthy participants. It is ascertained that the variables of salivary blood and salivary leucocytes correspond to the condition of gingivitis in children with diagnosed pyelonephritis and nephrotic syndrome. Conclusion: A definite impingement of excretory system disorders upon gingiva, especially among children suffering from nephrotic syndrome, is ascertained.

Salivary Leucocytes as In Vitro Model to Evaluate Nanoparticle-Induced DNA Damage.

Metal oxide nanoparticles (NPs) have a wide variety of applications in many consumer products and biomedical practices. As a result, human exposure to these nanomaterials is highly frequent, becoming an issue of concern to public health. Recently, human salivary leucocytes have been proposed as an adequate non-invasive alternative to peripheral blood leucocytes to evaluate genotoxicity in vitro. The present study focused on proving the suitability of salivary leucocytes as a biomatrix in the comet assay for in vitro nanogenotoxicity studies, by testing some of the metal oxide NPs most frequently present in consumer products, namely, titanium dioxide (TiO2), zinc oxide (ZnO), and cerium dioxide (CeO2) NPs. Primary and oxidative DNA damage were evaluated by alkaline and hOGG1-modified comet assay, respectively. Any possible interference of the NPs with the methodological procedure or the hOGG1 activity was addressed before performing genotoxicity evaluation. Results obtained showed an increase of both primary and oxidative damage after NPs treatments. These data support the use of salivary leucocytes as a proper and sensitive biological sample for in vitro nanogenotoxicity studies, and contribute to increase the knowledge on the impact of metal oxide NPs on human health, reinforcing the need for a specific regulation of the nanomaterials use.

Early genotoxic effects from exposure to environmental pollutants young people from South Italy

Abstract Children and young people are particularly sensitive to the environmental pollution which is closely related to degenerative diseases. Several studies show that a genotoxic damage during young age can increase the risk of chronic diseases in adulthood. The young people are more vulnerable than adults to the environmental pollutants because they spend more time outdoors, they have immaturity of some organs and of the mechanisms involved in the cellular repair. In the present study, the early biological effects of exposure to a particularly polluted area of Southern Italy were evaluated in 200 children (6-10 year-old) and 100 young people (18-25 year-old). This area, worldwide known as Sarno basin, is characterized by strong anthropization, many agro-food processing industries, massive use of fertilizers and pesticides in agricultural practices and a strong river pollution. The comet assay was chosen because it reflects cumulative exposure to a variety of environmental factors and it was performed on salivary leukocytes in the children selected for the survey, while in the young people the DNA damage was evaluated in human peripheral blood lymphocytes. As in previous studies were not find significant differences between salivary leukocytes and blood lymphocytes we preferred the sampling of saliva for the children to avoid bloody practices. Furthermore, before cell sampling the children's parents were interviewed using an ad hoc questionnaire designed to gather additional information about exposure sources. A questionnaire was administered also to the young people to have more information on their lifestyle and some characteristic of the area of exposure (vehicular traffic and so on). The results showed a clear damage from exposure in the children differently from young people. Key messages Comet assay was performed in vitro on lymphocytes of 200 children (6-10 year-old) and 100 young people (18-25 year-old) exposed to a particularly polluted area of Southern Italy. An evident DNA damage was observed in lymphocytes coming from children; no genetic material alterations were observed in young people.

Genotoxic damage in workers exposed to pigment-grade titanium dioxide (TiO2) during paint production

Abstract TiO2 is broadly used in a wide range of applications such as photocatalysis, pigments and additives. Despite the promising commercial opportunities, different studies have shown that TiO2 can induce oxidative stress, inflammatory and genotoxic effects. The European Union has classified certain titanium dioxide powders and powder mixtures containing TiO2 as a suspected carcinogen (Category 2) via inhalation. Considering that the information about the potential adverse health effects of TiO2 in occupational environment are still scarce and controversial, the purpose of the study was to investigate the genotoxic effects and oxidative stress in workers exposed to TiO2 during paint production. Biomarkers of early effect (DNA damage and micronuclei) were evaluated in the buccal cells and salivary leucocytes of 30 workers (15 production workers and 15 controls). To collect information about personal details, occupational history, medication, smoking, diet, physical activity, a questionnaire was administered to all workers. Personal and area monitoring have been carried out to determine airborne inhalable and respirable fraction of TiO2; a NanoTracer was used to monitor the presence of ultrafine particles. In spite of the low mass concentration and a mild percentage of nanoparticles, filter deposited TiO2 was in sub-micron size, thus accounting for its respirability. Preliminary results on salivary leucocytes show a slight higher DNA damage in the exposed workers as compared to the controls. Whereas biomarker assessment is still in progress, these preliminary findings show that workers are exposed to low but measurable levels of TiO2 able to induce a mild genotoxic damage. A combined approach using both personal exposure assessment and biomonitoring can improve the risk assessment in occupational settings in which TiO2 is handled. Moreover, this also suggest to take precautionary measure during specific activities and operational phases thus decreasing the risks for worker. Key messages The results of the present study may promote effective risk management practices in occupational environments that uses TiO2. The results obtained suggest the introduction of activities and operational phases with lower risks for the worker.

Gingival health in children with mixed and permanent dentition with diagnosed pyelonephritis

Introduction: The share of hospitalizations of children with diagnosed urinary tract infections amounts to 4.6 % to 10 % of all hospitalized children. The interest towards fast and minimally invasive diagnostic tests has been exponentially increasing during the last decade. Saliva has been implemented as a biological medium for multi-aspect diagnostic investigations. The determination of the degree of gingival inflammation based on quantitative verification of salivary white blood cells correlates positively to data obtained by application of conventional parameters for evaluation of the periodontal status. The indicator of salivary blood has been investigated as a significant factor for the assessment of the inflammatory process of gingival tissue. Aim: The aim of the study is related to exploration, analysis and assessment of gingival health of children with mixed and permanent dentition with diagnosed pyelonephritis in comparison to children without common health disorders. Materials and Methods: The participants in the study are divided into two main groups - children with pyelonephritis and healthy controls. Each of the 2 groups includes two subgroups based on the type of dentition. A combination of methods has been implemented - clinical, paraclinical, and statistical methods. Conclusion: Among children suffering from pyelonephritis, the indicator of salivary blood is associated with the degree of gingival inflammation. The significance of the PLI and GI indicators in their role as predictors of destructive inflammatory processes of the gingival tissue is verified. The concentration of salivary leucocytes correlates to the intensity and activity of gingival inflammatory disorders.

Steeping from green and black robusta coffee beans increase viability of Peripheral Blood Mononuclear Cells (PBMC) and salivary leukocytes which is induced by streptococcus mutans

Steeping from green and black robusta coffee beans increase viability of Peripheral Blood Mononuclear Cells (PBMC) and salivary leukocytes which is induced by streptococcus mutans

The role of Kuniran (U. moluccensis) and Gurami (O. goramy) fish thorns and scales in increasing salivary leukocyte and monocyte cells viability against Streptococcus mutans

Background: Kuniran thorns and Gurami fish scales are rich in protein and minerals such as dentin believed to increase cell viability against Streptococcus mutans (S. mutans) that causes dental caries. These, in turn, can cause systemic diseases if left untreated. Purpose: This study aims to analyze the influence of Kuniran thorns and Gurami fishes scales on the viability of monocytes and salivary leukocytes against S. mutans. Methods: Monocytes and leukocytes salivary cells were placed on a microtiter plate and treated according to the nature of each group. This study comprised the following groups: control group: untreated; S. mutans group: induced by S. Mutans; Gurami thorn group: thorns + S. Mutans; Gurami scales group: scales + S. Mutans; Kuniran thorn group: thorns + S. Mutans; Kuniran scales group: scales + S. mutans. Viability analysis involved staining with Tripan Blue. Furthermore, the number of viable cells (white) was calculated under an inverted microscope at 200 times magnification from five fields of view. Data was analyzed by means of an ANOVA test followed by LSD test. Results: The ANOVA and LSD tests confirmed significant differences (0.01<P). Kuniran thorns and Gurami fish scales increased the viability of monocytes and salivary leukocytes, but not significantly. The content of flavonoids, amino acids, omega 3, omega 6 and antioxidants increased leukocyte metabolism, thereby increasing cell resistance to S. mutans infection. Conclusion: Kuniran thorns (U. moluccensis) and Gurami (O. goramy) fish scales increase the viability of salivary leukocyte and monocyte cells against Streptococcus mutans.

Expression of Salivary Secretory Leukocyte Protease Inhibitor in Smokers, Leukoplakia and OSCC

Expression of Salivary Secretory Leukocyte Protease Inhibitor in Smokers, Leukoplakia and OSCC

Primary DNA damage in salivary leukocytes of children exposed to air pollutants. MAPEC_LIFE project

Primary DNA damage in salivary leukocytes of children exposed to air pollutants. MAPEC_LIFE project

Exercise does not increase salivary lymphocytes, monocytes, or granulocytes, but does increase salivary lysozyme

ABSTRACTAn increase in salivary leukocytes may contribute to the exercise-induced increase in salivary antimicrobial proteins (AMPs). However, exercise-induced changes in salivary leukocytes have not been studied. The purpose of the study was to describe salivary leukocyte changes with exercise. Participants (n = 11, 20.3 ± 0.8 years, 57.2 ± 7.6 ml kg−1 min−1 peak oxygen uptake ((VO) ̇2peak), 11.1 ± 3.9% body fat) ran for 45 min at 75% of VO2peak. Stimulated saliva (12 mL) was collected pre- and immediately post exercise. Saliva was filtered through a 30 µm filter before analysis of leukocytes (CD45+), granulocytes (CD45+CD15+), monocytes (CD45+CD14+), T-cells (CD45+CD3+), and B-cells (CD45+CD20+) using flow cytometry. Saliva was analysed for Lysozyme (Lys) using ELISA. Exercise did not alter any leukocyte subset. The major constituent of leukocytes pre-exercise were granulocytes (57.9 ± 30.3% compared with monocytes: 5.1 ± 2.7%, T-cells: 17.1 ± 8.9%, B-cells: 12.1 ± 10.2%) (P < 0.05). In a subset of n = 6, Lys secretion rate increased after exercise (pre: 5,170 ± 5,215 ng/min; post: 7,639 ± 4,140 ng/min) (P < 0.05). Exercise does not result in increased granulocytes, but does increase Lys. Further, these data suggest that an increase in salivary leukocytes is not needed to increase Lys.

Comet assay in salivary leukocytes: evaluation of early effects of air pollution exposure in pre-school children.

Comet assay in salivary leukocytes: evaluation of early effects of air pollution exposure in pre-school children.

Primary and oxidative DNA damage in salivary leukocytes as a tool for the evaluation of air pollution early biological effects in children: current status of the MAPEC (Monitoring Air Pollution Effects on Children for supporting public health policy) study

Primary and oxidative DNA damage in salivary leukocytes as a tool for the evaluation of air pollution early biological effects in children: current status of the MAPEC (Monitoring Air Pollution Effects on Children for supporting public health policy) study

Flow cytometry-based analysis by Sysmex-UF1000i® is an alternative method in the assessment of periodontal inflammation

BackgroundGingivitis is a common inflammatory condition. We explored the value of flow cytometry of saliva in patients with periodontal inflammation. MethodsA cohort of 249 healthy adults (age range: 18–81y; 2.5th to 97.5th percentile: 19–66y) was investigated for caries, dental plaque and gingivitis. Saliva was analyzed using flow cytometry on a Sysmex UF-1000i®. ResultsSysmex UF-1000i® is capable to reproduce reliable measurements of cellular components in saliva. A statistically significant lower number of salivary bacteria was found in patients with gingivitis in comparison with healthy adults (p<0.0001). A significant difference in salivary leukocyte count was found between patients with different gingival index scores (p<0.0005). The gingivitis score was strongly dependent on the number of salivary leukocytes, the age of the patient and the degree of caries and dental plaque (r2=0.60, p<0.001). At a cut-off level of 103 leukocytes/μl saliva, an area under the curve of 0.82 was obtained with a sensitivity of 76% and a specificity of 78% in patients (>35y) with a gingivitis score of 3. ConclusionFlow cytometry is an alternative method to evaluate local inflammatory processes in the mouth with a sensitivity of 76% and a specificity of 78%.

Assessing caries, dental plaque and salivary flow in asthmatic adolescents using inhaled corticosteroids

BackgroundA number of studies have reported that inhaled corticosteroids may cause a greater incidence of caries, reduced salivary flow, changes in saliva composition and an increased frequency of dental plaque, probably through alterations in the oral microbiota. The objective was to compare the frequency of caries, dental plaque and non-stimulated salivary flow rate among asthmatic adolescents using inhaled corticosteroids and non-asthmatic adolescents, as well as the salivary biochemical parameters (pH and leucocytes) in both groups. MethodsThis research has a descriptive cross-sectional design to compare dental health of 40 asthmatics on inhaled corticosteroids and 40 non-asthmatic adolescents (median age 13 years). ResultsThe findings were a higher number of tooth surfaces affected by dental caries (median 4 versus 1.5), and more dental plaques (median 70.5 versus 60.7) among asthmatic adolescents. They also had a significantly higher frequency of salivary leucocytes. The non-stimulated salivary flow was similar in both groups. ConclusionsThe results suggest an association between the use of inhaled corticosteroids and an increased risk of dental caries and bacterial plaque, which calls for special attention of these patients by doctors and dental health professionals.

Role of salivary leukocyte protease inhibitor in periodontal disease progression

Context:Proteases play a major role in the tissue destruction involved in periodontal disease. It is known that the balance between proteases and their inhibitors is a major determinant in maintaining tissue integrity. The association between the proteases and periodontitis is well established, but not many studies have been carried out to know the role played by a protease inhibitor like salivary leukocyte protease inhibitor (SLPI) in periodontitis.Aim:The aim of the present study was to correlate SLPI with periodontitis.Settings and Design:Case-control study.Materials and Methods:Seventy-five clinically confirmed cases of periodontitis and 20 controls were included in the study. A detailed case history and periodontal index (PI) were recorded. Two milliliters of unstimulated saliva samples was obtained and subjected to quantification of SLPI leaves using SLPI in enzyme-linked immunosorbent assay (ELISA) kit. Based on the periodontal index score of the individuals, the cases and controls were divided into groups A, B and C, and the obtained SLPI levels were compared among the groups.Statistical Analysis:Mann-Whitney U test and correlation coefficient test.Results:The results showed that in the initial stages of periodontitis there is a tendency of SLPI levels to be raised. The SLPI levels were found to be reduced in the terminal stages of periodontitis.Conclusion:It appears that SLPI accumulates in the local environment, at least in the initial stages of the periodontal disease, probably to inhibit the action of increased elastic activity.

Quercetin and the Glucosides Inhibit Nitration of a Salivary Component 4-Hydroxyphenylacetic Acid Catalyzed by Salivary Polymorphonuclear Leukocytes

Mixed whole human saliva contains 4-hydroxyphenylacetic acid (HPA), nitrite and polymorphonuclear leukocytes. Salivary leukocytes nitrated HPA to 4-hydroxy-3-nitrophenylacetic acid in the presence of nitrite, and phorbol myristate acetate stimulated the nitration. Quercetin and the glucosides, which are found in the oral cavity after ingestion of quercetin-rich foods, inhibited the leukocyte-dependent nitration. The inhibition by quercetin and the glucosides was in part due to the flavonol-dependent scavenging of nitrogen dioxide which was formed by myeloperoxidase-dependent oxidation of nitrite. Salivary components, SCN and uric acid, also inhibited the nitration. The above re-sults suggest that quercetin can cooperate with SCN and uric acid to prevent nitration in the oral cavity.

Flow cytometry as a new method to quantify the cellular content of human saliva and its relation to gingivitis

Background: Determining the cellular content of saliva by means of conventional microscopy chamber counting is a very time-consuming and operator-sensitive procedure. This study concentrated on the use of flow cytometry to examine the cellular content of saliva. Erythrocytes, leukocytes, epithelial cells and bacteria were quantified and the results were compared with caries experience and the presence of gingivitis. Methods: 258 uncentrifuged vortexed paraffin-stimulated saliva samples (112 males and 146 females) were analyzed with the UF-100® flow cytometer. Salivary reference values were established for erythrocyte, leukocyte, epithelial cell and bacterial count. Caries experience (DMF) and the presence of gingivitis were recorded. Results: Caries experience or caries risk could not be assessed with flow cytometry. However, salivary flow cytometry may be useful in determining an individual's risk for gingivitis: a significant increase in salivary leukocytes was observed in individuals with gingivitis. At a cut-off level of 10 3 leukocytes μl −1 saliva, a sensitivity of 76% and a specificity of 45% was obtained. Other analytes were not significantly different between individuals with and without gingivitis. Conclusion: Flow cytometry of paraffin-stimulated human saliva seems a promising diagnostic or predictive tool and further investigations of diseases of the oro-pharyngeal loge, such as tonsillitis and periodontitis, should be carried out in the future.