muramidase and Dental-Plaque

It is becoming increasingly apparent that the traditional clinical criteria are inadequate for: determining active disease sites in periodontitis, monitoring quantitatively the response to therapy or measuring the degree of susceptibility to future breakdown. In an attempt to develop objective measures, a wide variety of studies have been undertaken using saliva, blood, plaque and gingival crevicular fluid (GCF) as the specimen source. Examination has included: specific bacteria and their products; host cells and their products (enzymatic and antibacterial, both immunologic and non-immunologic); products of tissue injury derived from local epithelial and connective tissues and bone. Although most of the work to date has failed to provide reliable aids to the clinician, refinements in techniques for sampling and the availability of more sophisticated analytic techniques give cause for optimism. Methods proposed for detection of disease-associated bacteria in subgingival plaque vary in their sensitivity and specificity. Dark field microscopy shows some correlation with existing disease; however, the limited specificity of this method imposes severe restrictions on its usefulness. Highly specific polyclonal and monoclonal antisera to suspected pathogens Bacteroides gingivalis and Actinobacillus actinomycetemcomitans have been developed and improved methods of identification of these microbes in plaque by ELISA immunofluorescence and flow cytometry are under development. With respect to the host response, a strong correlation between antibody patterns to specific bacteria and periodontal disease categories appears to be emerging. Although most studies have focused on serum antibody derived from peripheral blood, a shift to detection of local antibody response appears to be likely. Techniques of measurement that are exquisitely sensitive have been developed for detection of major immune recognition proteins such as antibody and complement in crevicular fluid. Research efforts attempting to correlate local antibody response to local disease activity are underway. Measurement of GCF flow rate, endotoxin, H2S, butyrate and a variety of enzymes (e.g., collagenase, arylsulfatase, B-glucuronidase) show good correlation with levels of gingivitis. In periodontitis, the most promising markers of tissue breakdown are prostaglandins of the E series, the enzymes collagenase and aspartate aminotransferase, sulfated glycosaminoglycans, osteoclastic activating factor and bon

Topics: Animals; Antibodies, Bacterial; Bacteria; Bacteriological Techniques; Bone Resorption; Butyrates; Butyric Acid; Complement System Proteins; Dental Plaque; Electrolytes; Endotoxins; Evaluation Studies as Topic; Humans; Hydrogen Sulfide; Intracellular Fluid; Lactoferrin; Leukocytes; Muramidase; Periodontal Diseases; Polyamines; Propionates

A variety of components provide salivary secretions with an array of potentially effective means of combating cariogenic challenges. These defense factors range from a laissez-faire mechanical cleansing to exquisitely controlled production of highly specific antibodies. In between the two extremes are antibacterial systems whose operating characteristics are only beginning to be understood. These systems are well worth our attention. They may be the key to our understanding of variations in individual susceptibility, and could provide valuable leads for development of anticaries agents.

Topics: Adsorption; Agglutination; Animals; Bacteria; Cell Adhesion; Cell Aggregation; Dental Caries; Dental Enamel Solubility; Dental Plaque; Humans; Hydrogen-Ion Concentration; Hydroxyapatites; Lactoferrin; Lactoperoxidase; Muramidase; Saliva; Secretory Rate

Chlorhexidine (CHX)-containing mouthrinses are recommended as adjuncts to mechanical oral hygiene. The problem associated with side effects, however, has stimulated the search for alternative antiplaque agents. The aim of this study was to investigate the plaque inhibitory effects of two mouthrinses containing amine fluoride/stannous fluoride (ASF) and antimicrobial host proteins (lactoperoxidase, lysozyme and lactoferrin; LLL), respectively.. The study was an observer-masked, randomized 4x4 Latin square cross-over design balanced for carryover effects, involving 12 healthy volunteers in a 4-day plaque regrowth model. A 0.12% CHX mouthrinse and a saline solution served as positive and negative controls, respectively. On day 1, subjects received professional prophylaxis, suspended oral hygiene measures, and commenced rinsing with their allocated rinses. On day 5, subjects were scored for disclosed plaque.. The ASF rinse showed a significant inhibition of plaque regrowth in comparison to both saline and LLL solutions, but the lowest plaque indices were obtained with the CHX formulation (P<0.01). There were no significant differences between LLL rinse and saline (P>0.05). Such pattern of efficacy was the same in anterior and posterior teeth and in vestibular and lingual surfaces as well, with the exception of the lingual anterior surfaces. In these sites, differences between the CHX and ASF rinses were not significant (P>0.05). A significantly higher prevalence of side effects was found in subjects using the CHX product (P<0.0042).. Although the effect on plaque regrowth observed with 0.12% CHX rinsing was superior to that with ASF, the ASF rinse was not associated with side effects. These findings, together with those from long-term trials, suggest that the ASF rinse may represent an effective alternative to CHX rinse as an adjunct to oral hygiene. On the contrary, the LLL rinse did not significantly inhibit plaque regrowth.

Topics: Adult; Analysis of Variance; Chlorhexidine; Cross-Over Studies; Dental Plaque; Drug Combinations; Female; Fluorides, Topical; Humans; Lactoferrin; Lactoperoxidase; Male; Mouthwashes; Muramidase; Observer Variation; Patient Compliance; Single-Blind Method; Statistics, Nonparametric; Tin Fluorides

Student nurses (aged 20-26 years) were assigned to two groups that were matched for plaque levels and gingival health. For six months, one group used a standard fluoride dentifrice while the other used an identical dentifrice to which zinc citrate (1%, w/w) and Triclosan (0.2%, w/w) had been added. Levels of natural antimicrobial proteins (lysozyme, lactoferrin, salivary peroxidase and Immunoglobulin A) in whole, unstimulated saliva taken from the students at the start and on completion of the six months were measured. No statistically significant differences were found in the levels of antimicrobial proteins in saliva between the test and placebo groups.

Topics: Adult; Anti-Infective Agents; Citrates; Citric Acid; Dental Plaque; Dentifrices; Double-Blind Method; Female; Humans; Immunoglobulin A, Secretory; Lactoferrin; Muramidase; Peroxidases; Placebos; Saliva; Salivary Proteins and Peptides; Time Factors; Triclosan

Although the pathogenesis of periodontal lesions has not been sufficiently studied, recent studies show that plaque formation and host immune response are important factors. The purpose of this study was improving efficiency of plaque-induced gingivitis treatment in children with immunological correction of saliva by administration of polyvitamins and lysozyme tablets. We have examined 60 12-year-old children diagnosed with plaque-induced gingivitis and divided them into the main and control groups consisted of 30 children in each accordingly. The children of both groups were treated by sanitation and professional oral hygiene. The children of the main group besides were prescribed with multivitamins complex "Supervit" and tablets "Lizak". The efficiency of the introduced complex we have assessed by contain of immunoglobulins A (IgA), immunoglobulins G (IgG), secretory immunoglobulin A (s-IgA), interleukin 1β (IL-1β), interleukin 4 (IL-4) and lysozyme in saliva. After 6 month the treatment children from the main group showed a decline in concentration of IL-1β by 30,06 % (р<0,01), IgA by 33,34 %, IgG by 12,5 % (р<0,05). The present data support the high efficiency of the introduced treatment that has been proved by positive progress of immunological indexes in saliva taken within six and 12 month since the research.

Topics: Child; Dental Plaque; Female; Gingivitis; Humans; Immunoglobulin A, Secretory; Male; Muramidase; Saliva; Tablets; Treatment Outcome; Vitamins

The scientific community has definitely demonstrated the importance of the use of mouthwash in daily oral hygiene. In our pilot study, we tested the effectiveness of a novel mouth rinse containing sea salt, xylitol, and lysozyme.

Topics: Adolescent; Dental Plaque; Humans; Mouthwashes; Muramidase; Oral Hygiene; Pilot Projects; Sodium Chloride; Streptococcus mutans; Xylitol

In the Japan Ground Self-Defense Force (JGSDF), personnel periodically perform intensive training that mimics the conditions seen in battle and during natural disasters. Military training involves intensive, stressful conditions, and changes in immune responses have been found in personnel following training. Good oral condition is important for military personnel to fulfill their duties; however, they have difficulty performing daily oral care under training conditions. In this study, we investigated the impact of a 7-day field training on the oral health status of JGSDF personnel by comparing their oral condition before and just after training.. The participants were 59 male and 3 female JGSDF personnel undergoing a 7-day field training. All personnel provided informed written consent to participate, and this study was approved by the ethics committee of the Kagoshima University Graduate School of Medical and Dental Sciences. Oral health behaviors before and during the training period were surveyed using a self-administered questionnaire. Dental caries was assessed before training in terms of decayed, missing and filled teeth (DMFT), and periodontal condition was examined before and immediately after training using the community periodontal index (CPI). The presence of eight species of bacteria in dental plaque, including commensal streptococci that are early colonizers on the tooth surface, cariogenic bacteria, and periodontopathic bacteria, was determined using real-time polymerase chain reaction. We also assessed antibacterial factors and a stress marker in saliva samples. Sample collection was performed before and just after training. In addition to difference analysis between groups, logistic regression analysis was performed to examine the association between each health behavior and periodontal deterioration.. The frequency of toothbrushing decreased, and snacking increased during the training period. Thirty-five personnel (56.5%) showed an increase in individual CPI code, and 57 personnel (91.9%) showed deterioration in the CPI code in 1 or more sextants after training (Figure 1). Toothbrushing frequency was significantly associated with CPI deterioration; the odds ratio in subjects who did not brush their teeth was 7.51 compared to those who brushed at least once during the training period. Severe periodontal deterioration was observed in the high-DMFT group (Figure 2), and toothbrushing frequency during the training period decreased more in this group compared to the low-DMFT group. The percentages of Streptococcus sanguinis and Streptococcus gordonii increased significantly after the training period suggesting dental plaque maturation, and an increase in S. sanguinis was associated with toothbrushing frequency. The lactoferrin concentration in saliva increased significantly after training.. We demonstrated periodontal deterioration in JGSDF personnel after a 7-day training. Behavioral changes, especially discontinuation of regular toothbrushing, fostered dental plaque maturation, resulting in inflammatory changes in participants' periodontal condition. The results indicate the importance of performing toothbrushing at least once over a 7-day training period for prevention of periodontal deterioration. The regimen could be applicable to evacuees from disasters because they are under conditions of stress that may limit oral hygiene activity.

Topics: Adult; alpha-Amylases; Dental Caries; Dental Plaque; Enzyme-Linked Immunosorbent Assay; Female; Health Behavior; Humans; Japan; Logistic Models; Male; Middle Aged; Military Personnel; Muramidase; Oral Health; Real-Time Polymerase Chain Reaction; Statistics, Nonparametric; Surveys and Questionnaires; Teaching; Workforce

Intestinal lactobacilli have been successfully used as probiotics to treat gastrointestinal disorders, but only limited data are available for the probiotic properties of oral lactobacilli to combat oral diseases. We aimed to characterize oral lactobacilli for their potential probiotic properties according to the international guidelines for the evaluation of probiotics, and to select potential probiotic strains for oral health.. The study included 67 salivary and subgingival lactobacilli of 10 species, isolated from healthy humans. All strains were identified using amplified ribosomal DNA restriction analysis, tested for antimicrobial activity against oral pathogens, tolerance of low pH and bile content. Thereafter, the lysozyme tolerance and antibiotic susceptibility of 22 potential probiotic strains were assessed.. The majority of strains suppressed the growth of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, and Streptococcus mutans, but none inhibited Candida albicans. The lowest pH tolerated by lactobacilli following 4 h of incubation was pH 2.5, but none of the strains grew at this pH. All strains tolerated a high concentration of lysozyme (10 mg/ml) and half of the strains tolerated a high concentration of human bile [5% volume/volume (V/V)]. Four Lactobacillus plantarum and two Lactobacillus oris strains expressed resistance to tetracycline and/or doxycycline.. Strains of L. plantarum, Lactobacillus paracasei, Lactobacillus salivarius, and Lactobacillus rhamnosus expressed both high antimicrobial activity and high tolerance of environmental stress. The absence of transferable antibiotic-resistance genes in L. plantarum strains remains to be confirmed. These results suggest a potential for oral lactobacilli to be used as probiotics for oral health.

Topics: Anti-Bacterial Agents; Bile; Dental Plaque; Drug Resistance, Bacterial; Humans; Hydrogen-Ion Concentration; Lactobacillus; Muramidase; Probiotics; Saliva

To investigate bacterial populations in subgingival and supragingival plaque samples of patients with inflammatory periodontal diseases and activities of the lysosomal enzymes--lysozyme, alkaline phosphatase, and beta-glucuronidase--in peripheral venous blood, in gingival crevicular fluid, and mixed nonstimulated saliva.. The study included 60 patients with inflammatory periodontal diseases without any internal pathology and 24 periodontally healthy subjects. Molecular genetic assay (Micro-IDent plus, Germany) for complex identification of additional six periodontopathic bacteria was applied. The activity of lysozyme was determined turbidimetrically, the activity of alkaline phosphatase--spectrophotometrically with a "Monarch" biochemical analyzer, the activity beta-glucuronidase--according to the method described by Mead et al. and modified by Strachunskii.. A statistically significant association between clinical and bacteriological data was found in the following cases: gingival bleeding in the presence of Eubacterium nodatum, Eikenella corrodens, Capnocytophaga spp. (P<0.01); pathological periodontal pockets in the presence of Peptostreptococcus micros (alpha< or =0.05 and beta< or =0.2), Fusobacterium nucleatum (alpha< or =0.05 and beta< or =0.2), Campylobacter rectus (alpha< or =0.05 and beta< or =0.2), and Capnocytophaga spp. (P<0.05); and satisfactory oral hygiene in the presence of all microorganisms investigated (P<0.05). The activity of lysozyme in gingival crevicular fluid and mixed nonstimulated saliva indicates the severity of periodontal inflammation. Based on clinical data, in assessing the amount of lysozyme in mixed nonstimulated saliva, sensitivity and specificity of 100% was found. Increased activities of lysozyme, alkaline phosphatase, and beta-glucuronidase were found in peripheral venous blood of patients with inflammatory periodontal disease as compared to control group.. The main principles of the treatment of periodontal inflammatory diseases should be based on microorganism elimination, creation of individual treatment means affecting microflora in the mouth and immune system of macroorganisms.

Topics: Adult; Alkaline Phosphatase; Campylobacter rectus; Capnocytophaga; Chi-Square Distribution; Dental Plaque; Eikenella corrodens; Eubacterium; Fusobacterium nucleatum; Gingival Hemorrhage; Glucuronidase; Humans; Muramidase; Nephelometry and Turbidimetry; Peptostreptococcus; Periodontal Index; Periodontitis; Risk Factors; Saliva; Sensitivity and Specificity; Spectrophotometry

Extracellular polysaccharides (PS) synthesized by oral bacteria constitute one of their major virulence factors. The PS, synthesized from sucrose, facilitate adhesion and colonization by bacteria to tooth surfaces. The study was designed to test the effect of in situ production of extracellular PS by Streptococcus mutans on the bactericidal activity of human neutrophils. These effects were tested on bacteria pre-exposed to sucrose (PS-positive Strep. mutans) and compared to bacteria not exposed to sucrose (PS-negative Strep. mutans). The interactions between neutrophils and Strep. mutans were tested in suspension and on bacteria in an experimental model of dental plaque. Viability of Strep. mutans was measured by [3H]-thymidine incorporation into the bacteria. Degranulation of neutrophils was evaluated by the release of lysozyme, and the production of reactive oxygen products was measured by chemiluminescence. When neutrophils were incubated with suspended bacteria, the viability of PS-negative Strep. mutans was 20% of that of bacteria not incubated with neutrophils (control), while the viability of PS-positive Strep. mutans was 40% of the control. In the experimental dental-plaque model, 50% of the PS-negative Strep. mutans were killed by neutrophils while the viability of PS-positive Strep. mutans was not different than of the control. Degranulation of neutrophils was not affected by the presence of extracellular PS of Strep. mutans. Artificial stimulation of neutrophils with phorbol myristate acetate also did not enhance the bactericidal effect of neutrophils on PS-positive Strep. mutans. However, PS-positive Strep. mutans elicited oxygen-reactive products from neutrophils, 2-fold less than with PS-negative Strep. mutans. The results indicate that in situ production of bacterial extracellular polysaccharides might be a major virulence factor of Strep. mutans, enabling PS-positive Strep. mutans in the dental-plaque biofilm to evade killing by human neutrophils.

Topics: Anti-Bacterial Agents; Biofilms; Carcinogens; Cell Degranulation; Dental Plaque; Humans; Indicators and Reagents; Luminescent Measurements; Luminol; Lymphocyte Activation; Muramidase; Neutrophils; Polysaccharides, Bacterial; Radiopharmaceuticals; Reactive Oxygen Species; Streptococcus mutans; Sucrose; Tetradecanoylphorbol Acetate; Thymidine; Tritium; Virulence

Cell suspensions of Streptococcus sobrinus can be aggregated by high molecular-weight alpha-1,6 glucans. The aggregation depends on the fidelity of a cell wall-bound, glucan-binding lectin (GBL). It is thought that the lectin may play a part in the sucrose-dependent accretion of streptococci in dental plaques. Results showed that the anionic detergent, sodium dodecyl sulphate (SDS) was a potent inhibitor of the lectin. When cells were incubated in SDS and washed to remove the detergent, lectin activity was diminished. Following incubation of the cells with SDS in the presence of glucan T-10, a low molecular-weight alpha-1,6 glucan, the loss of activity was less pronounced, suggesting that the glucan afforded partial protection against denaturation. Urea and guanidine hydrochloride were good inhibitors of the lectin, but, unlike SDS, were not able to inhibit it irreversibly, except at very high concentrations. Cationic detergents, such as cetylpyridinium bromide (and chloride), also irreversibly denatured the streptococcal lectin, but were not as effective as SDS in abolishing its activity. The results suggest that alpha-1,6 glucan stabilizes the GBL of S. sobrinus, rendering it more resistant to the effect of chaotropes. This may be one reason why dental plaques tend to resist detergents in dentrifices.

Topics: Anti-Bacterial Agents; Bacterial Proteins; Cell Wall; Cetylpyridinium; Dental Plaque; Dentifrices; Detergents; Endopeptidases; Glucans; Guanidine; Humans; Lectins; Ligands; Molecular Weight; Muramidase; Octoxynol; Sodium Dodecyl Sulfate; Streptococcus sobrinus; Sucrose; Urea

Samples of whole saliva and dental plaque were collected from initially 10-year old subjects who participated in a 40-month cohort study investigating the effect of chewing gum usage on caries rates. The subjects represented nine cohorts of which one did not receive gum, while in eight cohorts the subjects received gum containing either xylitol, sorbitol, their mixtures, or sucrose as bulk sweeteners, the maximum sweetener consumption in the form of gums being up to 10.7 g/day, used in 3-5 daily chewing episodes. Gum usage had no significant effect on the levels of salivary protein, IgA, alpha-amylase, peroxidase, lysozyme, SCN and buffer capacity. At the endpoint, the group that received 100% xylitol pellet-shaped gum five times/day, had significantly lower levels of sucrase (p <0.05) and free sialic acid (p < 0.001) in whole saliva than at baseline. This group showed significantly (p <0.05) smaller plaque index scores at two cross-sectional measurements, and exhibited the lowest log(10) counts of salivary lactobacilli at endpoint than most other groups. The salivary levels of peptidase(s) (oligopeptidase B-like enzymes) hydrolyzing N-alpha-benzoyl-DL-arginyl-p-nitroaniline were significantly (p<0.05) or almost significantly lower in groups which received 100% xylitol pellet gums. All groups exhibited obviously an aging-related increase of salivary mutans streptococcus scores, except the above xylitol group in which the mean scores did not change.

Topics: alpha-Amylases; Aniline Compounds; Buffers; Cariostatic Agents; Chewing Gum; Child; Cohort Studies; Colony Count, Microbial; Cross-Sectional Studies; Dental Caries; Dental Plaque; Dental Plaque Index; Humans; Immunoglobulin A, Secretory; Lactobacillus; Muramidase; Peptide Hydrolases; Peroxidases; Saliva; Salivary Proteins and Peptides; Sialic Acids; Sorbitol; Streptococcus mutans; Sucrase; Sucrose; Sweetening Agents; Thiocyanates; Xylitol

A series of rat caries experiments was carried out to test the relative cariogenic potential and to identify the major carcinogenic elements of 22 popular snack foods. Parameters that were measured included rat caries, number of cariogenic bacteria in plaque, salivary parameters including flow rate, buffering capacity, total protein, lysozyme and amylase content, and composition of test foods including protein, fat, phosphorus, calcium, fluoride, galactose, glucose, total reducing sugar, sucrose, and starch. Many interesting relationships were observed between food components, numbers of plaque bacteria, salivary components, and specific types of carious lesions. Protein, fat, and phosphorus in foods were all associated with inhibition of both sulcal and buccolingual (smooth-surface) caries. Food fluoride was associated with inhibition of buccolingual caries, whereas calcium was related to inhibition of sulcal caries. Glucose, reducing sugar, and sucrose in foods were all related to promotion of both sulcal and smooth-surface caries. The numbers of Streptococcus sobrinus in plaque were associated with promotion of smooth-surface caries only, whereas lactobacilli, non-mutans bacteria, and total viable flora were related to promotion of both smooth-surface and sulcal caries. The salivary flow rate was associated with inhibition of both buccolingual and sulcal caries. Salivary buffering capacity (at pH 7) and salivary lysozyme delivery were associated with inhibition of number and severity of sulcal caries, while the salivary amylase content was related to the promotion of the number of sulcal lesions.

Topics: Amylases; Animals; Bacteria; Bacterial Physiological Phenomena; Buffers; Calcium; Cariogenic Agents; Colony Count, Microbial; Dental Caries; Dental Plaque; Dietary Carbohydrates; Dietary Fats; Dietary Proteins; Food; Food Analysis; Lactobacillus; Muramidase; Phosphorus; Rats; Rats, Sprague-Dawley; Saliva; Salivary Proteins and Peptides; Secretory Rate; Streptococcus sobrinus

The occurrence of opportunistic pathogens and the concentration of some antimicrobial factors in the oral cavity of both acute and chronic leukaemia patients were studied. Enterobacteria were isolated from both dental plaque and crevicular fluid of all the groups examined, with few differences between healthy volunteers and leukaemic subjects; yeasts were found in both the crevicular fluid and the dental plaque samples of chronic leukaemia patients, but only in the plaque of healthy volunteers. Acute leukaemia patients did not have yeasts, but they were the only group colonized by the pseudomonads. IgA and N-acetyl-D-glucosaminidase (NAGase) significantly increased in chronic leukaemia patients compared with controls, whilst lysozyme seemed to present no marked differences for all groups. A further increase in NAGase concentration and an elevation in lysozyme content of saliva was observed for chronic leukaemia patients with severe periodontal lesions.

Topics: Acetylglucosaminidase; Dental Plaque; Enterobacteriaceae; Gingival Crevicular Fluid; Humans; Immunoglobulin A; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Leukemia, Myeloid, Acute; Mouth; Muramidase; Opportunistic Infections; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Pseudomonas; Saliva; Yeasts

Many studies have attempted to relate levels of antimicrobial proteins in saliva to oral health; results have been inconsistent, and one reason might be inconsistency of measures of plaque and saliva within subjects. This study investigated associations between plaque and salivary variables in longitudinal data. Whole saliva, and 8-h plaque pooled from buccal first permanent molars, was obtained from 32 dental students on Tuesdays from 3:00-6:00 p.m. over 4 weeks. Salivary flow rate was determined, and samples were assayed for lysozyme, lactoferrin, total peroxidase, myeloperoxidase, OSCN-, sIgA and total protein. Colonies on mitis-salivarius agar were assigned to Streptococcus sanguis, Strep. mutans or Strep. salivarius on the basis of morphology, supplemented by the API Rapid Strep identification system. Consistency of values within subjects across weeks was evaluated by repeat-measures analysis of variance and intraclass correlation; data were transformed to reduce skewness. Pearson's r was used to determine associations between plaque and salivary variables. Significant intraclass correlations (alpha = 0.05) were found for all salivary variables except myeloperoxidase, and for total flora, total streptococci, Strep. sanguis and Strep. sanguis as a proportion of total streptococci. Significant Pearson correlations with Strep. sanguis as a proportion of total streptococci were found for total protein (r = -0.24), sIgA (r = -0.22), lactoferrin (r = -0.19) and OSCN- (r = 0.20) when data from all weeks were pooled (n = 128). Strep. sanguis proportions tended to be low in subjects with high values for salivary proteins; the range of proportions was wider in subjects with low salivary values. These findings suggest some consistency of weekly values for many plaque and salivary variables. They also support previous cross-sectional data which suggested that salivary antimicrobial proteins may have some effect on plaque composition. This study was made before recent revisions in streptococcal taxonomy, and further research is needed to clarify interactions of salivary proteins with currently defined species.

Topics: Bacteria; Circadian Rhythm; Colony Count, Microbial; Cross-Sectional Studies; Dental Plaque; Female; Humans; Immunoglobulin A, Secretory; Lactoferrin; Longitudinal Studies; Male; Muramidase; Peroxidase; Peroxidases; Saliva; Salivary Proteins and Peptides; Secretory Rate; Streptococcus; Streptococcus mutans; Streptococcus sanguis; Thiocyanates

Resting and stimulated whole saliva was collected from 94 children aged 12-14 years and analyzed for thiocyanate, hypothiocyanite, 'free' and 'total' lysozyme, lactoferrin and secretory IgA. Clinical assessments of the amounts of plaque and gingival inflammation were made, and plaque was collected for determination of dry weight. An inverse relationship was observed between salivary thiocyanate concentrations in both resting and stimulated saliva and the amounts of plaque and gingival inflammation in these subjects (p < 0.05). Lactoferrin concentration in stimulated saliva was directly related to the amounts of plaque and gingivitis (p < 0.05). 'Total' lysozyme concentration in stimulated saliva was directly related to the amount of plaque (p < 0.05), and the 'free' lysozyme concentration in the same saliva was directly related to the amount of gingivitis (p < 0.05). The direct relationship observed between clinical measurements and both lysozyme and lactoferrin concentrations in saliva may have been due to contributions from gingival crevicular fluid. Cluster analysis identified three groups of subjects with different profiles in resting whole saliva, and in particular with different levels of secretory IgA. A statistically significant difference was observed in the quantity of plaque collected from subjects in two of these groups (p < 0.05). These results from cluster analysis using resting whole saliva from children confirmed the findings of a previous study with young adults.

Topics: Adolescent; Child; Cluster Analysis; Dental Plaque; Gingivitis; Humans; Immunoglobulin A, Secretory; Lactoferrin; Multivariate Analysis; Muramidase; Saliva; Salivary Proteins and Peptides; Secretory Rate; Thiocyanates

Samples of resting and stimulated whole saliva and stimulated parotid saliva were collected from 40 young adults. One week later, after 48 h on a standardized diet without oral hygiene, all available plaque was collected for dry weighing. An inverse relationship was found between the 'free' lysozyme concentration in stimulated parotid saliva and plaque dry weight (r = -0.46, p less than 0.01). There were no other statistically significant correlation coefficients between concentrations of individual salivary constituents and plaque dry weight. However, cluster analysis of constituents in resting whole saliva revealed three groups of subjects with different salivary profiles, and in particular with different concentrations of both IgA and hypothiocyanite. Subsequent analysis revealed differences in plaque dry weight between the groups, demonstrating the potential biological significance of cluster membership based on salivary factors.

Topics: Adult; Anti-Infective Agents; Dental Plaque; Female; Humans; Immunoglobulin A, Secretory; Lactoferrin; Male; Muramidase; Parotid Gland; Regression Analysis; Saliva; Salivary Proteins and Peptides; Secretory Rate; Thiocyanates; Time Factors

Saliva antimicrobial proteins may interact in a common system to influence the oral ecology. Clinical studies of antimicrobial protein action thus may require a multiple-protein approach. Multivariate statistical methods have been used to describe possible patterns of interaction for lysozyme, lactoferrin, salivary peroxidase and secretory IgA in stimulated parotid saliva. However, oral microbes are most likely to encounter antimicrobial proteins in mixed resting saliva. Relationships among levels of lysozyme, lactoferrin, salivary peroxidase, and secretory IgA therefore were investigated in whole saliva from 216 subjects, and an attempt made to relate interperson variation in those proteins to differences in health and status, and dental plaque accumulation and composition. All proteins were significantly (alpha = 0.05) correlated with each other (r = 0.38-0.52, p less than 0.001). There was only one axis of common variation among proteins, and that axis was significantly correlated (p less than 0.001) with total protein (r = 0.84) and flow rate (r = -0.56). That pattern deviated from the previous finding that proteins of acinar origin tended to vary independently from proteins of ductal origin in stimulated parotid saliva. The difference between parotid and whole saliva may reflect constitutive secretion of all proteins at low levels of stimulation. Common variation of unstimulated saliva proteins suggests that antimicrobial actions can be compared in subjects at population extremes. There were no significant associations between antimicrobial proteins in whole saliva and measures of health status or plaque accumulation. However, the proportions of Streptococcus sanguis were significantly correlated with lysozyme (r = -0.26), lactoferrin (r = -0.34), peroxidase (r = -0.30), total protein (r = -0.37), flow rate (r = 0.24) and principal-components scores (r = -0.33) in a subset of subjects (n = 85) where commercial biochemical tests were used to supplement species identification by colony morphology. Those findings may indicate that saliva antimicrobial proteins can affect the composition of dental plaque.

Topics: Adolescent; Adult; Bacteria; Bacterial Physiological Phenomena; Dental Plaque; Dental Plaque Index; Female; Health Status; Humans; Immunoglobulin A, Secretory; Lactoferrin; Male; Muramidase; Oral Health; Peroxidases; Saliva; Salivary Proteins and Peptides; Secretory Rate; Streptococcus; Streptococcus mutans; Streptococcus sanguis; Thiocyanates

Human polymorphonuclear leukocytes (PMN) were able to generate and release superoxide anions upon stimulation of Streptococcus mutans, Bacteroides gingivalis, and Capnocytophaga ochracea when incubated aerobically but not when incubated anaerobically. Lysozyme release and phagocytosis by PMN were independent of oxygen, and no difference between PMN incubated aerobically or anaerobically was observed (PMN stimulated by B. gingivalis released 7.6% total lysozyme when aerobic and 6.9% when anaerobic). There were variations in lysozyme release and phagocytosis for the three organisms, particularly for phagocytosis. B. gingivalis and C. ochracea yielded lower phagocytosis values than those for S. mutans, e.g., at 1 h 67% of the initial inoculum of S. mutans was phagocytosed (versus only 40% for B. gingivalis). Transmission electron microscopy showed that both S. mutans and B. gingivalis were internalized into classical phagolysosomes. In contrast, C. ochracea showed two forms of internalization; C. ochracea either formed a classical phagolysosome or was tightly bound in the cytoplasm with no surrounding cell membrane. Intracellular killing of S. mutans and C. ochracea was unaffected by anaerobiosis, but killing of C. ochracea was much lower than that of S. mutans (1 x 10(7) to 2 x 10(7) bacteria killed compared with 5.1 x 10(7) bacteria killed at 6 h). In contrast, a greater number of B. gingivalis was killed in the presence of oxygen (5.3 x 10(7) bacteria were killed when aerobically incubated and 1.9 x 10(7) bacteria were killed when anaerobically incubated). These results suggest that the ability to survive anaerobically may enable some bacteria to evade PMN killing; however, abnormal phagocytosis may represent a more efficient way to evade both oxygen-dependent and -independent killing mechanisms, leading to enhanced virulence of the organism.

Topics: Aerobiosis; Anaerobiosis; Bacteroides; Capnocytophaga; Dental Plaque; Humans; Muramidase; Neutrophils; Opsonin Proteins; Oxygen; Phagocytosis; Streptococcus mutans; Superoxides

Topics: Adolescent; Adult; Dental Plaque; Female; Humans; Male; Middle Aged; Muramidase

Topics: Dental Plaque; Humans; Immunoglobulin A, Secretory; Muramidase; Salivary Proteins and Peptides

The effect of lysozyme-inactivation on L(+)-lactic acid (LA) production in dental plaque suspensions was evaluated. From 10 children 24-h plaque was collected and lysozyme activity inhibited by addition of goat antiserum to human lysozyme. Acid production was stimulated by addition of glucose. The results showed significantly increased LA levels (50-150%) in lysozyme-inactivated plaque suspensions from 8 of the subjects compared to untreated controls. The increase in acid production activity was not related to plaque lysozyme levels. The findings indicate that the presence of lysozyme may be limiting on acid production in the early dental plaque.

Topics: Adolescent; Albumins; Child; Dental Plaque; Glucose; Humans; Immune Sera; Isomerism; Lactates; Lactic Acid; Muramidase

The contribution of proteins from saliva and gingival crevicular fluid (GCF) to plaque was determined by comparing extracts of supragingival plaque of unknown age formed on normal teeth with plaque formed on artificial teeth in complete or partial dentures where crevicular fluid is absent. There was a total absence of albumin and a virtual absence of IgG from denture plaque samples, confirming their crevicular origin. The concentration of lactoferrin was much higher than that of lysozyme in all supragingival but not in the denture plaque samples, suggesting that GCF provided more lactoferrin than lysozyme to plaque. Amylase was a component in both denture and supragingival plaque, present in similar amounts in both deposits. Cysteine-containing phosphoproteins from saliva were in low concentration but present in all plaque samples; proline-rich proteins were virtually absent, reflecting the high vulnerability to proteolysis of these proteins. Salivary proteins in plaque extracts do not correspond with their relative concentrations in saliva.

Topics: Adult; Albumins; Amylases; Dental Plaque; Dentures; Gingiva; Humans; Immunoglobulin G; Lactoferrin; Middle Aged; Muramidase; Salivary Proteins and Peptides

Compared with anion-activated cell lysis of oral bacteria damaged with either lysozyme or trypsin, cells which were treated with both of these enzymes showed a far greater degree of lysis. This was true regardless of whether turbidimetric, DNA release, or electron microscopic assays were used to monitor the lytic process. At an acidic pH of 5.2 and an NaHCO3 concentration of 100 mM, the kinetics of lysis for two different serotype c strains of Streptococcus mutans were similar. At 0 to 100 mM bicarbonate, however, differences in the lytic susceptibilities of the two strains were evident. At pH 5.2, NaHCO3, but not NaSCN, NaCl, or NaF, was effective in promoting cell lysis of the oral bacteria. At apparent sublytic concentrations of NaHCO3, lysis was achieved by adding appropriate concentrations of NaSCN, NaCl, or NaF to the lysozyme-protease-damaged cells. In in vivo studies, hamsters given a combination of NaHCO3, NaCl, and NaSCN were found to have significantly reduced levels of S. mutans on their molar teeth compared with that found in controls or animals exposed to any one of the salts alone or to a combination of chloride and thiocyanate only. The results suggest that bicarbonate is an essential anion which, together with the other major salivary inorganic monovalent anions, plays an active role in the lysis and ultimate elimination of cariogenic bacteria.

Topics: Actinomyces; Animals; Bicarbonates; Cricetinae; Dental Plaque; Fluorides; Hydrogen-Ion Concentration; Lacticaseibacillus casei; Muramidase; Streptococcus mutans; Thiocyanates; Trypsin

Topics: Adolescent; Adult; Dental Caries; Dental Plaque; Female; Gingivitis; Humans; Immunoglobulin A, Secretory; Immunoglobulin G; Male; Muramidase; Saliva

Veillonella alcalescens subsp. dispar was grown in a synthetic medium containing either radiolabeled thymidine or uridine to monitor cell lysis by assay of the release of deoxyribonucleic acid or ribonucleic acid (RNA), respectively. Biochemical analyses demonstrated that, although human or hen egg white lysozymes alone did not release deoxyribonucleic acid or RNA, the nucleic acids were liberated in equal amounts from lysozyme-treated cells by the addition of low concentrations of the sodium salts of HCO-3, SCN-, Cl-, and F-, RNA release was dependent on enzyme and anion concentration. Human lysozyme was more potent than hen egg white lysozyme, and bicarbonate was the most effective anion in promoting bacteriolysis. Surprisingly, ultrastructural analyses differed from biochemical results. Lysozyme alone caused lysis in approximately 40% of the cell population. Detailed ultrastructural examination revealed aggregated cytoplasmic components which appeared as small clumps, explaining why nucleic acids were not measurable in the biochemical assays. In reaction mixtures containing lysozyme plus inorganic salts, electron microscopy results were compatible with biochemical data. Ultrastructural studies demonstrated that the addition of inorganic salts to lysozyme-treated cells resulted in the solubilization of the protoplasmic aggregates of lysed cells, presumably freeing the complexed RNA, and in the rapid lysis of the remaining cells (approximately 60%). These data suggest that electron microscopy must be used in conjunction with biochemical assays to assess lytic damage of bacterial cells.

Topics: Anions; Bacteriolysis; Dental Plaque; DNA, Bacterial; Humans; In Vitro Techniques; Microscopy, Electron; Muramidase; RNA, Bacterial; Saliva; Veillonella

Separate samples of supragingival dental plaque overtly free of blood were centrifuged to obtain the free fluid phase (plaque fluid). Bound protein was eluted from the plaque bacteria and matrix by washing the plaque with a low-pH buffer. The plaque fluid, low pH eluate, and whole saliva were assayed for immunoglobulins A, G, and M, the third component of complement, lysozyme, lactoferrin, and lactoperoxidase. Concentrations of total protein and albumin were also determined. Antibody reactive with specific plaque bacteria was detected by indirect immunofluorescent microscopy. Specific and nonspecific immune proteins were present in plaque fluid from adult subjects at significantly greater concentrations than in their saliva, which suggests that these proteins are concentrated in dental plaque. The results indicate that both saliva and gingival exudate contribute to the immunological proteins found in the free fluid phase of dental plaque. The observation that immunoglobulin A antibody reactive with plaque bacteria could be detected in plaque fluid suggests that a wide variety of immunological reactions may occur in the dental plaque. These potential interactions between host, plaque bacteria, and their products could serve to influence the plaque flora and its ability to induce disease.

Topics: Aged; Aging; Antibodies, Bacterial; Child; Complement C3; Dental Plaque; Humans; Immunoglobulins; Lactoferrin; Lactoperoxidase; Middle Aged; Muramidase; Saliva

Topics: Acid Phosphatase; Adolescent; Adult; Alkaline Phosphatase; Amylases; Body Fluids; Child; Child, Preschool; Dental Plaque; Glucosyltransferases; Humans; Muramidase; Peptide Hydrolases; Saliva; Sucrase

Topics: Amylases; Dental Plaque; Humans; Muramidase

In the present study we identified a gram-negative anaerobic rod referred to as Y4 which was cytotoxic for human polymorphonuclear leukocytes. Y4 was isolated from dental plaque of a patient with juvenile periodontitis and presented most of the taxonomic characteristics of Actinobacillus species. Under experimental conditions, viable Y4 were cytotoxic for human peripheral blood polymorphonuclear leukocytes in serum-free cultures. Cytotoxicity was dependent on bacterial concentrations and was enhanced in the presence of a fresh or heat-inactivated (56 degrees C, 30 min) autologous serum. Leukotoxicity was independent of phagocytosis. Y4 leukotoxic effect was abolished when bacteria were heat treated (56 degrees C, 30 min) or when incubations were carried out at 4 degrees C instead of at 37 degrees C. The leukotoxicity was monitored by electron microscopy and biochemically by measuring lactate dehydrogenase indicator of cell viability. No cytotoxic effects of Y4 on human mononuclear cells, chicken fibroblasts, or mouse macrophages were detected under the conditions studied. Polymorphonuclear leukocytes may play an important role in the host defense against bacteria in periodontal disease. The cytotoxic effect of Y4 for polymorphonuclear leukocytes presented in this study is the first report of a direct offensive microbial vector in a plaque-derived microorganism and may prove to be relevant in the pathogenesis of juvenile periodontitis.

Topics: Dental Plaque; Fibroblasts; Humans; L-Lactate Dehydrogenase; Lysosomes; Macrophages; Muramidase; Neutrophils; Periodontitis; Peroxidase; Phagocytosis; Species Specificity

Topics: Adult; Cells, Cultured; Dental Plaque; Gingivitis; Humans; Lactoferrin; Lysosomes; Male; Muramidase; Neutrophils; Peroxidase

Topics: Antibodies, Bacterial; Child; Complement C3; Dental Caries; Dental Plaque; Glucosyltransferases; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Muramidase; Saliva; Streptococcus mutans; Teichoic Acids

The deposition of bacterial plaques on tooth surfaces appears to be responsible for the initiation and progression of periodontal disease. In this study, human peripheral blood polymorphonuclear leukocytes (PMNs) actively released lysosomal constituents upon in vitro exposure to either viable or irradiated, supragingival or subgingival dental plaque. Plaques were obtained from the PMN donors (autologous plaque) or from pooled samples (homologous plaque) secured from patients with periodontal lesions. Fresh sera from PMN donors amplified the release reactions to supragingival and subgingival plaques. Heated (56 degrees C, 30 min) sera also enhanced release reactions, but not as consistently as fresh serum. It was postulated that modulation of PMN release by serum is mediated by complement components and/or antibodies to plaque bacteria. Electron microscopic observations indicated that degranulation and discharge of PMN lysosomal enzymes may be associated with phagocytosis of gram-positive and gram-negative plaque bacteria and with reverse endocytosis of lysosomes from cells contacting relatively large masses of aggregated plaque bacteria. These data suggest that PMN lysosome release in response to plaque may serve as a potential mechanism of tissue injury in the pathogenesis of gingival and periodontal inflammation.

Topics: Bacteria; Dental Plaque; Endocytosis; Gingivitis; Glucuronidase; Hot Temperature; Humans; Immune Sera; In Vitro Techniques; Lysosomes; Muramidase; Neutrophils; Periodontitis; Peroxidase; Peroxidases; Phagocytosis

Topics: Adsorption; Dental Plaque; Dental Porcelain; Muramidase

Topics: Chewing Gum; Dental Plaque; Humans; Middle Aged; Muramidase; Periodontal Pocket; Periodontitis

Rabbit polymorphonuclear leukocytes were incubated with a sonically treated suspension of pooled dental plaque to determine if the plaque would induce release of lysosomal enzymes from the polymorphonuclear leukocytes. Cells incubated with plaque at 37 degrees C released significantly greater amounts of the lysosomal enzymes, beta-glucuronidase and lysozyme, than did cells incubated with plaque at 0 degrees C or without plaque at 37 degrees C. This response was both dose and time dependent. Release of the cytoplasmic enzyme lactate dehydrogenase was minimal, and there were no significant differences in lactate dehydrogenase release between cells at 0 and 37 degrees C, or without plaque. These results indicate that dental plaque can induce the selective release of lysosomal enzymes, which could be involved in the periodontal injury produced by dental plaque.

Topics: Animals; Cells, Cultured; Cytoplasm; Dental Plaque; Exocytosis; Gingivitis; Glucuronidase; Humans; L-Lactate Dehydrogenase; Lysosomes; Muramidase; Neutrophils; Periodontitis; Rabbits; Temperature

The studies reviewed in this chapter provide further evidence that the secretion of lysosomal enzymes and other hydrolases is a constitutive function of certain cells whereas in other cells is an inducible process probably contributing to the pathology of a variety of diseases. Little is known of the mechanisms mediating the secretion of lysosomal enzymes. We have summarized evidence suggesting a role of microfilaments and microtubules in controlling enzyme release, but further studies of the biochemical mechanisms which control the activity of these subcellular structures are required. The fusion of lysosomes with the plasma membrane has been observed in several situations and the mechanisms underlying processes of this nature have been studied in lower organisms (Satir et al. 1973; Plattner 1974). Agents, such as concanavalin A, which interfere with the fusion of endosomes with lysosomes (Goldman 1974; Edelson and Cohn 1974a, b) should also be useful in determining the chemical nature of membrane components involved in the fusion process. New information on the fate of secreted acid hydrolases has been obtained from studies of the uptake of lysosomal enzymes by fibroblasts. Clearly, the mechanisms by which these cells endocytose secreted lysosomal enzymes will be a subject for detailed study in view of the important of directing enzymes and drugs into lysosomes (De Duve et al. 1974). The mechanisms by which extracellular inhibitors inactivate hydrolytic enzymes, particularly proteinases, is also being clarified (for review see Davies 1975) and this should aid in finding new ways for preventing tissue damage caused by the excessive secretion of these enzymes. Further investigation concerning the secretion of lysosomal enzymes should establish the essential physiological role which these enzymes play at both extracellular and intracellular sites. Also, a close examination of the interaction of both endogenous and exogenous stimuli of inflammation with cells resulting in the secretion of hydrolytic enzymes, will clarify the mechanisms underlying the initiation and progression of the inflammatory process in its diverse forms.

Topics: Animals; Anti-Inflammatory Agents; Antigen-Antibody Complex; Bone Resorption; Carrageenan; Cell Transformation, Neoplastic; Cell Wall; Complement System Proteins; Cytochalasin B; Dental Plaque; Glucuronidase; Humans; Hydrolases; Inflammation; Lysosomes; Macrophages; Microbial Collagenase; Muramidase; Neutrophils; Nucleotides, Cyclic; Plasminogen Activators; Pneumoconiosis; Streptococcus pyogenes

Topics: Amino Acids; Amylases; Antibodies, Bacterial; Dental Deposits; Dental Plaque; Humans; Immunoglobulin A; Immunoglobulin M; Muramidase; Proteins; Saliva; Streptococcus

Topics: Actinomyces; Animals; Bacteria; Bacterial Physiological Phenomena; Cathepsins; Cell Count; Dental Plaque; Female; Glucuronidase; Hydrolases; In Vitro Techniques; Leukocytes; Lysosomes; Muramidase; Rabbits; Streptococcus mutans; Streptococcus sanguis

Human supragingival dental plaque was collected from patients with various degrees of caries and periodontal disease. Plaque extracts, prepared in five different solutions (four varied from pH 1.8 to 12.7; one contained urea), were analyzed by polyacrylamide gel electrophoresis, and tested for amylase and lysozyme enzyme activity. Because no qualitative or quantitative advantages of using the extremes of pH or urea were observed, all subsequent extracts were prepared in phosphate buffered saline at pH 7.3. Concentrated extracts were fractionated by gel filtration and characterized by polyacrylamide gel electrophoresis, peptide mapping, molecular weight estimation, determination of enzymatic activities and amino acid and carbohydrate analyses. Regions of similarity among the gels were revealed by comparing the electrophoretic patterns of pooled plaque extract, normal serum and whole saliva. The elution pattern of pooled plaque extract from a standardized Sephadex G-200 column indicated the presence of both high and low molecular weight proteins that might have correlated with the components of normal serum and saliva. A predominant and dialyzable third fraction had no correlate in either serum or saliva. The small peptides in this fraction were subjected to amino acid, carbohydrate and peptide map analyses. The most abundant amino acids were alanine, glutamic acid, glycine, valine, leucine, lysine and serine. These small components contained no neutral or amino sugars. Pooled plaque extract and the small peptides exhibited similar peptide maps.

Topics: Alanine; Amylases; Betaine; Dental Caries; Dental Plaque; Glutamates; Glycine; Humans; Leucine; Muramidase; Periodontal Diseases; Saliva; Serine; Valine

Separate plaque samples (collected from 13 patients who had experienced caries and various degrees of periodontal disease) were each dispersed in 1 ml of distilled water, homogenized and lyophilized. Each lyophilisate was extracted in 1 ml of buffered saline, concentrated and analyzed. Enzyme activity studies revealed amylase in all plaque samples. Lysozyme was present occasionally. By radial immunodiffusion, IgG and IgA were shown to be in plaque extracts of some patients. Immunofluorescence examination of the sediments of individual plaque samples revealed that IgG and IgA occurred frequently. Polyacrylamide gel electrophoresis patterns of the extracts of plaque from different individuals exhibited marked variability despite some zones of similarity. Peptide maps of the dialysable material of the individual plaque extracts were remarkably similar.

Topics: Amylases; Dental Caries; Dental Plaque; Humans; Immunoglobulin A; Immunoglobulin G; Muramidase; Periodontal Diseases; Proteins

Topics: Dental Plaque; Dextranase; DNA; Drug Synergism; Microscopy, Electron, Scanning; Muramidase; Streptomyces; Trypsin

Topics: Adult; Dental Plaque; Female; Gingivitis; Humans; Hyaluronoglucosaminidase; Isoelectric Focusing; Male; Muramidase; Peptide Hydrolases; Time Factors

Topics: Centrifugation; Chitin; Dental Plaque; Glycols; Hot Temperature; Humans; Hydrogen-Ion Concentration; Muramidase; Time Factors; Viscosity