Healing of apical periodontitis after endodontic treatment: a comparison between a silicone-based and a zinc oxide-eugenol-based sealer.

AIM: To assess the treatment results up to 1 year after endodontic treatment of apical periodontitis using a silicone-based sealer in comparison with Grossman's sealer, and to compare the results at 3 months after treatment with the 12-month follow-up to assess the prognostic value of a 3-month control. METHODOLOGY: A total of 199 teeth were treated at three centres. The sealer was randomly chosen at the time of filling. Treatment results were evaluated clinically and radiographically 3 and 12 months after root-canal filling. The periapical status was evaluated using the periapical index (PAI). RESULTS AND CONCLUSIONS: Average PAI scores decreased from 3.43 at start to 2.21 at 12 months for Grossman's sealer and from 3.40 to 2.26 for the silicon-based material. No significant difference between the groups at start or any of the follow ups was seen. The 3-month control was adequate in establishing significant healing in both groups. The improvement of the periapical condition continued at the 12-month examination.

Lactoperoxidase inhibits glucosyltransferases from Streptococcus mutans in vitro.

This study examines the possible effect of the antimicrobial peroxidase system on the activity of streptococcal glucosyltransferases B, C and D (GtfB, GtfC and GtfD), either in solution (GtfB and GtfC) or when adsorbed to hydroxyapatite (GtfC and GtfD) at pH 6.5. The lactoperoxidase (LP) system (LP, H(2)O(2), SCN(-)) had no effect on the activity of dissolved GtfC, but the activity of dissolved GtfB was enhanced. The LP system, however, strongly inhibited the activities of both GtfC and GtfD in their adsorbed form. LP enzyme, without its substrates, inhibited all three Gtf enzymes: GtfB and GtfC in concentrations between 10 and 100 microg/ml in liquid phase and adsorbed GtfC and GtfD in concentrations between 25 and 50 microg/ml. This inhibition was in part abolished in liquid phase, but not in solid phase, if the substrates of LP were added. This study shows that the lactoperoxidase system can exert inhibitory activity against streptococcal Gtfs without generating oxidizing agents.

Saliva flow rate, amylase activity, and protein and electrolyte concentrations in saliva after acute alcohol consumption.

OBJECTIVE: The aim of our study was to evaluate the effects of acute alcohol consumption on saliva secretion rate and selected salivary parameters in healthy nonalcoholic volunteers. STUDY DESIGN: Twenty-four volunteers (37.7 +/- 9.6 years, mean +/- SD) consumed 0.6 g or 0.7 g alcohol/kg of body weight (for women and men, respectively) in a soft drink. Saliva samples were collected, first (S0) before any alcohol was consumed, 45 minutes after consumption (S1) and, finally, 60 minutes after S1 (S2). Flow rates of both resting whole saliva and paraffin-stimulated (SWS) whole saliva were assessed. SWS was assessed for amylase, total protein, inorganic phosphate (PO4(3-)), sodium (Na+), potassium (K+), and calcium (Ca2+) content. RESULTS: SWS, but not resting whole saliva (in milliliters/minute), decreased significantly after consumption of alcohol. Amylase activity (P =.010) and the concentrations of Na+ (P =.000) and Ca2+ (P =.002) decreased significantly between S0 and S1. When SWS was analyzed for output, the total protein concentration (S0 to S1, P =.000; S0 to S2, P =.033) and amylase activity (S0 to S1, P =.000) decreased significantly. Further, the output of all the studied electrolytes decreased significantly as blood alcohol concentration increased. CONCLUSIONS: We conclude that acute alcohol consumption causes a decrease in SWS flow rate. The decrease in flow rate also results in impaired output of total protein and amylase, as well as in a decrease in the output of electrolytes.

Dental diseases and loss of teeth in a group of Finnish alcoholics: a radiological study.

A total of 85 Finnish alcohol-dependent subjects and 53 controls were studied with panoramic radiography. The aim was to study the possible associations between prolonged alcohol consumption and dental health. The mean number of teeth, caries lesions, endodontic treatments, periapical lesions, marginal bone loss, and periodontal infrabony pockets was studied. The subjects met the diagnostic criteria of alcohol dependence as set out in DSM-IV and ICD-10. The control group comprised social drinking volunteers with an AUDIT score < or =8. For the final results the subjects were divided into groups on the basis of sex and age. The social backgrounds of the subjects were similar, except for employment and smoking. The results show significantly fewer teeth and more caries in the alcoholic group. There was a tendency for the alcoholics <45 years of age to have more endodontically treated teeth than the controls, but no difference in the number of periapical lesions in endodontically treated teeth was found. Horizontal bone loss and the presence of calculus were more frequent in alcoholic men than in alcoholic women. Significantly more horizontal bone loss was observed in the group of alcoholic nonsmokers than in nonalcoholic nonsmokers. In the nonsmoking groups alcoholics had significantly more periodontal destruction than the nonsmoking controls. We conclude that radiological dental health among individuals dependent on alcohol is weakened by more caries, more horizontal bone loss, and more numerous vertical infrabony pockets than social drinkers.

Changes in whole saliva in patients with coeliac disease.

Many systemic diseases impair salivary flow rate and composition and therefore incite oral pathological processes. This study analyses the composition of whole saliva in patients with diagnosed coeliac disease (CD) and in healthy controls, and monitors possible changes in saliva composition after a short oral gluten challenge. Paraffin-stimulated whole saliva was collected from 128 CD patients and 55 healthy controls. In a separate study, paraffin-stimulated whole saliva samples were collected from 33 CD patients and 10 controls both before and 24 h after an oral mucosal and submucosal gluten challenge. No difference in saliva flow rate was observed, but total protein (P

Saliva and dental caries.

Caries is a unique multifactorial infectious disease. Our understanding of etiological factors, the progress of the disease, and the effectiveness of prophylactic procedures have led us to believe that we understand the disease. However, we still have too few answers to many questions: "Why can we not predict who will get the disease?" "Why do we not become immunized?" "How much saliva is enough?" or "Which salivary components are protective?" and "Which salivary components predispose for caries?" It is generally accepted, however, that saliva secretion and salivary components secreted in saliva are important for dental health. The final result, "caries to be or not to be", is a complex phenomenon involving internal defense factors, such as saliva, tooth surface morphology, general health, and nutritional and hormonal status, and a number of external factors-for example, diet, the microbial flora colonizing the teeth, oral hygiene, and fluoride availability. In this article, our aim is to focus on the effects of saliva and salivary constituents on cariogenic bacteria and the subsequent development of dental caries.

Effect of lactoperoxidase system on glucosyltransferase D of Streptococcus mutans.

OBJECTIVE: To investigate the effects of the peroxidase system (LP, H2O2, SCN) and its components on the activity of glucosyltransferase D (GtfD) originating from Streptococcus mutans (S. mutans). METHODS: GtfD was incubated in buffered-KCl assay mixture that contained 20 microM dextran and 100 mM sucrose including 14C-glucose. The activity of GtfD was measured with a scintillation counter. The effects of lactoperoxidase system and its components on GtfD activity were examined by incubating different components separately and together with GtfD. RESULTS: The lactoperoxidase system-generated hypothiocyanite (OSCN) had no effect on the activity of streptococcal GtfD, while the lactoperoxidase enzyme inhibited GtfD. The ratio between the 2 enzymes, LP and GtfD, was important for the inhibition. However, if LP was combined with its substrates, SCN and/or a high concentration of H2O2, it enhanced the activity of GtfD. CONCLUSION: Peroxidase-generated antimicrobial agent HOSCN/OSCN is not inhibitory against GtfD, whereas low concentrations of the LP-enzyme inhibit GtfD.

Effects of the common cold and intranasal fluticasone propionate treatment on mucosal host defense assessed by human saliva.

OBJECTIVE: The purpose of this investigation was to study the effect of a potent topical steroid, fluticasone propionate, on patients with early signs and symptoms of the common cold. To characterize the mucosal inflammatory response, salivary defense factors and flow rate in these patients were analyzed. STUDY DESIGN: Forty patients with symptoms of the common cold were randomized into 2 groups to receive either high-dose fluticasone propionate (100 microg per nostril) or placebo 4 times daily for 6 days. Paraffin-stimulated whole saliva was collected on day 1 (before the onset of medication), day 7 (posttreatment), and day 21 (follow-up). RESULTS: Salivary flow rate, innate host defense factors, and total protein content were not affected by the common cold. IgA increased between day 7 and day 21 (P < or = .01; Student 2-tailed t test), and the relative proportions of salivary peroxidase and IgA increased on day 7 (P = .01) and day 21 (P= .05). In patients receiving fluticasone, saliva flow rate was lower on day 21 (P < or = .05) than on days 1 and 7. The innate salivary defense factors were not affected, but IgA increased both on day 7 (P < or = .001) and on day 21 (P < or = .001) in comparison with day 1. CONCLUSIONS: Of the oral mucosal defense factors, only IgA is activated during the common cold. Intranasally administrated fluticasone propionate does not have a suppressive effect on salivary antimicrobial capacity.

The effects of different (pseudo)halide substrates on peroxidase-mediated killing of Actinobacillus actinomycetemcomitans.

Actinobacillus actinomycetemcomitans is a Gram-negative bacterium which has an important role in localized juvenile and in progressive periodontitis. It is sensitive to killing by the myeloperoxidase (MP)-hydrogen peroxide (H2O2)-chloride system which is part of the innate host defense mediated by polymorphonuclear leukocytes. Since it has been recently suggested that thiocyanate, instead of chloride, could serve as a main substrate for MP as for lactoperoxidase (LP) and salivary peroxidase, we investigated in this study the effect of both LP and MP systems on A. actinomycetemcomitans with different (pseudo)halide substrates, thiocyanate, chloride and iodide. The concentrations of the substrates were physiological for oral fluids, as was the concentration range of H2O2. Both peroxidases produced end products with identical antibacterial activity with thiocyanate and iodide. The oxidation of iodide resulted in the highest antimicrobial efficiency followed by chloride and thiocyanate. Addition of thiocyanate into either MP-H2O2-chloride or MP/LP-H2O2-iodide system abolished the bactericidal activity of the oxidized halide. However, the chloride did not affect the bactericidality of the MP-H2O2-iodide system, but when all 3 (pseudo)halide substrates were present no antimicrobial effect was recorded. Our study shows that the presence of thiocyanate in physiological amounts is able to prevent the bactericidal activity of halide-peroxidase systems in low H2O2 concentrations. These results explain why thiocyanate-peroxidase systems of either innate origin (saliva, crevicular fluid) or introduced by commercial oral hygiene products are most probably ineffective against A. actinomycetemcomitans in vivo. Further studies of halide/thiocyanate ratio are needed to develop products which are also effective against oral anaerobes.

Growth of xylitol-resistant versus xylitol-sensitive Streptococcus mutans strains in saliva.

Five Streptococcus mutans pairs (serotype c S. mutans 10449 and four clinical isolates of S. mutans: 123.1, LG1, OMFA, T10B) were used to find out if the xylitol-resistant (XR) natural mutants of the corresponding xylitol-sensitive (XS) S. mutans parental strains differ in their growth patterns in saliva. The isogenic X natural mutants of the parental S. mutans strains were selected after sequential cultivations in the presence of xylitol and glucose. The XR/XS strains pairs were grown in individual and pooled glucose-supplemented filter-sterilized salivas (one to five sequential cultivations). The two salivas used represented subjects with good or poor support of the growth of S. mutans in vivo. Protease and peptidase activities were determined from the saliva growth media and cell suspensions. Salivary protein profiles were analyzed using SDS-PAGE and native IEF before and after the cultivations. The growth properties of the XR/XS S. mutans pairs were similar in both individual and pooled salivas. Sequential cultivation of all strains did not show any differences in growth patterns. XS strains were inhibited by the presence of xylitol (2% w/v) in pooled saliva, as shown for other glucose-supplemented media. Protease and peptidase activities of the XR/XS S. mutans pairs were low and of similar magnitude. Also, the general hydrolytic properties of most XR/XS S. mutans pairs appeared similar as judged by the small growth-induced changes in salivary protein profiles. In conclusion, saliva, the source of nutrients for salivary microorganisms in vivo, favored neither the XR nor the XS strains of S. mutans.

Stimulated salivary flow rate and composition in asthmatic and non-asthmatic adults.

The number of decayed, missed and filled permanent teeth (DMFT), the degree of periodontal inflammation (Periodontal Status Index, PSI), stimulated salivary flow rate and the concentrations of total protein, lactoferrin, lysozyme, myeloperoxidase, salivary peroxidase, calcium, potassium, sodium and thiocyanate in whole saliva of 26 adult asthma patients were compared with those of 33 non-asthmatic controls. The saliva was also analysed for mutans streptococci, lactobacilli, total anaerobic flora and Candida spp. The mean PSI (p < 0.05; 95% confidence interval for the difference between means (95% CI) 2.47-25.30) was higher and the mean stimulated salivary flow rate (p < or = 0.05; 95% CI 0.57-0.55) was lower in the asthmatic group than in the control group. No differences were found between the groups in non-immune defense factors, except for myeloperoxidase. The myeloperoxidase concentrations were higher in asthmatics than in non-asthmatics (p < 0.05; 95% CI 4.4-134.0 ng/ml). No differences in microbial counts were found. It was concluded that stimulated salivary flow rates decrease while myeloperoxidase concentrations increase in adult asthmatic patients compared with non-asthmatic adults. The higher concentrations of myeloperoxidase are explained by a higher PSI in asthmatics.

Combined inhibitory effect of fluoride and hypothiocyanite on the viability and glucose metabolism of Streptococcus mutans, serotype c.

The separate and combined effects of peroxidase-generated hypothiocyanite (HOSCN/OSCN-) and F- ions on glucose uptake and growth of Streptococcus mutans ATCC 25175 were investigated. S. mutans cells were grown to late exponential or stationary growth phase, harvested, washed and suspended in 2.0 ml of sterilized human whole saliva supplemented with 10 mM D-glucose. This saliva-bacteria mixture was supplemented with 5-150 microM H2O2 at pH 5.0 or 6.5. At pH 5.0, up to 103 +/- 21 microM HOSCN/OSCN- was generated. After 20 h of incubation at 37 degrees C, the saliva-bacteria suspension exposed to HOSCN/OSCN- were plated on mitis salivarius agar plates and incubated anaerobically for 2 days. Identical experiments were made with F- ions (0.5, 1.0 and 5.0 mM). Both HOSCN/OSCN- and F- caused a significant dose-dependent growth inhibition at pH 5.0, whereas no inhibition was observed at pH 6.5. When F- and HOSCN/OSCN- were added simultaneously at pH 5.0, an additive effect of growth inhibition was observed. In glucose incorporation experiments the bacteria-saliva mixture was exposed to 1 microM HOSCN/OSCN-, 0.5 mM F- or both. F-, HOSCN/OSCN- or their combination in sterilized whole saliva at pH 5.0 caused 14.2, 67.8 and 74.2% inhibition, respectively. These observations indicate that F- and HOSCN/OSCN- ions have an additive inhibitory effect on S. mutans and therefore their combination is likely to be more antibacterial than either agent alone.

Release of cariostatic agents from a new buffering fluoride- and xylitol-containing lozenge to human whole saliva in vivo.

A new buffering lozenge (sucking tablet) was developed for patients susceptible to dental caries and erosion, in particular for those with reduced salivary secretion. As active ingredients this lozenge comprises of a combination of xylitol, fluoride, calcium, phosphate, zinc and buffering compounds. To test the lozenge's activity in vivo, the release of ingredients was monitored in 19 healthy subjects for 22 min after sucking the lozenge was completed. In subjects with a normal salivary secretion rate the lozenge caused only a slight stimulation of saliva flow, but a significant elevation both in salivary pH and buffer effect was observed. Furthermore, fluoride, calcium and phosphate were effectively released into whole saliva with peak values 2-4 min after use. The same salivary parameters were also quantitated after 1 month's regular use (3 lozenges/day) but no consistent long-term changes were found. Salivary mutans streptococci and total anaerobic microflora did not change significantly during the long-term use. The results show that the buffering fluoride- and xylitol-containing lozenge, which also releases calcium and phosphate, is active in vivo but its serviceability as a remineralizing agent, in particular for elderly patients with reduced salivary flow rate, has to be analysed separately.

Effects of oral hygiene products containing lactoperoxidase, lysozyme, and lactoferrin on the composition of whole saliva and on subjective oral symptoms in patients with xerostomia.

This study evaluates the effects of two oral hygiene products containing nonimmunoglobulin antimicrobial agents on whole saliva and on subjective oral symptoms in patients with xerostomia. Twenty patients used a lactoperoxidase-system-containing toothpaste (Biotene) combined with the use of a mouthrinse (Biotene), comprising also lysozyme and lactoferrin, for 4 weeks. Saliva samples were collected at base line, after 4 weeks' use of the products, and at the end of a 4-week washout period. Samples were analyzed for selected biochemical and microbiologic factors. The effects on subjective oral symptoms were also recorded. A 4-week daily use of toothpaste and mouthrinse relieved the symptoms of oral dryness in 16 patients. The levels of salivary hypothiocyanite, lysozyme, lactoferrin, or myeloperoxidase activity did not change, but there was a significant decrease in salivary pH (P < 0.05), total peroxidase activity (P < 0.05), and total protein content (P = 0.01). In patients with the lowest salivary flow rates (n = 5) a significant (P > or = 0.04) increase was detected in salivary hypothiocyanite concentrations. No major changes occurred in salivary microflora. The products relieved subjective oral symptoms in most xerostomic patients, but this was not necessarily related to the presence of antimicrobial agents.

Effects of delmopinol on antimicrobial peroxidase systems and lysozyme in vitro and in human whole saliva.

Delmopinol is a new surface-active agent which can reduce plaque formation and gingivitis. This study was aimed to analyze whether delmopinol (0.0032-0.65 mM) interferes with the activity of two surface-active oral antimicrobial enzymes, salivary peroxidase and lysozyme. In addition to human whole saliva (pH 5.0 and 6.0), the experiments were done in 0.1 M phosphate buffer (pH 6.0) with purified lactoperoxidase (LPO) and myeloperoxidase (MPO). LPO and MPO were significantly inhibited in buffer by delmopinol concentrations > 6.5 mM and > or = 3.2 mM, respectively. No such inhibition was found for total peroxidase activity in mixed saliva. In vitro, delmopinol was found to desorb surface-bound peroxidases in an active form to the liquid phase. In further analyses, the possible effect of delmopinol on peroxidase-generated hypothiocyanite (HOSCN/OSCN-) was studied in saliva and buffer. No effect was found in buffer, but salivary HOSCN/OSCN- declined significantly with 6.5 mM delmopinol. This was obviously due to an enhanced decay of hypothiocyanite, rather than its reduced rate of formation. No delmopinol-related inhibition of lysozyme occurred in saliva or buffer. The results suggest that high concentration (6.4 mM -0.2%) of delmopinol may lower the concentrations of antimicrobial HOSCN/OSCN- in saliva but has no effect on human lysozyme.

Effect of saliva composition on growth of Candida albicans and Torulopsis glabrata.

Candida albicans and Torulopsis glabrata are the most prevalent yeasts in humans. The majority harbor C. albicans in the oral cavity, but only a few develop oral candidiasis. We have sought a possible relationship between indigenous salivary constituents, including antimicrobial and nutritive factors, and the growth rate and/or viability of inoculated fungi in glucose-supplemented sterilized saliva. Stimulated whole saliva was collected from 30 healthy donors. Saliva samples were sterilized, supplemented with glucose and inoculated with C. albicans or T glabrata. After incubation of the inoculates for 20 h, the number of viable cells were counted. All saliva samples were analyzed for different indigenous salivary components and Candida before as well as after sterilization. Besides a 4% reduction in calcium (Ca2+) and thiocyanate (SCN-) concentrations, sterilization did not affect the concentrations of saliva electrolytes, but the proteins were significantly reduced (19-85%). Indigenous candidal carriage (n=19) correlated with neither the growth of inoculated fungi nor any of the analyzed components in saliva. The growth of C. albicans and T. glabrata was similar at pH 5 but, at pH 6, C. albicans had a remarkably slower growth rate than T. glabrata. Statistical analysis showed that the 5-h growth of C. albicans at pH 5 was associated with water and electrolyte secretion, whereas the growth after 20 h was associated with variations in protein-glycoprotein content. The growth of T. glabrata was not related to variations in the salivary variables analyzed.

Newer saliva collection methods and saliva composition: a study of two Salivette kits.

UNLABELLED: Saliva is frequently used as a diagnostic fluid and several collection devices have been developed. OBJECTIVE: The aim of the present study was to investigate the validity and reliability of two types of Salivette collection kits (non-covered cotton roll and polypropylene covered polyether roll) relative to conventional collection of saliva using paraffin wax chewing stimulation. MATERIALS AND METHODS: Whole saliva samples were collected from 16 healthy volunteers. Following a cross-over design saliva was collected in a standardized way. The flow rate was determined and saliva samples were analyzed for pH, buffer capacity, electrolytes and protein/glycoprotein content. RESULTS: We find that Salivette methods do not allow evaluation of flow rate. pH was unaffected but buffer capacity was lower in Salivette collected than in paraffin wax-stimulated saliva. The non-covered cotton rolls reduced the content of Na+, K+, Cl-, as well as glycoprotein markers (hexosamines, fucose, sialic acid), lysozyme, lactoferrin, salivary- and myeloperoxidase but increased the concentrations of Ca2+, PO4(3)- and SCN-. Polypropylene covered polyether rolls affected saliva composition less than the non-covered cotton rolls. Thus, SCN- and sIgA concentrations were higher and lysozyme activity lower in the former (covered roll) saliva than in paraffin wax saliva. The reliability of the Salivette kits was good. CONCLUSION: We conclude that the Salivette method generates data significantly different from conventional paraffin wax-stimulated saliva such as buffer capacity and several electrolytes and organic components. Care should be taken in interpreting the results when such methods are employed.

Effects of a lactoperoxidase-system-containing toothpaste on dental plaque and whole saliva in vivo.

The effects of a lactoperoxidase-system-containing toothpaste. Biotene, on saliva and dental plaque were studied. In a double-blind crossover study 20 healthy volunteers used an experimental (comprising the complete peroxidase system) or a placebo (without lactoperoxidase, KSCN, and glucose oxidase) toothpaste twice daily for 2 weeks separated by a 2-week washout period. At base lines and at the end of both test periods saliva and plaque samples were collected, and plaque pH changes were monitored. Saliva was analyzed for hypothiocyanite (HOSCN/OSCN-) and thiocyanate (SCN-) concentrations and salivary peroxidase activity. The amount of total streptococci, mutans streptococci, lactobacilli, and total anaerobic flora was determined both in saliva and in plaque samples. The accumulation and the acidogenicity of plaque were also quantitated. A 2-week daily use of Biotene had no effect on salivary flow rate, peroxidase activity, HOSCN/OSCN-, SCN-, or any of the monitored bacterial counts compared with the placebo toothpaste. The accumulation of dental plaque was not affected by the lactoperoxidase-system-containing toothpaste. The acidogenicity of plaque did not change significantly, nor did the two test dentifrices differ in their ability to inhibit the plaque pH drop caused by sucrose in subjects with normal salivary flow rate.

Lysozyme and lactoperoxidase inhibit the adherence of Streptococcus mutans NCTC 10449 (serotype c) to saliva-treated hydroxyapatite in vitro.

Lysozyme, lactoperoxidase and salivary peroxidase inhibit the metabolism and growth of mutans streptococci, but any possible effects on the adherence of these bacteria are unknown. In this study the effects of lysozyme and lactoperoxidase on the adhesion of 3H-labelled Streptococcus mutans (NCTC 10449, serotype c strain) to saliva-coated hydroxyapatite were studied at pH 5.0 and 7.0. Human whole saliva was either lysozyme-depleted and centrifuged, or sterilized and dialysed to achieve no detectable lysozyme and peroxidase activities; this modified saliva was used to form experimental pellicles. The incorporation of lysozyme (50-200 micrograms/ml) to the pellicle caused a significant (p < 0.01) reduction in the adherence of S. mutans without any loss of bacterial viability. Pretreatment of either saliva-coated apatite or S. mutans cells with lysozyme did not change the results but lysozyme bound more readily to bacteria than to the experimental pellicles. Also, lactoperoxidase (10-200 micrograms/ml) reduced significantly (p < 0.001) the adherence of S. mutans but, in contrast to lysozyme, in a dose-dependent way. The strongest inhibition of adhesion was found when both saliva-coated apatite and bacteria were pretreated with lactoperoxidase. This enzyme bound to experimental pellicles in preference to streptococci. A non-specific protein control, albumin, did not block the inhibition by lysozyme or lactoperoxidase. The inhibition of adherence of a serotype c strain of S. mutans to saliva-coated hydroxyapatite is a novel antibacterial mechanism for both lysozyme and lactoperoxidase.