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Study of root caries in an artificial mouth.

The object of this project was to develop a laboratory model system to study root and enamel caries. Model plaque biofilms containing consortia of four (Streptococcus mutans, Streptococcus sobrinus, Actinomyces naeslundii, and Lactobacillus rhamnosus) or six (S mutans, S sobrinus, S mitis, A naeslundii, L rhamnosus, and Veillonella dispar) species of caries pathogens were developed in a artificial mouth. Caries lesions similar to those found in vivo were produced on enamel and dentine blocks and in intact root tissues by these consortia more effectively than biofilms of the individual species. Fluoride supplied at 5 ppm suppressed enamel caries effectively, less so dentinal lesion formation, but much more so than F incorporated into the mineralised surface of enamel and roots. This is the first direct comparison of the effect on caries of environmental F and tooth mineral F induced by a plaque-like bacterial biofilm.

Actinomyces↗

Effects of resin composite components on glucosyltransferase of cariogenic bacterium.

The aim of our study was to investigate the effect of resin composite components on glucan synthesis by glucosyltransferase (GTase) derived from a cariogenic bacterium, Streptococcus sobrinus B13. The eluates from cured composites stored in 0.05 mol/L potassium phosphate buffer (KPB) (pH 6.8) for 2 weeks at 37 degrees C stimulated the formation of water-insoluble glucan, whereas those from amalgam inhibited it. This finding suggests that the eluates from the resin composites enhance GTase activity and contribute to plaque formation. In the individual resin components, a diglycidyl methacrylate and triethylene glycol dimethacrylate significantly enhanced GTase activity. However, most polymerization inhibitors and accelerators were potent inhibitors of enzyme activity. Thus, GTase stimulation by eluates from resin composites is hypothesized to be heavily dependent on the effect of the eluated resin monomers, even though the other eluting components had inhibitory effects on GTase.

Composite Resins↗

Immunogenic and protective potential of mutans streptococcal glucosyltransferase peptide constructs selected by major histocompatibility complex class II allele binding.

Mutans streptococcal glucosyltransferases (GTF) have been demonstrated to be effective components of dental caries vaccines. We had previously selected peptide subunits of GTF for vaccine development based on putative functional significance and conservation of GTF primary structure among enzyme isoforms. In this study, 20 20-mer linear GTF peptides were synthesized, 17 identified on the basis of the highest potential major histocompatibility complex (MHC) class II-binding activity using computer-generated algorithms (Epimatrix and ProPred) and 3 with previously demonstrated functional significance. The immunoreactivities of these peptides were explored with rodent systems. Sera from GTF-immunized rats, assessed for binding to linear peptides by enzyme-linked immunosorbent assay, demonstrated immunoglobulin G antibody reactivity with peptides 6 and 11 and a T-cell proliferation response to peptides 6, 9, 11, and 16. Multiple antigenic peptide (MAP) constructs were synthesized from promising linear sequences. Rats that were immunized with MAP 7, 11, or 16, respectively, responded well to the immunizing MAP. Most importantly, a robust immune response (antibody and T-cell proliferation) was observed to native GTF following MAP 11 (amino acids 847 to 866; VVINNDKFVSWGITDFEM) immunization. This response inhibited GTF enzyme function. Two dental caries pathogenesis experiments were performed wherein rats were immunized with MAP constructs 11, 16, and/or 11 plus 16, followed by infection with cariogenic Streptococcus sobrinus. In both experiments cariogenic bacterial recoveries were reduced relative to total streptococci in the MAP 11- and MAP 11 plus 16-immunized groups, and the extent of dental caries was also significantly reduced in these groups. Thus, we have identified a peptide with projected avid MHC-binding activity that elicited immunoreactivity with native GTF and demonstrated protection against dental caries infection after immunization, implying that this peptide may be important in a subunit dental caries vaccine.

Animals↗

Effects of copper, iron and fluoride co-crystallized with sugar on caries development and acid formation in deslivated rats.

The purpose was to explore the effects of combinations of copper, iron and fluoride (Cu, Fe and F) incorporated in sucrose by co-crystallization on caries development in the deslivated rat model and to examine acid formation by bacteria in the rat mouth. Ninety-six Sprague-Dawley rats were infected with Streptococcus sobrinus 6715 and desalivated when aged 26 days. Eight groups were placed in a König-Höfer programmed feeder and received 17 meals daily at hourly intervals, and essential nutrition (NCP No. 2) by gavage twice daily for 21 days. The groups received (1) plain sucrose, (2) F (8 parts/10(6)) co-crystallized with sucrose, (3) Fe (88 parts/10(6)) sucrose, (4) Cu (75 parts/10(6)) sucrose, (5) Cu + F sucrose, (6) Cu + L Fe sucrose, (7) F + Fe sucrose, and (8) Cu + Fe + F sucrose. At death the jaws were removed and sonicated in 0.9% saline solution for microbial assessment. In addition, organic acid assays were performed for each animal. Keyes smooth-surface and sulcal caries scores were lowest in the Cu + Fe + F sucrose group, but not statistically significantly different from those of the other Cu groups. The numbers of Strep. sobrinus found in the groups that received Cu, Cu + Fe, Cu + F, F + Fe and Cu + Fe + F sugar were lower than in the control group. Lactic acid was found in lower concentrations in Fe, Cu, Cu + F, Cu + Fe and F + Fe groups than in the other groups. It appears that combinations of Cu; Fe and F co-crystallized with sugar may have an additive effect in reducing the cariogenic potential of sugar by affecting lactic acid formation and reducing bacterial colonization.

Acetates↗

Oolong tea polyphenols inhibit experimental dental caries in SPF rats infected with mutans streptococci.

An extract of oolong tea (semifermented tea leaves of Camellia sinensis) and its chromatographically isolated polyphenolic compound was examined for in vitro inhibitory effects on glucosyltransferases (GTases) of mutans streptococci and on caries development in Sprague-Dawley rats infected with mutans streptococci. The samples showed no detectable effect on the growth of mutans streptococci. However, insoluble glucan synthesis from sucrose by the GTases of Streptococcus mutans MT8148R and Streptococcus sobrinus 6715 was markedly inhibited, as was sucrose-dependent cell adherence of these mutans streptococci. The administration of the oolong tea extract and the isolated polyphenol compound into diet 2000 and drinking water resulted in significant reductions in caries development and plaque accumulation in the rats infected with mutans streptococci. The active components in the oolong tea extract were presumptively identified as polymeric polyphenols which were specific for oolong tea leaves. These results indicate that the oolong tea polyphenolic compounds could be useful for controlling dental caries.

Animals↗

Dye-mediated bactericidal effect of He-Ne laser irradiation on oral microorganisms.

Little attention has been given to the bactericidal effect of laser irradiation, particularly using low-power energy lasers. It has been demonstrated that He-Ne laser light has an inhibitory action on dental plaque. The purpose of this study was to investigate the bactericidal effect of He-Ne laser irradiation on cariogenic microorganisms. The bactericidal effect was determined by the formation of a growth-inhibitory zone or by the counting of viable bacterial colonies. Streptococcus sobrinus AHT that is a Gram-positive microorganism was sensitive to He-Ne laser light, but Escherichia coli, a Gram-negative microorganism, was resistant. The effect of several dyes necessary to instigate a bactericidal action was also examined. A growth-inhibitory zone was observed using 10 kinds of blue, purple, or green dyes, which were mainly phenylmethane dyes. The leakage of potassium from S. sobrinus AHT following laser irradiation was determined using an atomic absorption spectrophotometer. The leakage began to increase following irradiation for 2 min, and reached a plateau following irradiation for 30-60 min. Moreover, to examine some changes in the dye itself following laser irradiation in the absence of bacteria, ultraviolet-visible absorption spectra and 1H NMR spectra were recorded. In this study, it was indicated that the bactericidal effect on cariogenic bacteria by He-Ne laser irradiation was efficient only in the presence of specific dyes. It is suggested that this laser may be suitable for clinical applications in preventive dentistry.

Colony Count, Microbial↗

Aspiration pneumonia: dental and oral risk factors in an older veteran population.

OBJECTIVES: To investigate the importance of medical and dental factors in aspiration pneumonia in an older veteran population. DESIGN: Prospective enrollment of subjects with retrospective analysis of data. SETTING: Department of Veterans Affairs outpatient clinic, inpatient ward, and nursing home. PARTICIPANTS: 358 veterans age 55 and older; 50 subjects with aspiration pneumonia. MEASUREMENTS: Demographic and medical data; functional status; health-related behaviors; dental care utilization; personal oral hygiene; comprehensive dental examination; salivary assays including IgA antibodies; and cultures of saliva, throat, and dental plaques. RESULTS: Two logistic regression models produced estimates of significant risk factors. One model using dentate patients included: requiring help with feeding (odds ratio (OR) = 13.9), chronic obstructive pulmonary disease (COPD) (OR = 4.7), diabetes mellitus (OR = 3.5), number of decayed teeth (OR = 1.2), number of functional dental units (OR = 1.2), presence of important organisms for decay, Streptococcus sobrinus in saliva (OR = 6.2), and periodontal disease, Porphyromonous gingivalis in dental plaque (OR = 4.2), and Staphylococcus aureus presence in saliva (OR = 7.4). The second model, containing both dentate and edentulous patients included: requiring help with feeding (OR = 4.7), COPD (OR = 2.5), diabetes mellitus (OR = 1.7), and presence of S. aureus in saliva (OR = 8.3). CONCLUSION: This study supports the significance of oral and dental factors while controlling for established medical risk factors in aspiration pneumonia incidence.

Activities of Daily Living↗

Tansmission, diversity and virulence factors of Sreptococcus mutans genotypes.

Dental caries is an infectious and transmissible disease, in which many genetic, environmental and behavioral risk factors interact. The mutans streptococci (MS), mainly Streptococcus mutans and Streptococcus sobrinus are the microorganisms most strongly associated with this disease. The main virulence factors associated with MS cariogenicity include adhesion, acidogenicity and acid tolerance. These properties work together to modify the physico-chemical properties of the biofilm, resulting in ecological changes in the form of increased proportions of S. mutans and other acidogenic and aciduric species. In addition, reports of higher numbers of S. mutans genotypes with increased virulence in caries-active subjects suggest the importance of microenvironmental factors in increasing the risk of caries. This review focuses on the transmission and establishment of different genotypes of S. mutans and the role they play in the development of dental caries.

Animals↗

The effect of propranolol on salivary gland function and dental caries development in young and aged rats.

Medications commonly used in elderly people cause hyposalivation and are associated with an enhanced prevalence of dental caries. Propranolol (a beta-adrenergic antagonist) is a commonly used antihypertensive agent that is prescribed for long-term use. The purpose of this investigation was to compare the effects of this drug on salivary composition and flow rate, and on caries, in young and aged rats. Forty young (28-day) and 36 aged (20-month) female Sprague-Dawley rats were infected with Streptococcus sobrinus 6715 and fed a cariogenic diet for 28 days. Propranolol was given in high (20 mg/kg/day) and low (10 mg/kg/day) doses via osmotic pumps. Unoperated and desalivated animals served as controls. Smooth-surface caries scores in the young animals receiving propranolol at 20 mg/kg/day were statistically higher than in the young intact rats (p < or = 0.05). Increased smooth-surface and sulcal caries scores were recorded in the aged propranolol-treated animals, but the differences were not statistically significant when compared with those in intact aged animals. Propranolol in aged animals did not affect the amount of alveolar bone loss but increased the risk of development of root caries. Young animals harboured greater populations of Strep. sobrinus and total cultivable flora than did all aged groups except the desalivated group. Salivary flow rates, induced by pilocarpine, were not decreased by the chronic administration of propranolol. Although the total protein concentration in parotid and submandibular saliva from drug-treated animals was reduced, differences were not observed in their SDS-PAGE profile when compared with unoperated animals. The findings demonstrate that chronic use of propranolol reduced the total protein concentration in saliva of all animals, increased caries susceptibility, but did not reduce the stimulated salivary flow rate.

Aging↗

Pilocarpine alters caries development in partially-desalivated rats.

This study examined the effect of pilocarpine on caries and saliva composition in rats with compromised salivary gland function. Eight litters of specific-pathogen-free female Sprague-Dawley rats were divided into five groups for surgery. Partial desalivation was performed in either of three ways: (1) Both parotid ducts were ligated; (2) both submandibular/sublingual (SM/SL) glands were excised, or (3) one parotid duct was ligated, and submandibular/sublingual glands were removed unilaterally. Completely desalivated animals and unoperated animals served as positive and negative controls, respectively. One week following surgery, groups were subdivided so that half of each group had osmotic pumps implanted which delivered pilocarpine HCl (24 mg/kg/day). All animals were infected with Streptococcus sobrinus 6715 and fed cariogenic diet ad libitum for 28 days. Caries scores, microbiological data, and saliva flow rate and composition were determined for all animals. Animals which had both SM/SL glands removed and received pilocarpine developed significantly lower sulcal caries (p < or = 0.05) compared with the animals that had both SM/SL removed but did not receive pilocarpine. The concentration of protein in parotid saliva in these pilocarpine-treated animals was unaffected, and no differences were observed in the electrophoretic profiles on SDS-PAGE. Pilocarpine appears to exert its greatest caries-protective effect when the parotid glands remain intact.

Analysis of Variance↗

The effect of ovariectomy on dentin formation and caries in adult rats.

Since acute osteoporosis is known to enhance bone remodeling and osteoid formation, it may also affect dentin apposition. We induced osteoporosis in 15-week-old Wistar rats by ovariectomy and dental caries by Streptococcus sobrinus infection in the presence of either a high sucrose (cariogenic) diet or a non-cariogenic diet. Intact animals with the same diets served as controls. After 99 days of cariogenic challenge the rats were killed, and the success of ovariectomy was confirmed by failure to detect ovarian tissue and observation of marked atrophy of the uterine horns. Areas of dentinal apposition during the experiment and carious lesions were quantified with a tetracycline method. Ovariectomy activated dentin formation significantly in rats fed either a high-sucrose or a non-cariogenic diet, indicating enhanced odontoblast function. The rate of dentinal apposition in adult rats was smaller than reported in young animals. The effect of ovariectomy on caries remained negligible.

Animals↗

Coimmunization with complementary glucosyltransferase peptides results in enhanced immunogenicity and protection against dental caries.

Peptide constructs from the catalytic (CAT) and glucan-binding (GLU) regions of the mutans streptococcal glucosyltransferase enzymes (GTF) can provide immunity to dental caries infection. A strategy of coimmunization was tested to determine whether protection could be enhanced. Rats were immunized with one of the previously described peptide constructs from the CAT or GLU region of the GTF of mutans streptococci or coimmunized with a combination of these constructs (CAT-GLU). Coimmunized animals demonstrated significantly higher serum immunoglobulin G (IgG) and salivary IgA antibody levels to CAT or GTF than rats immunized with either construct alone. To assess the functional significance of coimmunization with these constructs, animals were immunized as above or with Streptococcus sobrinus GTF and then infected with S. sobrinus to explore the effects of immunization on immunological, microbiological, and disease (dental caries) parameters. Serum antibody from the communized group inhibited S. sobrinus GTF-mediated insoluble glucan synthesis in vitro above that of the individual-construct-immunized groups. Immunization with CAT or GLU constructs resulted in significantly reduced dental caries after infection with S. sobrinus compared with sham-immunized animals. Coimmunization produced greater reductions in caries than after immunization with either CAT or GLU. Also, significant elevations in lymphocyte proliferative responses to CAT, GLU, and GTF were observed after coimmunization with CAT-GLU compared with the responses after immunization with the individual constructs. The results suggested that increased numbers of memory T cells, which could proliferate to CAT, were generated by coimmunization. The experiments support the functional significance of these GTF domains in dental caries pathogenesis and present coimmunization as a simple alternative to intact GTF to enhance protective immunity against cariogenic microorganisms.

Amino Acid Sequence↗

Trehalulose does not induce dental caries in rats infected with mutans streptococci.

The effects of trehalulose, a structural isomer of sucrose, and a syrup (TP syrup) rich in trehalulose and palatinose on caries development were examined in specific pathogen-free Sprague-Dawley rats. Streptococcus mutans MT8148R and Streptococcus sobrinus 6715 fermented the syrup which resulted in acid production, while both strains were found not to utilize trehalulose. Furthermore, trehalulose did not serve as a substrate for glucosyltransferases of these mutans streptococci to synthesize water-insoluble glucan, and it inhibited the sucrose-dependent adherence of mutans streptococci to a glass surface. Although trehalulose induced no significant dental caries in specific pathogen-free rats infected with either MT8148R or 6715, TP syrup was found to induce significant but low dental caries. Furthermore, replacement of the dietary sucrose content with trehalulose resulted in a significant reduction of caries development in rats infected with strain 6715.

Animals↗

Does social deprivation in 1, 2, 3 and 4-year-old Scottish infants influence the frequency isolation of caries-associated micro-organisms?

OBJECTIVES: The aim of this study was to determine if there are associations between the level of social deprivation/affluence and the frequency isolation of caries-associated micro-organisms (Streptococcus mutans, Streptococcus sobrinus, lactobacilli and yeasts) in a large cohort of infants examined annually from 1 to 4 years of age. METHODS: DEPCAT was used to measure the socio-economic status of all consented infants (n=1099--1392) born in Dundee during a 1 year period (total n=1974). Caries-associated micro-organisms were cultured from saliva when the infants were 1, 2, 3 and 4 years of age. Standardised dental examinations were also carried out annually. Log linear analysis, which controlled for caries, was used to look for associations between DEPCAT and the isolation frequency of caries-associated micro-organisms. RESULTS: When controlling for caries, there was an association between DEPCAT and the isolation frequency of yeasts when the infants were 1 and 2 but not when 3 and 4 years old, whereas lactobacilli were associated only when the infants were 3 and 4 years old. Correlations between S. mutans and social deprivation were usually dependent on the caries status of the infants. CONCLUSIONS: The relationship between social deprivation and the isolation frequencies of caries-associated micro-organisms is complex with lactobacilli developing an association when the infants were 3 and 4 years old in contrast to yeasts which were only associated when the infants were 1 and 2 years old. Streptococcus mutans was associated with social deprivation when the infants were 2 years old and older, but dependent on caries status in the 3 and 4 year olds.

Age Factors↗

Adhesion in vitro of oral streptococci to porcelain, composite resin cement and human enamel.

We investigated the initial adhesion in vitro of oral streptococci to porcelain inlays. Four strains of streptococci, Streptococcus mitis NCTC12261, Streptococcus oralis ATCC9811, Streptococcus sanguis ATCC10556 and Streptococcus sobrinus OMZ176, were used in this study. The disc specimens were made of porcelain, composite resin cement and human enamel. These specimens, with or without a saliva-coat, were immersed in a suspension of each streptococci strain at 37 degrees C for 1 hr, and the numbers of cells adhering to specimens were counted after staining. The saliva-coat significantly decreased the numbers of adhering cells in all strains tested (t-test, p < 0.05). The adhesion to the porcelain and resin cements could be explained by a thermodynamic approach, although the adhesion to enamel could not be explained. These results indicated the possibility that the mechanism is different in initial adhered strains of streptococci between enamel and dental restorative materials.

Bacterial Adhesion↗

Composition and flow rate of saliva and caries development in young rats following administration of lithium.

Lithium carbonate (Li2CO3), an antipsychotic medication, was investigated to determine the effects on the stimulated flow and composition of saliva and on caries development in rats. Lithium carbonate was delivered via osmotic pumps at 10, 15 and 60 mg/kg/day. All animals were infected with Streptococcus sobrinus 6715 and fed a cariogenic diet. Lithium administration exerted a caries-protective effect on sulcal surfaces, though the reduction in caries scores was statistically significant only for the groups receiving Li2CO3 at 10 and 60 mg/kg/day compared to control animals (p +/- 0.05, ANOVA). There were no detectable alterations in protein concentration, electrophoretic profiles, or flow rates of stimulated parotid and submandibular saliva associated with the decreased sulcal caries incidence. Weight gain, water consumption and the populations of total cultivable flora recovered were not influenced by the administration of lithium. Investigation into the mechanisms of the sulcal caries reduction will require further exploration of changes caused by lithium on salivary gland function.

Animals↗

Salivary IgA to cariogenic bacteria in HIV-positive children and its correlation with caries prevalence and levels of cariogenic microorganisms.

The interrelationship of HIV infection, dental caries and mucosal immune responses remains controversial. In our study population of 40 HIV-infected and 40 healthy control children (ages 2-5 years) there was a significantly higher prevalence of dental caries in HIV-infected children (P<0.05). The extent of caries correlated with the severity of HIV disease. To determine whether the immunosuppression that ensues after HIV infection could contribute to the increased caries prevalence, the concentrations of total IgA and IgA specific to cariogenic bacteria (Streptococcus mutans, Streptococcus sobrinus and Lactobacillus acidophilus) were determined in whole saliva by enzyme-linked immunosorbent assay. Levels of the same bacteria were also quantified in saliva using checkerboard DNA-DNA hybridization. A significantly increased level of total salivary IgA was found in the HIV-positive population (P < 0.05), but there were comparable titers of specific IgA to cariogenic bacteria in HIV-positive and healthy controls. The microbiological assessment also demonstrated similar levels of cariogenic microorganisms in both groups. We conclude that HIV-positive children appear to maintain the capacity to mount a mucosal immune response to cariogenic microorganisms, at least until late stages of disease.

Analysis of Variance↗

Remote glucosyltransferase-microparticle vaccine delivery induces protective immunity in the oral cavity.

Intranasally administered dental caries vaccines show significant promise for human application. Alternate mucosal routes may be required, however, to induce caries-protective salivary IgA antibody in children with respiratory diseases. Since rectal mucosa contains inductive lymphoid tissue, we hypothesized that the rectal route could be used to induce salivary immunity to mutans streptococcal glucosyltransferase (GTF), resulting in protective immunity to experimental dental caries. We first explored the ability of glucosyltransferase, incorporated into polylactide-co-glycolide (PLGA) microparticles (MP), and administered rectally together with mucosal adjuvant, to induce a salivary IgA antibody response. Groups of Sprague-Dawley rats (6/group) were immunized rectally on days 0, 7, 14 and 21 with a) GTF-MP alone, b) GTF-MP with cholera toxin, c) GTF-MP with detoxified mutant Escherichia coli toxin (dLT), or d) sham immunized with PLGA and cholera toxin. An additional group was immunized intranasally with GTF-MP alone. Saliva and nasal washes of all intranasally immunized rats contained IgA antibody to glucosyltransferase on day 28. Salivary IgA antibody was also detected in 7/12 rats rectally immunized with GTF-MP and cholera toxin or dLT, although responses were lower than those obtained by intranasal immunization. Most fecal extracts from rectally delivered GTF-MP plus cholera toxin or dLT rats contained IgA antibody to GTF-MP. Low levels of fecal IgA antibody were detected in 3/6 intranasally immunized rats and 2/6 rats rectally immunized with GTF-MP alone. We then examined the extent to which salivary IgA antibody induced by the rectal route could be protective. At 25, 31 and 38 days of age, two groups of female Sprague-Dawley rats (13/group) were rectally immunized with GTF-MP and cholera toxin or with empty microparticles and cholera toxin (sham group). A third group was intranasally immunized with GTF-MP alone. After demonstrating salivary IgA responses to GTF in most GTF-immunized rats, all animals were infected with streptomycin-resistant Streptococcus sobrinus and placed on diet 2000. After 79 days of infection, total caries on molar surfaces were lower in both rectally (7.9 +/- 1.0) and intranasally (7.1 +/- 0.9; P < 0.0.03) immunized groups compared with the sham-immunized group (11.9 +/- 1.6). Smooth surface caries were significantly lower (P < 0.05) in both rectally and intranasally immunized groups. These results support the interconnectedness of the mucosal immune system and indicate that rectal immunization with GTF-MP, together with adjuvant, or intranasal immunization with GTF-MP alone, can induce protective levels of salivary antibody in rats.

Administration, Rectal↗