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PubMed2000

Caries inhibitory activity of cacao bean husk extract in in-vitro and animal experiments.

Cacao bean husk extract (CBH) was examined for inhibitory effects on the caries-inducing properties of mutans streptococci in vitro and on caries development in specific pathogen-free Sprague-Dawley rats infected with mutans streptococci. CBH reduced the growth rate of almost all oral streptococci examined, which resulted in the reduction of acid production. Furthermore, insoluble glucan synthesis by the glucosyltransferases from Streptococcus mutans MT8148R and Streptococcus sobrinus 6715 was significantly inhibited by CBH. Hence, the sucrose-dependent cell adherence of mutans streptococci was also depressed by CBH. The administration of CBH in drinking water resulted in significant reductions of caries development and dental plaque accumulation in rats infected with either Strep. sobrinus 6715 or Strep. mutans MT8148R, and the minimum cariostatic concentration was 1.0 mg/ml. These results indicate that CBH possesses powerful anticariogenic potential.

Analysis of Variance↗

PubMed1999

Effects of topical application of free and liposome-encapsulated lactoferrin and lactoperoxidase on oral microbiota and dental caries in rats.

Four groups of rats were inoculated with Streptococcus sobrinus ATCC 33478 and fed a cariogenic diet for 42 days. Topical treatment with either distilled water, sodium fluoride (0.2%), a solution containing lactoferrin and lactoperoxidase, or a solution containing liposome-encapsulated lactoferrin and liposome-encapsulated lactoperoxidase was applied at intervals for 35 days. Caries incidence in groups treated with liposome-encapsulated lactoferrin and lactoperoxidase was significantly lower than in control groups. The number of viable Strep. sobrinus and the proportion of Strep. sobrinus in the total counts were significantly higher in liposome-treated groups. Free lactoferrin and lactoperoxidase did not cause a significant reduction in caries incidence.

Administration, Oral↗

PubMed1995

Anticariogenicity of calcium phosphate complexes of tryptic casein phosphopeptides in the rat.

Casein phosphopeptides (CPP) stabilize calcium phosphate through the formation of casein-phosphopeptide amorphous calcium-phosphate complexes (CPP-CP). The ability of CPP-CP to reduce caries activity was investigated by use of specific-pathogen-free rats inoculated with Streptococcus sobrinus. The animals consumed a defined cariogenic diet free of dairy products. Solutions (100 microL) of the CPP-CP (0.1, 0.2, 0.5, 1.0% w/v) were applied to the animals' molar teeth twice daily. Other groups of animals received solutions containing 500 ppm F, the non-phosphorylated peptides of a casein tryptic digest (0.5% w/v), or the calcium-phosphate complex of a synthetic octapeptide, Ac-Glu-Ser(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-NHMe, corresponding to the common sequence in the CPP. The CPP-CP significantly reduced caries activity in a dose-response fashion, with 1.0% CPP-CP producing 55% and 46% reductions in smooth surface and fissure caries activity, respectively, being similar to that of 500 ppm F. The anticariogenic effects of CPP-CP and fluoride were additive, since animals receiving 0.5% CPP-CP plus 500 ppm F had significantly lower caries activity than those animals receiving either CPP-CP or fluoride alone. The tryptic digest of casein with the phosphopeptides selectively removed showed no anticariogenic activity. The synthetic octapeptide-calcium phosphate complex significantly reduced caries activity, confirming that this calcium-phosphate-stabilizing portion of the casein phospho-peptides is associated with anticariogenicity. The CPP-CP did not significantly affect the level of S. sobrinus in fissure plaque.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed2003

Activity of human beta-defensin 3 alone or combined with other antimicrobial agents against oral bacteria.

The in vitro activities of human beta-defensin 3 (hBD-3) alone or combined with lysozyme, metronidazole, amoxicillin, and chlorhexidine were investigated with the oral bacteria Streptococcus mutans, Streptococcus sanguinis, Streptococcus sobrinus, Lactobacillus acidophilus, Actinobacillus actinomycetemcomitans, and Porphyromonas gingivalis. hBD-3 showed bactericidal activity against all of the bacterial species tested. The bactericidal effect was enhanced when the peptide was used in combination with the antimicrobial agents mentioned above.

Aggregatibacter actinomycetemcomitans↗

PubMed1993

Acquisition of manganous ions by mutans group streptococci.

The cariogenic bacteria Streptococcus sobrinus and S. cricetus were shown to have an absolute requirement for manganous ion in order to bind glucans or to adhere to glass in the presence of sucrose. The bacteria possessed a reasonably high affinity transport system for 54Mn2+, yielding a Km of about 12 microM. The Vmax for uptake of 54Mn2+ in S. sobrinus was increased when the bacteria were grown in Mn-depleted medium, but the Km remained the same. There was no evidence for two Mn2+ uptake systems, commonly observed for many bacteria. Ions such as Ca2+, Co2+, Co3+, Cu2+, Fe2+, Fe3+, Hg2+, Mg2+, Ni2+, and Zn2+ did not inhibit the uptake of 54Mn2+ by the bacteria, although Cd2+ was a potent inhibitor. Fractionation experiments showed that manganese was distributed in protoplasts (67%) and in the cell wall (33%). Approximately 80% of the 54Mn2+ in S. sobrinus was rapidly exchangeable with nonradioactive Mn2+. Electron spin resonance experiments showed that all of the manganese was bound or restricted in mobility. Proton motive force-dissipating agents increased the acquisition of 54Mn2+ by the streptococci, probably because the wall became more negatively charged when the cell could no longer produce protons.

Bacterial Adhesion↗

PubMed1992

Noncariogenicity of maltitol in specific pathogen-free rats infected with mutans streptococci.

The effect of maltitol on caries development was examined in an experimental caries system employing specific pathogen-free (SPF) Sprague-Dawley rats. Fourteen strains of oral streptococci, including mutans streptococci, did not utilize the maltitol nor produce sufficient acid to demineralize tooth enamel. Furthermore, maltitol did not serve as a substrate for glucosyltransferases of either Streptococcus mutans MT8148R or Streptococcus sobrinus 6715 to synthesize water-insoluble glucan. Maltitol induced no significant dental caries in SPF rats infected with these mutans streptococci, and replacement of the dietary sucrose content with maltitol resulted in a trend towards caries reduction in SPF rats.

Animals↗

PubMed1999

Effect of matrix placement on furcation perforation repair.

Furcation perforations are a serious complication during endodontic treatment, and a matrix to aid placement of repair material has been recommended. This study tested the sealing ability of amalgam and Ketac silver placed with and without plaster of Paris as a matrix. A bacterial penetration technique was used to test the seal. Perforations created in the pulpal floor of extracted human mandibular molars were repaired as follows: group 1, amalgam; group 2, amalgam plus plaster as a matrix; group 3, Ketac silver; and group 4, Ketac silver plus plaster (17 teeth/group). Leakage was measured by placing bacteria (Streptococcus sobrinus) in the pulp chamber, and recording the time taken for bacterial growth in a medium bathing the root surface. A plaster of Paris matrix improved the seal with amalgam, but not with Ketac silver. Ketac silver provided the best seal, but all materials showed complete leakage within 22 days.

Calcium Sulfate↗

PubMed2002

Cariogenicity of different types of milk: an experimental study using animal model.

This study evaluated the cariogenic potential of infant formulas and cow's milk, using a high cariogenic challenge in the animal model. Sixty female Wistar rats infected with Streptococcus sobrinus and desalivated were randomly divided into 6 groups, which received ad libitum: 1) sterilized deionized distilled water (SDW) with 5% sucrose; 2) cow's milk; 3) Nan 2; 4) Nestogeno 2; 5) Ninho growth supporting; 6) SDW. Groups 1 and 6 also received essential diet NCP#2 by gavage, twice a day. After 21 days, the animals were killed and evaluated according to recovered oral microbiota and caries score by using a modified Keyes method. The analysis of the carbohydrates in the milk samples was performed using HPLC. The results were analyzed by Shapiro-Wilk and Kruskal-Wallis tests. Cow's milk had the lowest cariogenic potential compared to the other test groups, but it was not statistically different from group 6. The percentage of S. sobrinus obtained from the oral cavity of the animals was not statistically different among the groups studied, except for the SDW group. It was concluded that cow's milk was not cariogenic and infant formulas showed some cariogenic potential.

Animals↗

PubMed2003

Nested PCR for detection of mutans streptococci in dental plaque.

AIMS: Mutans streptococci such as Streptococcus mutans and Streptococcus sobrinus have been implicated in human dental caries. In an attempt to develop a rapid and sensitive method for detecting Strep. mutans and Strep. sobrinus in dental plaque, a nested PCR amplification based on the 16S rRNA gene was employed. METHODS AND RESULTS: A universal set of PCR primers for bacterial 16S rRNA gene was introduced for the first PCR, and then two sets of primers specific for the 16S rRNA gene sequences of either Strep. mutans or Strep. sobrinus were used for the second PCR. Eighteen plaque samples were analyzed, and a nested PCR was shown to be more sensitive for detecting Strep. mutans and Strep. sobrinus than direct PCR. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The 16S rRNA gene-based nested PCR method is a rapid and sensitive method for the detection of mutans streptococci, and may also be suitable for carrying out large-scale studies on the cariogenicity of mutans streptococci.

Adult↗

PubMedDate not supplied

Effect of Apis mellifera propolis from two Brazilian regions on caries development in desalivated rats.

The purpose of the present study was to evaluate the effect of Apis mellifera propolis collected from two regions of Brazil on caries development in desalivated rats. Ethanolic extracts of propolis (EEP) were prepared from crude propolis samples collected in Minas Gerais state (MG), southeastern Brazil, and Rio Grande do Sul state (RS), southern Brazil. The flavonoid composition of EEP was analyzed by high-performance thin-layer chromatography (HPTLC) and reversed-phase high-performance liquid chromatography (HPLC). For the animal study, 30 specific pathogen-free Wistar rats were infected with Streptococcus sobrinus 6715 and surgically desalivated. The rats were randomly divided into three groups which were treated with 80% ethanol (control), EEP from MG and EEP from RS. The animals were placed in a König-Höfer programmed feeder and received 17 meals of diet 2000 daily at hourly intervals. The solutions were applied on the rat molars (25 microl on molars of each quadrant) twice a day, by using graduate syringes. After 3 weeks, the animals were killed by CO(2) asphyxiation. For microbial assessment, the left jaw was removed and sonicated in 154 mM NaCl solution. Dental caries was evaluated according to Larson's modification of Keyes' system. The HPTLC patterns and HPLC profiles demonstrated that both quality and quantity of flavonoid aglycones of EEP from MG were different compared to EEP from RS. In general, it is apparent that EEP from RS contained the highest concentrations of pinocembrin, chrysin, acacetin and galangin. The group of animals treated with EEP from RS showed the lowest smooth-surface and sulcal caries scores as well as less caries severity in smooth-surface and sulcal lesions, and these data were statistically different when compared with the control group. The group treated with EEP from MG only demonstrated a significant difference in the severity of sulcal lesions when compared to the control group. The percentage of S. sobrinus was lower in the groups treated with EEP, but did not differ statistically from the control group. The results showed that the cariostatic effect of propolis depends on its composition, and consequently the region of collection of propolis samples.

Analysis of Variance↗

PubMed1998

Rapid isolation of chromosomal DNA from oral streptococci and polymerase chain reaction-oriented restriction fragment-length polymorphism analysis for genetic heterogeneity.

We have extensively modified the published method for the lysis of gram-positive bacteria to isolate chromosomal DNA from only 1 ml of oral streptococcal overnight culture. Cells were incubated with lysozyme and R Nase A in the presence of polyethylene glycol. After centrifugation, cells were lysed with sodium dodecyl sulfate and proteinase K. Following ethanol precipitation, sodium dodecyl sulfate solution was added to the residue, and the pellet was completely dispersed by incubating at 65 degrees C. The chromosome was purified by extraction over phenol and chloroform. Two regions corresponding to the ribosomal RNA (rrn) operon and the glucosyltransferase gene were amplified using the chromosome from Streptococcus mutans and Streptococcus sobrinus by polymerase chain reaction (PCR). Genetic heterogeneity was assessed by restriction fragment-length polymorphism (PCR-RFLP). The PCR-RFLP analysis readily allowed us to subtype each strain, suggesting that the strategy presented here will provide a useful tool to verify epidemiological studies at the molecular level.

Base Sequence↗

PubMed1992

Evaluation of the cariogenic potential of the intense natural sweeteners stevioside and rebaudioside A.

Stevioside and rebaudioside A, two intense natural sweeteners, that are constituents of the South American plant Stevia rebaudiana, were tested for cariogenicity in albino Sprague-Dawley rats. Sixty rat pups colonized with Streptococcus sobrinus were divided into four groups and fed stevioside, rebaudioside A or sucrose added to basal diet 2000 as follows: group 1, 30% sucrose; group 2, 0.5% stevioside; group 3, 0.5% rebaudioside A, and group 4, no addition. All four groups were sacrificed after 5 weeks. S. sobrinus counts were made and caries was evaluated according to Keyes' technique. There were no differences in food and water intake and weight gains between the four groups. There were significant differences in sulcal caries scores (p < 0.02) and S. sobrinus counts (p < 0.05) between group 1 and the other three groups. There were no significant differences between the stevioside, rebaudioside A and no-addition groups. It was concluded that neither stevioside nor rebaudioside A is cariogenic under the conditions of this study.

Animals↗

PubMed1991

Cariogenicity of Milchschnitte (Ferrero GmbH) and apple in program-fed rats.

The cariogenicity of Ferrero Milchschnitte and apple was tested in rats using a frequency-controlled feeding machine. Twelve litters of Osborne-Mendel rats orally infected with Streptococcus sobrinus OMZ-176 and Actinomyces viscosus Ny-1 were used. Four treatment groups were tested: (1) cooked starch/sucrose 1:1 as positive control; (2) Ferrero Milchschnitte; (3) apple; (4) gastric intubation only as negative control. Groups 1-3 were additionally fed by gastric intubation with essential nutrients. Rats on Milchschnitte and apple had less fissure and smooth-surface caries than rats on a starch/sucrose diet, but significantly more than the negative controls. The Milchschnitte group exhibited less plaque extent than the positive and the negative control. The apple group showed a higher plaque extent and massively extended erosions on molar surfaces. The CFU means of S. sobrinus were decreasing from treatment 1-4. Although differences between treatments were not significant, they were associated by concomitantly decreasing caries incidence. The test foods were also assessed using intraoral plaque-pH telemetry in 6 human volunteers. Both Milchschnitte and apple were acidogenic in human telemetry. They can therefore not be regarded as safe for teeth.

Animal Feed↗

PubMed2003

Mutans streptococci strains prevalence before and after cavity preparation during Atraumatic Restorative Treatment.

Critics argue that all carious dentine is not removed from the hand-prepared cavity during the Atraumatic Restorative Treatment (ART) procedure, and that the caries process is soon resumed. The aim of this study was to determine the effectiveness of ART in removing carious tissue, by investigating the numbers of mutans streptococci and lactobacilli, with emphasis on the prevalence of Streptococcus mutans and Streptococcus sobrinus strains before, and after ART treatment of dental caries. Two microbiology samples were collected. The first sample was removed from the centre of the carious lesion at the enamel-dentine junction, and the second was collected from the centre of the hard cavity wall above the pulp, after the soft infected dentine had been manually removed. A total of 71 mutans streptococci isolates from 31 children and 40 carious teeth were subcultured, biochemically characterised and genotyped by the arbitrarily primed polymerase chain reaction (AP-PCR). Results showed a significant decrease in TVC (P<0.0001), mutans streptococci (P < 0.0001) and lactobacilli (P = 0.0002) after cavity preparation. AP-PCR identified S. mutans strains that were undetectable during biotyping, and divided clinical isolates into two main clusters. In all, 63% (45/71) of isolates from the carious lesions comprised S. mutans strains. After cavity preparation, this was reduced to 35% (25/71), of which 30% (21/71) were S. mutans and the remaining 6% (4/71) S. sobrinus strains. The number of mutans streptococci strains was below detectable levels in 19 of the prepared cavities. The significant decrease in bacteria after manual cavity preparation demonstrates the reliability of a standardized ART technique, yet the presence of S. mutans strains shows that the effectiveness of the ART procedure can vary during treatment and between dental practitioners.

Child↗

PubMed1997

Inhibition by hop bract polyphenols of cellular adherence and water-insoluble glucan synthesis of mutans streptococci.

The inhibitory effect of hop bract polyphenols (HBP) on cariogenic streptococci was investigated. It was found that the high molecular weight polyphenol (estimated about 36,000-40,000) inhibited the cellular adherence of Streptococcus mutans MT8148 (serotype C) and Streptococcus sobrinus ATCC 33478 (serotype g) at much small concentrations than the polyphenols extracted from oolong tea or green tea leaves. Furthermore, HBP also inhibited the action of glucosyltransferase, which was involved in the water-insoluble glucan synthesis, but did not suppress the growth and the acid production of the bacteria. These results suggest that HBP would be a candidate to act against dental caries caused by Mutans Streptococci.

Bacterial Adhesion↗

PubMed1997

Cariostatic effect of aspartame in rats.

This study examined the effects of aspartame (APM) on caries and the Streptococcus sobrinus population. Six groups of rats (20 in each group) colonized with S. sobrinus were fed varying amounts of sucrose (SU) and APM in their diets as follows: 1-0.15% APM, 2-0.30% APM, 3-30% SU, 4-30% SU + 0.15% APM, 5-50% SU and 6-50% + 0.15% APM. Ten from each group were sacrificed at 6 weeks and 10 at 12 weeks. S. sobrinus populations at both intervals were negligible in groups 1 and 2 and had no differences between groups 3, 4, 5 and 6. No caries were found in groups 1 and 2 Animals fed SU plus APM had significantly lower caries than animals fed the same amounts of SU (p = 0.001-0.0002). We conclude that APM is noncariogenic and anticariogenic.

Analysis of Variance↗

PubMed1996

Incorporation of caseinoglycomacropeptide and caseinophosphopeptide into the salivary pellicle inhibits adherence of mutans streptococci.

The protective effects of milk and milk products against dental caries have been demonstrated in many animal studies. We have shown that this effect was mediated by micellar casein or caseinopeptide derivatives. A reduction in the Streptococcus sobrinus population in the oral microbiota of animals fed diets supplemented with these milk components was consistently observed. A possible explanation for these findings is that milk components are incorporated into the salivary pellicle, thereby reducing the adherence of S. sobrinus. This hypothesis was tested in vitro by the incubation of bovine enamel discs with unstimulated saliva. The resulting pellicle was washed and incubated with caseinoglycomacropeptide (CGMP) and/or caseinophosphopeptide (CPP) labeled with 17- and 12-nm gold particles. All samples were prepared for electron microscopy by high-pressure freezing followed by freeze-substitution. It was demonstrated by high-resolution scanning electron microscopy with back-scattered electron imaging, as well as by transmission electron microscopy, that both peptides were incorporated into the pellicle in exchange for albumin, confirming previous findings. This protein was identified with a mouse anti-human serum albumin followed by goat anti-mouse IgG labeled with 25-nm gold particles. Incorporation of CGMP and/or CPP into salivary pellicles reduced the adherence of both S. sobrinus and S. mutans significantly. It is suggested that the calcium and phosphate-rich micellar casein or caseinopeptides are incorporated into the pellicle. The resulting ecological shifts, together with the increased remineralization potential of this biofilm, may explain its modified cariogenic potential.

Adult↗

PubMed1996

[The effects of lipoteichoic acid and calcium on the adherence of oral streptococci in vitro].

In this study, the effects of lipoteichoic acid (LTA) and calcium on adherence of oral streptococci in vitro were investigated. The results revealed that the adherence of streptococcus sobrinus to a glass surface was increased by the addition of calcium, in the condition of the same calcium ion concentration, LTA inhibited adherence, suggesting that LTA and the bacterial cell surface LTA competed for the same site of calcium ion, providing some evidence to support calcium-bridging theory. The result also showed that calcium ion not only influenced sucrose-independent adherence, but also infulenced sucrose-dependent adherence. The effect of calcium ion on the adherence has a threshold, over which the adherence was reduced.

Bacterial Adhesion↗