Antimicrobial activity of calcium hydroxide liners on Streptococcus sanguis and S. mutans

The antimicrobial activity of 10 calcium hydroxide liners on Streptococcus sanguis and S. mutans was studied. One hundred plates of tryticose soy agar-sheep red blood cells (10%), each with four distinct 3 mm diameter wells, were divided into two groups of 50. One group was inoculated with S. sanguis and the second group with S. mutans. A 4 mg liner was placed in each of three wells; the fourth well was left empty as a control. Plates were incubated at 37 degrees C and observed at 24 hours, 48 hours, and 7 days. All liners inhibited growth of both organisms. The inhibitory zones for S. mutans were larger than inhibitory zones for S. sanguis for all tested liners. Life Fast Set material had significantly less inhibitory effect on S. mutans. VLC Dycal, Life, and Life Fast Set materials had significantly less inhibitory effect on S. sanguis at p = 0.05.     

Antimicrobial activity of four root canal sealers against endodontic pathogens

The antibacterial effects of various types of widely used endodontic sealers have not been compared systematically on facultative or obligate anaerobic endodontic pathogens. The aim of this study was to evaluate the antimicrobial properties of four commonly used endodontic sealers: two epoxy-resin-based sealers (AH26, AH plus), one zinc-oxide eugenol-based sealer (N2), and one calcium hydroxide-based sealer (Sealapex). The testing microbes were four facultative anaerobic species (Streptococcus mutans, Streptococcus sanguis, Escherichia coli, and Staphylococcus aureus) and four obligate anaerobic species (Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum, and Prevotella intermedia). The freshly mixed sealers were placed into the prepared wells of agar plates inoculated with the test microorganisms. After varying periods of incubation (2 days for facultative anaerobic species and 7 days for obligate anaerobic species), the zones of growth inhibition were observed and measured. All the sealers were distinctly different from each other in their antimicrobial activity. The sealers showed different inhibitory effects depending on the types and bacterial strains. N2 containing formaldehyde and eugenol proved to be the most effective against the microorganisms. The extreme antimicrobial potency of this root canal sealer must be weighted against its pronounced tissue toxic effect.     

The antimicrobial effect of various endodontic sealers

The antimicrobial activity of an endodontic sealer can be helpful in destroying any remaining root canal microbes. Therefore it was decided to test the antimicrobial activity of several commonly used endodontic sealers. The sealers used were Grossman's sealer, Tubliseal, Calciobiotic, Sealapex, Hypocal, eucapercha, Nogenol, and AH26. Also tested were dry calcium hydroxide powder, calcium hydroxide mixed with saline, and a Teflon formulation. The microbes used were Streptococcus mutans (a gram-positive microaerophile). Staphylococcus aureus (a gram-positive facultative anaerobe), and Bacteroides endodontalis (a gram-negative obligate anaerobe). The freshly mixed sealers were placed into the prepared wells of agar plates inoculated with the test microorganisms. After varying periods of incubation, the zones of inhibition of bacterial growth were observed and measured. Grossman's sealer had the greatest overall antibacterial activity. However, AH26 had the greatest activity against B. endodontalis. The zinc oxide-eugenol-based sealers had more antimicrobial activity than either the calcium hydroxide-based sealers or eucapercha.     

Antineoplastic agents inhibit the growth of Streptococcus mutans and Streptococcus sanguis in vitro

The effect of methotrexate (MTX) and doxorubicin on the growth, metabolism and ultrastructure of Streptococcus mutans and Streptococcus sanguis was studied in vitro. Both anticancer drugs exerted an inhibitory effect on the oral streptococci. MTX was more inhibitory than doxorubicin. The minimum inhibitory concentrations (MICs) of MTX to S. mutans were 0.25-2.5 micrograms/ml and that of doxorubicin 0.2 mg/ml. The MICs of MTX and doxorubicin to S. sanguis were 0.025 micrograms/ml and 2.0-0.02 mg/ml, respectively. When saliva samples of patients with malignant tumors receiving various doses of MTX were analyzed, MTX was found to be secreted into the oral cavity at concentrations ranging from 0.014 to 4.486 micrograms/ml. The saliva of these patients was also found to inhibit the growth of S. mutans, and the inhibition zones were in accordance with the MIC values observed. The results suggest that anticancer therapy must be taken into account when the salivary microbiological findings of cancer patients are interpreted.     

Effect of penicillin on Streptococcus mutans, Streptococcus sanguis and lactobacilli in hamsters and in man

The effect of penicillin on the number of oral Streptococcus mutans, Streptococcus sanguis and lactobacilli in hamsters and in man was investigated. This is of interest as S. mutans and lactobacilli are involved in the carious process while S. sanguis is not. Hamsters infected with both S. mutans and S. sanguis or only S. sanguis received penicillin in their drinking water for 14 d. The treatment reduced the proportion of S. mutans and S. sanguis in dental plaque to undetectable levels. After the penicillin treatment the population of S. mutans and S. sanguis gradually increased. In man, the effect of oral penicillin therapy was examined in 21 adults with more than 2 X 10(5) S. mutans per ml saliva. The penicillin treatment had almost no effect on the numbers of S. sanguis and lactobacilli, but a pronounced decrease in the number of S. mutans was observed. The duration of this effect, however, was short. Consequently, such treatment alone is of limited value for the control of the oral infection of these microorganisms.     

Growth inhibition of Streptococcus anginosus (milleri) by three calcium hydroxide sealers and one zinc oxide-eugenol sealer

The inhibition of growth of Streptococcus anginosus (milleri) by three calcium hydroxide sealers--calciobiotic root canal sealer (Hygienic Corporation, Akron, OH), Sealapex (Kerr Division, Sybron Corporation, Romulus, MI), and Apexit (Vivadent, USA, Amherst, NY)--was compared with a zinc oxide-eugenol sealer: Roth (Roth International, Chicago, IL). Sixteen brain heart infusion agar plates were inoculated with S. anginosus. Each plate was divided into five separate areas. In each area, a 0.1 ml droplet of a given sealer was placed such that each plate had five areas with the same sealer. There were four plates (for each of the four sealers) for a total of 20 observation areas to evaluate each sealer. The plates were incubated at 37 degrees C in a 5% CO2 atmosphere. After varying periods of incubation, the zones of inhibition of bacterial growth were observed and measured. Roth sealer had a statistically significant larger mean zone of inhibition than the calcium hydroxide sealers. However, all of the sealers exhibited clinically relevant antimicrobial activity. It is likely that the eugenol in the Roth sealer is responsible for its greater antimicrobial activity.     

Technical note: antimicrobial action of glass-ionomer lining cement on S. sanguis and S. mutans

There are a number of glass-ionomer lining cements available to the profession. It is generally agreed that the glass-ionomer cements demonstrate reasonably good biocompatibility. The purpose of this study was to evaluate the antimicrobial activity of six glass-ionomer cements on S. sanguis and S. mutans. Forty-eight plates of TSA-SRBC (10%), each with 3.0-mm-diam. wells, were divided into two groups (1 and 2) of 24 each. Group 1 was inoculated with S. sanguis (10558) and Group 2 with S. mutans (6715-13 w.t.). Each well was filled with 4.0 mg of one of the following liners: Glassic (A), Shofu lining cement (B), GC lining cement (C), Ever Bond (D), Gingiva Seal (E), and Ketac Bond (F); the empty wells served as control. Plates were incubated under microaerophilic conditions at 37 degrees C, and checked daily over seven days for inhibition zones. Average inhibition zones (mm) at 7 days were as follows: Group 1-1A = 10.00, 1B = 7.83, 1C = 18.50, 1D = 13.00, 1E = 8.67, and 1F = 9.33; Group 2-2A = 3.08, 2B = 10.25, 2C = 12.75, 2D = 15.58, 2E = 8.83, and 2F = 3.20. All control wells were 0.0 GC liner and Ever Bond showed significantly greater overall inhibition of microbial growth than did other tested liners (p less than 0.5). A one-way ANOVA and SNK test showed that all tested liners were significantly more toxic toward S. sanguis than toward S. mutans (p less than 0.05).     

An agar diffusion study comparing the antimicrobial activity of Nanoseal with some other endodontic sealers

The aim of this study was to evaluate the antimicrobial activity of a new experimental nano‐hydroxyapatite epoxy resin‐based sealer (Nanoseal) with several other commercially available sealers; AH26, Tubliseal, Sealapex and Roekoseal against Enterococcus faecalis, Pseudomonas aeruginosa, Streptococcus mutans, Streptococcus sobrinus and Escherichia coli for up to 7 days. Agar diffusion was used in this study. Fifty Muller‐Hinton agar plates were prepared and divided into five experimental groups (n = 10), for each micro‐organism. Another 10 agar plates were used as positive and negative controls. Endodontic sealers were tested against each micro‐organism. Inhibition zones produced were recorded. The results of this study showed that all test materials exhibited inhibition zones towards the tested micro‐organisms for 7 days except for Roekoseal, which showed no inhibition zones. Nanoseal and AH26 exhibited similar zones of inhibition. Significant difference was found between Nanoseal and the other tested sealers (P < 0.001).     

Effect of penicillin on Streptococcus mutans, Streptococcus sanguis and lactobacilli in hamsters and in man

Abstract – The effect of penicillin on the numbers of oral Streptococcus mutans, Streptococcus sanguis and lactobadlii in hamsters and in man was investigated. This is of interest as S. mutans and lactobacilli are involved in the carious process while S. sanguis is not. Hamsters infected with both S. mutans and S. sanguis or only S. sanguis received penicillin in their drinking water for 14 d. The treatment reduced the proportion of S. mutants and S. sanguis in dental plaque to undetectable levels. After the penicillin treatment the population of S. mutans and S. sanguis gradually increased. In man, the effect of oral penicillin therapy was examined in 21 adults with more than 2 × 10⁵ S. mutans per ml saliva. The penicillin treatment had almost no effect on the numbers of S. sanguis and lactobacilli, but a pronounced decrease in the number of S. mutans was observed. The duration of this effect, however, was short. Consequently, such treatment alone is of limited value for the control of the oral infection of these microorganisms.     

The effect of newly developed root canal sealers on rat dental pulp cells in primary culture

Cytotoxicity was compared between new root canal sealers (New A, New B, and New B-2) and conventional root canal sealers (AH26, Diaket, Canals, Tubi-Seal, and Sealapex). Dental pulp cells of rats, obtained through primary cell culture, were used. In the experiments with the freshly prepared sealers, the incorporation of [3H]thymidine into cellular DNA was analyzed. In the experiments with the set sealers, the morphological changes of cells were microscopically examined. In the experiments with the fresh sealers, AH26 and Diaket showed strong inhibitive effects on the DNA synthesis, whereas no such effects were noted with the new sealers and Sealapex. On the other hand, in the experiments with the set sealers, the new sealers and Diaket exerted little influence on the cells. However, other sealers had considerable toxicity. These results suggest that the new sealers were the least toxic in vitro, compared with five conventional sealers.     

Selective effects of histidine-rich polypeptides on the aggregation and viability of Streptococcus mutans and Streptococcus sanguis

Enriched preparations of histidine-rich polypeptides (HRPs) and isolated HRP pairs (1-2, 3-4 and 5-6) degrade in the presence of fresh autologous whole saliva to a series of low-molecular-weight cationic peptides (HRPs 6a-c and 7). Analysis of the HRPs during degradation indicates that: HRP 1 is not the parent molecule of the HRPs; the HRP pairs do not convert to each other in a cascade-like sequence in saliva; and the HRPs can be separated into 2 groups consisting of HRPs 1-2 and 3-7. Preparations containing HRPs 1-7, 1-2, and 3-7 were obtained by fractionation and separation on Bio-Rex 70, and tested for aggregating and antibacterial effects against Streptococcus mutans BHT, S. mutans GS-5 and Streptococcus sanguis G9B. HRPs 1-2 had significant aggregating effects on all 3 strains but the other HRPs had little to no agglutinating ability. The HRPs did not inhibit the growth of S. sanguis, and HRPs 1-2 enhanced its growth. No growth enhancement by the HRPs was observed for the 2 S. mutans strains. However, significant bacterial inhibition of the S. mutans strains was noted after incubation with HRPs 3-7. The data suggest that the dissimilar effects of HRPs 1-2 and 3-7 may be of importance in the colonization and growth of S. mutans and S. sanguis in vivo.     

Effect of chlorhexidine on the relative proportions of Streptococcus mutans and Streptococcus sanguis in hamster plaque

abstract — The effect of chlorhexidine on the proportions of Streptococcus mutans and Streptococcus sanguis in plaque was studied in hamsters fed a diet containing 28% sucrose. In animals given chlorhexidine in their drinking water for 10 d a decrease in the population of S. mutans and an increase of S. sanguis occurred in the plaque. Following the removal of chlorhexidine the population of S. mutans increased again in the presence of sucrose and the number of S. sanguis returned to initial values. When animals were given a sucrose-free diet the low proportion of S. mutans observed following the short-term chlorhexidine period persisted. These data indicate that there is an inverse relationship between the number of S. sanguis and S. mutans in plaque and that the sensitivity in vivo of S. mutans to chlorhexidine can be used to suppress the population of S. mutans with a concomitant rise in the proportion of S. sanguis .     

Effect of chlorhexidine on the relative proportions of Streptococcus mutans and Streptococcus sanguis in hamster plaque.

The effect of chlorhexidine on the proportions of Streptococcus mutans and Streptococcus sanguis in plaque was studied in hamsters fed a diet containing 28% sucrose. In animals given chlorhexidine in their drinking water for 10 d a decrease in the population of S. mutans and an increase of S. sanguis occurred in the plaque. Following the removal of chlorhexidine the population of S. mutans increased again in the presence of sucrose and the number of S. sanguis returned to initial values. When animals were given a sucrose-free diet the low proportion of S. mutans observed following the short-term chlorhexidine period persisted. These data indicate that there is an inverse relationship between the number of S. sanguis and S. mutans in plaque and that the sensitivity in vivo of S. mutans to chlorheximide can be used to suppress the population of S. mutans with a concomitant rise in the proportion of S. sanguis.     

Water-soluble glucans from Streptococcus mutans strain Ing-Britt as an energy source for bacterial growth

Two fractions of water-soluble glucans with different molecular weight were produced by Streptococcus mutans IB. The larger glucan had a molecular weight of about 40,000. The molecular weight of the small glucan was estimated to be 4,100 by gel filtration chromatography and by biochemical methods. These glucans were tested for their ability to initiate and support growth of S. mutans IB and Streptococcus sanguis 903. S. sanguis 903 could grow with the low molecular weight glucan at a reduced rate compared with glucose. S. mutans IB could not utilize any of the glucans. No endo- or exo-glucanase activities could be detected in culture supernatants of any of the strains. In addition to maltose S. sanguis 903 could also utilize maltotriose, maltopentaose and maltoheptaose for growth while S. mutans IB could not grow with maltopentaose or maltoheptaose.     

The effect of tubular penetration of root canal sealers on dye microleakage

An in vitro study of a possible correlation between penetration of dentinal tubules by four root canal sealers and microleakage of external fluids into the canal was done using a dye leakage method and scanning electron microscopy. The root canals of 45 teeth were instrumented and the smear layer removed prior to obturation of root canals with gutta-percha and one of four sealers: Diaket, Endomethasone, CRCS or Ketac-Endo. The extent of leakage was scored after immersion in India ink for 72 h. The same specimens were also used for scanning electron microscopic evaluation. There was a statistical difference in leakage patterns between the groups ( P <0.05). Diaket had lower microleakage scores than the other sealers ( P <0.05). When the scores for penetration of sealers into the tubules were analysed, Ketac-Endo demonstrated the least penetration ( P <0.01). There appeared to be a converse relation between tubular penetration and dye leakage, but the correlation was not statistically significant ( P <0.05).     

In-vitro adherence of oral streptococci in the presence of sucrose and its relationship to cariogenicity in the rat

Streptococcus sanguis I gave a significantly greater percentage coverage (cell-pellicle attachment) of saliva-coated glass in the presence of sucrose than did Strep. sanguis II (p less than 0.025), and both these gave greater percentage coverages than the other species tested, between which no significant differences were noted. There was a large number of significant differences in clump size (cell-cell attachment) between species-pairs. Among the mutans streptococci, there were significant differences in the percentage coverage between Streptococcus rattus/Streptococcus mutans (p less than 0.05) and Strep. rattus/Streptococcus sobrinus (p less than 0.01), and in clump size between all species-pairs with the exception of Streptococcus cricetus or Strep. sobrinus and Strep. mutans. The rank order of species in relation to fissure caries was mutans streptococci greater than Streptococcus salivarius greater than Streptococcus milleri greater than Strep. sanguis greater than Streptococcus faecalis greater than Streptococcus mitis greater than Streptococcus lactis. There was a significant correlation between percentage coverage in vitro and fissure caries in vivo for strains of Strep. sanguis (p less than 0.05) and pooled strains of Strep. mitis and Strep. sanguis (p less than 0.01). On comparing data for adherence in the presence or the absence of sucrose, the sugar had no effect on the percentage coverage of the seven species tested, but significantly increased the clump size of the mutans streptococci (p less than 0.01) and Strep. sanguis (p less than 0.05).     

Metabolism of 17B-estradiol by oral Streptococcus mutans, Streptococcus sanguis, Bacillus cereus and Candida albicans

The ability of oral isolates of Streptococcus mutans, Streptococcus sanguis, Bacillus cereus and Candida albicans to metabolize 17 beta-estradiol was analyzed by thin-layer chromatography using 4-14C-estradiol as the substrate. All test microorganisms metabolized 17 beta-estradiol. Estrone was the main metabolic product, except for B. cereus, which converted 17 beta-estradiol mainly to unidentified, more polar metabolites. S. sanguis and B. cereus had the most active metabolism but S. mutans and C. albicans were also active.     

A simplified diagnostic system for cultural detection and enumeration of Streptococcus mutans

A simple dip-slide test (Cariescreen SM) based on MSB selective agar was devised for detection and quantitation of Streptococcus mutans in oral samples. For this test, a bacitracin tablet is dissolved in a vial containing buffered saline diluent. Paraffin-stimulated saliva is collected in this diluent vial. A dip slide which incorporates a modified MSB agar (minus bacitracin) is immersed briefly in the diluted saliva. After addition of a CO2-generating tablet, the screw-cap dip slide is closed tightly in the vial and incubated for two days at 37 degrees C and one day at room temperature. S. mutans populations in saliva are estimated by comparison with a colony density chart. Growth of reference strains of S. mutans was equivalent on Cariescreen SM dip slides and on MSB agar plates. Reference strains of Streptococcus sanguis, Streptococcus salivarius, Streptococcus mitis, and Streptococcus milleri did not grow on Cariescreen SM dip slides. Aliquots of saliva from 50 schoolchildren and 51 adults were tested by the dip-slide method and by conventional plating methods in MSB agar. Very good correlation (0.93) between the two methods was obtained. This simplified S. mutans detection system is suitable for use by clinical personnel in dental clinics or other non-laboratory settings for identification of subjects potentially at risk for caries.     

Effects of acidification on growth and glycolysis of Streptococcus sanguis and Streptococcus mutans

After carbohydrate intake, pH in dental plaque decreases rapidly and reaches about 4 within a few minutes. The acidification not only promotes demineralization of tooth surface but can also cause damage to bacteria in dental plaque. We, therefore, investigated the effect of acidification on the dental plaque bacteria Streptococcus sanguis and Streptococcus mutans. At pH 4.0 and 4.2, both growth and glycolytic activities in these streptococci were repressed. Prolonged acidification (for 60 min at pH 4.0) not only repressed both growth and glycolytic activities but also impaired them in S. sanguis cells with concomitant inactivation of the glycolytic enzymes, hexokinase, phosphofructokinase, glyceraldehyde-phosphate dehydrogenase and enolase. The impaired abilities of glycolysis and growth recovered following incubation at pH 7.0 for 80–90 min, and this was accompanied by reactivation of the glycolytic enzymes. On the other hand, these impairments were not observed in S. mutans cells exposed to prolonged acidification. These results indicate that the low pH frequently occurring in dental plaque may transiently impair streptococcal glycolysis and growth and that S. mutans is more durable to the acidification than S. sanguis.     

Streptococcus mutans in plaque and saliva after mouthrinsing with SnF2

Abstract – Mouthrinses with SnF2 in 11 subjects significantly reduced ( P <0.01) the total colony forming units (CFU) count and the numbers of Streptococcus mutans and Streptococcus sanguis in plaque. The numbers of S. mutans and S. sanguis were significantly more reduced than was the total CFU count. After rinsing with SnF2 S. sanguis was present in 97% and S. mutans in only 42% of plaque samples from tooth surfaces where they were detected after rinsing with water. SnF2 also significantly reduced ( P <0.01) the number of S. mutans in saliva. Mouthrinses with NaF did not markedly affect the number of S. mutans either in plaque or in saliva.