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In this narrative review, the potential reasons for the high occurrence of enterococci in filled root canals are explored. The pulpless root canal appears to be a habitat for these bacteria, particularly for Enterococcus faecalis. However, re-surveying the literature in caries research, it can be concluded that, contrary to earlier belief, enterococci are rare if ever found at the advancing front of dentinal lesions. The same is the case for true primary endodontic infections, but some uncertainty remains, because the coronal seal and the history of teeth harbouring enterococci have rarely been accurately investigated. Furthermore, from longitudinal studies with a known infection at the initiation of treatment, which was carried out under controlled asepsis, it is questionable whether enterococci are as difficult to eliminate from the canal system as is commonly held. A more likely explanation for the high occurrence of enterococci in filled root canals is that they enter after treatment, but from which source? The intriguing finding in this context is that enterococci do not appear to be colonizers of the oral cavity. They are merely transient oral bacteria, unless there is a predilection site such as the unsealed necrotic or filled root canal. The origin of this infection is most likely food. Using the example of enterococci in filled root canals, this paper highlights the possible importance of transient microorganisms in the oral cavity and changes in a microenvironment that can create favourable conditions for infection.

The development of indirect immunofluorescence assays using MAbs as specific probes has made it possible to detect a variety of suspected periodontal pathogens in subgingival plaque, with exquisite sensitivity. The studies demonstrate high reproducibility of the results if different MAbs are used to assess the same bacterial species. Bacteroides forsythus, Bact. gingivalis, Bact. intermedius and Wolinella recta, but not Actinobacillus actinomycetemcomitans were found in sites of adult periodontitis with high prevalence but quite distinct degrees of colonization. The colonization levels of Bact. forsythus and Bact. gingivalis were significantly associated with the probing depth of the lesions. The investigations demonstrate the usefulness of serological analysis of plaque and indicate that several of the organisms implicated in the aetiology of periodontal diseases may be tolerated in high numbers in subgingival lesions, without periodontal destruction.

A dual-chamber dentifrice, which contains sodium fluoride (NaF) in one component and dicalcium phosphate dihydrate (dical) in the other, has been developed. The dentifrice is packaged in a dual-chamber tube and is formulated to deliver 1100 ppm F. A series of studies consisting of in vitro fluoride uptake, in vivo calcium labeling, intraoral remineralization-demineralization, and animal caries studies were performed to support the improved anticaries efficacy of this product in comparison to a sodium fluoride/silica dentifrice (NaF/silica). An in vitro fluoride uptake study comparing 1100 ppm F NaF/dical dentifrice to 1100 ppm F NaF/silica showed that NaF/dical delivered significantly more fluoride than NaF/silica, 3.72 +/- 0.36 micrograms/cm2 versus 2.41 +/- 0.10 micrograms/cm2. A 6-day in vivo brushing study with a 44Ca labeled NaF/dical dentifrice showed that calcium from dical penetrated demineralized enamel and was present in plaque up to 18 hrs since the last brushing. An intra-oral remineralization-demineralization study was performed to evaluate NaF/dical's ability to promote remineralization in comparison to three silica-based dentifrices containing 0, 250, and 1100 ppm F as NaF. The percent mineral changes after treatment were +20.44 +/- 17.14 for NaF/dical, +9.27 +/- 19.53 for 1100 ppm NaF/silica, -1.43 +/- 20.57 for 250 ppm NaF/silica, and -12.36 +/- 32.76 for 0 ppm F/silica. A statistical analysis showed that the dual-chamber NaF/dical dentifrice was significantly more effective than the 1100 ppm NaF/silica dentifrice at promoting remineralization. A rat caries study was performed to evaluate NaF/dical ability to prevent caries in comparison to 1100 ppm F NaF/silica, 250 ppm F NaF/silica, silica, and dical dentifrices. The mean smooth surface caries scores were 1.6 +/- 2.8 for NaF/dical, 5.5 +/- 6.2 for 1100 ppm F NaF/silica, 10.6 +/- 6.2 for 250 ppm F NaF/silica, 13.7 +/- 4.7 for 0 ppm F/silica, and 9.5 +/- 7.8 0 ppm F/dical. A statistical analysis showed that the the dual-chamber NaF/dical dentifrice was superior to all other treatments tested in preventing caries in rats. The dical dentifrice was significantly superior to the silica dentifrice in preventing caries, which indicates that dical alone exhibits anticaries efficacy. In conclusion, individual and cumulative results from the fluoride uptake, intra-oral remineralization-demineralization, and rat caries studies from the dual chamber NaF/dical dentifrice support the improved anticaries efficacy of this product.

This study validates an in situ model for ecological studies of dental plaque exposed to various antimicrobial agents with different modes of action on plaque bacteria. Eleven subjects wore two acrylic appliances, each containing two bovine enamel discs, during two 1-wk test periods. Using a split-mouth crossover design, the appliances were dipped twice daily for 1 min into water (control; treatment A), fluoride (26.3 mM NaF; B), zinc acetate (20.0 mM; C), or fluoride plus zinc acetate (D). Four of the subjects used also chlorhexidine diacetate (2.2 mM; E) and chlorhexidine plus fluoride (F). At the end of each period, plaque was collected from the discs, after which the microbiota were analyzed by culture, automated quantitative immunofluorescence, and a viability fluorescence stain. As compared to control, treatments B, C, and D resulted in a significant reduction of individual taxa as detected by immunofluorescence, whereas similar bacterial viability and total bacterial numbers were observed. In contrast, chlorhexidine significantly reduced bacterial viability, total cell numbers, and the abundance of most of the enumerated taxa. We conclude that this in situ model is well suited to study effects of antimicrobial agents on dental plaque ecology. Combined with viability testing, immunofluorescence is obviously superior to culture in detecting taxa-specific shifts caused by antimicrobial agents.

The present study investigated a recently developed automated image analysis technique for its applicability to the enumeration of selected bacteria in supragingival dental plaque. Following initial calibration, the system is capable to count fluorescence-labeled target cells in up to 48 samples without user interference. Test samples contained a characteristic mixture of planktonic bacteria, small almost planar bacterial aggregates, and large, virtually indisruptable clumps with cells from multiple species. Due to their complex composition, these samples provided a challenging validation step for the image analysis system. Automated enumeration of target bacteria was compared with visual counting of the fluorescence-labeled bacteria. Results are shown for six taxa (Actinomyces naeslundii, Fusobacterium nucleatum, Prevotella intermedia/Prev. nigrescens, Streptococcus gordonii/Strep. oralis/Strep. sanguis, Strep. sobrinus, and Veillonella dispar/ V. parvula) with characteristic differences in abundance, cell morphology and aggregation behavior. Results revealed good correspondence between the two enumeration techniques (correlation coefficients ranging from 0.77 to 0.92) provided that the portion of target bacteria exceeded 0.05% of the total bacterial cell number. This work demonstrates the applicability and usefulness of fully automated immunofluorescence to analyze such complex ecosystems as supragingival dental plaque.

The purpose of this study was the clinical and microbiological re-examination of dental hygienists, who, 30 months before, had shown remarkably high supragingival levels of periodontitis-associated micro-organisms. Interdental plaque was collected from the same molar sites and investigated by the same immunofluorescence assay with taxa-specific monoclonal antibodies as at the initial examination. On average, the 15 re-examined subjects showed slightly increased plaque levels but unchanged bleeding on probing scores (0.3-1.4). Pocket formation was restricted to a single subject. Prevotella intermedia/P. nigrescens and Peptostreptococcus micros were present in every plaque sample. Prevalences of Actinobacillus actinomycetemcomitans, Bacteroides forsythus and Campylobacter rectus were again between 20-40%, but some fluctuation within subjects was noted. The data confirm supragingival plaque as a natural habitat for periodontitis-associated bacteria in periodontially healthy persons, and indicate that colonization with A. actinomycetemcomitans, B. forsythus or C. rectus is mostly stable in spite of better than average personal plaque control.

Most evidence suggests that only a finite number of bacteria are responsible for dental caries and periodontal diseases. This knowledge led to the development of microbial tests which can identify suspected pathogens. Current evaluation of the diagnostic power of microbial tests has shown that they have a low sensitivity and a low prognostic value. Despite these shortcomings, there are valid indications for microbiological-based diagnosis. Salivary microbial tests for the detection of mutans streptococci and lactobacilli may be useful, for example, in young children, oligosialic patients, and orthodontic patients. These tests can be used to monitor the success of chemopreventive measures or compliance with dietary recommendations. Microbial diagnosis may also be valuable in the treatment of early-onset periodontitis or in subjects who respond poorly to periodontal therapy. The use of microbial tests to monitor the efficacy of chemotherapy or mechanical treatment is of particular interest.

Analysis of cholesterol levels in 34 forensic patients indicated significant associations with measures of the patients' perceptions of the emotional content in auditory stimuli. In particular, patients with low cholesterol showed hypersensitivity in the detection of anger and sadness The authors hypothesize the involvement of serotonin in the hypersensitivity effects. Organicity and age were ruled out as contributing factors.

The histopathology of arrested root caries lesions was examined in extracted human teeth. The main structural characteristics of arrested lesions were the completely mineralized surface area and the formation of a distinct sclerosis of the dentinal tubules. Intertubular dentin was, with the exception of the dentinal tubules, fully mineralized up to the surface. Dentinal tubules near the surface were either filled with ghosts of microorganisms or with crystals of different shapes. Sclerosis of the dentinal tubules was characterized by the presence of three different patterns of intratubular mineralization that occur in distinct regions of the zone of tubular sclerosis. The patterns were distinguishable by the type of crystals and their association with organic structures such as collagenous fibrils or odontoblast processes. It is suggested that arrested lesions are based on (1) the formation of an inner barrier that interrupts the diffusion of substrata from the pulp to invaded bacteria, (2) the formation of an outer barrier by a compact, highly mineralized surface region which blocks the diffusion of products of bacterial metabolism into dentin, and (3) an area of mineralization which extends from the outer barrier toward the root canal within demineralized dentin. The present study demonstrates the considerable potential of caries lesions in dentin to become arrested, and subsequently partially remineralized. These phenomena seem to depend on the severity of an active lesion and its location on the root surface. This should be taken into account when diagnosing root caries lesions. The potential of root caries lesions to become arrested indicates that the treatment concept of active root caries lesions should be reconsidered.

Despite intensive research, water fluoridation and, where practiced, diet control, remain the most effective methods of preventing caries. In this paper, a number of approaches discussed appear to hold promise that they may enhance the caries preventive effect of fluoride, and others, when developed and applied, may hasten the day when dental caries will cease to be a public health problem.

By using a sensitive enzyme-linked immunosorbent assay, 200 randomly selected sera from Red Cross blood donors were screened for immunoglobulin G (IgG), IgA, and IgM levels against Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius. A subgroup of 79 blood donors was clinically examined for type and extent of periodontal destruction, and serological and clinical data were subjected in all possible dual combinations to correlation analyses. The results revealed that the majority of the blood donors suffered from moderate to severe adult periodontitis, often coupled with severe gingival inflammation. No cases of localized juvenile periodontitis or rapidly progressive periodontitis were observed. The extent of periodontal destruction proved to be significantly correlated only to the IgG response levels against B. gingivalis. Corresponding correlation tests assessing the relationships of loss of attachment, bone loss, pocket depth, and papillary bleeding index with the IgG responses to A. actinomycetemcomitans were of marginal significance, while the IgG responses to B. intermedius revealed no relationship to the periodontal health status. The specific IgM responses proved to be unrelated to the clinical parameters, but interestingly, they were found to be highly correlated with each other. Specific IgA levels were frequently too low for enzyme-linked immunosorbent assay testing and, therefore, had to be exempted from statistical analyses. Assessments of the serotype specificity of strongly elevated IgG responses to A. actinomycetemcomitans disclosed no evidence for an association of a particular serotype-specific IgG response with the occurrence of adult periodontal destruction. In contrast to results of earlier studies, a number of sera were found to contain strongly elevated IgG levels against two or even all three serotypes. Although derived by an alternative approach, the reported results largely corroborate earlier observations linking only the occurrence of elevated anti-B. gingivalis IgG responses to the presence of marked periodontal lesions in adults.

Dental plaque was collected from approximately 3,500 schoolchildren, and immediately pooled and frozen. The lyophilized plaque was processed in several batches as follows:After an aqueous extraction the water-insoluble portion was further extracted with 1 N KOH. Both aqueous and alkaline extracts were further subfractionated by precipitation at different ethanol concentrations. Nature and composition of carbohydrates in sub-fractions were investigated using acid hydrolysis and end products were identified by gas liquid partition chromatography.29.6% of plaque dry weight, containing 6.9% carbohydrates, 1.2% nitrogen and more than 4% proteins, were water-soluble. The water-insoluble portion (67.1%), contained 11.3% carbohydrates and 7.4% nitrogen; in addition, 30.7% constituents insoluble in 1 N KOH were found. In all subfractions prepared by ethanol precipitations, substantial amounts of material other than carbohydrates were found. The sugar composition of hydrolysates showed glucose to be the main sugar constituent; however, small amounts of pentoses, other hexoses and disaccharides were also present. The water-insoluble matrix polysaccharides containing predominantly α-1,3 linkages were calculated to account for 1.35% of the plaque dry weight/ 5.6% of the plaque dry weight consisted of low molecular water-soluble carbohydrates. Glucose and oligosaccharides forming the bulk of this fraction were assumed to represent intermediates resulting from enzymatic breakdown of α-1,6-linked dextran. The high content of fermentable carbohydrates in dental plaque suggests that microbial activity is not limited by the supply of fermentable substrates.

Occasional clinical observations have shown extensive caries lesions in small children between 12 and 36 months of age, in some of which cases, while sugar intake history was negative, continuous use of teething jellies, lozenges or syrups was established. This prompted the hypothesis that such products were the source of the hidden fermentable sugars, and the sugar content of a number of products available on the Swiss market was therefore analyzed. Sucrose was determined enzymatically using the Boehringer sucrose/glucose UV-test. The content of other fermentable carbohydrates was checked by GLC. The use of such products should not be recommended for real or assumed pain related to the eruption of primary teeth. Among the eight products tested only one was found to be free of fermentable sugars. All other preparations contained 31-93% fermentable sugars, chiefly sucrose. Declaration of fermentable sugars in such products should be enforced by food and drug legislation.

The histopathology of human root caries was examined in extracted teeth by different optical methods. The present part of the study details the sequential stages of initial caries in both cementum and peripheral dentin. Significant differences are shown between the mechanisms operating on the various dental hard tissues during development of caries. Histologically distinguishable reaction patterns depended on both the degree of the cariogenic challenge and the respective structural features of cementum and peripheral dentin. Earliest lesions, histologically visible as small clefts traversing cementum and extending into peripheral dentin, were clinically not detectable. Cementum was stepwise destroyed until peripheral dentin became exposed. The exposed peripheral dentin was sclerosed and tubule-free. Demineralization of dentin preceded the degradation of the organic matrix. Frequently, a hypermineralized layer occurred in both cementum and exposed dentin. Its localization was not identical with the anatomical surface. Initial penetration of bacteria into peripheral dentin occurred along small clefts. Breakdown of the organic matrix was the final step in the destructive phase of the carious process.

An investigation was carried out to detect antibodies in human serum and parotid saliva against an extract showing glucosyltransferase activity derived from Streptococcus mutans OMZ 176. Antibodies were assayed by the passive haemagglutination, complement fixation and double diffusion precipitation tests. The series of 87 subjects was divided into a caries-free and treated caries (CFTC) group and an active caries (AC) group. Antibody titres were correlated with the decayed, missing and filled teeth index (DMF). Serum haemagglutinating antibodies were detected in all subjects, with titres between 1:10 and 1:640; a negative correlation was found in the CFTC group (p < 0.01) and a positive correlation in the AC group (p < 0.02). Salivary haemagglutinating antibodies were detected in titres of up to 1:64 and a positive correlation was revealed in the CFTC group between the DMF index and antibody titre (p < 0.05). In the AC group, antibody titres were significantly lower than in the CFTC group (p < 0.02) but they showed no relationship with the DMF index. Functional immune inhibition of glucosyltransferase activity was found with a rabbit antiserum and significantly lower rates of glucan formation were detected in human sera with high antibody titres than in those with low antibody titres (p < 0.05).

Small oral spirochaetes with a strict dependence on either glucuronic acid (GluA) or galacturonic acid (GalA) were isolated from European patients with periodontitis and from Chinese patients with either gingivitis or acute necrotizing ulcerative gingivitis (ANUG). Thirteen such isolates were similar phenotypically to Treponema pectinovorum ATCC 33768T and this classification was confirmed by 16S rRNA sequencing. However, four isolates differed from T. pectinovorum by their small cell size, by a prominent beta-glucuronidase activity, by a distinct protein and antigen profile, by an inability to grow on pectin as sole source of carbohydrate and by a markedly enhanced growth rate when supplied with a second carbohydrate (L-arabinose, D-galactose, D-glucose, D-fructose, D-mannitol, D-mannose, pectin, D-ribose or D-xylose) in addition to the essential GluA/GalA. By 16S rRNA sequencing these four isolates clustered in the recently described phylotype 'Smibert-2'. T. pectinovorum (14 strains) and 'Smibert-2' (four isolates with beta-glucuronidase activity) could each be subdivided into two serotypes based on immunoblot reactivity with two mAbs. Representatives of the two groups, including T. pectinovorum ATCC 33768T, showed a 1:2:1-type periplasmic flagellar arrangement. 'Smibert-2' is described as a novel species, Treponema parvum sp. nov., with isolate OMZ 833T (= ATCC 700770T) proposed as the type strain and OMZ 842 (= ATCC 700773) as reference strain for a second serotype.

RECENT evidence strongly suggests that dental caries is an infective disease caused predominantly by Streptococcus mutans1–3. This organism has two major characteristics responsible for its cariogenicity: first, in the presence of fermentable carbohydrates it is capable of rapidly producing acid to below the pH required for dissolving enamel, and second, the organism produces glucosyltransferase which is a constitutive enzyme responsible for the synthesis of extracellular polysaccharides4. These polysaccharides are glucans which are commonly referred to in the literature as dextran; they form a major component of the bacterial plaque matrix and may be responsible for adhesion of the bacterial plaque to tooth enamel5.