I. Kleinberg

Oral mucosal wetness in hypo- and normosalivators

After a person swallows, a film of residual saliva covers the oral hard- and soft-tissue surfaces. Here, the thickness of this film was measured at 11 selected mucosal surfaces on each side of the mouth (22 sites total) in two groups of dry-mouth and one group of normal individuals. Each group contained 25 individuals; one of the dry-mouth groups had resting flow rates < or = 0.1 ml/min while the other and the normal had flow rates above that. Residual saliva thickness was determined by placing frying-pan-shaped filter-paper strips (Sialopaper) against the mucosa at each site for 5 s and measuring the saliva volume collected with a modified Periotron 6000 micro-moisture meter; the thickness was then calculated by dividing the collected saliva volume by the strip area. The two groups with dry-mouth symptoms had mean resting (unstimulated) saliva flow rates of 0.04 and 0.19 ml/min and mean mucosal saliva thicknesses of 22.4 and 27.8 microns, respectively. The control group had a higher mean saliva flow rate of 0.39 ml/min and mucosal saliva thickness of 41.8 microns. As was observed in a previous study on normosalivators, the various sites had a characteristic pattern of wetness, with the hard palate and lips the least moist regions. In this study, these observations, were also true in the two dry-mouth groups. Lower resting saliva flow rates were associated with lower mucosal thickness of saliva and with dryness symptoms becoming evident when hyposalivation was below about 0.1-0.2 ml/min. The characteristic pattern of mucosal wetness was not affected by saliva flow rate. As saliva readily collects in the floor of the mouth and is then spread over other mucosal surfaces upon swallowing, it was suggested that hyposalivation could also lead to the dryness symptoms because there was not enough saliva to cover the various oral surfaces, especially the palate and the lips. In this regard, a critical level of moisture was proposed as necessary to protect vulnerable mucosal surfaces from becoming dry. Lower resting saliva flow rates and correspondingly lower mucosal wetness were also associated with a more acidic salivary pH, which was shown earlier to be associated with lower dental plaque pH.

Bacteria in human mouths involved in the production and utilization of hydrogen peroxide

Earlier studies have demonstrated that pure cultures of oral streptococci produce hydrogen peroxide but none has found any free peroxide in dental plaque or salivary sediment despite streptococci being major components of their mixed bacterial populations. The absence of peroxide in plaque and sediment could be due to the dominance of its destruction over its formation by bacterial constituents. To identify which of the oral bacteria might be involved in such a possibility, pure cultures of 27 different oral bacteria were surveyed (as well as dental plaque and sediment) for their peroxide-forming and peroxide-removing capabilities. Peroxide production was measured for each of the pure cultures by incubation with glucose at low and high substrate concentrations (2.8 and 28.0 mM) for 4 h and with the pH kept at 7.0 by a pH-stat. Removal of hydrogen peroxide was assessed in similar experiments where peroxide at 0, 29.4, 147.2 or 294.4 mM [0, 0.1, 0.5 and 1% (w/v)] replaced the glucose. Hydrogen peroxide formation was seen with only three of the bacteria tested, Streptococcus sanguis I and II (sanguis and oralis), and Strep. mitior (mitis biotype I); levels of hydrogen peroxide between 2.2 and 9.8 mM were produced when these micro-organisms were grown aerobically and 1.1 and 3.9 mM when grown anaerobically. Earlier reports indicate that such levels were usually sufficient to inhibit the growth of many plaque bacteria. The amounts formed were similar at the two glucose levels tested, suggesting that maximum peroxide production is reached at low glucose concentration. None of the three peroxide-producing organisms was able to utilize hydrogen peroxide but five of the other 24 tested, Neisseria sicca, Haemophilus segnis, H. parainfluenzae, Actinomyces viscosus and Staphylococcus epidermidis, could readily do so, as could the mixed bacteria in salivary sediment and dental plaque, both of which contain relatively high numbers of these peroxide-utilizing micro-organisms. The ability of the bacteria in plaque and sediment to degrade hydrogen peroxide was considerable and extremely rapid; peroxide removal and usually complete within the first 15 min of the incubation even when its initial level was as high as 294.4 mM. This almost overwhelming ability to remove peroxide was confirmed when peroxide-producing and -using cultures were mixed and when each of eight salivary sediments was incubated with glucose and with peroxide at concentrations up to 294.4 mM. In the glucose incubations, no hydrogen peroxide was observed, indicating dominance of microbial peroxide removers over hydrogen peroxide producers.(ABSTRACT TRUNCATED AT 400 WORDS)

Effect of orthodontic band placement on the chemical composition of human incisor tooth plaque

Abstract The placement of orthodontic bands reduced the pH, calcium and phosphorus levels and increased carbohydrate levels in the incisor plaques of 13 adolescent subjects. Maxillary plaques showed lower pH, Ca and P levels and a tendency for higher carbohydrate levels than corresponding mandibular plaques. The results are consistent with the view that pH influences plaque composition. They also provide an additional explanation concerning the trapping of food and plaque which increases dental caries susceptibility after the placement of orthodontic bands.

Arginolytic and ureolytic activities of pure cultures of human oral bacteria and their effects on the pH response of salivary sediment and dental plaque in vitro.

Thirty-nine different microorganisms commonly found in supragingival plaque and salivary sediment were screened for their ability to raise the pH by producing base from arginine, lysylarginine and urea. Only Actinomyces naeslundii and Staphylococcus epidermidis showed significant pH-rise activity with all three compounds. Eleven bacteria demonstrated such activity with arginine and lysylarginine but not with urea. Only one, Actinomyces viscosus, produced a pH-rise with urea but not with the two arginine compounds. The remaining 26 bacteria showed little or no base-forming activity with any of the three test substrates. The ability of the different oral bacteria to produce base (especially from urea) was a less universal function than their ability to produce acid from fermentable carbohydrate. Substituting pure cultures of arginolytic or non-arginolytic bacteria for portions of the mixed bacterial populations of plaque or sediment in test incubations containing glucose and arginine altered their ability to produce pH-fall-pH-rise responses shaped like those of the Stephen curve in vivo. In general, addition of arginolytic bacteria made these in vitro pH responses less acidic, whereas addition of non-arginolytic bacteria made the responses more acidic. Because of the relatively high arginolytic activity of the plaque harvested in this study, the effect of adding non-arginolytic bacteria was more readily seen than the converse. Similar changes in levels of ureolytic microorganisms and incubation with glucose and urea had little effect on sediment or plaque being able to produce a pH-fall-pH-rise type of response. When increasing proportions of the mixed bacteria in salivary sediment were replaced with the highly cariogenic Lactobacillus casei or Streptococcus mutans, the pH minimum became slightly more acidic and then slightly more alkaline, whereas the pH-rise became progressively and significantly less. Thus arginolytic bacteria have a different and greater effect on shaping the pH response of plaque or sediment than ureolytic bacteria. A large change in the proportions of arginolytic or non-arginolytic microorganisms may be needed to make a plaque microflora potentially non-cariogenic or cariogenic, respectively.

A comparison of the microbial compositions of pooled human dental plaque and salivary sediment

Abstract The comparison in 12 subjects included Actinomyces, Veillonella, fusobacteria, Neisseria, yeasts, lactobacilli, total streptococci, Streptococcus mitis, Streptococcus salivarius, Streptococcus mutans, Streptococcus sanguis and total bacteria grown aerobically and anaerobically. Plaque contained more bacteria than salivary sediment per unit mass which was attributed mainly to the presence of more non-viable epithelial cells in salivary sediment than in plaque. The relative numbers of each species were similar except that Strep, sanguis was higher in plaque than sediment. The contribution of plaque to salivary sediment during its collection was estimated by comparing the volume of sediment collected before and after tooth-brushing or dental prophylaxis. Brushing reduced the sediment collected by 30 per cent and the dental prophylaxis decreased it by about 20 per cent. It was thus evident that (i) dental plaque is a significant but not the major source of bacteria for salivary sediment and (ii) pooled dental plaque and salivary sediment have microbial compositions much closer than previously thought.

Measurement and comparison of the residual saliva on various oral mucosal and dentition surfaces in humans

Using a paper-strip absorption method, the amounts of residual saliva on 20 soft-tissue sites in different regions of the mouths of 20 individuals were surveyed once in the morning after a 12-h fast and again approx. 1-2 h after lunch. After swallowing, saliva at each site was immediately collected on filter-paper strips in a dipstick fashion for 5 s and the volumes were measured electronically with a Periotron micro-moisture meter. A clear pattern of wetness was evident and was almost identical for fasting and postprandial determinations. The hard palate and labial mucosa were covered with the least residual saliva; the floor of the mouth and back of the tongue were the wettest. In the same 20 participants, the amounts of residual saliva on various dentition sites were next measured and, as expected, much higher residual amounts were found in approximal embrasures and occlusal fossae than on adjacent facial or lingual smooth areas. Molars gave higher values than premolar and incisor embrasures. To relate residual saliva dipstick volumes to saliva thickness values, filter-paper strips were applied flat against the same mucosal or dentition surfaces in 10 of the participants, and the volume of the saliva absorbed was measured electronically as before. As the areas of the strips used were known, saliva thicknesses could be calculated. These ranged from 0.01 mm on the hard palate to 0.07 mm on the posterior of the dorsum of the tongue. For the incisor teeth, the calculated residual saliva thickness determined in the same way was about 0.01-0.02 mm. Blotting values plotted against dipstick values for oral sites where blotting could be readily performed showed a linear relation, which could be used as a standard curve to enable the easily done dipstick measurements in microlitres to be converted to saliva thicknesses in millimeters. As blotting could not be done in embrasures and occlusal fossae, this paper-strip absorption method was unsuitable for similar quantification of residual saliva in these sites but was done in another way described elsewhere. Overall, the results indicated that variations in dental morphology, and in the saliva secreted and available to the different oral regions, are the basic factors responsible for the wide variations in residual amounts of saliva seen on the diverse hard- and soft-tissue surfaces of human mouths. Also, finding that the hard palate and inner lips are covered by very thin films of residual saliva suggested that only a small reduction in their quantity would be needed to trigger the dry mouth sensation in hyposalivators.

Catabolism of arginine by the mixed bacteria in human salivary sediment under conditions of low and high glucose concentration

The catabolism of glucose by the oral mixed bacteria results in a lowering of the pH whereas arginine degradation favours a rise. In the mouth, low and high levels of glucose cause different plaque pH conditions which, in turn, might affect the rate and mode of degradation of arginine. This possibility was examined in the suspended salivary-sediment system where these pH conditions can be simulated. With the pH, the metabolic parameters examined were arginine utilization, ammonia, carbon dioxide and putrescine formation, utilization of glucose and changes in levels of L(+)- and D(-)-lactic acid. At the lower glucose concentration, the pH rapidly fell and then slowly rose whereas, with the higher glucose level, the pH showed a greater fall and no subsequent rise. The more acidic pH conditions favoured by the higher glucose level inhibited arginine degradation and the appearance of its various end-products and intermediates. Arginine degradation with arginine-[U-14C] and paper chromatography-autoradiography showed successive appearance of citrulline, ornithine and putrescine and, depending upon the pH, some succinate. When the pH was held constant at several different values, arginine degradation was optimal when the pH was near neutrality. In supplementary experiments, arginine had little effect on the ability of the oral mixed bacteria to utilize glucose and produce and utilize lactic acid, whereas the arginine peptide, arginylisoleucine and saliva supernatant stimulated these processes. Thus glycolysis enhancement and a more rapid clearance of fermentable carbohydrate by the oral bacteria would accompany pH-rise activity with arginine peptide and saliva but would not accompany pH-rise activity with arginine.

A comparison of the acid-base metabolisms of pooled human dental plaque and salivary sediment

The acid-base metabolisms of the mixed bacteria in pooled dental plaque and salivary sediment sampled from the same subjects were compared in vitro. Plaque at a suspension concentration of 8.3 per cent (v/v) was found to produce pH responses like those of sediment at 16.7 per cent (v/v) with all substrates and under all incubation conditions tested. The substrates examined included several carbohydrates (glucose, sucrose and starch) and several nitrogenous substrates (urea, arginine and the arginine peptide glycyl-glycyl-lysyl-arginine also called sialin). Also examined were the effects of endogenous substrates and of salivary supernatant and fluoride. A difference in suspension concentration was necessary to achieve similarity in pH response which was attributed to the presence of more non-viable epithelial cells in sediment than in plaque. Under these conditions, salivary sediment showed a slightly greater buffering capacity than plaque, a difference that was not evident if salivary supernatant was present. It was clear from this study that salivary sediment and pooled dental plaque from the same subjects have similar acid-base metabolisms and that the more abundant and readily available sediment could be used to study such metabolism in dental plaque.

Measurement of tooth hypersensitivity and oral factors involved in its development

The various methods of measurement of dentinal hypersensitivity are based upon the types of stimuli used to elicit a pain response in teeth, which include thermal, tactile, evaporative, electrical and osmotic. Pulpal inflammation in its early stages reduce the threshold of pain response to these stimuli but electrical stimulation may make it possible to assess the possible contribution of such inflammation to sensitivity determinations. Although the magnitude of each stimulus is quantifiable, patient response is subjective, which necessarily makes measurements of dentinal sensitivity semisubjective. Various methods of testing dentinal sensitivity are discussed, along with their advantages and disadvantages. The teeth most suited for measurement in clinical studies are the canines and premolars. This is because approx. 80% of the sensitivity lesions are associated with these teeth, which have similar thicknesses of root dentine. Data from several studies involving the same subjects indicate that individual measurements readily return to baseline and that the commonly seen placebo effect is probably due to some as yet unidentified factor in desensitizing formulations. Possible roles of salivary and plaque environmental factors in the development of dentinal sensitivity are discussed, as well as methods for their measurement.

Acid-base pH curves in vitro with mixtures of pure cultures of human oral microorganisms

Pure cultures of microorganisms commonly found in supragingival plaque were incubated alone and in combinations to determine the bacterial contribution to the pH-fall-pH-rise that is the central characteristic of the Stephan-curve pH change seen in plaque in vivo after brief exposure to a sugar solution. To avoid the complicating conditions of saliva flow and plaque diffusion, experiments were done with bacterial suspensions in incubations in vitro. In an initial experimental series where each microorganism was incubated only with glucose, all but a few produced the initial pH fall. Some also showed a subsequent small, sharp rise in the pH which then quickly levelled off; this was due to metabolism of endogenous substrate accumulated by most microorganisms during their growth in culture. When arginolytic and non-arginolytic bacteria were each then incubated with both glucose and arginine present (the glucose substrate to stimulate a pH fall and the arginine to stimulate a pH rise), the non-arginolytic gave a progressively more acidic pH response with progressive increase in the cell concentration, whereas the arginolytic bacteria produced a much smaller and variable pH decrease with similar cell concentration increase. Mixing pure cultures of either arginolytic or non-arginolytic bacteria gave acid-base pH responses similar to those of their respective pure cultures, whereas mixing arginolytic with non-arginolytic bacteria resulted in an approximate averaging of their different curves. The organisms present in highest proportion in a mixture had the greatest effects. The outcome of mixing the most numerous streptococcal and actinomyces species found normally in supragingival plaque indicated that the well-established difference in the acidity level of the Stephan pH response of caries-active and caries-inactive plaques could be due to differences in the proportions of their arginolytic and non-arginolytic members.