Dorthea A. Johnson

Impact of aging on human salivary gland function: A community-based study

A comprehensive evaluation of salivary flow rates and composition was undertaken in an age- and community-stratified population. A nonmedicated subpopulation was used to assess the effect of “primary aging” on salivary gland function. Unstimulated whole, parotid and submandibular/sublingual (SMSL) saliva, as well as citrate-stimulated parotid and SMSL saliva were collected from 1006 subjects. Flow rates were determined, and the total protein concentrations measured. Height and caloric intake were documented. Subjects were divided into six age groups from 35 to 75+ years old. Significant age-related decreases in the secretion rates of unstimulated whole (p<0.001), stimulated parotid (p<0.01) and unstimulated and stimulated SMSL (both p<0.0001) saliva were observed in the total popu lation. In the non-medicated subpopulation, agerelated decreases in salivary secretions were observed in unstimulated whole (p<0.01) and unstimulated and stimulated SMSL (p<0.01 and p<0.0001, respectively). Multiple regression analysis revealed that, as well as age, caloric intake was related to unstimulated SMSL and stimulated SMSL saliva in the whole population, and height was a contributor to unstimulated whole saliva and unstimulated parotid saliva flow rate variances. In the non-medicated popu lation, caloric intake was the significant independent variable for unstimulated and stimulated parotid secretion, as was height for unstimulated whole and SMSL flow rates. Age-related increases in the total protein concentration of unstimulated parotid (p<0.001) and unstimulated SMSL (p<0.05) saliva were evident in the whole population, but not in the non-medicated subgroup. These data suggest that there are significant age-related alterations in salivary function.

Effect of donor age on the concentrations of histatins in human parotid and submandibular/sublingual saliva

Histatins are small proteins of human glandular saliva that have antifungal properties. Recent studies show that oral candidal infections increase with age, suggesting an age-associated compromise in oral host defence. Here, the effect of age and of physiological gland stimulation on the concentration and secretion of salivary histatins was investigated. Parotid and submandibular/sublingual salivas were collected from six young adults under unstimulated, mechanical (chewing) and gustatory (0.025 M and 0.1 M citric acid) stimulation, and the concentration and secretion of histatins was measured by cationic polyacrylamide gel electrophoresis with subsequent densitometric scanning of the stained gels. With gland stimulation, parotid saliva showed no significant increase in histatin concentration (microg/ml); however, histatin secretion (microg/min) increased up to 26-fold (p<0.005; ANOVA). Stimulation of submandibular/sublingual saliva resulted in significant increases in both histatin concentration (p<0.005) and secretion (p<0.0005). Ageing effects on salivary histatins were determined in citric acid (0.1 M)-stimulated parotid and submandibular/sublingual saliva samples collected from 80 individuals (divided into four age groups having approximately equal numbers of males and females: 35-44 years; 45-54 years; 55-64 years and 65-76 years). None of the patients was taking medications or wore dentures. ANOVA showed no sex differences in histatins. Regression analysis showed significant age-associated decreases for parotid saliva histatin concentration (p<0.002) and secretion (p<0. 002) as well as for submandibular/sublingual saliva histatin concentration (p<0.0001) and secretion (p<0.0001). Both saliva types showed significant (p<0.0001) decreases in the histatin concentration per mg of total protein, suggesting a preferential decrease in salivary histatins compared to total salivary protein. These results suggest that the salivary histatin component of the oral host defence system is compromised with increasing age.

Changes in rat parotid salivary proteins associated with liquid diet-induced gland atrophy and isoproterenol-induced gland enlargement

The composition of secretory proteins in the parotid saliva of untreated rats, fed a stock pelleted diet (CON), was compared to that of rats maintained on a liquefied diet (LIQ), so as to reduce gland secretory activity, and to rats treated chronically with isoproterenol (ISO), so as to enhance activity. Each experimental treatment resulted in marked changes in protein composition. In CON rats, the basic and acidic proline-rich proteins accounted for 25 per cent of all secretory protein. In LIQ rats, the proportion was 13 per cent. Several basic proline-rich proteins were absent and the major acidic proline-rich protein was markedly reduced. Amylase was reduced whereas DNase and a leucine-rich protein (fraction I) were increased. The proportion of cystine-rich protein (fraction V) was not different from CON rats. The changes observed in the saliva of ISO rats were in marked contrast to these findings. Basic and acidic proline-rich proteins were increased and accounted for 90 per cent of all secretory protein, amylase was responsible for the remaining 10 per cent. Thus the composition of secretory proteins in the parotid saliva of rats can be altered by experimental conditions which affect gland secretory activity. The mechanisms by which these changes occur is not known.

Parotid saliva protein profiles in caries-free and caries-active adults

OBJECTIVE The objective of this study was to determine if there were any differences in the parotid saliva output and composition related to caries activity. STUDY DESIGN Stimulated parotid saliva samples were collected from 85 healthy young adults, caries-active or caries-free. Flow rates were determined, and samples were analyzed for pH and buffer capacity, total protein, electrolytes, proteins with a high performance liquid chromatography method, and histatins. RESULTS There were no differences in flow rates or pH, but buffer capacity was higher in women than in men, and K+ and Cl- were both slightly higher in the caries-active group. The women had a significantly higher total protein concentration, as well as higher concentrations of each of the individual protein components assayed. There were no differences attributable to caries activity. CONCLUSIONS Significant sex differences in salivary protein concentrations exist. Caries activity may be related to some salivary electrolyte alterations, but not to protein composition.

A population-based study of salivary lysozyme concentrations and candidal counts

The relationship between salivary lysozyme concentration and oral candida load was examined in 595 adults. Unstimulated whole saliva, and citrate-stimulated parotid and submandibular/sublingual saliva were collected from each participant. Candida colony-forming units (c.f.u.) in unstimulated whole saliva were determined. An enzyme-linked immunosorbent assay for lysozyme using commercially available antibodies was developed. This assay showed a linear relation of salivary lysozyme concentrations from 0.5 to 4.0 ng/ml. Significant negative relations were observed between lysozyme concentration and flow rate: r = -0.16 (p < 0.001) for stimulated parotid and r = -0.22 (p < 0.0001) for stimulated submandibular/sublingual saliva. The lysozyme concentration in stimulated submandibular/sublingual saliva was higher in males than in female, but no sex difference was observed for stimulated parotid saliva. The lysozyme concentration of stimulated parotid saliva was positively correlated with candida counts (r = 0.18: p < 0.005). Further study of groups according to their levels of candida in whole saliva revealed that lysozyme concentrations were higher in the high candida (> or = 1000 c.f.u./ml) group than in the zero and moderate candida categories in stimulated parotid saliva (p < 0.001): there were no concentration differences in stimulated submandibular/sublingual saliva. These results suggest that parotid lysozyme concentration increases as candida load increases.

Measuring short-term γ-irradiation effects on mouse salivary gland function using a new saliva collection device

A restraining device was designed specifically for the collection of whole saliva from mice without using anesthesia. As the procedure does not involve surgical cannulation of the salivary glands, saliva can be collected from the same mouse at different times. The time between the injection of a secretory stimulant (pilocarpine) and the appearance of saliva in the mouth (lag time) was 100.5 +/-8.5 s (mean+/-S.E.M., n=10) for control mice. The volume of saliva collected in the first 5 min was three times greater than that collected between 15 and 20 min. The average flow rate for a collection period of 15 min was 16.7 +/-1.8 microl/min (n=10). The flow rate was decreased 50% (P<0.005) whereas the lag time was increased more than 300% (P<0.05) at 24 h after irradiation. The concentrations of a 23.5-kDa protein and a mucin were decreased after irradiation whereas there was no significant effect on the concentration of amylase or peroxidase.

Interaction of age and specific saliva component output on caries

Background and aims: The purpose of this study was to explore the relationship between individual salivary components, dental caries and age, utilizing the data from the Oral Health: San Antonio Longitudinal Study of Aging (OH:SALSA). Methods: The study population comprised a well-defined stratified sample of 811 dentate men and women. Subjects were divided into four age groups from 35 to 75+ years old. Unstimulated and stimulated submandibular/sublingual saliva flow rates, unstimulated and stimulated parotid saliva flow rates, total protein, 6 individual proteins and 4 inorganic constituents were measured. Specific salivary components were lactoferrin, secretory IgA, albumin, lysozyme, mucin, cystatin, K+, Ca+2, Na+ and Cl−. Caries measurements were the DMFT Index for crowns and for roots, Tooth Health Index for crowns and roots, Tooth caries, Root caries and Tooth restoration. The data on saliva components were square root transformed for linearity prior to analysis. Analysis was carried out in two stages. Partial correlation was performed, in order to identify significant relationships between specific salivary components and caries measurements, controlling for age group. In the second stage, using caries measurement as the dependant variable, the effects of age, flow rate and specific salivary component output (product of flow rate and concentration) were examined. Results: Significant associations were found between caries, age and specific individual submandibular/sublingual salivary proteins (lactoferrin, albumin, lysozyme, mucin and cystatin) and specific inorganic constituents (K+, Ca+2, Na+ and Cl− ). Conclusions: Changes in submandibular/sublingual salivary component output during aging are correlated with high caries prevalence. These changes in saliva components over age may represent caries risk indicators.

Differences in basic proline-rich proteins in rat parotid saliva following chronic isoproterenol treatment or maintenance on a liquid diet

The basic proline-rich proteins (BPRP) in the stimulated parotid saliva of rats treated for 8 days with isoproterenol and rats fed a liquid diet for 2 weeks were compared to those in the stimulated parotid saliva of untreated rats fed a stock pelleted-diet (control). In the control, the BPRP were separated into 5 groups designated Peak A (the basic proline-rich glycoprotein), SP-1, SP-2, SP-3 and SP-4. The percentage of BPRP in each group was as follows: Peak A, 6.5 per cent; SP-1, 37 per cent; SP-2, 6.5 per cent; SP-3, 32.4 per cent; SP-4, 17.6 per cent. In the parotid saliva of rats fed the liquid diet, proteins corresponding to Peak A and SP-2 were not present, the proportion of BPRP in SP-4 was increased to almost 90 per cent while the proportions of material in SP-1 and SP-3 were reduced to 3 and 8 per cent, respectively. In the saliva of rats subjected to chronic isoproterenol treatment, a protein corresponding to SP-4 was not present; proteins corresponding to Peak A, SP-1 and SP-3 were present and in amounts similar to their proportion in untreated rats although material in SP-2 increased to 36 per cent.

Influence of mastication on saliva, plaque pH and masseter muscle activity in man.

An earlier study showed that frequent gum chewing may enhance parotid gland function and reduce the acidogenicity of dental plaque. The aim now was to determine whether these effects would be observed after a 2-week period of diet altered to increase masticatory effort, and secondarily to assess the effects of chewing gum on masseter muscle activity. Ten subjects took part in the first experiment. Saliva was collected before and after the diet change and the plaque pH response to a sucrose challenge was measured. Subjects completed 3-day diet histories and wore electromyographic (EMG) devices to record masseter activity. In the second experiment, 10 subjects wore EMG devices for 3 days to record masseter activity on three daily regimens: baseline (no gum chewing), hourly gum chewing (sugar-free gum chewed for 10 min every hour) and chewing five sticks of gum each for 20 min during the day. Data were analysed by paired t test or repeated-measures analysis of variance. For the first experiment, EMG data indicated significant increases in chewing activity (p < 0.05), although there were no changes in salivary flow rates or the plaque pH response to sucrose. The second experiment showed that total EMG activity increased significantly on both gum-chewing regimens (p < 0.01), the magnitude of the increase being greater for hourly chewing. Overall, masseter EMG activity was increased 41% by diet alteration, compared to increases of 96 and 152% on the five-stick and hourly gum-chewing regimens, respectively.

Effects of Food Mastication on Rat Parotid Gland Adrenergic and Cholinergic Cell Surface Receptors

Adult male rats were fed diets of differing texture (liquid, powder, standard pelleted, or bulk pelleted) to alter food mastication. After 2 weeks, the parotid glands were removed and adrenergic and muscarinic-cholinergic cell surface receptor density (fM bound/mg protein) and ligand binding dissociation constants (Kd in nM) were determined by radioligand binding techniques on a crude membrane fraction. For all diets, gland weight increased as the requirement for food mastication increased (i.e., liquid < powder < standard pelleted < bulk pelleted). Among the diets, neither beta-two nor alpha-two receptor density was altered. Beta-one receptor density was directly related to dietary mastication. Compared with the standard pelleted diet, beta-one receptor density was reduced 21% for the liquid diet and 7% for the powdered diet; for the bulk-pelleted diet, beta-one receptor density was increased 11%. With respect to alpha-one receptor density, it was not affected by the liquid or powdered diet when compared with the standard pelleted diet, but alpha-one receptors were increased 14% with the bulk-pelleted diet. Muscarinic-cholinergic receptor density for the liquid diet fed rats was 27% less than for the standard-pelleted diet; powdered diet did not differ from standard pelleted, while that for the bulk-pelleted diet was increased 6%. With but minor exceptions, ligand binding affinity was unaffected by the changes in diet texture. These studies demonstrate that dietary mastication as well as affecting parotid gland weight, cell size, and saliva production also influences autonomic cell surface receptor density.