Effects of homeopathic treatment on salivary flow rate and subjective symptoms in patients with oral dryness: a randomized trial.

Twenty-eight patients with xerostomia participated in a blind, placebo-controlled longitudinal study of the possible effects of homeopathic medicines on oral discomfort. All patients were first divided in two groups according to their medication. After that the two groups were randomly assigned according to a coin-toss to the experimental or control group. Most patients had systemic diseases, such as rheumatoid arthritis and/or Sjögren's syndrome, and frequent daily medications. The randomly selected experimental group (n=15) got an individually prescribed homeopathic medicine and the control group (n=13) a placebo substance (sugar granules), both for 6 weeks. Neither group knew of the nature of the medicine. Oral dryness was evaluated by measurement of unstimulated and wax-stimulated salivary flow rates and visual analogue scale. With only two exceptions, the experimental group experienced a significant relief of xerostomia whereas no such effect was found in the placebo group. Stimulated salivary flow rate was slightly higher with homeopathy than placebo but no consistent changes occurred in salivary immunoglobulin (IgA, IgG) levels. In an open follow-up period those receiving homeopathic medicine continued treatment and the placebo group patients were treated with individually prescribed homeopathic medicines. The symptoms of xerostomia improved in both groups. Our results suggest that individually prescribed homeopathic medicine could be a valuable adjunct to the treatment of oral discomfort and xerostomic symptoms.

Clinical applications of antimicrobial host proteins lactoperoxidase, lysozyme and lactoferrin in xerostomia: efficacy and safety.

Innate human salivary defence proteins, lysozyme, lactoferrin and peroxidase, are known to exert a wide antimicrobial activity against a number of bacterial, viral and fungal pathogens in vitro. Therefore, these proteins, alone or in combinations, have been incorporated as preservatives in foods and pharmaceuticals as well as in oral health care products to restore salivas' own antimicrobial capacity in patients with dry mouth. These antimicrobials used in oral health care products, such as dentifrices, mouth-rinses, moisturizing gels and chewing gums, have been purified from bovine colostrum. In this review I critically evaluate the clinical efficacy and safety of this kind of preventive approach against various oral diseases and symptoms.

Efficacy of a slow-release device containing fluoride, xylitol and sorbitol in preventing infant caries.

A novel slow-release administration device, the "Fall-Asleep Pacifier" (FAP), was studied as a prophylactic measure against mutans streptococcal oral infection and dental caries in a risk group of 1-year-old children by comparing the test (T, n = 34) and control (C, n = 88) groups in a prospective cohort study. In the T group the children received their fluoride tablets (Fludent, containing NaF corresp. 0.25 mg F0- , xylitol 159 mg and sorbitol 153 mg) in the evenings in FAP. In the C group the children received the same dose of Fludent crushed in food in the evenings. The proportion of children, whose plaque samples from the upper incisors were mutans streptococcus positive at the age of 24 months, was significantly (P < 0.05) greater in group C (25%) than in group T (9%). The children in the T group developed significantly (P < 0.001) less (none) new dentinal carious lesions in their primary dentitions than the children in the C group between 2 and 3 1/2 years of age. Fifty-four percent of the children to whom the FAP was offered complied with regular use of it. The beneficial effect observed in the T group compared with the C group was apparently mostly due to the administration mode via FAP, which could prolong the intra-oral bioavailability of the prophylactic preparation.

Colostral proteins from cows immunised with Streptococcus mutans/S. sobrinus support the phagocytosis and killing of mutans streptococci by human leucocytes.

Passive immunisation, based on bovine colostral preparations, is an area of active research. Specific bovine antibodies inhibit the virulence factors of target pathogens but the interactions between whey preparations and human immune defence cells are not well known. Bovine colostrum inhibits the phagocytic activity of bovine leucocytes and this may reflect the biological activity of immunoglobulins in it. Therefore, this study aimed to examine the effects of bovine whey protein preparations from the colostrum of Streptococcus mutans/S. sobrinus-immunised and sham-immunised cows on binding, ingestion and killing of these bacteria by human leucocytes. Binding and ingestion of FITC-labelled bacteria were estimated by flow cytometry and leukocyte activation was measured as chemiluminescence. Killing rate was estimated by plate counting and by measuring bioluminescence from S. mutans- containing the insect luciferase gene. Colostral whey protein preparation from hyperimmunised cows activated human leucocytes by opsonising specific bacteria. Neutrophils, eosinophils and monocytes weakly phagocytosed non-opsonised bacteria and bacteria opsonised with control product. On the contrary, binding and ingestion were efficient in the presence of the preparation from immunised cows. Thus, these results show that bovine colostral whey proteins are able to support the activation of human phagocytes against pathogenic microbes and that this property is related to specific antibodies in whey preparations. These whey proteins may also be clinically useful, especially in preventing the colonisation of newly erupted teeth by mutans streptococci.

Effects of bovine immune and non-immune whey preparations on the composition and pH response of human dental plaque.

Colostral products from non-immunized cows (CP) and cows immunized with mutans streptococci (IP) were used as mouth rinses in a short-term human study. The acidogenic potential of the products was tested and found to be negligible in vivo before application to subsequent rinsing tests. At first, all the participants received a professional tooth cleaning, after which they rinsed with one of the solutions (IP; CP; water) three times per day for 3 d. After each rinsing period, the resting pH and decrease in plaque pH after sucrose challenge were determined, the amount of plaque was estimated, and all available plaque was collected. No significant differences were recorded in the composition or in the amounts of accumulated plaque. The resting pH values of plaques with low "innate" pH were increased after the IP rinsing period. Surprisingly, the lowest pH values after the sucrose challenge were recorded in IP plaques. The number of cultivable facultative flora or total streptococci were not affected by different rinsings, but the relative number of mutans streptococci significantly decreased after the IP rinsing period when compared to the CP period. Thus, the short term rinsing indicates favourable effects of bovine immune whey on human dental plaque.

Concentrated bovine colostral whey proteins from Streptococcus mutans/Strep. sobrinus immunized cows inhibit the adherence of Strep. mutans and promote the aggregation of mutans streptococci.

The aim of this study was to examine the effect of bovine colostral whey proteins from cows immunized with Streptococcus mutans/Strep. sobrinus on the adherence and aggregation of caries-inducing bacteria, i.e., mutants streptococci. Both adherence and aggregation are important phenomena in the bacterial colonization of the human oral cavity. In all adherence experiments there was a significant difference between treatments by immune product (IP; from immunized cows) and a control product (CP; a similar product from non-immunized cows). The adherence of 35S-labelled Strep. mutans cells (serotype c) to parotid saliva-coated hydroxyapatite (SHA) was dose-dependently inhibited by both IP and CP if SHA was coated with either product before exposure to bacteria, but markedly lower concentrations of IP than CP were effective. When instead of SHA the bacterial cells were pretreated with IP or CP, only IP strongly and dose-dependently inhibited streptococcal adherence. When bacteria, IP or CP, and SHA were incubated simultaneously, a significant difference between IP and CP treatments was again found. Further, IP effectively aggregated both Strep. mutans and Strep. sobrinus cells, whereas hardly any effect was seen with CP. Both IP and CP aggregated the control bacterium Strep. sanguis, which affected the adherence of the pretreated bacteria.

Generation of bovine immune colostrum against Streptococcus mutans and Streptococcus sobrinus and its effect on glucose uptake and extracellular polysaccharide formation by mutans streptococci.

Due to potential side-effects of active immunization by cariogenic mutans streptococci, oral administration of passively-derived antibodies could be a more acceptable way to reduce colonization and virulence of these microorganisms in human dentition. The aim of this study was to produce antistreptococcal immunoglobulins into bovine colostrum and explore the possible antibacterial mechanisms of these immunoglobulins against mutans streptococci. Specific serum IgG antibodies to whole cell antigens of both Streptococcus mutans and Streptococcus sobrinus increased rapidly in cows during immunization and were high also in the final whey-product. Low concentration (0.5% w/v) of bovine immune preparation inhibited significantly the incorporation of [14C]glucose by both S. mutans and S. sobrinus. Higher concentration (> 1%) was needed to inhibit the glucosyltransferase or fructosyltransferase activities of these bacteria. No such inhibitory effects were observed with the control preparation from the non-immunized cows. Our results indicate that bovine immune colostrum has a significant inhibitory potential against mutans streptococci, apparently dependent on the presence of specific IgG antibodies against S. mutans and S. sobrinus.

Effects of low-dose oral contraceptives on female whole saliva.

The composition and flow rate of paraffin-stimulated whole saliva were analysed in 22 women, of whom 11 used oral contraceptives and 11 did not. Ten men served as the controls. The salivary samples were collected during one month (oral contraceptive users and men), or during one menstrual cycle (non-users). The saliva analyses included flow rate, pH, buffer effect, sialic acid, thiocyanate, peroxidase, lysozyme, amylase, immunoglobulins A, G and M, total protein, mutans streptococci, lactobacilli, yeasts and total numbers of aerobic bacteria. The salivary buffer effect of oral contraceptive users was significantly (p less than 0.005) higher than that of non-users. All the other constituents showed intra- and interindividual variation in all groups, but with no apparent hormone-dependency.

PIP: The flow rate and composition of whole saliva were analyzed in 11 women using low dose oral contraceptives in comparison with 11 menstruating women and 10 men. Paraffin-stimulated whole saliva samples were collected Monday, Wednesday and Friday mornings for 1 cycle or 1 month in all subjects, checked for pH and buffer effect (Dentobuff method, Orion Diagnostics, Espoo, Finland, a measure of bicarbonate content) immediately, and frozen for later assay of salivary lysozyme, amylase, peroxidase, thiocyanate, sialic acid, total protein, IgA, IgG, IgM, Mutans streptococci, Lactobacilli, yeasts and aerobic bacteria. The oral contraceptives taken were Marvelon (Organon, Holland) by 4 subjects, Microgynon (Leiras, Finland) by 1, and Trikvilar (Leiras) by 6. The only significant differences between subject groups of cycle phases was a higher salivary buffer effect in oral contraceptive users than that seen in non-users, who resembled male controls. There was a wide individual variation in most values, but less variation in pH and buffer effect. Salivary buffer effect, which is correlated with HCO3-content and salivary flow, is also higher in late pregnancy.

The protective effect of peroxidase and thiocyanate against hydrogen peroxide toxicity assessed by the uptake of [3H]-thymidine by human gingival fibroblasts cultured in vitro.

The hypothiocyanite ion (OSCN-) is the principal oxidation product of the salivary peroxidase-thiocyanate (SCN-)-hydrogen peroxide antimicrobial system. Supplementation of human saliva in vitro and in vivo with low amounts (less than 1.0 mM) of hydrogen peroxide increase the concentration of salivary OSCN- (in vivo up to 0.3 mM). Elevated concentrations of OSCN- are strongly antimicrobial and may therefore be protective against dental caries. However, as OSCN- is a highly-reactive oxidizing agent, its possible toxic effect on human cells was studied using gingival fibroblasts as target cells. Concentrations of OSCN- (up to 300 microM) had no effect on [3H]-thymidine incorporation into the cells. However, fibroblasts were sensitive to peroxide so that 100 microM of H2O2 caused over 80 per cent reduction in [3H]-thymidine incorporation. The toxicity of H2O2 could be entirely prevented by adding lactoperoxidase and SCN- to the cell culture before the addition of peroxide. Thus, conversion of toxic H2O2 to non-toxic OSCN- in fibroblast culture by lactoperoxidase and SCN- suggests a dual role for the salivary peroxidase system: protection of human cells from H2O2 toxicity and antimicrobial action against oral pathogens. Furthermore, the elevated concentrations of OSCN- which produce inhibition of bacterial metabolism did not damage human cells.

Human milk peroxidase is derived from milk leukocytes.

Peroxidase enzymes present in human colostrum, saliva, polymorphonuclear leukocytes, and bovine milk were compared with respect to their molecular exclusion chromatographic behavior and immunological cross-reactivity. Human milk peroxidase gave an elution profile similar to myeloperoxidase derived from blood polymorphonuclear leukocytes. Human salivary peroxidase reacted with an antibody directed against bovine lactoperoxidase, but with the same antibody preparation no reaction was detected either with human milk peroxidase or leukocyte myeloperoxidase. We conclude that the peroxidase enzyme in human milk is different from the human salivary and the bovine enzymes and is probably derived from milk leukocytes.

Interaction of specific and innate factors of immunity: IgA enhances the antimicrobial effect of the lactoperoxidase system against Streptococcus mutans.

The antimicrobial effect of the lactoperoxidase (LPO) system (enzyme with the thiocyanate ion and hydrogen peroxide) on Streptococcus mutans NCTC 10449 (serotype c) was significantly enhanced when the system was combined with secretory IgA. Similar enhancement was observed with LPO-myeloma IgA1 or IgA2 combinations. This enhancement of the antimicrobial efficiency was not dependent on the presence of specific antibodies to S. mutans in the IgA preparation, but seemed to require binding between LPO and immunoglobulin. However, neither human polyclonal nor myeloma IgG or IgM nor rabbit IgG enhanced the antibacterial activity of the LPO system. None of the immunoglobulins, when added alone, produced antimicrobial effects. LPO was shown to bind to colostral secretory IgA, myeloma IgA1, IgA2, and to a lesser degree to monoclonal and polyclonal IgG and monoclonal IgM. This binding had a stabilizing effect on the enzyme activity. Our results suggest that IgA significantly enhances the antibacterial efficiency of one of the innate immune factors--the LPO system.

The amino acid composition of Streptococcus mutans and its culture medium supplemented with xylitol.

The amino and keto acid composition of the cells of Streptococcus mutans, strain Ingbritt, maintained and grown on a Trypticase-Phytone based medium without any added carbohydrate or supplemented with xylitol or glucose, was analyzed. The results showed no remarkable differences in the portions of individual amino acids liberated by acid hydrolysis from the cellular proteins of cells grown in the above mentioned media. However, the amount of free amino acids in the water extracts of the cells grown in the glucose medium differed considerably from those obtained from cells grown in the two other media. The amounts of free amino acids of the medium at the end of the growth period were higher in the glucose containing medium than in the two other media. The content of keto acids was lower in the cells grown in the presence of xylitol or without added carbohydrate when compared to those cells grown in glucose containing medium.