Marianne Lenander-Lumikari

Fungicidal effect of human lactoferrin against Candida albicans

Human lactoferrin (LF) in its iron-free state (apo LF), killed Candida albicans in a time- and dose-dependent way. The lethal effect was stronger at pH 7.0 than at pH 5.5 and maximum inhibition at neutral pH was achieved in 25 min when the fungal cells were exposed to LF in 0.05 mM KCl at 37 degrees C. Fe(3+)-saturated LF had no fungicidal activity. Apo LF-mediated killing was also temperature-dependent with enhanced inhibition at higher temperatures (37 degrees, 42 degrees C). The presence of 1 mM D-glucose did not affect the candidacidal activity of apo LF but both phosphate and bicarbonate ions at physiological salivary concentrations completely blocked the anti-fungal effect. Therefore it seems unlikely that LF belongs to the major host defence factors against oral candidosis.

Lysozyme and Lactoperoxidase Inhibit the Adherence of Streptococcus mutans NCTC 10449 (Serotype c) to Saliva-Treated Hydroxyapatite in vitro

Lysozyme, lactoperoxidase and salivary peroxidase inhibit the metabolism and growth of mutans streptococci, but any possible effects on the adherence of these bacteria are unknown. In this study the effects of lysozyme and lactoperoxidase on the adhesion of 3H-labelled Streptococcus mutans (NCTC 10449, serotype c strain) to saliva-coated hydroxyapatite were studied at pH 5.0 and 7.0. Human whole saliva was either lysozyme-depleted and centrifuged, or sterilized and dialysed to achieve no detectable lysozyme and peroxidase activities; this modified saliva was used to form experimental pellicles. The incorporation of lysozyme (50-200 micrograms/ml) to the pellicle caused a significant (p < 0.01) reduction in the adherence of S. mutans without any loss of bacterial viability. Pretreatment of either saliva-coated apatite or S. mutans cells with lysozyme did not change the results but lysozyme bound more readily to bacteria than to the experimental pellicles. Also, lactoperoxidase (10-200 micrograms/ml) reduced significantly (p < 0.001) the adherence of S. mutans but, in contrast to lysozyme, in a dose-dependent way. The strongest inhibition of adhesion was found when both saliva-coated apatite and bacteria were pretreated with lactoperoxidase. This enzyme bound to experimental pellicles in preference to streptococci. A non-specific protein control, albumin, did not block the inhibition by lysozyme or lactoperoxidase. The inhibition of adherence of a serotype c strain of S. mutans to saliva-coated hydroxyapatite is a novel antibacterial mechanism for both lysozyme and lactoperoxidase.

Effects of a lactoperoxidase-system-containing toothpaste on dental plaque and whole saliva in vivo

The effects of a lactoperoxidase-system-containing toothpaste. Biotene, on saliva and dental plaque were studied. In a double-blind crossover study 20 healthy volunteers used an experimental (comprising the complete peroxidase system) or a placebo (without lactoperoxidase, KSCN, and glucose oxidase) toothpaste twice daily for 2 weeks separated by a 2-week washout period. At base lines and at the end of both test periods saliva and plaque samples were collected, and plaque pH changes were monitored. Saliva was analyzed for hypothiocyanite (HOSCN/OSCN-) and thiocyanate (SCN-) concentrations and salivary peroxidase activity. The amount of total streptococci, mutans streptococci, lactobacilli, and total anaerobic flora was determined both in saliva and in plaque samples. The accumulation and the acidogenicity of plaque were also quantitated. A 2-week daily use of Biotene had no effect on salivary flow rate, peroxidase activity, HOSCN/OSCN-, SCN-, or any of the monitored bacterial counts compared with the placebo toothpaste. The accumulation of dental plaque was not affected by the lactoperoxidase-system-containing toothpaste. The acidogenicity of plaque did not change significantly, nor did the two test dentifrices differ in their ability to inhibit the plaque pH drop caused by sucrose in subjects with normal salivary flow rate.

The sensitivity of Porphyromonas gingivalis and Fusobacterium nucleatum to different (pseudo)halide-peroxidase combinations compared with mutans streptococci

Previous studies have shown that the peroxidase system with iodide is particularly effective against Actinobacillus actinomycetemcomitans. In the present study, the effects of iodide, chloride and thiocyanate in combinations with lactoperoxidase (LP) and myeloperoxidase (MP) on the viability of Porphyromonas gingivalis, Fusobacterium nucleatum, Streptococcus mutans and S. rattus were analysed. Bacteria were incubated in buffer solution containing peroxidase, substrate(s) and H2O2 (all in oral physiological concentrations), and plated after 0, 0.5 and 1 h. The oxidation product of iodide was the most bactericidal against all the bacteria tested. The effect was significantly weaker on mutans streptococci. Physiological concentrations of thiocyanate abolished the effects of LP-H2O2-iodide and MP-H2O2-iodide/chloride combinations. Thiocyanate-peroxidase systems have already been used in oral hygiene products. The incorporation of iodide into these products could make them much more potent against periodontal pathogens, and also help to prevent transmission of these pathogens from person to person via saliva.

Growth of xylitol-resistant versus xylitol-sensitive Streptococcus mutans strains in saliva

Five Streptococcus mutans pairs (serotype c S. mutans 10449 and four clinical isolates of S. mutans: 123.1, LG1, OMFA, T10B) were used to find out if the xylitol-resistant (XR) natural mutants of the corresponding xylitol-sensitive (XS) S. mutans parental strains differ in their growth patterns in saliva. The isogenic X natural mutants of the parental S. mutans strains were selected after sequential cultivations in the presence of xylitol and glucose. The XR/XS strains pairs were grown in individual and pooled glucose-supplemented filter-sterilized salivas (one to five sequential cultivations). The two salivas used represented subjects with good or poor support of the growth of S. mutans in vivo. Protease and peptidase activities were determined from the saliva growth media and cell suspensions. Salivary protein profiles were analyzed using SDS-PAGE and native IEF before and after the cultivations. The growth properties of the XR/XS S. mutans pairs were similar in both individual and pooled salivas. Sequential cultivation of all strains did not show any differences in growth patterns. XS strains were inhibited by the presence of xylitol (2% w/v) in pooled saliva, as shown for other glucose-supplemented media. Protease and peptidase activities of the XR/XS S. mutans pairs were low and of similar magnitude. Also, the general hydrolytic properties of most XR/XS S. mutans pairs appeared similar as judged by the small growth-induced changes in salivary protein profiles. In conclusion, saliva, the source of nutrients for salivary microorganisms in vivo, favored neither the XR nor the XS strains of S. mutans.

Lactoperoxidase Inhibits Glucosyltransferases from Streptococcus mutans in vitro

This study examines the possible effect of the antimicrobial peroxidase system on the activity of streptococcal glucosyltransferases B, C and D (GtfB, GtfC and GtfD), either in solution (GtfB and GtfC) or when adsorbed to hydroxyapatite (GtfC and GtfD) at pH 6.5. The lactoperoxidase (LP) system (LP, H2O2, SCN–) had no effect on the activity of dissolved GtfC, but the activity of dissolved GtfB was enhanced. The LP system, however, strongly inhibited the activities of both GtfC and GtfD in their adsorbed form. LP enzyme, without its substrates, inhibited all three Gtf enzymes: GtfB and GtfC in concentrations between 10 and 100 µg/ml in liquid phase and adsorbed GtfC and GtfD in concentrations between 25 and 50 µg/ml. This inhibition was in part abolished in liquid phase, but not in solid phase, if the substrates of LP were added. This study shows that the lactoperoxidase system can exert inhibitory activity against streptococcal Gtfs without generating oxidizing agents.