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journal of dentistry 36 (2008) 977–983

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Influence of saliva substitute films on initial Streptococcus
mutans adhesion to enamel and dental substrata
Sebastian Hahnel *, Martin Rosentritt, Gerhard Handel, Ralf Bu¨rgers
Department of Prosthetic Dentistry, Regensburg University Medical Center, Regensburg D-93042, Germany

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Article history:

Objectives: The aim of this in vitro study was to evaluate whether saliva substitutes contain-

Received 19 May 2008

ing antimicrobial agents influence the initial adhesion of Streptococcus mutans to bovine

Received in revised form

enamel and various dental materials.

20 July 2008

Methods: Specimens of a denture base resin, a veneering composite and a dental ceramic

Accepted 7 August 2008

were prepared according to the manufacturers instructions and polished. Standardized
bovine enamel slabs were prepared for reference. Surface roughnesss and surface free
energy were determined. Fifteen specimens of each substratum were rinsed with four saliva


substitutes (Salinum, Aldiamed, Saliva natura and Saliva Orthana), a negative (PBS) and a

Saliva substitute

positive control (protein mixture) for 2 h at 37 8C in a flow chamber, and were subsequently


exposed to S. mutans NCTC 10449 suspension for 4 h at 37 8C. Adherent bacteria were


quantified using a fluorometric assay. Statistical analysis was performed using one- and

Streptococcus mutans

two-way ANOVA ( p < 0.05), and post hocs were analyzed using the Tukey-Kramer multiple


comparison test ( p < 0.05).
Results: Substrata as well as saliva substitutes influenced fluorescence intensities decisively. No significant differences in fluorescence intensities indicating similar adhesion of S.
mutans were found between substrata that had been exposed to the negative control, the
positive control, Saliva Orthana and Aldiamed. On substrata with high surface free energy
(ceramic and bovine enamel), significantly higher fluorescence intensities indicating higher
adhesion of streptococci were found to specimens that had been exposed to Saliva natura
and Salinum.
Conclusions: The influence of saliva substitutes on initial S. mutans adhesion appears to be
dependent on the substratum surface properties. Only little influence of antimicrobial
agents was found.
# 2008 Elsevier Ltd. All rights reserved.



Xerostomia is a common disease and an increasing number of
people are affected by salivary gland dysfunctions. Reasons
for hyposalivation are manifold, ranging from radiation
therapy of malignant head tumors1 and medical remedies2
to immune mediated diseases like the Sjo¨gren Syndrome.3

Xerostomia may also occur as a symptom of diabetes,
hypertension or cardiovascular diseases.2 There have been
reports that people suffering from xerostomia incur a greater
risk of the accumulation of oral diseases associated with
pathogenic microorganisms such as dental caries, periodontal diseases or fungal infections,2 due to the insufficient
saliva secretion reducing the self-cleaning mechanisms of

* Corresponding author. Tel.: +49 941 944 6059; fax: +49 941 944 6171.
E-mail address: (S. Hahnel).
0300-5712/$ – see front matter # 2008 Elsevier Ltd. All rights reserved.

and one of their common primary goals is the lubrication of dental and mucous tissues. Rodgau. 3M Espe.6 Several artificial saliva and mouth rinse solutions containing proteins. Roughness values higher than 0.17 The aim of this in vitro study was to examine whether films formed by commercial saliva substitutes on bovine enamel and dental restorative materials.2 mL) were examined on three specimens for each material. G). Peak-to-valley surface roughness was determined at three spots on each specimen (one in central position. mutans to surfaces treated with saliva substitutes containing antimicrobial agents is lower than its adhesion to surfaces treated with conventional saliva substitutes. Biofac A/ S. Ivoclar Vivadent) and conventional polishers. . and since the 1990s. lysozyme (lysozyme from hen egg white. Sigma–Aldrich). G) or linseed extract (Salinum.1.11 and indirectly by chelating available iron.2 mm. Fluka Biochemika) and mucin (mucin from porcine stomach Type III. Fluka Biochemika. MO.978 journal of dentistry 36 (2008) 977–983 the oral cavity and to the limited antimicrobial effects of the residual saliva. Biomedica GmbH.14 however. two at the margins) using a profilometric contact surface measurement device (Perthen S6P. It was hypothesized that the adhesion of S. Go¨ttingen.11 Xylitol. Buehler Ltd. lactoperoxidase and its substrates (thiocynate and hydrogen peroxide).10. aggregate bacteria. G). diiodomethane (Sigma–Aldrich. FL) were manufactured according to the manufacturers’ instructions. Numerous artificial saliva substitutes for the treatment of xerostomia have been developed in the past. Kastrup.5 % chloramine solution until use. a-amylase (a-amylase from hog pancreas. Du¨sseldorf. Another property of saliva substitutes for preventing caries is antimicrobial activity. All specimens were smoothed using a rotating grinding disk apparatus (Motopol 8. and the left and right contact angles of each drop were averaged. little effort has so far been made to evaluate the potential effects of saliva substitute films on dental materials and tooth surfaces on the initial adhesion of oral bacteria.2 mm have been found to foster the adhesion of microorganisms. Heraeus Kulzer.. or xylitol7 have been introduced. effects on microbial growth and metabolism have also been described. 2. G). Therapeutical approaches for relieving xerostomia are most often restricted to symptomatic therapy. St. Ivoclar Vivadent. Rabel and Kaelble (OWRK) method.8–10 Berlutti et al. three liquids differing in hydrophobicity were used: deionized water. Lysozyme has been found to hydrolyse peptidoglycans in the bacterial cell wall. 2 mm thick) of a denture base resin (Palapress vario. and were stored in 0. initial microbial adhesion is important as it has been found to influence later plaque formation decisively by means of coadherence and coaggregation. Materials Round standardized specimens (diameter 10 mm.2. Godalmy. Sigma–Aldrich. CH). water in combination with hydroxyethyl cellulose for water binding and enhanced adhesion (Aldiamed. 2.6 this appears to be a promising approach for the prevention of microbial-related diseases. G) and a lithium disilicate ceramic (Empress 2. St. IL.13 This phenomenon has most commonly been attributed to its lacking fermentation by oral microorganisms. another goal of saliva substitutes was to allow for remineralization of both the enamel and dentin. Materials and methods 2.4.16 Surprisingly. Medac GmbH. as its film forming properties have been evaluated in previous. Darmstadt. Bovine. mutans. Saliva Orthana. G). Louis. DataPhysics Instruments GmbH. the integration of antimicrobial agents against oral microorganisms into artificial saliva and other healthcare products has been proposed. Filderstadt. Lemgo. Surface free energy of the various substrata was calculated from contact angle measurements using the sessile drop method and an automated contact angle measurement device (OCA 15plus. UK) were examined (cf. Buchs. Schaan. which is an essential nutrient for bacteria. newly developed saliva substitute designed for the in vitro evaluation of bacterial adhesion to dental materials consisting of PBS supplemented with albumin (Albumin.15 Saliva substitutes are intended to form films on oral surfaces to ensure the lubrication of dental and mucous tissues. unpublished studies. USA) and ethylene glycol (Merck KgaA. Feinpru¨f-Perthen. Phosphate-buffered saline (PBS. In addition. G). For measurements. Table 1). Seefeld. Ten drops for each liquid (0. Standardized enamel slabs (diameter 10 mm. Roughness values up to 0. USA) was used as negative control. lactoferrin. Wedel.12. which differ in surface free energy. Buehler GmbH. specimens with higher surface roughness were rejected for keeping surface roughness far below the threshold value at 0. carbohydrates and lipids would not allow any film formation. and were subsequently polished to high gloss using polishing paste (universal polishing paste.08 mm were tolerated. Sinclair Pharmaceuticals Ltd. Saliva substitutes Four commercial saliva substitutes either based on mucin (Saliva natura. Coventry. activate bacterial autolysins. DK). as it was assumed that the lack of proteins. has been found to reduce in vivo caries and plaque formation. Hanau. Louis. UK) and grinding paper (grain 1000 and 4000. as a constituent of chewing gums. 2 mm thick) were prepared from the vestibular surface of freshly extracted anterior bovine teeth using conventional diamond burs (Brasseler. a veneering composite (Sinfony. and inhibit bacterial adherence as well as glucose uptake and acid production. G). Wendt. influence the initial adhesion of S. Sigma–Aldrich) was used as a ‘‘positive’’ control. An additional. enzymes and carbohydrates featuring antimicrobial properties such as lysozyme. The surface free energy and the polar and disperse components were calculated according to the Owens.18 All specimens were stored in distilled water for 6 days prior to the experiments to minimize the influence of residual monomers or toxic constituents on cell viability.5 particularly for the prevention of the development and progress of dental caries. yet it has been demonstrated that the film-forming properties differ considerably between various commercial saliva substitutes. reported that lactoferrin may influence Streptococcus mutans aggregation and biofilm formation directly by inhibiting the adherence to abiotic surfaces. G..

It has been found that the relative fluorescence intensity correlates linearly with the amount of adherent bacteria. 30 mL Resazurin (Resazurin. BMG Labtech. Louis. ascorbic acid. sodium benzoate. 2. Statistical analysis Means and standard deviations were calculated using SPSS 13. the artificial saliva solution was pumped down. 19 8C. G Salinum Linseed extract Sinclair Pharmaceuticals Ltd. CA. USA – PBS – Saliva Orthana Pig gastric mucin Biofac A/S. sorbitol. NY. sodium benzoate and potassium sorbate Aqua. poloxamer 407. sodium benzoate. St. Berkeley Heights.875 mL/min for 4 h. phosphatebuffered saline) Positive control – Sigma–Aldrich. the Resazurin reduction assay features the advantage that only viable cells are subject to quantification. G) was established.24 2. USA). linseed extract. mutans NCTC 10449 (DSMZ. and incubated for 12 h as described above. Cells were harvested by centrifugation (2200 rpm.6. Quantification of adherent bacteria Resazurin reduction (Alamar Blue) was used for viable cell quantification as described earlier.21. San Rafael. propylene glycol. Subsequently. St. methylparabene. In prior experiments.65  108 cells/ mL. UK Aldiamed Mundspu¨llo¨sung Water in combination with hydroxyethylcellulose Biomedica GmbH.19 and the optical density was adjusted to 0. lactoferrin. MO. propylparabene. mutans suspension prior to incubation.20 2. aroma. fluorescent pigment Resorufin (maximum absorbance at 573 nm). unless stated otherwise. The saliva substitutes Salinum and Saliva natura are distributed in the US as Numoisyn (Align Pharmaceuticals. sodium hydroxide and citric acid Aqua. G).5 L of S. xylitol. Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH. Louis. a-amylase (1 mg/mL). this cell quantification method is based on the reduction of the blue. Tuttlingen. In this study. The day before the experiment. and relative fluorescence intensities were measured using an automated multifunctional plate reader (Fluostar Optima. Eriodictyon crassifolium. and subsequently kept at 4 8C. hydroxyethylcellulose. Wedel. non-fluorescent redox indicator Resazurin (maximum absorbance at 605 nm) into the violet. Chicago. The optical density was measured and readjusted continuously during the experiment to provide similar conditions for cell adhesion over the whole experiment. USA) and MouthKote (Parnell Pharmaceuticals. the S. USA) were added to 2. as only metabolically active cells are capable of the reduction process.3. which corresponds to a microbial concentration of 3. Under continuous stirring. 2. sorbitol. specimens were removed from the chambers. lysozyme (10 mg/mL) and mucin (850 mg/L) Sodium chloride. xylitol. Hettich Rotixa P. magnesium chloride.. IL. the bacteria suspension was adjusted to a constant temperature of 37 8C in a tempered water bath. UK) were used to ensure that saliva substitute films have formed on the various specimens after 2 h of rinsing.5.4. The cell suspension was subsequently subjected to low intensity ultrasonic energy in order to disperse bacterial chains. aloe barbadensis and EDTA CI42090 Ingredients as stated by the manufacturer. washed twice with PBS and resuspended in the same buffer. After 4 h. chondrus crispus. USA). Cambridge. Cambridge Instruments. statistical analysis was performed using one-way analysis of variance (ANOVA). samples were rinsed with bacterial suspension at a constant flow rate of 1. Kastrup. potassium thiocyanate and peppermint flavour Aqua. A single colony was incubated with sterile DSMZ-medium 92 (Trypticase Soy Yeast Extract Medium) at 37 8C for 16 h.875 mL/min at 37 8C. After 2 h. G Albumin (40 mg/mL). the specimens were rinsed with the various artificial saliva solutions at a constant flow rate of 1. G). and the system was rinsed with PBS for 5 min. Godalmy. Braunschweig. NJ. 1 mL of bacteria suspension was inoculated with 250 mL of sterile medium. For the evaluation of differences in surface free energies.3 at 550 nm (Genesys 10-S. USA). and bacteria were transferred onto an agar plate and incubated at 37 8C for 48 h. SEM micrographs (Stereoscan 240. Thermo Spectronic. Offenburg. dipotassium phosphate.979 journal of dentistry 36 (2008) 977–983 Table 1 – Saliva substitutes used in this study Saliva substitute Base Manufacturer Ingredients Negative control (PBS.23 In contrast to numerous other quantification assays.22 In brief. For quantification of adherent bacteria. Test assay An artificial mouth system as described in a previous study was used for the simulation of oral conditions. aroma.21 For simulating the formation of artificial pellicles.0 for Windows (SPSS Corporation. All experiments were performed twice for validation of test results. mutans suspension was removed and the system was rinsed with PBS solution for 5 min in order to remove any unbound bacteria. DK Saliva natura Herbal mucin Medac GmbH. sorbitol. Bacteria A frozen ( 60 8C) preculture of the strain S. 5 min. For the evaluation of differences in fluorescence . lysozyme. Rochester. Rodgau. Sigma–Aldrich.

No statistical differences were found between specimens that had been treated with the negative control. p = 0. Regardless of the artificial saliva coating. Fluorescence intensities measured for specimens treated with Saliva natura were significantly higher than values measured for ceramic specimens that had been exposed to Salinum ( p < 0. No statistical differences were observed between specimens treated with the negative control. significantly higher fluorescence intensities ( p < 0.008).001) and the saliva substitute ( p < 0. one-way ANOVA revealed significant differences in fluorescence intensities between the specimens treated with the various saliva substitutes.05.001) except for the veneering composite ( p = 0. were found for specimens made of the veneering composite (Fig. indicating no differences in S. one-way and two-way analysis of variance (ANOVA) were used. p = 0. Aldiamed (1025) or Saliva natura (1208).001 for all specimens) were recorded for specimens that had been exposed to Saliva natura (25624) and Salinum (18843) than for specimens treated with the negative control (7705). Fluorescence intensities measured for Fig.015).002) or the positive control (890. Saliva Orthana (384) or Aldiamed (1588).112). Results 3. Surface properties of the evaluated materials One-way ANOVA results revealed significant differences in surface free energy between the various substrata ( p = 0. Mean fluorescence intensities ranged from 737 (specimens exposed to Saliva natura) to 1725 (specimens exposed to the Saliva Orthana). means and standard deviations. For bovine enamel (Fig. Adhesion values were similar to specimens that had been exposed to Saliva Orthana (1156). the positive control (4692). 3). p = 0. Fig. 1 – Relative fluorescence intensities (no units) of the denture base resin after exposure to the various saliva substitutes and incubation with Streptococcus mutans. Saliva Orthana (4440) or Aldiamed (5459).84 mJ/m 2. For specimens made of the denture base resin (Fig.2. p = 0. significantly higher fluorescence intensities ( p < 0. specimens exposed to Saliva natura were significantly higher than values measured for specimens that had been exposed to Salinum ( p = 0.62 mJ/m 2). For all substrata ( p < 0. the positive control.027) and the veneering composite (31. mutans adhesion. p = 0. which was significantly higher than the surface free energy values calculated for the denture base resin (33. 2 – Relative fluorescence intensities (no units) of the veneering composite after exposure to the various saliva substitutes and incubation with S. 3. .980 journal of dentistry 36 (2008) 977–983 intensity. similar fluorescence intensities. the positive control (1372). 4). 3.93 mJ/m 2. For all tests.001 for all specimens) were observed for specimens that had been treated with Saliva natura (7435) and Salinum (5108) than for specimens treated with the negative control (495). Saliva Orthana or Aldiamed.148). the veneering composite and the ceramic.005) but not significantly different from values for ceramic (36.022).1. Post hoc analysis was performed using the TukeyKramer multiple comparison test.08 mJ/m 2. the level of significance was set to a = 0.001) influenced fluorescence intensities. Post hoc analyses found no statistical differences in surface free energy between the denture base resin.001). mutans. the positive control. The highest surface free energy was observed for bovine enamel (mean surface free energy 43. 2). significantly higher fluorescence intensities were found for specimens that had been exposed to Salinum (1858) compared with specimens that had been exposed to the negative control (717. 1). Saliva Orthana and Aldiamed (Fig. means and standard deviations. For ceramic specimens. Bacterial adhesion Two-way ANOVA results showed that both the substratum ( p < 0.

journal of dentistry 36 (2008) 977–983 Fig.27 some researchers maintain that the original substratum surface properties are still transferred through the salivary protein layer28 and still influence microbial adhesion. 3 – Relative fluorescence intensities (no units) of the ceramic after exposure to the various saliva substitutes and incubation with S. Furthermore. It was hypothesized that films formed by saliva substitutes containing antibacterial enzymes reduce initial S. 4 – Relative fluorescence intensities (no units) of bovine enamel after exposure to the various saliva substitutes and incubation with S. no differences in bacterial adhesion were observed between the treatments with the various saliva substitutes. Fig. a broad range of dental materials as well as bovine enamel differing in surface free energy has been used in this study. Films formed by the various saliva substitutes as well as those formed by the positive control did not reduce adhesion of S. or microbial adherence over a longer period of time.21 and may thus serve as a suitable device for simulating bacterial adherence to solid surfaces.18. It would have been interesting to evaluate differences in substratum surface free energy after exposure to the various saliva substitutes. On the veneering composite. mutans adhesion.29 and is regarded as one of the major causative agents for the formation of dental caries. The influence of the various saliva substitutes on S. phosphate buffered saline solution supplemented with albumin. much greater adhesion was found for specimens treated with Saliva natura and Salinum than for those treated with the other saliva substitutes or the control. specimens used in this study were polished to high gloss to exclude potential influences of substratum surface roughness on adhesion values. It is desirable that saliva substitutes reduce the adherence of pathogenic microorganisms. mutans to various substrata differing in surface 981 free energy. mutans. However. Substratum surface roughness and surface free energy have been found to decisively influence microbial adhesion to oral surfaces. yet the findings of this study were not consistent with that hypothesis.29 Dental caries is a disease that is very common among patients suffering from xerostomia. S. was introduced as a solution of proteins with known composition and protein concentration. which yielded lowest values for surface free energy.25 thus. This in vitro assay simulates merely one aspect of microbial adhesion and the performance of saliva substitutes. means and standard deviations. mutans adhesion appeared to be influenced heavily by the underlying substratum. mutans. With regard to this aspect. amylase. mutans to the various substrata compared with the negative control. Discussion The aim of this in vitro study was to evaluate the potential of commercial saliva substitute films in influencing initial adhesion of S. A ‘‘positive’’ control. means and standard deviations. lysozyme and mucin. further studies should investigate aspects such as cell viability by directly incubating microorganisms with saliva substitutes. which might indicate a higher .2 which justifies the selection of S. mutans has been chosen as a representative oral bacterium as it has been found in early plaque. thus. The artificial mouth model used in this study simulates important parameters such as permanent shear stress and continuous saliva flow. mutans as a representative strain for this in vitro study. The formation of saliva substitute films was simulated prior to the adhesion assay by rinsing the specimens with the various saliva substitutes. 4. it has been found that in vivo substrata with high surface free energy harbour substantially more plaque than substrata with low surface free energy.26 Although saliva coating equalizes differences in substratum surface free energy. for substrata with higher surface free energies such as bovine enamel and ceramic. yet some of the saliva substitutes yielded very sticky films on the specimens allowing no proper evaluation and measurement of contact angles.

[8] Iacono VJ. Lumikari M. Oral Diseases 2002. similar adhesion was found for high surface free energy substrata treated with Saliva Orthana and Aldiamed. In this study. Schulte-Mo¨nting J. New concepts in the pathogenesis of Sjo¨gren syndrome: many questions. Acknowledgments The saliva substitutes were kindly provided by their manufacturers. lactoferrin and peroxidases: antibacterial effects on cariogenic bacteria and clinical applications in preventive dentistry. finding high sensitivity for Streptococcus rattus and for Streptococcus cricetus but not for S. yet increased adhesion of S. Sugiura et al. after an incubation of 24 h. Aldiamed). they found much lower adsorbed amounts on hydrophilic than on hydrophobic surfaces. The effects of xylitol on S. mutans to specimens with high surface free energy was observed after exposure to some of the investigated saliva substitutes. Kielbassa AM. The effect of commercially available saliva substitutes on predemineralized bovine dentin in vitro.16 This phenomenon might serve as an explanation for the different adhesion of S.7 However. 24:737–47. this studies findings indicate that their antibacterial effects in saliva substitute films on the very initial bacterial adhesion are rather poor based on the flow chamber assay. mutans adhesion to the various substrata. Clinical applications of antimicrobial host proteins lactoperoxidase. however. Head & Neck 2002. too. mutans strains are necessary to validate these considerations. clinicians might consider saliva substitutes that do not influence S. the present investigation yielded ambiguous data concerning the performance of mucin-based saliva substitutes. Dorner T. mutans appears to be dependent on the substratum. detailed investigations using various S.8:192–8. specimens with high surface free energy treated with Saliva natura. [4] Kielbassa AM.31.982 journal of dentistry 36 (2008) 977–983 risk for dental caries in vivo. fewer answers. Xerostomia: etiology. Thus. Concerning the performance of lysozyme. Pollock JJ. Parichereh-Shohadai S. Oral Diseases 2002. [7] Tenovuo J. Furthermore. this phenomenon highlights that in vivo investigations are necessary to clarify the impact of antimicrobial agents on salivary levels of caries inducing microorganisms. Germaine GR. Visch LL. recognition and treatment. Schulte-Mo¨nting J. Soukka T. However. mutans to surfaces coated with Saliva natura and Salinum. 5. with porcine gastric mucin as their major constituent. mutans adherence appeared to be poor.134:61–9. they did not confer significant decreases in S. however. Lipsky PE. Lumikari and Tenovuo reported that the susceptibility of mutans streptococci to lysozyme is strain-dependent. and cationic polypeptides against Streptococcus sanguis and Streptococcus . which is based on herbal mucins. Selective antibacterial properties of lysozyme against oral microorganisms. However. references [1] Wijers OW. DiRienzo S. [6] Tenovuo J. Moore PA. yet this might be attributable to distinct test conditions and incubation time. It has been reported that salivary lysozyme and salivary lactoferrin are incorporated into the in vivo salivary pellicle on enamel and dental restorative surfaces. [3] Hansen A. Conclusions Within the limitations of an in vitro study it can be concluded that the influence of films formed by saliva substitutes on the initial adhesion of S.87:197–208. Bactericidal activity of human lysozyme.9:40–7. For specimens treated with Saliva Orthana and the positive control. Salivary lysozyme.34 Both lysozyme and lactoferrine have been found to inhibit the adherence of oral bacteria to tooth surfaces. Levendag PC. it is obvious that further extensive in vitro and in vivo analysis is necessary to clarify the influence of saliva substitutes on initial microbial adhesion. mutans adhesion. and lactoferrin against several microorganisms from the oral mucosa using a modified agar diffusion test. Boonzaaijer M. Journal of the American Dental Association 2003.35 Their results differ from the outcomes of this study. Proceedings of the Finnish Dental Society 1991. showed decisively higher adhesion. Infection and Immunity 1980. compared with either of the controls. It is important to consider that the amount of enzymes that are bound to the substratum surfaces is rather low. MacKay BJ.32 and that the enzymatic activity of these proteins is maintained when they are bound to the substratum surfaces. which may be due to the short incubation time as well as the rather low xylitol concentrations used for the supplementation of saliva substitutes. Braaksma MMJ. lysozyme (Aldiamed) or lactoferrin (Aldiamed).30 similar adhesion was observed. These considerations indicate that further studies are necessary to elucidate this aspect more thoroughly by means of ellipsometry to correlate bacterial adhesion and absorption of saliva constituents directly. [5] Meyer-Lueckel H. [2] Guggenheimer J. Supportive Care in Cancer 2000.29:623–32. Although some of the saliva substitutes used in this study contain antibacterial proteins and enzymes such as xylitol (Saliva natura. mutans. investigated the antimicrobial effects of a saliva substitutes containing lactoperoxidase.15:563–70.10 This observation might serve as another explanation for the results of this study. muramidase-inactive lysozyme. mutans adhesion to prevent dental caries formation. Schmitz PIM. Little information is available in the literature concerning the efficiency of antimicrobial enzymes in saliva substitutes. observed that saliva substitutes do not adsorb equally to hydrophilic and hydrophobic surfaces.8:23–9.33. which indicates that the influence on initial microbial adhesion might also be low. however. Effect of saliva substitutes on mineral content of demineralized and sound dental enamel. they found significant zones of inhibition around the saliva substitute for all bacteria but not for Candida albicans. lysozyme and lactoferrin in xerostomia: efficacy and safety. antimicrobial agents in saliva substitutes showed no significant influence on S. lysozyme. [9] Laible NJ. Patients with head and neck cancer cured by radiation therapy: a survey of the dry mouth syndrome in long-term survivors. Christersson et al. Current Opinion in Rheumatology 2003.

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Nyvad B. Influence of different restorative materials on lysozyme and amylase activity of the salivary pellicle in situ. Sliepen I. Kho HS. Film-forming properties and viscosities of saliva substitutes and human whole saliva. Morikawa A. Caton MC. Journal of Biomedical Materials Research Part B Applied Biomaterials 2008. Detection of salivary a-amylase and lysozyme exposed on the pellicle formed in situ on different materials. Petrucca A.13:258–69. Gagnon D. Archives of Oral Biology 2005. Adherence of Streptococcus sanguis to salivary mucins bound to glass.69:1771–5. Fejerskov O. Hannig C. Okuda K. Foster JS.74:1904–13. et al. Journal of Dental Research 1998. Journal of Clinical Periodontology 1987. Clinical Oral Investigations 2007. Becker K. Clinical Oral Implants Research 2006. Nakayama GR. Herkstro¨ter F. Journal of Oral Rehabilitation 1991. D’Haen J. Journal of Biomedical Materials Research Part A 2007. Wilson I. 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