Journal Mobile Options
Table of Contents
Vol. 46, No. 2, 2012
Issue release date: April 2012
Caries Res 2012;46:130–139

Enzyme Activities in the Oral Fluids of Patients Suffering from Bulimia: A Controlled Clinical Trial

Schlueter N. · Ganss C. · Pötschke S. · Klimek J. · Hannig C.
aDepartment of Conservative and Preventive Dentistry, Dental Clinic, Justus Liebig University, Giessen, and bClinic of Conservative Dentistry, Faculty of Medicine ‘Carl Gustav Carus’, Technical University of Dresden, Dresden, Germany

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Patients with bulimia nervosa are at high risk for dental erosion. However, not all bulimic patients suffer from erosion, irrespective of the severity of their eating disorder. It is often speculated that differences in the saliva are important, however, little is known about salivary parameters in bulimic patients, particularly directly after vomiting. The aim of the clinical trial was to compare different salivary parameters of subjects suffering from bulimia with those of healthy controls. Twenty-eight subjects participated (14 patients with bulimia nervosa, 7 of them with erosion; 14 matched healthy controls). Resting and stimulated saliva of all participants was analysed as well as saliva collected from bulimic patients directly and 30 min after vomiting. Parameters under investigation were flow rate, pH, buffering capacity and the enzyme activities of proteases in general, collagenase, pepsin, trypsin, amylase, peroxidase, and lysozyme. Regarding flow rate, pH and buffering capacity only small differences were found between groups; buffering capacity directly after vomiting was significantly lower in bulimic subjects with erosion than in subjects without erosion. Differences in enzymatic activities were more pronounced. Activities of proteases, collagenase and pepsin in resting and proteases in stimulated saliva were significantly higher in bulimic participants with erosion than in controls. Peroxidase activity was significantly decreased by regular vomiting. Proteolytic enzymes seem to be relevant for the initiation and progression of dental erosion directly after vomiting, maybe by both hydrolysis of demineralized dentine structures as well as modulation of the pellicle layer.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Apajalathi S, Sorsa T, Railavo S, Ingman T: The in vivo levels of matrix metalloproteinase-1 and -8 in gingival crevicular fluid during initial orthodontic tooth movement. J Dent Res 2003;82:1018–1022.

    External Resources

  2. Bardow A, Lagerlöf F, Nauntofte B, Tenovuo J: The role of saliva; in Fejerskov O, Kidd E, Nyvad B, Baelum V (eds): Dental Caries. The Disease and Its Clinical Management. Oxford, Blackwell Munksgaard, 2008, pp 189–208.
  3. Bardow A, Moe D, Nyvad B, Nauntofte B: The buffer capacity and buffer systems of human whole saliva measured without loss of CO2. Arch Oral Biol 2000;45:1–12.
  4. Bartlett DW, Coward PY: Comparison of the erosive potential of gastric juice and a carbonated drink in vitro. J Oral Rehabil 2001;28:1045–1047.
  5. Birkhed D, Heintze U: Salivary secretion rate, buffer capacity, and pH; in Tenovuo JO (ed): Human Saliva: Clinical Chemistry and Microbiology. Boca Raton, CRC Press, 1989, vol 1, pp 25–73.
  6. Davis GE: Identification of an abundant latent 94-kDa gelatin-degrading metalloprotease in human saliva which is activated by acid exposure: implications for a role in digestion of collagenous proteins. Arch Biochem Biophys 1991;286:551–554.
  7. DenBesten PK, Punzi JS, Li W: Purification and sequencing of a 21 and 25 kDa bovine enamel metalloproteinase. Eur J Oral Sci 1998;106:345–349.
  8. Descamps FJ, Martens E, Ballaux F, Geboes K, Opdenakker G: In vivo activation of gelatinase B/MMP-9 by trypsin in acute pancreatitis is a permissive factor in streptozotocin-induced diabetes. J Pathol 2004;204:555–561.
  9. Frydrych AM, Davies GR, McDermott BM: Eating disorders and oral health: a review of the literature. Aust Dent J 2005;50:6–15.
  10. Furuyama M, Koshika S, Kitamura Y, Nakayama Y: Trypsin-like protease and glucose-6-phosphate dehydrogenase in the human submandibular salivary gland. Arch Oral Biol 1987;32:761–762.
  11. Ganss C, Hardt M, Blazek D, Klimek J, Schlueter N: Effects of toothbrushing force on the mineral content and demineralized organic matrix of eroded dentine. Eur J Oral Sci 2009;117:255–260.
  12. Ganss C, Hardt M, Lussi A, Cocks A-K, Klimek J, Schlueter N: Mechanism of action of tin-containing fluoride solutions as anti-erosive agents in dentine – an in vitro tin-uptake, tissue loss and scanning electron microscopy study. Eur J Oral Sci 2010;118:376–384.
  13. Ganss C, Klimek J, Starck C: Quantitative analysis of the impact of the organic matrix on the fluoride effect on erosion progression in human dentine using longitudinal microradiography. Arch Oral Biol 2004;49:931–935.
  14. Ganss C, Lussi A: Diagnosis of erosive tooth wear; in Lussi A (ed): Dental Erosion: From Diagnosis to Therapy. Monogr Oral Sci. Basel, Karger, 2006, pp 32–43.
  15. Garant PR: Dentin: Oral Cells and Tissues. Berlin, Quintessence Publishing, 2003, pp 25–52.
  16. Gonçalves R, Soares S, Mateus N, de Freitas V: Inhibition of trypsin by condensed tannins and wine. J Agric Food Chem 2007;55:7596–7601.
  17. Hannig C, Attin T, Hannig M, Henze E, Brinkmann K, Zech R: Immobilisation and activity of human alpha-amylase in the acquired enamel pellicle. Arch Oral Biol 2004a;49:469–475.
  18. Hannig C, Becker K, Häusler N, Hoth-Hannig W, Attin T, Hannig M: Protective effect of the in situ pellicle on dentin erosion – an ex vivo pilot study. Arch Oral Biol 2007;52:444–449.
  19. Hannig C, Hannig M, Attin T: Enzymes in the acquired enamel pellicle. Eur J Oral Sci 2005;113:1–12.

    External Resources

  20. Hannig C, Spitzmüller B, Al-Ahmad A, Hannig M: Effects of Cistus-tea on bacterial colonization and enzyme activities of the in situ pellicle. J Dent 2008a;36:540–545.
  21. Hannig C, Spitzmüller B, Hannig M: Characterisation of lysozyme activity in the in situ pellicle using a fluorimetric assay. Clin Oral Invest 2009;13:15–21.

    External Resources

  22. Hannig C, Spitzmüller B, Knausenberger S, Hoth-Hannig W, Hellwig E, Hannig M: Detection and activity of peroxidase in the in situ formed enamel pellicle. Arch Oral Biol 2008b;53:849–858.
  23. Hannig C, Spitzmüller B, Miller M, Hellwig E, Hannig M: Intrinsic enzymatic crosslinking and maturation of the in situ pellicle. Arch Oral Biol 2008c;53:416–422.
  24. Hannig M, Balz M: Protective properties of salivary pellicles from two different intraoral sites on enamel erosion. Caries Res 2001;35:142–148.
  25. Hannig M, Fiebiger M, Güntzer M, Döbert A, Zimehl R, Nekrashevych Y: Protective effect of the in situ formed short-term salivary pellicle. Arch Oral Biol 2004b;49:903–910.
  26. Hannig M, Joiner A: The structure, function and properties of the acquired pellicle. Monogr Oral Sci 2006;19:29–64.
  27. Hara AT, Lussi A, Zero DT: Biological factors; in Lussi A (ed): Dental Erosion – From Diagnosis to Therapy. Monogr Oral Sci. Basel, Karger, 2006, vol 20, pp 88–99.
  28. Ingman T, Tervahartiala T, Ding Y, Tschesche H, Haerian A, Kinane DF, Konttinen YT, Sorsa T: Matrix metalloproteinases and their inhibitors in gingival crevicular fluid and saliva of periodontitis patients. J Clin Periodontol 1996;23:1127–1132.
  29. Joiner A, Schwarz A, Philpotts CJ, Cox TF, Huber K, Hannig M: The protective nature of pellicle towards toothpaste abrasion on enamel and dentine. J Dent 2008;36:360–368.

    External Resources

  30. Kim TH, Lee KJ, Yeo M, Kim DK, Cho SW: Pepsin detection in the sputum/saliva for the diagnosis of gastroesophageal reflux disease in patients with clinically suspected atypical gastroesophageal reflux disease symptoms. Digestion 2008;77:201–206.
  31. Kleter GA, Damen JJ, Everts V, Niehof J, ten Cate JM: The influence of the organic matrix on demineralization of bovine root dentin in vitro. J Dent Res 1994;73:1523–1529.
  32. Lamanda A, Cheaib Z, Turgut MD, Lussi A: Protein buffering in model systems and in whole human saliva. PLoS One 2007;2:e263.

    External Resources

  33. Lilienthal B: An analysis of the buffer systems in saliva. J Dent Res 1955;34:516–530.
  34. Lindstad RI, Sylte I, Mikalsen SO, Seglen PO, Berg E, Winberg JO: Pancreatic trypsin activates human promatrix metalloproteinase-2. J Mol Biol 2005;350:682–698.
  35. Lussi A, Jaeggi T: Erosion – diagnosis and risk factors. Clin Oral Invest 2008;12:S5–S13.
  36. Lussi A, Schlueter N, Rakhmatullina E, Ganss C: Dental erosion – an overview with emphasis on chemical and histopathological aspects. Caries Res 2011;45(suppl 1):2–12.
  37. Maeda H: A new lysozyme assay based on fluorescence polarization or fluorescence intensity utilizing a fluorescent peptidoglycan substrate. J Biochem 1980;88:1185–1191.
  38. Mäkelä M, Salo T, Uitto VJ, Larjava H: Matrix metalloproteinases (MMP-2 and MMP-9) of the oral cavity: cellular origin and relationship to periodontal status. J Dent Res 1994;73:1397–1406.
  39. Moon PC, Weaver J, Brooks CN: Review of matrix metalloproteinases’ effect on the hybrid dentin bond layer stability and chlorhexidine clinical use to prevent bond failure. Open Dent J 2010;4:147–152.
  40. Morishita Y, Iinuma Y, Nakashima N, Majima K, Mzuguchi K, Kawamura Y: Total and pancreatic amylase measured with 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltoside. Clin Chem 2000;46:928–933.
  41. Nascimento FD, Minciotti CL, Geraldeli S, Carrilho MR, Pashley DH, Tay FR, Nader HB, Salo T, Tjaderhane L, Tersariol IL: Cysteine cathepsins in human carious dentin. J Dent Res 2011;90:506–511.
  42. Nekrashevych Y, Hannig M, Stosser L: Assessment of enamel erosion and protective effect of salivary pellicle by surface roughness analysis and scanning electron microscopy. Oral Health Prev Dent 2004;2:5–11.

    External Resources

  43. Proctor GB, Chan KM: A fluorometric assay of peroxidase activity utilizing 2′,7′-dichlorofluorescein with thiocyanate: application to the study of salivary secretion. J Biochem Biophys Methods 1994;28:329–336.
  44. Richter C, Tanaka T, Yada RY: Mechanism of activation of the gastric aspartic proteinases: pepsinogen, progastricsin and prochymosin. Biochem J 1998;335:481–490.
  45. Robb ND, Smith BG, Geidrys LE: The distribution of erosion in the dentitions of patients with eating disorders. Br Dent J 1995;178:171–175.
  46. Sarosiek J, Rourk RM, Piascik R, Namiot Z, Hetzel DP, McCallum RW: The effect of esophageal mechanical and chemical stimuli on salivary mucin secretion in healthy individuals. Am J Med Sci 1994;308:23–31.
  47. Sas R, Dawes C: The intra-oral distribution of unstimulated and chewing-gum-stimulated parotid saliva. Arch Oral Biol 1997;42:469–474.
  48. Scheutzel P, Gerlach U: Alpha-amylase isoenzymes in serum and saliva of patients with anorexia and bulimia nervosa. Z Gastroenterol 1991;29:339–345.
  49. Schlueter N, Ganss C, Hardt M, Schegietz D, Klimek J: Effect of pepsin on erosive tissue loss and the efficacy of fluoridation measures in dentine in vitro. Acta Odontol Scand 2007;65:298–305.
  50. Schlueter N, Hardt M, Klimek J, Ganss C: Influence of the digestive enzymes trypsin and pepsin in vitro on the progression of erosion in dentine. Arch Oral Biol 2010;55:294–299.
  51. Shafik A, El Sibai O, Shafik AA, Mostafa R: Effect of topical esophageal acidification on salivary secretion: identification of the mechanism of action. J Gastroenterol Hepatol 2005;20:1935–1939.

    External Resources

  52. Sorsa T, Tjäderhane L, Salo T: Matrix metalloproteinases (MMPs) in oral diseases. Oral Dis 2004;10:311–318.
  53. Stuchell RN, Mandel ID: A comparative study of salivary lysozyme in caries-resistant and caries-susceptible adults. J Dent Res 1983;62:552–554.
  54. Tatsuguchi A, Fukuda Y, Ishizaki M, Yamanaka N: Localization of matrix metalloproteinases and tissue inhibitor of metalloproteinases-2 in normal human and rabbit stomachs. Digestion 1999;60:246–254.
  55. Tjäderhane L, Larjava H, Sorsa T, Uitto VJ, Larmas M, Salo T: The activation and function of host matrix metalloproteinases in dentin matrix breakdown in caries lesions. J Dent Res 1998;77:1622–1629.
  56. Uitto VJ: Human gingival proteases. I: Extraction and preliminary characterization of trypsin-like and elastase-like enzymes. J Periodontal Res 1987;22:58–63.
  57. van Strijp AJ, Jansen DC, DeGroot J, ten Cate JM, Everts V: Host-derived proteinases and degradation of dentine collagen in situ. Caries Res 2003;37:58–65.
  58. Veerman EC, van den Keybus PA, Vissink A, Nieuw Amerongen AV: Human glandular salivas: their separate collection and analysis. Eur J Oral Sci 1996;104:346–352.
  59. Vilen ST, Nyberg P, Hukkanen M, Sutinen M, Ylipalosaari M, Bjartell A, Paju A, Haaparanta V, Stenman UH, Sorsa T, Salo T: Intracellular co-localization of trypsin-2 and matrix metalloprotease-9: possible proteolytic cascade of trypsin-2, MMP-9 and enterokinase in carcinoma. Exp Cell Res 2008;314:914–926.
  60. Vray B, Hoebeke J, Saint-Guillain M, Leloup R, Strosberg AD: A new quantitative fluorimetric assay for phagocytosis of bacteria. Scand J Immunol 1980;11:147–153.
  61. Vuotila T, Ylikontiola L, Sorsa T, Luoto H, Hanemaaijer R, Salo T, Tjäderhane L: The relationship between MMPs and pH in whole saliva of radiated head and neck cancer patients. J Oral Pathol Med 2002;31:329–338.
  62. Zheng X, Pan H, Wang Z, Chen H: Real-time enzymatic degradation of human dentin collagen fibrils exposed to exogenous collagenase: an AFM study in situ. J Microsc 2011;241:162–170.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50