The level of beta defensin-2 in saliva and its expression in parotid gland epithelial cells after probiotic (Lactobacillus reuteri) induction to inhibit Streptococcus mutans in caries

ABSTRACT

Objective: The aim of this study was to prove that administrating L. reuteri probiotics can increase the level of BD-2 saliva and BD-2 expression in the epithelial parotid glands of Wistar rats. Materials and Methods: Experimental design in this study was randomized control group post test only. Twenty-four white male Rattus norvegicus Wistar strain rats were divided into four groups. The negative control group included rats not induced by S. mutans whereas the positive control group included rats induced by S. mutans. The two treatment groups are as follows: treatment 1 (T1), the group that is induced for 14 days by L. reuteri and 7 days by S. mutans and treatment 2 (T2), the group which is induced simultaneously by S. mutans and L. reuteri for 14 days. L. reuteri culture at a concentration of 108 colony-forming unit/ ml and S. mutans culture at a concentration of 1010 are induced in the oral cavity of the Wistar rats. The Elisa technique is used to examine the salivary level of BD-2, whereas the immunohistochemical technique is used to examine the BD-2 expression in the epithelial salivary glands. Results: The study shows the increasing levels of BD-2 and BD-2 expression in the epithelial parotid glands after the administration of L. reuteri probiotics. Besides, there is a relationship between the increasing expression of BD-2 in the epithelial parotid glands with the decreasing amount of S. mutans. Conclusion: Giving L. reuteri probiotic scan increases the level of saliva of BD-2 and the expression of BD-2 in the parotid glands.

Key words:

Beta defensin-2 - caries - Lactobacillus reuteri probiotics - Streptococcus mutans

• REFERENCES

• 1 Shifa S, Muthu MS, Amarlal D, Rathna PrabhuV. Quantitative assessment of IgA levels in the unstimulated whole saliva of caries-free and caries-active children. J Indian Soc Pedod Prev Dent 2008; 26: 158-61
• 2 National Report. Basic Health Research. Jakarta: Research and Development Agency, Ministry of Health of Indonesian Republic; 2007: p. 140
• 3 Basic Health Research of East Java Province. Jakarta: Research and Development Agency, Ministry of Health of Indonesian Republic; 2007: p. 143
• 4 Tao R, Richard J, Jurevic, Kimberly K. Coulton. Salivary antimicrobial peptide expression and dental caries experience in children. Ant Agent and Chem 2005; 49: 3883-8
• 5 Dale BA, Fredericks LP. Antimicrobial peptides in the oral environment: Expression and function in health and disease. Curr Issues Mol Biol 2005; 7: 119-33
• 6 Dale BA, Tao R, Kimball JR, Jurevic RJ. Oral antimicrobial peptides and biological control of caries. BMC Oral Health 2006; 6 (Suppl. 01) S13
• 7 Wallace TC, Guarner F, Madsen K, Cabana MD, Gibson G, Hentges E. et al. Human gut microbita and its relationship to health and disease. Nutrition Reviews Vol 2011; 69: 392-403
• 8 Lamont RJ, Burne RA, Lantz MS, Leblanc DJ. The Oral Environment-Oral Microbiology and The Immune response, American Society for Microbiology Press. 2006: 201-29
• 9 Valeur N, Engel P, Carbajal N, Connolly E, Ladefoged K. Colonization and immunomodulation by Lactobacillus reuteri ATCC 55730 in the human gastrointestinal tract. Appl Environ Microbiol 2004; 70: 1176-81
• 10 Ooshima T, Sumi N, Izumitani A, Sobue S. Effect of inoculum size and frequency on the establishment of Streptococcus mutans in the oral cavities of experimental animals. J Dent Res 1988; 67: 964-8
• 11 Gresz V, Tae-Hwan K, Hong G, Agre P, Steward MC, King LS. et al. Immunolocalization of AQP-5 in Rat Parotid and Submandibular Salivary Glands After Stimulation of Secretion in vivo. Am J Physiol Gastrointes Liver Physiol 2004; 287: 151-G161
• 12 Sutton S. Measurement of microbial cells by optical density. J Validation Technol 2011; 17: 46-9
• 13 Soini Y, Pääkkö P, Lehto VP. Histopathological evaluation of apoptosis in cancer. Am J Pathol 1998; 153: 1041-53
• 14 Pizem J, Cor A. Detection of apoptosis cells in tumour paraffin sections. Radiol Oncol 2003; 37: 225-32
• 15 Nishimura E, Eto A, Kato M, Hashizume S, Imai S, Nisizawa T. et al. Oral streptococci exhibit diverse susceptibility to human beta-defensin-2: Antimicrobial effects of hBD-2 on oral streptococci. Curr Microbiol 2004; 48: 85-7
• 16 Yin C, Dang HN, Gazor F, Huang GT. Mouse salivary glands and human β-defensin-2 as a study model for antimicrobial gene therapy: Technical considerations. Int J Antimicrob Agents 2006; 28: 352-60
• 17 Ganz T. Defensins: Antimicrobial peptides of innate immunity. Nat Rev Immunol 2003; 3: 710-20
• 18 Ghosh D, Porter E, Shen B, Lee SK, Wilk D, Drazba J. et al. Paneth cell trypsin is the processing enzyme for human defensin-5. Nat Immunol 2002; 3: 583-90
• 19 Abiko Y, Nishimura M, Kaku T. Defensins in saliva and the salivary glands. Med Electron Microsc 2003; 36: 247-52
• 20 Schlee M, Wehkamp J, Altenhoefer A, Oelschlaeger TA, Stange EF, Fellermann K. Induction of human beta-defensin 2 by the probiotic Escherichia coli Nissle 1917 is mediated through flagellin. Infect Immun 2007; 75: 2399-407
• 21 Uehara A, Takada H. Synergism between TLRs and NOD1/2 in oral epithelial cells. J Dent Res 2008; 87: 682-6