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Anti-Inflammatory Effects of Lactobacillus Brevis (CD2) on Periodontal Disease

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Anti-Inflammatory Effects of Lactobacillus Brevis (CD2) on Periodontal Disease See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/6260480 Anti-inflammatory effects of Lactobacillus brevis (CD2) on periodontal disease Article in Oral Diseases · August 2007 DOI: 10.1111/j.1601-0825.2006.01291.x · Source: PubMed CITATIONS READS 78 88 7 authors, including: Mariagrazia Perilli Antonella Polimeni Università degli Studi dell'Aquila Sapienza University of Rome 123 PUBLICATIONS 2,475 CITATIONS 205 PUBLICATIONS 2,068 CITATIONS SEE PROFILE SEE PROFILE Gianfranco Amicosante Maria Grazia Cifone Università degli Studi dell'Aquila Università degli Studi dell'Aquila 243 PUBLICATIONS 6,197 CITATIONS 229 PUBLICATIONS 4,952 CITATIONS SEE PROFILE SEE PROFILE Some of the authors of this publication are also working on these related projects: Probiotics and their Metabolites View project NDM-1 mutants View project All content following this page was uploaded by Maria Grazia Cifone on 17 October 2016. The user has requested enhancement of the downloaded file. All in-text references underlined in blue are added to the original document and are linked to publications on ResearchGate, letting you access and read them immediately. Oral Diseases (2007) 13, 376–385. doi:10.1111/j.1601-0825.2006.01291.x Ó 2007 The Authors. Journal compilation Ó 2007 Blackwell Munksgaard All rights reserved http://www.blackwellmunksgaard.com ORIGINAL ARTICLE Anti-inflammatory effects of Lactobacillus brevis (CD2) on periodontal disease DN Della Riccia1, F Bizzini1, MG Perilli2, A Polimeni3, V Trinchieri4, G Amicosante2 and MG Cifone1 1Department of Experimental Medicine, University of L’Aquila, L’Aquila, Italy; 2Department of Biomedical Sciences and Technologies, University of L’Aquila, L’Aquila, Italy; 3Department of Odontostomatologic Sciences, University ÔLa Sapienza’, Rome, Italy; 4Department of Infectious and Tropical Diseases, University ÔLa Sapienza’, Rome, Italy OBJECTIVES: To analyze the anti-inflammatory effects Introduction of Lactobacillus brevis extracts on periodontitis patients and to investigate the involved mechanisms in vitro on Lactic acid bacteria (LAB) belong to a variety of genera activated macrophages. used for milk fermentation. The primary metabolic end METHODS: Eight healthy subjects and 21 patients with product from carbohydrate metabolism is lactic acid, chronic periodontitis were enrolled to analyze the effect which in turn preserves milk by providing the acidity of L. brevis-containing lozenges on periodontitis-associ- necessary for a tart flavor and for changes in the ated symptoms and signs. Before and after the treat- structure of casein to achieve syneresis and desired ment, the patients received a complete periodontal functional characteristics. The extensive knowledge on examination. Saliva samples, collected before and after LAB has opened new possibilities for their application treatment, were analyzed for metalloproteinase and ni- (for reviews see Konings et al, 2000; C¸aglar et al, 2005). tric oxide synthase (NOS) activity, immunoglobulin-A Reports of many investigators have confirmed that LAB and their products have beneficial effects on the health (IgA), prostaglandin E2 (PGE2) and c-interferon (IFN-c) levels. Arginine deiminase (AD) and NOS activities were of animals and humans, i.e., protection against enteric determined through a radiometric assay. Metallopro- infections, use as an oral adjuvant, immunopotentiation teinases were assayed by zymogram and Western blot- in malnutrition, and prevention of chemically induced tumors (Bodana and Rao, 1990; Roberfroid, 1998; ting, whereas IgA, PGE2 and IFN-c were assayed by enzyme-linked imunosorbent assay tests. Dugas et al, 1999; Gorbach, 2000). Almost all genera of RESULTS: The treatment led to the total disappearance LAB are able to produce bacteriocins that are primarily or amelioration of all analyzed clinical parameters in all lethal to other strains and species of bacteria form short- patients. This was paralleled to a significant decrease of lived pores in biologic membranes by interactions with different compounds including nisin, lipid II and nitrite/nitrate, PGE2, matrix metalloproteinase, and IFN- c levels in saliva samples. phospholipids, thereby killing the target bacteria (Kon- CONCLUSION: Our results suggest that the anti- ings et al, 2000). In the attempt to better characterize the inflammatory effects of L. brevis could be attributed to metabolic features of some LAB strains, our group has the presence of AD which prevented nitric oxide gen- recently shown the presence of high levels of arginine eration. Our findings give further insights into the deiminase (AD) in Lactobacillus brevis (Di Marzio et al, knowledge of the molecular basis of periodontitis and 2001). As bacterial AD may compete with nitric oxide have a potential clinical significance, giving the experi- synthase (NOS) by using the same substrate, arginine, mental ground for a new innovative, simple and effica- we decided to investigate the ability of L. brevis extracts cious therapeutical approach of periodontal disease. to inhibit NOS activity as well other inflammatory Oral Diseases (2007) 13, 376–385 parameters [c-interferon (IFN-c), prostaglandin E2 (PGE2) and metalloproteinases], known to be associated Keywords: L. brevis; nitric oxide; metalloproteinases; prostaglan- with NOS induction (Cifone et al, 1999; Goodwin et al, din E ; c-interferon 1999; Di Marzio et al, 2001; Ishii et al, 2003). In 2 particular, the chronic periodontal disease was chosen as the known inflammatory model to test the potential Corresponding Author: Dr MG Cifone, Department of Experimental in vivo anti-inflammatory effects of L. brevis. Medicine, University of L’Aquila, Via Vetoio, 10, Coppito 2, 67100 Periodontal disease is a major cause of tooth loss in L’Aquila, Italy. Tel: 0862 433503, Fax: 0862 433523, E-mail: pres- humans and is one of the most prevalent diseases [email protected] associated with bone loss. Following bacterial coloniza- Received 19 November 2006; revised 9 January 2006; accepted 30 January 2006 tion, the gingiva becomes inflamed leading, in some cases, Anti-inflammatory effects of L. brevis DN Della Riccia et al 377 to the destruction of the alveolar bone. Periodontitis has (Centricon 10) from Amicon (Amicon, Millipore Co., two distinct but interconnected etiologic components: Bedford, MA, USA). IFN-c (Endogen, Inc., Walburn, periodontopathic bacteria adjacent to the periodontal MA, USA) and PGE2 Elisa kits (Assay Designs Inc., tissues, and host-mediated connective tissue-destructive Ann Arbor, MI, USA) were purchased from TEMA responses to the causative bacteria and their metabolic ricerca srl (Bologna, Italy). IgAs Elisa kits were products (Golub et al, 1998). Treatment strategies obtained from IPR (Catania, Italy). against periodontal diseases have evolved with the aim of eliminating specific pathogens or suppressing destruc- Preparation of bacterial extracts tive host response. Although the pathogenesis of the Lactobacillus brevis, obtained from VSL Pharmaceuti- various forms of this disease is not completely under- cals (Gaithersburg, MD, USA) in a pure lyophilized stood, several factors are believed to play an important form (108 colony-forming units per gram), was routinely role, including inflammatory cytokines (McGee et al, grown in MRS broth medium under aerobic conditions. 1998; Graves, 1999), PGE2 (Offenbacher, 1996; Tsai et al, For the determination of AD activity and for in vitro 1998; Kornman, 1999), nitric oxide (Matejka et al, 1999; experiments on macrophages, L. brevis cultures were Takeichi et al, 2000) and salivary immunoglobulin-A washed and resuspended in 10 ml phosphate-buffered (IgA) antibodies (Schenk et al, 1993). Moreover, many of solution (PBS), sonicated (30 min, alternating 10 s the characteristics of periodontal diseases such as inflam- sonication and 10 s pause) with a Vibracell sonicator mation and attachment loss are associated with proteo- (Sonic and Materials Inc., Danbury, CT, USA), centri- lytic events. Indeed, oral damage can result from the fuged at 7500 g for 20 min at 4°C and the supernatants release of an array of proteolytic enzymes by colonizing stored at )20°C until used (bacterial extract). For bacteria as well as host-derived proteases (Henskens et al, the in vitro experiments, the bacterial extract was 1996; Baron et al, 1999; Korostoff et al, 2000; Travis and added to cell cultures at 0.5 mg/105cells/1 ml)1 (final Potemba, 2000). Selection of an appropriate delivery concentration). system is an important factor and, periodontitis being a For gene expression studies, genomic DNA was Ôlocalized’ disease condition, local drug treatment should isolated from an overnight culture of L. brevis using be preferred. a WIZARD genomic DNA purification kit (Promega, The aims of this study were firstly to analyze the Milan, Italy), according to the manufacturer’s effects of AD-expressing L. brevis-containing lozenges, instructions. both on the clinical signs of chronic periodontitis patients and on the NOS activity levels, matrix metal- Arginine deiminase activity loproteinase (MMP) levels, eicosanoid generation, IgA Arginine deiminase activity was determined in L. brevis 3 levels, IFN-c levels of the saliva. The mechanisms extracts by measuring the conversion of [ H]-L-arginine to 3 underlying the anti-inflammatory effect of L. brevis [ H]-L-citrulline. Samples were homogenized by sonica- extract were investigated in vitro on rat peritoneal tion in homogenization ice-cold buffer (25 mM Tris-HCl, macrophages activated
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