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Salivary levels of tumor necrosis factor-alpha in periodontitis.


Periodontal inflammation results in gingival bleeding, pocket formation, destruction of alveolar bone, and eventually loss of teeth. The gingival bleeding is a clinical system of both scurvy and periodontitis, the two conditions are distinct disease entities. Unlike for scurvy, which is caused by vitamin C deficiency, the etiological agents in periodontitis are dental plaque bacteria, especially gram negative micro-organisms. The continued local or systemic bacterial stimulus causes release of proinflammatory mediators, which may have a role in the pathogenesis of atherosclerosis and stroke [1-3]. Lower vitaminosis is associated primarily with defective collagen synthesis, causing tissue dysfunction such as impaired wound healing and ruptured capillaries because of insufficient support of the capillary walls by the connective tissues[3]. In this report we present data characterizing the pattern of TNF-alpha production in whole saliva of patients with periodontitis. We wanted to evaluate whether the salivary concentrations of TNF-alpha were changed in periodontitis.

Materials and methods

The 25 patients (M:F, 13:12) age group 17-30 years were selected. A clinical periodontal examination was performed on all the patients and controls by two calibrated examiners using a calibrated periodontal probe (Hu-friedy, Chicago, IL, USA). The following variables were determined: oral hygiene status, gingival inflammation, probing depth, and clinical attachment level measurements. Oral hygiene status was assessed as the percentage of surface demonstrating plaque. Probing depth and attachment level measurements were performed at six sites on each tooth. Gingival bleeding was assessed on the sites at which probing depth was measured. Gingival redness was determined on two gingival units per tooth. After informent consent had been obtained and, each individual expectorated 10 ml of unstimulated whole saliva into a sterile centrifuge tube. The saliva was centrifuged for 3 min at 8000/g and the clarified supernatant was filtered through a 0.45 mm low protein binding membrane, separated into 0.5 ml aliquots and frozen at _/808C until use. Concentrations of TNF-a were determined using the human TNF-a ELISA kit purchased from SIGMA ImmunoChemicals (St Louis, MO, USA) (4). The assay was performed according to the manufacturer's instructions, and the results are expressed in pg/ml. The detection limit for TNF-a was 4.4 pg/ml, respectively. Data were analylased by SPSS and Students' t test were applied.

Results and discussion

Sufficient quantities of TNF-a (1.45 pg/ml)were present in all saliva sample obtained from healthy subjects. Salivary levels of TNF-a were elevated in all patients presenting symptomatic periodontitis. The median values (2.83 pg/ml)) of TNF-a in saliva of the patients with the periodontitis, of the disease were significantly (p</0.001) higher in comparison with the values obtained from control subjects.

We found a significantly higher amount of TNF-a in all saliva samples obtained from patients with periodontitis in comparison with the healthy controls. A permanent or prolonged presence of high TNF-alpha amounts in saliva may contribute to progression of periodontitis [4-8]. As a diagnostic fluid, saliva is underused. It offers distinctive advantages over serum because it can be collected non-invasively by individuals with modest training. Further more, saliva analysis demonstrated comparable values of various biochemical and immunological parameters with those that are routinely evaluated in blood [6]. Saliva has more diagnostic value that is generally appreciated and has promise as an aid in the diagnosis of systemic and especially oral pathologies. Our data suggest an important role of high concentrations of TNF-alpha in perpetuation of the pathogenetical events in periodontitis. Inhibiting TNF-alpha activities in this disease may provide therapeutic benefits but more experimental and clinical studies are required for possible efficacy of drugs that can interfere with excess TNF-alpha [5-9]. Based on the presence of TNF-alpha in whole saliva we think that saliva analysis is a useful, worthwhile tool in the diagnosis and follow-up of periodontal disease, particularly in patients with gingivitis and periodontitis affections of oral mucosa.


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[2.] Rai, B., S.C. Anand, S. Kharb, 2006. The panoramic radiograph as a detective of cardiovascular risk. World J. Med. Sci., 1(2): accepted.

[3.] Rai, B., S.C. Anand, R. Jain, S. Kharb. Salivary vitamin C and E in oral cancer. Redox Report (accepted).

[4.] Abram, M., D. Vuckovic, B. Wraber, M. Doric, 2000. Plasma cytokine response in mice with bacterial infection. Mediat Inflamm, 9: 229-234.

[5.] Nakao, S., T. Kuwano, T. Ishibashi, M. Kuwano, M. Ono, 2003. Synergistic effect of TNF-alpha in soluble VCAM-1-induced angiogenesis through alpha 4 integrins. J. Immunol., 170: 5704-5711.

[6.] Gerstenfeld, L.C., T.J. Cho, T. Kon, et al, 2003. Impaired fracture healing in the absence of TNF-alpha signaling: the role of TNF-alpha in endochondral cartilage resorption. J. Bone Miner Res., 18: 1584-1592.

[7.] Sugermann, P.B., N.W. Savage, G.J. Seymour, L.J. Wals, 1996. Is there a role for tumor necrosis factor-alpha (TNF-alpha) in oral lichen planus? J. Oral Pathol. Med., 25: 219-224.

[8.] Yammamoto, T., K. Yoneda, E. Ueta, T. Osaki, 1994. Serum cytokines, interleukin-2 receptor, and soluble intercellular adhesion molecule-1 in oral disorders. Oral Surg. Oral Med. Oral Pathol., 78: 727-735.

[9.] Nagler, R.M., O. Hershkovich, S. Lischinsky, E. Diamond, A.Z. Reznick, 2002. Saliva analysis in the clinical setting: revisiting an under used diagnostic tool. J. Investig Med., 50: 214-225.

Dr. Balwant Rai

Editor In Chief International Journal of Dental science, P.D.M. Dental College and Research Institute, Haryana

Corresponding Author: Dr. Balwant Rai, Editor In Chief International Journal of Dental science, P.D.M. Dental College and Research Institute, Haryana E-mail:
Table I: Basic clinical parameter and Salivary levels of tumor
necrosis factor-alpha level in periodontally healthy and with
periodontal disease.

Parameters Periodontal healthy

Number of teeth 26.7 [+ or -] 1.3
percentage of sites with plaque 32.0 [+ or -] 12.8
Bleeding on probing 14.9 [+ or -] 7.3
Gingival redness 4.3 [+ or -] 1.8
Probing depth [greater than
 or equal to] 5 mm 1.5 [+ or -] 1.3
Attachment level sites > 3 mm 0.4 [+ or -] 0.2
Attachment level sites [greater
 than or equal to] 3 mm 1.1 [+ or -] 0.9
Mean probing depth (mm) 2.0 [+ or -] 0.8
Mean attachment level (mm) 0.8 [+ or -] 0.8
Salivary tumor necrosis factor
 alpha (pg/ml) 1.45 [+ or -] 0.19

 With periodontal
Parameters disease p value

Number of teeth 24.8 [+ or -] 1.6 p < 0.001
percentage of sites with plaque 73.4 [+ or -] 12.0 p < 0.001
Bleeding on probing 54.3 [+ or -] 17.4 p < 0.001
Gingival redness 52.8 [+ or -] 27.0 p < 0.001
Probing depth [greater than
 or equal to] 5 mm 28.6 [+ or -] 13.8 p < 0.001
Attachment level sites > 3 mm 13.2 [+ or -] 11.8 p < 0.001
Attachment level sites [greater
 than or equal to] 3 mm 28.7 [+ or -] 21.6 p < 0.001
Mean probing depth (mm) 4.3 [+ or -] 0.8 p < 0.001
Mean attachment level (mm) 1.9 [+ or -] 1.1 p < 0.001
Salivary tumor necrosis factor
 alpha (pg/ml) 2.83 [+ or -] 0.14 p < 0.001
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Article Details
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Title Annotation:Original Article
Author:Rai, Balwant
Publication:Advances in Medical and Dental Sciences
Article Type:Clinical report
Geographic Code:9INDI
Date:May 1, 2008
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