TUMOUR NECROSIS FACTOR ALPHA LEVELS IN GINGIVAL CREVICULAR FLUID OF PERIODONTITIS PATIENTS WITH/WITHOUT MTDNA DELETION IN GINGIVAL TISSUE

Yıl 2014, Cilt: 24 Sayı: 2, 170 - 177, 11.02.2015

Cenk Çanakcı Gülnihal Doğan

https://doi.org/10.17567/dfd.11462

Cited By: 1

Öz

Aim: There are strong evidences to suggest that Tumour necrosis factor alpha (TNF-α) plays important role in the pathogenesis of destructive periodontal pathologies such as periodontitis. Excessive amounts increase reactive oxygen species (ROS) production by mitochondria, which are likely to damage cellular macromolecules, including mtDNA. The 5-kbp mtDNA deletion of gingival tissue was detected in destructive periodontal pathologies. In this study we aimed to investigate the gingival crevicular fluid (GCF) levels of TNF-α and the 5-kbp mtDNA deletion of gingival tissue in patients with periodontitis and healthy controls. Also, we examined correlations between the GCF levels of TNF-α and the 5-kbp mtDNA deletion of gingival tissue and clinical parameters of periodontitis patients. Material and Method: The study was carried out in 64 subjects: 32 patients with chronic periodontitis (CP) and 32 periodontally healthy controls. Samples were collected from the diseased gingival tissues with ≥5mm periodontal pockets, gingival inflammation, and bone loss during periodontal flap surgery from CP group and healthy gingival tissues were collected during crown lengthening procedures. TNF-α levels in GCF samples were assayed by commercial ELISA kits. Determination of the deleted mtDNA was employed a semiquantitative PCR method. Results: All clinical parameter scores of the tissue sampling areas were significantly higher in the CP group compared to the control group. The mean GCF TNF-α level in CP group was significantly higher than that of the controls. In CP group, the mean GCF TNF-α level of patients with deleted mtDNA was significantly higher than those of patients with non-deleted mtDNA Conclusion: According to this study findings, we speculated that overexpression of TNF-α, which is detected in GCF, in gingival tissue of periodontitis patients, may induces elevated ROS production in mitochondria and may lead to oxidative mitochondrial injury such as mtDNA deletion of neighboring cells

Anahtar Kelimeler

TNF-α, mtDNA deletion, gingiva, gingival crevicular fluid, destructive periodontal pathology

GINGIVAL DOKULARINDA 5-KBP MTDNA DELESYONU OLAN VEYA OLMAYAN PERIODONTITIS HASTALARININ DIŞETİ OLUĞU SIVISINDAKİ TÜMÖR NEKROZ FAKTÖRÜ ALFA SEVİYELERİ

Öz

 Amaç: Tümör nekroz faktörü alfa (TNF-α) ’nın yıkıcı periodontal hastalıklardan olan periodontitisin patogenezinde önemli rolleri olduğuna dair güçlü deliller sunulmuştur. Mitokondrilerce reaktif oksijen türleri (ROT) üretiminin aşırı miktarlarda artması sonucu hücresel makromoleküllerden olan mtDNA’ ya zarar verebilir. Yıkıcı periodontal hastalıklarda gingival dokulardan 5-kbp mtDNA delesyonu tespit edilmiştir. Bu çalışmada periodontitisitli ve sağlıklı hastaların dişeti oluğu sıvısında TNF-α ve dişeti dokularında 5-kbp mtDNA delesyonunun araştırılması amaçlanmıştır. İlaveten periodontitis hastalarınının klinik parametreleri ile DOS TNF-α seviyesi ve dişeti dokularında 5-kbp mtDNA delesyonu arasındaki korelasyon durumu araştırılmıştır.  Material ve Metod: Bu çalışmaya 32 periodontal sağlıklı, 32 kronik periodontitisli (KP) olmak üzere 64 hasta dahil edildi. Hastalıklı doku numunesi KP’li gruptan ≥5mm periodontal cebi, gingival inflamasyonu ve kemik kaybı olan hastalardan periodontal flep cerrahisi sırasında, sağlıklı dokular ise kontrol grubundan kron boyu uzatma işlemi sırasında toplandı. DOS’taki TNF-α seviyeleri ELISA kitleri ile belirlendi. mtDNA delesyonunun tespiti semikantitatif PCR metodu ile yapıldı. Bulgular: KP grubunda doku örneklerinin alındığı bölgelerde tüm klinik parametreler kontrol grubuna göre daha yüksekti. KP grubunun DOS TNF-α seviyesi kontrol grubuna göre daha yüksekti. KP grubunda mtDNA olan hastaların DOS TNF-α seviyeleri mtDNA olmayan hastalara göre daha yüksekti.  Sonuç:  Çalışmamızın sonuçlarına göre periodontitisli hastaların gingival dokularında ve DOS’ında artış gösteren TNF-α mitokondrilerde yükselen ROT üretimini indükleyebilir ve komşu hücrelerde mtDNA delesyonu gibi oksidatif mitokondriyal yaralanmalara sebebiyet verebilir.

Material and Method: The study was carried out in 64 subjects: 32 patients with chronic periodontitis (CP) and 32 periodontally healthy controls. Samples were collected from the diseased gingival tissues with ≥5mm periodontal pockets, gingival inflammation, and bone loss during periodontal flap surgery from CP group and healthy gingival tissues were collected during crown lengthening procedures. TNF-α levels in GCF samples were assayed by commercial ELISA kits. Determination of the deleted mtDNA was employed a semiquantitative PCR method.

Results: All clinical parameter scores of the tissue sampling areas were significantly higher in the CP group compared to the control group. The mean GCF TNF-α level in CP group was significantly higher than that of the controls. In CP group, the mean GCF TNF-α level of patients with deleted mtDNA was significantly higher than those of patients with non-deleted mtDNA

Conclusion: According to this study findings, we speculated that overexpression of TNF-α, which is detected in GCF, in gingival tissue of periodontitis patients, may induces elevated ROS production in mitochondria and may lead to oxidative mitochondrial injury such as mtDNA deletion of neighboring cells.

Anahtar Kelimeler

mtDNA, delesyonu, gingiva, dişeti oluğu sıvısı, yıkıcı periodontal hastalık

Kaynakça

• Genco RJ, Zambon JJ, Christersson LA. The origin of periodontal infections. Adv Dent Res 1988; 2: 245-59.
• Özcan E, Özdemir A, Çanakçı CF. The Role Of Reactive Oxygen Species In Periodontal Tissue Destruction. J Dent Fac Atatürk Uni 2011; 21: 255-61.
• Genco RJ. Host responses in periodontal diseases: current concepts. J Periodontol 1992; 63: 338-55.
• Hirose K, Isogai E, Miura H, Ueda I. Levels of gingivalis inflammatory cytokines in gingival crevicular fluid from adult human subjects. Microbiol Immunol 1997; 41: 21-6. Fimbriae and
• Lee HJ, Kang IK, Chung CP, Choi SM. The subgingival microflora and gingival crevicular fluid cytokines in refractory periodontitis. J Clin Periodontol 1995; 22: 885-90.
• Rossomando EF, Kennedy JE, Hadjimichael J. Tumour necrosis factor alpha in gingival crevicular fluid as a possible indicator of periodontal disease in humans. Arch Oral Biol 1990; 35: 431-4.
• Beutler B, Cerami A. Cachectin and tumour necrosis factor as two sides of the same biological coin. Nature 1986; 320: 584-8.
• Fiers W. Tumor necrosis factor. Characterization at the molecular, cellular and in vivo level. FEBS Lett 1991; 285: 199-212.
• Feingold KR, Soued M, Grunfeld C. Tumor necrosis factor stimulates DNA synthesis in the liver of intact rats. Biochem Biophys Res Commun 1988; 153: 576-82.
• Garcia-Ruiz C, Colell A, Mari M, Morales A, Fernandez-Checa JC. Direct effect of ceramide on the mitochondrial electron transport chain leads to generation of reactive oxygen species. Role of mitochondrial glutathione. J Biol Chem 1997; 272: 11369-77.
• Lo YY, Cruz TF. Involvement of reactive oxygen species in cytokine and growth factor induction of c-fos expression in chondrocytes. J Biol Chem 1995; 270: 11727-30.
• Yakes FM, Van Houten B. Mitochondrial DNA damage is more extensive and persists longer than nuclear DNA damage in human cells following oxidative stress. Proc Natl Acad Sci U S A 1997; 94: 514-9.
• Wallace DC, Shoffner JM, Trounce I, Brown MD, Ballinger SW, Corral-Debrinski M, Horton T, Jun AS, Lott MT. Mitochondrial DNA mutations in human degenerative diseases and aging. Biochim Biophys Acta 1995; 1271: 141-51.
• Lee HC, Pang CY, Hsu HS, Wei YH. Differential accumulations of mitochondrial DNA of various tissues in human ageing. Biochim Biophys Acta 1994; 1226: 37-43.
• Canakci CF, Tatar A, Canakci V, Cicek Y, Oztas S, Orbak R. New evidence of premature oxidative DNA damage: mitochondrial DNA deletion in gingival tissue of patients with periodontitis. J Periodontol 2006; 77: 1894-900.
• Sugano N, Kawamoto K, Numazaki H, Murai S, Ito K. Detection of mitochondrial DNA mutations in human gingival tissues. J Oral Sci 2000; 42: 221-3.
• Armitage GC. Development of a classification system for periodontal diseases and conditions. Ann Periodontol 1999; 4: 1-6.
• Silness J, Loe H. Periodontal Disease in Pregnancy. Ii. Correlation between Oral Hygiene and Periodontal Condtion. Acta Odontol Scand 1964; 22: 121-35.
• Loe H, Silness J. Periodontal Disease in Pregnancy. I. Prevalence and Severity. Acta Odontol Scand 1963; 21: 533-51.
• Morales A, Garcia-Ruiz C, Miranda M, Mari M, Colell A, Ardite E, Fernandez-Checa JC. Tumor necrosis factor increases hepatocellular glutathione by transcriptional regulation of the heavy subunit chain of gamma-glutamylcysteine synthetase. J Biol Chem 1997; 272: 30371-9.
• Hennet T, Richter C, Peterhans E. Tumour necrosis factor-alpha induces superoxide anion generation in mitochondria of L929 cells. Biochem J 1993; 289: 587-92.
• Rubin BY, Smith LJ, Hellermann GR, Lunn RM, Richardson NK, Anderson SL. Correlation between the anticellular and DNA fragmenting activities of tumor necrosis factor. Cancer Res 1988; 48: 6006- 10.
• Zimmerman RJ, Chan A, Leadon SA. Oxidative damage in murine tumor cells treated in vitro by recombinant human tumor necrosis factor. Cancer Res 1989; 49: 1644-8.
• Park YM, Han MY, Blackburn RV, Lee YJ. Overexpression of HSP25 reduces the level of TNF alpha-induced oxidative DNA damage biomarker, 8-hydroxy-2'-deoxyguanosine, in L929 cells. J Cell Physiol 1998; 174: 27-34.
• Canakci V, Canakci CF, Yildirim A, Ingec M, Eltas A, Erturk A. Periodontal disease increases the risk of severe pre-eclampsia among pregnant women. J Clin Periodontol 2007; 34: 639-45.
• Schulze-Osthoff K, Bakker AC, Vanhaesebroeck B, Beyaert R, Jacob WA, Fiers W. Cytotoxic activity of tumor necrosis factor is mediated by early damage of mitochondrial functions. Evidence for the involvement of mitochondrial radical generation. J Biol Chem 1992; 267: 5317-23.
• Nagakawa Y, Williams GM, Zheng Q, Tsuchida A, Aoki T, Montgomery RA, Klein AS, Sun Z. Oxidative mitochondrial DNA damage and deletion in hepatocytes of rejecting liver allografts in rats: role of TNF-alpha. Hepatology 2005; 42: 208-15.
• Yen TC, Su JH, King KL, Wei YH. Ageing-associated 5 kb deletion in human liver mitochondrial DNA. Biochem Biophys Res Commun 1991; 178: 124-31.
• Lim PS, Cheng YM, Wei YH. Large-scale mitochondrial DNA deletions in skeletal muscle of patients with end-stage renal disease. Free Radic Biol Med 2000; 29: 454-63.
• Katsumata K, Hayakawa M, Tanaka M, Sugiyama S, Ozawa T. Fragmentation of human heart mitochondrial DNA associated with premature aging. Biochem Biophys Res Commun 1994; 202: 102-10.
• Lee HC, Lim ML, Lu CY, Liu VW, Fahn HJ, Zhang C, Nagley P, Wei YH. Concurrent increase of oxidative DNA damage and lipid peroxidation together with mitochondrial DNA mutation in human lung tissues during aging--smoking enhances oxidative stress on the aged tissues. Arch Biochem Biophys 1999; 362: 309-16.
• Suzuki S, Hinokio Y, Komatu K, Ohtomo M, Onoda M, Hirai S, Hirai M, Hirai A, Chiba M, Kasuga S, Akai H, Toyota T. Oxidative damage to mitochondrial DNA and its relationship to diabetic complications. Diabetes Res Clin Pract 1999; 45: 161-8.