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

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

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