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THE INFLUENCES OF PERIODONTAL
STATUS AND PERIODONTAL PATHOGEN
QUANTITY ON SALIVARY
8-HYDROXYDEOXYGUANOSINE AND
INTERLEUKIN17 LEVELS
Yang et al
J Periodontol.2015
PRESENTED BY
BIBINA GEORGE
INTRODUCTION
• Periodontitis is one of the most prevalent oral diseases and a major cause of tooth
loss in adults.
• Among many pathogenic factors, plaque is well known to be an initial contributor
to periodontitis.
• Socransky SS et al. detected the subgingival microbiota using checkerboard
DNA-DNA hybridization and observed 5 complexes.
• Red complex:
- Porphyromonas gingivalis (Pg),
- Tannerella forsythia (Tf) and
-Treponema denticola (Td)
BIOMARKERS OF INFLAMMATION
8-OHdG
• Most stable product of ROS
• Marker of mitochondrial DNA
damage caused by premature
oxidation in the gingival tissues of
periodontitis patients
{Canakci CF et al}
IL-17
• Proinflammatory cytokine produced by
T-helper 17 cells
• Stimulate various cell types to produce
other inflammatory cytokines and
chemokines
• Preserves immune homeostasis
• Supports immune responses (Th1) and
combines with receptor activator of
RANK and RANKL, resulting in
osteoclastic bone resorption
AIM
• To investigate the influences of clinical and microbial parameters on
variation of 8-OHdG and IL-17 levels in saliva.
MATERIALS AND METHODS:
• SOURCE OF DATA:
Patients who attended the Department of Periodontics, School of
Stomatology, China Medical University .
• Ethical clearance:
Reviewed and approved by Ethics Committee of China Medical
University.
• Duration of study: Feb. 2013 – June 2014
SELECTION CRITERIA:
INCLUSION CRITERIA
• ≥ 20 teeth
• PD ≥ 4mm
• CAL ≥ 2mm
• Radiograph showing > 30 %
bone destruction
EXCLUSION CRITERIA
• Habits
• Systemic illness
• Pregnancy
EXPERIMENTAL DESIGN:
Periodontal Examination
• Simplified Oral Hygiene Index
(OHS) (Greene and Vermillion)
• Sulcular Bleeding Index (SBI)
• Probing pocket depth (PPD)
• Clinical attachment loss
measurements
Clinical Examination
• Measured four sites per tooth:
• Mesio-buccal
• Buccal
• Disto-buccal
• lingual
SAMPLE COLLECTION
• DNA Extraction and
Real time PCR
96 participants:
48 CP + 48 Healthy
SAMPLE COLLECTION
SUBGINGIVAL PLAQUE
SAMPLE
SALIVA
COLLECTION
Sterile Eppendorf tubes
Right
Quadrant
Left
Quadrant
MesiolingualMesiobuccal
500 µl phosphate
buffered saline
solution
5 ml sterile plastic
centrifuge tube
LAB
Stored at -80°C
DNA Extraction Real time - PCR
Saliva Sample ELISA
P. gingivalis
T. forsythia
T. denticola
8- OHdG
IL-17
STATISTICAL ANALYSIS
• Software: SPSS 13.0
• Tests:
Students t test
ANOVA
Pearson’s correlation analysis
P value ≤ 0.05
RESULTS
:
Comparison of Demographic and Clinical Parameters
Between CP and Control Groups at Baseline
Comparison of Salivary 8-OHdG and IL-17 Levels With Clinical
and Microbial Parameters at Baseline
Comparison of Clinical and Microbial Parameters Before and
After Nonsurgical Treatment
DISCUSSION
• Salivary 8-OHdG level in CP group
was higher than that in control group.
• 8-OHdG levels were significantly
decreased after treatment at both 1 and
3 months
• Variations in the clinical parameters
impacted the variation in the salivary 8-
OHdG level, while variations in the
microbial parameters affected the
variation in the salivary IL-17 level
8OHdG
Clinical
Parameters
CP
IL-17
Microbial
Parameter
CP
REVIEW OF LITERATURE:
• Settem RP et al. A bacterial glycan core linked to surface (S)-layer
proteins modulates host immunity through Th17 suppression. Mucosal
Immunol.2013 Mar;6(2):415-26. demonstrated that the surface
glycosylation of T.forsythia acted to ensure its persistence in the host, likely
through the suppression of Th17 responses. In addition, suggested that the
bacterium then induces the Toll-like receptor 2-Th2 inflammatory axis to cause
bone destruction.
• Dede FO et al. 8-hydroxy-deoxyguanosine levels in gingival crevicular fluid
and saliva in patients with chronic periodontitis after initial periodontal
treatment. J Clin Periodontol. 2013 Jun;84(6):821-8. evaluated the effects of
initial periodontal treatment on the gingival crevicular fluid (GCF) and
salivary levels of 8-hydroxy-deoxyguanosine (8-OHdG) as a marker of
oxidative deoxyribonucleic acid (DNA) damage in patients with chronic
periodontitis (CP) and revealed that DNA injury and oxidative stress increased in
tissue cells and especially in periodontal pockets in patients with CP, and the
periodontal treatment resulted in a significant decrease of 8-OHdG levels in the GCF
samples.
• Shaker OG et al. IL-17 and IL-11 GCF Levels in Aggressive and Chronic
Periodontitis Patients: Relation to PCR Bacterial Detection.Himdawi.
2012(2012)/174764. evaluated IL-17 and IL-11 in gingival crevicular fluid (GCF)
of generalized chronic periodontitis (GCP) and generalized aggressive periodontitis
(GAgP) patients in relation to periodontopathic bacteria and found that IL-17 level
was more in GCF of GAgP, suggesting a potential role in the aetiopathogenesis, and
the decreased ratio of IL-11/IL-17 might reflect an imbalance between the
proinflammatory and anti-inflammatory cytokines in different periodontal diseases.
The high positivity of P. gingivalis in the dental plaque was associated with
significantly increased GCF levels of IL-17 in GCP and GAgP patients.
• Ozcaka O et al. Interleukin-17 and interleukin-18 levels in saliva and
plasma of patients with chronic periodontitis. J Periodontol Res.
2011:Oct;46(5):592-8.investigated whether patients with chronic periodontitis
exhibited different salivary and/or plasma concentrations of interleukin (IL)-17 and
IL-18 compared with clinically healthy subjects and found IL-17 level significantly
lower than IL-18 in chronic periodontitis patient but higher than healthy patients .
• Pabon MCM et al.Detection of Treponema denticola in saliva obtained
from patients with various periodontal conditions. Clin Investigations.
March 2008;12(1):73–81. determined the prevalence of T. denticola in saliva
samples from patients with chronic periodontitis, aggressive periodontitis, and healthy
subjects by assessing the probing depth, clinical attachment loss, and extent of
periodontal breakdown. They concluded that the prevalence of T. denticola in CP
patients was significantly higher than those in healthy and AgP subjects. Furthermore,
all clinical measurements were significantly greater (P < 0.05) for T. denticola-positive
subjects compared to T. denticola-negative subjects.
CONCLUSION
• The salivary 8-OHdG and IL-17 levels in CP patients were significantly reduced
after non-surgical treatment.
• Variation in the salivary 8-OHdG level arose due to variations in the PD, CAL
and OHI-S.
• However, variation in the salivary IL-17 level was attributed to variations in the
PD, subgingival quantity of T. forsythia, and salivary quantities of P. gingivalis,
T. forsythia and T.denticola.
• These findings may be of great significance in facilitating the rapid evaluation
of clinical periodontal status.
CRITICAL APPRAISAL:
• Title of the article is self explanatory as it defines the necessity of
the study.
• The study does not precisely establish the possible relation between
the periodontal pathogens and 8-OHdG and IL-17.
• Study focuses more on 8-OHdG and IL-17 rather than the
implications of effects of the periodontopathogens.
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Jc3

  • 1. THE INFLUENCES OF PERIODONTAL STATUS AND PERIODONTAL PATHOGEN QUANTITY ON SALIVARY 8-HYDROXYDEOXYGUANOSINE AND INTERLEUKIN17 LEVELS Yang et al J Periodontol.2015 PRESENTED BY BIBINA GEORGE
  • 2. INTRODUCTION • Periodontitis is one of the most prevalent oral diseases and a major cause of tooth loss in adults. • Among many pathogenic factors, plaque is well known to be an initial contributor to periodontitis.
  • 3. • Socransky SS et al. detected the subgingival microbiota using checkerboard DNA-DNA hybridization and observed 5 complexes. • Red complex: - Porphyromonas gingivalis (Pg), - Tannerella forsythia (Tf) and -Treponema denticola (Td)
  • 4. BIOMARKERS OF INFLAMMATION 8-OHdG • Most stable product of ROS • Marker of mitochondrial DNA damage caused by premature oxidation in the gingival tissues of periodontitis patients {Canakci CF et al} IL-17 • Proinflammatory cytokine produced by T-helper 17 cells • Stimulate various cell types to produce other inflammatory cytokines and chemokines • Preserves immune homeostasis • Supports immune responses (Th1) and combines with receptor activator of RANK and RANKL, resulting in osteoclastic bone resorption
  • 5. AIM • To investigate the influences of clinical and microbial parameters on variation of 8-OHdG and IL-17 levels in saliva.
  • 6. MATERIALS AND METHODS: • SOURCE OF DATA: Patients who attended the Department of Periodontics, School of Stomatology, China Medical University . • Ethical clearance: Reviewed and approved by Ethics Committee of China Medical University. • Duration of study: Feb. 2013 – June 2014
  • 7. SELECTION CRITERIA: INCLUSION CRITERIA • ≥ 20 teeth • PD ≥ 4mm • CAL ≥ 2mm • Radiograph showing > 30 % bone destruction EXCLUSION CRITERIA • Habits • Systemic illness • Pregnancy
  • 8. EXPERIMENTAL DESIGN: Periodontal Examination • Simplified Oral Hygiene Index (OHS) (Greene and Vermillion) • Sulcular Bleeding Index (SBI) • Probing pocket depth (PPD) • Clinical attachment loss measurements Clinical Examination • Measured four sites per tooth: • Mesio-buccal • Buccal • Disto-buccal • lingual SAMPLE COLLECTION • DNA Extraction and Real time PCR 96 participants: 48 CP + 48 Healthy
  • 9. SAMPLE COLLECTION SUBGINGIVAL PLAQUE SAMPLE SALIVA COLLECTION Sterile Eppendorf tubes Right Quadrant Left Quadrant MesiolingualMesiobuccal 500 µl phosphate buffered saline solution 5 ml sterile plastic centrifuge tube LAB Stored at -80°C
  • 10. DNA Extraction Real time - PCR Saliva Sample ELISA P. gingivalis T. forsythia T. denticola 8- OHdG IL-17
  • 11. STATISTICAL ANALYSIS • Software: SPSS 13.0 • Tests: Students t test ANOVA Pearson’s correlation analysis P value ≤ 0.05
  • 12. RESULTS : Comparison of Demographic and Clinical Parameters Between CP and Control Groups at Baseline
  • 13. Comparison of Salivary 8-OHdG and IL-17 Levels With Clinical and Microbial Parameters at Baseline
  • 14. Comparison of Clinical and Microbial Parameters Before and After Nonsurgical Treatment
  • 15. DISCUSSION • Salivary 8-OHdG level in CP group was higher than that in control group. • 8-OHdG levels were significantly decreased after treatment at both 1 and 3 months • Variations in the clinical parameters impacted the variation in the salivary 8- OHdG level, while variations in the microbial parameters affected the variation in the salivary IL-17 level 8OHdG Clinical Parameters CP IL-17 Microbial Parameter CP
  • 16. REVIEW OF LITERATURE: • Settem RP et al. A bacterial glycan core linked to surface (S)-layer proteins modulates host immunity through Th17 suppression. Mucosal Immunol.2013 Mar;6(2):415-26. demonstrated that the surface glycosylation of T.forsythia acted to ensure its persistence in the host, likely through the suppression of Th17 responses. In addition, suggested that the bacterium then induces the Toll-like receptor 2-Th2 inflammatory axis to cause bone destruction.
  • 17. • Dede FO et al. 8-hydroxy-deoxyguanosine levels in gingival crevicular fluid and saliva in patients with chronic periodontitis after initial periodontal treatment. J Clin Periodontol. 2013 Jun;84(6):821-8. evaluated the effects of initial periodontal treatment on the gingival crevicular fluid (GCF) and salivary levels of 8-hydroxy-deoxyguanosine (8-OHdG) as a marker of oxidative deoxyribonucleic acid (DNA) damage in patients with chronic periodontitis (CP) and revealed that DNA injury and oxidative stress increased in tissue cells and especially in periodontal pockets in patients with CP, and the periodontal treatment resulted in a significant decrease of 8-OHdG levels in the GCF samples.
  • 18. • Shaker OG et al. IL-17 and IL-11 GCF Levels in Aggressive and Chronic Periodontitis Patients: Relation to PCR Bacterial Detection.Himdawi. 2012(2012)/174764. evaluated IL-17 and IL-11 in gingival crevicular fluid (GCF) of generalized chronic periodontitis (GCP) and generalized aggressive periodontitis (GAgP) patients in relation to periodontopathic bacteria and found that IL-17 level was more in GCF of GAgP, suggesting a potential role in the aetiopathogenesis, and the decreased ratio of IL-11/IL-17 might reflect an imbalance between the proinflammatory and anti-inflammatory cytokines in different periodontal diseases. The high positivity of P. gingivalis in the dental plaque was associated with significantly increased GCF levels of IL-17 in GCP and GAgP patients.
  • 19. • Ozcaka O et al. Interleukin-17 and interleukin-18 levels in saliva and plasma of patients with chronic periodontitis. J Periodontol Res. 2011:Oct;46(5):592-8.investigated whether patients with chronic periodontitis exhibited different salivary and/or plasma concentrations of interleukin (IL)-17 and IL-18 compared with clinically healthy subjects and found IL-17 level significantly lower than IL-18 in chronic periodontitis patient but higher than healthy patients .
  • 20. • Pabon MCM et al.Detection of Treponema denticola in saliva obtained from patients with various periodontal conditions. Clin Investigations. March 2008;12(1):73–81. determined the prevalence of T. denticola in saliva samples from patients with chronic periodontitis, aggressive periodontitis, and healthy subjects by assessing the probing depth, clinical attachment loss, and extent of periodontal breakdown. They concluded that the prevalence of T. denticola in CP patients was significantly higher than those in healthy and AgP subjects. Furthermore, all clinical measurements were significantly greater (P < 0.05) for T. denticola-positive subjects compared to T. denticola-negative subjects.
  • 21. CONCLUSION • The salivary 8-OHdG and IL-17 levels in CP patients were significantly reduced after non-surgical treatment. • Variation in the salivary 8-OHdG level arose due to variations in the PD, CAL and OHI-S. • However, variation in the salivary IL-17 level was attributed to variations in the PD, subgingival quantity of T. forsythia, and salivary quantities of P. gingivalis, T. forsythia and T.denticola. • These findings may be of great significance in facilitating the rapid evaluation of clinical periodontal status.
  • 22. CRITICAL APPRAISAL: • Title of the article is self explanatory as it defines the necessity of the study. • The study does not precisely establish the possible relation between the periodontal pathogens and 8-OHdG and IL-17. • Study focuses more on 8-OHdG and IL-17 rather than the implications of effects of the periodontopathogens.

Editor's Notes

  1. Early colonizers- Yellow, purple 2°-green, orange and red