EFFECT OF INITIAL PERIODONTAL THERAPY ON GINGIVAL CREVICULAR FLUID CYTOKINE PROFILE IN SUBJECTS WITH CHRONIC PERIODONTITIS.

1. EFFECT OF INITIAL PERIODONTAL THERAPY ON
GINGIVAL CREVICULAR FLUID CYTOKINE PROFILE
IN SUBJECTS WITH CHRONIC PERIODONTITIS.
Zekerdou A et al
Clin Exp Dent Res. 2017;3:62–68.
Presented by,
Dr. Bibina George

2. INTRODUCTION
› Periodontitis ------- Clinical Diagnosis
– induced by bacteria that activate the host's innate and eventually adaptive immune
systems (Kinane,2001)
› levels of biomarkers involved in interactions between bacteria and the host defense
(Buduneli & Kinane, 2011; Salvi & Lang, 2015).
› Levels of some constituents of saliva or gingival crevicular fluid (GCF)
correlate with disease activity or the risk for disease progression
(Buduneli & Kinane, 2011)

3. GCF
› Cytokines
• Mogi et al., 1999;
• Kurtis, et al., 2015
Il- 1beta
• Mogi et al., 1999;
• Zhang, Chen, Zhu, & Yan, 2016
IL- 6, IL-8 and TNF- alpha
• Padma,
• Sreedhara, Indeevar, Sarkar, & Kumar,
2014).
vegf

4. Biomarker Type Function
IL- 1beta Cytokine Osteoclast activation, Immune cell
recruitment, Vascular smooth
muscle cell contraction
Interferon gamma Cytokine Macrophage Activation,Th2
suppression
IL-1 ra (Receptor Antagonist) Cytokine Antagonist of IL-1
IL-6 Cytokine Regulator of t and B cell growth,
Acute phase protein production
IL-8 Cytokine Recruitment and activation of
neutrophils
TNF - alpha Cytokine Delays neutrophil apoptosis

5. AIM:
› To simultaneously assess GCF levels of 12‐selected cytokines and to analyze
to what extent these molecules discriminate sites with periodontitis from
healthy sites in diseased persons
› To compare levels in periodontally diseased and healthy individuals, and to
monitor longitudinal changes in healthy and diseased sites after initial
periodontal treatment.

6. MATERIALS AND METHODS:
› SOURCE OF DATA:
Patients seeking treatment from the University of
Geneva, School of Dental Medicine ---- October 2013 – March 2016 aged
between 30- 70 years.
› Ethical clearance:
Reviewed and approved by the ethical committee of the
University Hospitals of Geneva .

7. SELECTION CRITERIA
INCLUSION CRITERIA
› For Group H :
› Did not have any periodontal sites :
Probing depth (PD) > 4 mm, clinical
attachment loss ≥1 mm or radiographic
evidence of bone loss.
› For Group P :
› At least two teeth with probing PD ≥ 5 mm,
clinical attachment loss ≥ 1 mm, and
radiographic evidence of marginal bone
loss.
EXCLUSION CRITERIA
› H/o systemic illness
› Fever / Other signs of infection
› Antibiotic therapy taken 2
months prior to surgery.

8. 2
periodontally
diseased
sites (PD ≥
5mm)
HH
(PD ≤ 3mm)
2
periodontally
healthy sites
(PD ≤ 3mm)
24 Periodontitis
patients (Group P)
20 Healthy
Patients (Group H)
ISOLATE AND COLLECT GCF
SAMPLE FROM GINGIVAL
MARGIN
Transferred to microtube
Stored at - 20° C
Durapore membrane
strip

9. Experimental Design:
Clinical Parameters
• Plaque Index (Silness and
Loe )
• Gingival Index (Loe and
Silness)
• Probing pocket depth (PPD)
• BOP
• Gingival Recession (GR)
• Tooth Mobility
Clinical Examination
• Measured 6 sites
per tooth:
• Mesio-buccal
• Buccal
• Disto-buccal
• Mesiolingual
• Lingual
• Distolingual
TIME PERIOD
• 1 month
• 3 months

10. LABORATORY PROCEDURES
Specimen Assay buffer 96 well filter plates
Microsphere beadsIncubated 30 min
Washed and mixed with
secondary antibiotics

11. STATISTICAL ANALYSIS :
› Fisher’s exact test
› Wilcoxon signed – rank test
› Spearman’s rank correlation test

12. RESULTS:

14. P < 0.05

16. DISCUSSION :
› Significant differences for eight biomarkers (il‐1ra, il‐6, il‐8, il‐17, b‐fgf, gm‐csf,
mip‐1β, and tnf‐α) between samples from patients with untreated chronic
periodontitis and healthy persons were seen in this study.
› The presence of a generalized inflammatory state in persons with periodontal
disease that is not limited to clinically diseased sites .
› There were significant changes in GCF from both PP and PH sites for il‐1ra and
gm‐csf, further reinforcing the notion that GCF samples to a large‐part reflect
patterns of inflammatory mediators at the level of the person rather than the site.

17. Review of Literature :
› Gilowski et al,2014. reported higher levels of various biomarkers in the
GCF of periodontitis patients compared to healthy persons
› Sakai et al 2006, Yucel et al 2008, Liu et al 1996, Homlund et al 2004,
Engebretson et al.2002, Ozmeric 1998, Toker et al 2008, Tsai et al 2007
reported increased GCF and salivary levels of proinflammatory cytokines
like IL 1, IL6, IL8 in periodontal disease.

18. › A longitudinal study by Engebretson et al. evaluated IL-1 levels in GCF
from patients with varying degrees of periodontal disease following scaling
and root planing. An important observation was that, compared to shallow
sites in patients with mild/moderate periodontitis, shallower sites in patients
with severe periodontitis demonstrated increased levels of IL-1. This
suggests that GCF IL-1 expression is genetically influenced and not solely a
result of local clinical parameters.

19. › Gurlek et al,2017. in his cross- sectional study assessed cytokine levels in
peri-implant crevicular fluid (PICF)/ gingival crevicular fluid (GCF) and a
selection of subgingival/submucosal plaque bacteria from clinically healthy
or diseased sites in the same individuals and concluded that there were
many similarities but crucially some differences in biomarker levels (IL-1β
and sRANKL) and bacterial species between peri-implant and periodontal
sites in the same individuals suggesting similar pathogenic mechanisms

20. LIMITATIONS:
› Healthy participants are not matched for age and gender due to difficulty
of recruiting persons with no periodontal problems and same
characteristics as the test group
› Inability to identify distinct candidate markers in GCF to disclose active
disease at the level of the site.

21. CONCLUSION :
› The results of this study reflect the complexity of the host response in
periodontal diseases.
› IL‐1ra, IL‐6, IL‐8, IL‐17, b‐fgf, gm‐csf, mip‐1β, and tnf‐α identified
patients with chronic periodontitis, rather than diseased sites, suggesting a
generalized inflammatory state that is not limited to clinically diseased
sites.

22. THANK YOU