Evaluation of Gingival Crevicular Fluid and Saliva SOST and TWEAK, Gingival Crevicular Fluid RANKL and OPG Levels in Smokers and Non-smokers With Periodontitis
Conditions: Periodontitis, Smoking
Sponsor: Gazi University

 
 
 
Evaluation of Gingival Crevicular Fluid and Saliva SOST and TWEAK, Gingival 
Crevicular Fluid RANKL and OPG Levels in Smokers and Non-smokers with 
Periodontitis 
 
 
 
14.09.2023 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 
Study Protocol 
Study Population 
Participants who applied to Gazi University Faculty of Dentistry Department of 
Periodontology for periodontal treatment or control, who were systemically healthy and who 
agreed to participate in the study were included in the study. 
Participants who agreed to participate in the study were evaluated clinically and radiologically 
and diagnosed in accordance with the Classification of Periodontal and Peri-implant Diseases 
and Conditions 2017 Workshop criteria. Accordingly, the individuals who will take part in the 
study are; Systemically healthy, non-smoking periodontitis group (Group 1, number = 26), 
systemically healthy, smoker periodontitis group (Group 2, number = 26) and systemic and 
periodontal healthy group (Group 3, number = 26).  
Clinical examination 
In order to evaluate the clinical conditions of the participants in the study; Plaque index, 
gingival index, percentage of bleeding on probing, pocket depth and clinical attachment loss 
were recorded for each tooth from four regions (mesiobuccal, buccal, distobuccal, 
lingual/palatinal) using the Williams probe. Clinical indices were measured and recorded by a 
single calibrated investigator. Stages and grades in the classification of periodontitis patients; 
Radiographic bone loss was determined according to the parameters of pocket depth, amount 
of attachment loss, number of teeth lost due to periodontitis, bone loss%/age and case 
phenotype. 
GCF collection 
GCF samples were collected from all individuals participating in the study with paper strips. 
GCF samples were placed in 2 Eppendorf tubes with paper strips in fours. After the tubes 
were closed with parafilm, they were stored at -30°C until ELISA examination to determine 
cytokine levels. 
Saliva collection 

Total saliva samples were collected from the individuals participating in the study by spitting 
the saliva accumulated at the bottom of the mouth into a test tube every 60 seconds for a 
period of 5 minutes. The collected saliva samples were stored at -30°C until ELISA 
examination to determine cytokine levels. 
Laboratory analysis  
Gingival crevicular fluid and saliva samples were examined by the ELISA method in 
accordance with the manufacturer's instructions (Bioassay Technology Laboratory Human 
Sklerostin SOST BT-LAB ELISA Kit, Bioassay Technology Laboratory Human 
Osteoprotegerin OPG OPG BT-LAB Kit ELISA kit, Bioassay Technology Laboratory Human 
Tumour necrosis related weak inducer of apoptosis, TWEAK BT LAB ELISA kit, Human 
Receptor Activator of Nuclear Factor Kappa B Ligand RANKL BT-LAB ELISA kit) to 
determine biomarker levels.