The aim of this study is to evaluate and compare the levels of bone resorption (RANKL, OPG) and oxidative stress (Nitrotyrosine, NQO1) markers, along with pro-inflammatory cytokines (IL-1β), in the saliva and peri-implant sulcular fluid (PISF) of patients with peri-implantitis, periodontitis, and healthy individuals. The study seeks to determine the diagnostic value of these biomarkers in the early detection and progression of peri-implant diseases.
Patients applying to the University of Health Sciences, Gulhane Faculty of Dentistry, Department of Periodontology, will be screened according to the inclusion and exclusion criteria. Following clinical and radiographic examinations, participants will be categorized into four groups (n=20 per group): Periodontitis, Peri-implantitis, Periodontal Health, and Peri-implant Health. All clinical parameters, including Plaque Index (PI), Gingival Index (GI), Bleeding on Probing (BOP), Probing Depth (PD), and Clinical Attachment Loss (CAL), will be recorded at six sites per tooth/implant using a Williams periodontal probe. The biological sample collection protocol will be conducted as follows: Saliva Collection: To minimize circadian rhythm variations, unstimulated whole saliva samples (3 ml) will be collected between 9:00 AM and 10:00 AM. Patients will be instructed to refrain from eating, drinking, or oral hygiene procedures for at least one hour prior to collection. Samples will be centrifuged at 1,000g for 10 minutes to remove cellular debris, and the supernatant will be stored at -80°C until laboratory analysis. Peri-implant Sulcular Fluid (PISF) and Gingival Crevicular Fluid (GCF) Collection: Samples will be obtained from the site with the deepest probing depth in each participant. The area will be isolated with cotton rolls and gently dried with an air syringe to prevent salivary contamination. Periopaper strips will be inserted into the sulcus/pocket for 30 seconds. Strips contaminated with blood or visible saliva will be discarded. Laboratory Analysis: The concentrations of Interleukin-1 beta (IL-1β), Nitrotyrosine, Receptor Activator of Nuclear Factor Kappa-B Ligand (RANKL), Osteoprotegerin (OPG), and NAD(P)H: Quinone Oxidoreductase 1 (NQO1) will be quantified in both saliva and PISF/GCF samples using commercially available Enzyme-Linked Immunosorbent Assay (ELISA) kits according to the manufacturer's instructions.
Study Type
OBSERVATIONAL
Enrollment
80
Salivary and Peri-implant Sulcular Fluid (PISF) IL-1β Levels.
The concentration of Interleukin-1 beta (IL-1β), a pro-inflammatory cytokine, will be measured in picograms per milliliter (pg/mL) using ELISA kits to evaluate the severity of inflammation and bone destruction.
Time frame: At the baseline clinical examination (Day 0).
Salivary and Peri-implant Sulcular Fluid (PISF) Nitrotyrosine Levels
Concentration of Nitrotyrosine will be measured in saliva and PISF samples using ELISA kits. Nitrotyrosine serves as a biochemical marker of nitrosative stress to evaluate oxidative damage.
Time frame: Baseline (at the time of clinical examination)
Salivary and Peri-implant Sulcular Fluid (PISF) NQO1 Levels
NAD(P)H: Quinone Oxidoreductase 1 (NQO1) levels will be measured using ELISA kits to assess the local and systemic antioxidant defense response.
Time frame: Baseline (at the time of clinical examination)
Salivary and Peri-implant Sulcular Fluid (PISF) RANKL Levels
Receptor Activator of Nuclear Factor Kappa-B Ligand (RANKL) levels will be measured to evaluate osteoclast activation and bone destruction potential.
Time frame: Baseline (at the time of clinical examination)
Salivary and Peri-implant Sulcular Fluid (PISF) OPG Levels
Osteoprotegerin (OPG) levels will be measured. OPG acts as a decoy receptor for RANKL, and its level indicates the body's attempt to inhibit bone resorption.
Time frame: Baseline (at the time of clinical examination)
Clinical Attachment Loss (CAL) and Probing Depth (PD)
CAL and PD will be measured in millimeters (mm) at six sites per tooth/implant using a Williams periodontal probe to assess the severity of tissue destruction.
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Time frame: Baseline (at the time of clinical examination)
Plaque Index (PI) and Gingival Index (GI)
PI and GI will be recorded using Silness-Löe and Löe-Silness indices (scored 0-3) to evaluate oral hygiene status and gingival inflammation.
Time frame: Baseline (at the time of clinical examination)