Evaluation of Oxidative Stress and Methylated Arginine Levels in Individuals With Periodontitis

NCT06973148 | Completed DMC

• 2 other identifiers: 04/1117.02.2021, 04/11
• Study Type: observational
• Target: 40
• Locations: 1 country
• Sites: 1

Brief Summary

You are being invited to participate in a scientific research study. Before you decide to take part, it is essential that you understand why the study is being conducted and what your participation will involve. Please take the time to read the following information carefully. Feel free to ask if anything is unclear or need more details. This study aims to evaluate saliva composition, specifically focusing on amino acid and protein building blocks, oxidative stress, and the Nitric Oxide metabolism system, including levels of ADMA (Asymmetric Dimethylarginine) and SDMA (Symmetric Dimethylarginine). By examining these parameters, we aim to contribute to a better understanding of the biological mechanisms involved in gum health and systemic responses. You have been selected to participate in this study based on your periodontal (gum) health status. A total of 40 individuals, 20 diagnosed with periodontitis and 20 diagnosed as periodontally healthy, who meet the study criteria, will be included. If you choose to participate, you will receive non-surgical periodontal treatment as part of your routine care. Before the treatment, biological samples, including saliva and serum, will be collected. These samples will be used solely for scientific analysis and will not affect the outcome of your dental treatment. Although the total duration of the study is 2 years, your involvement will conclude at the beginning of periodontal therapy. There are no anticipated risks associated with participation beyond those commonly encountered during dental examinations and treatments. You will not benefit directly; however, your contribution will help advance scientific knowledge about the factors influencing oral and systemic health. All information collected from you during the study will remain confidential. Your identity will be protected, and personal data will not be disclosed in any reports or publications resulting from this study. Participation in this study is entirely voluntary. You are free to decline or withdraw at any time, without needing to give a reason. Your decision will not affect the quality of care you receive. All tests, examinations, and procedures performed as part of this study will be provided at no cost to you. Participation will not result in any financial cost or compensation.

Conditions

Periodontitis; Oxidative Stress

Keywords

periodontitis; oxidative stress; antioxidante; methylated arginines

Outcome Measures

Primary Outcomes (7)

• Power analysis

  Sample size analysis was performed a priori using G\*Power software before the initiation of the study. The analysis applied a large effect size (Cohen's d = 0.8) for independent two-group comparisons, with a significance level (α) of 0.05 and a statistical power of 80%, resulting in a calculated total sample size of 40 participants.

  Baseline

• Plaque Index (PI)

  Separate intergroup comparisons will be conducted for each clinical outcome measure to allow precise and independent reporting. Thea plaque index (PI), assessed using the Silness and Löe plaque index, which evaluates the thickness of plaque along the gingival margin on a scale from 0 (no plaque) to 3 (abundant plaque). All cl

  Baseline

• Probing depth (PD)

  The probing depth (PD), measured in millimeters at six sites per tooth (mesiobuccal, midbuccal, distobuccal, mesiolingual, midlingual, and distolingual) using a standardized manual periodontal probe (e.g., UNC-15 probe) with markings at 1 mm increments to ensure accuracy.

  Baseline

• Bleeding on probing (BOP)

  The bleeding on probing (BOP), recorded as the presence or absence of bleeding within 15 seconds of probing at each site and reported as the percentage of positive sites per participant.

  Baseline

• Clinical attachment loss (CAL)

  Separate intergroup comparisons will be conducted for each clinical outcome measure to allow precise and independent reporting. The clinical attachment loss (CAL), also measured in millimeters at the same six sites per tooth, calculated as the distance from the cementoenamel junction (CEJ) to the base of the periodontal pocket.

  Baseline

• Determination of salivary and serum oxidative stress index

  Total antioxidant status (TAS) and total oxidant status (TOS) were measured at Baseline in saliva and serum samples using Erel's validated method. TAS was based on the reduction of the ABTS⁺ (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radical by antioxidants, causing a decrease in color intensity read at 600 nm with a Varioskan Multimode Reader. Calibration used ascorbic acid; results were expressed in mmol ascorbic acid equivalents/L. Higher TAS indicates stronger antioxidant capacity. TOS involved oxidation of Fe²⁺ to Fe³⁺, forming a colored complex with xylenol orange, read at 530 nm; results were in μmol H₂O₂ equivalents/L. Higher TOS reflects elevated oxidant levels. The oxidative stress index (OSI) was calculated as (TOS/TAS) × 10, a unitless index where higher values indicate greater oxidative stress. All assays were performed in duplicate for reliability.

  Baseline

• Determination of Salivary and Serum Methylated Arginine Metabolites

  Salivary and serum levels of arginine, asymmetric dimethylarginine (ADMA), and symmetric dimethylarginine (SDMA) were measured at the baseline time point. Quantification was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with an electrospray ionization (ESI) source (Thermo Scientific Accessmax). Samples underwent protein precipitation and derivatization before injection. Chromatographic separation was achieved on a reversed-phase column under optimized gradient conditions, and mass detection was performed in multiple reaction monitoring (MRM) mode for high specificity. Calibration curves with known standards ensured quantitative accuracy, and results were expressed in micromoles per liter (µmol/L). All measurements were performed in duplicate for reliability.

  Baseline

Study Arms (2)

periodontitis group

Stage III Grade B periodontitis

Healthy group

periodontally healthy

Eligibility Criteria

• Age: 18 Years - 64 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)
• Sampling Method: Non-Probability Sample

Study Population

The study population consisted of individuals recruited from the Department of Periodontology, Ankara University Faculty of Dentistry, between June 2021 and December 2022.

You may qualify if:

• Having at least 16 natural teeth
• Aged between 18 and 64 years
• Systemically healthy (no known systemic diseases)

You may not qualify if:

• Having diabetes, cancer, cardiovascular diseases, and/or any autoimmune diseases such as rheumatoid arthritis
• Pregnant and lactating mothers 3-Tobacco users
• Patients using antibiotics, anti-inflammatory medications, immunosuppressive drugs, and contraceptives within three months
• Patients who had periodontal treatment during the last six months

Sponsors & Collaborators

• Ankara Medipol University: lead
• Ankara University: collaborator
• Medipol University: collaborator

Study Sites (1)

Ankara University

Ankara, 06500, Turkey (Türkiye)

Location

Related Publications (1)

• Balkan B, Guney Z, Balkan E, Balkan EP, Kurgan S, Balci N, Serdar MA, Gunhan M. Interaction between methylated arginine metabolites and oxidative stress in periodontitis: a cross-sectional study. BMC Oral Health. 2025 Nov 14;25(1):1781. doi: 10.1186/s12903-025-06972-6.

  PMID: 41239382 DERIVED

Biospecimen

Retention: SAMPLES WITH DNA

Saliva and serum samples

MeSH Terms

Conditions

Periodontitis

Condition Hierarchy (Ancestors)

Periodontal Diseases; Mouth Diseases; Stomatognathic Diseases

Study Officials

• Zeliha Guney

  Ankara Medipol University

  PRINCIPAL INVESTIGATOR

• Sivge Kurgan, Prof.

  Ankara University

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: CROSS SECTIONAL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Assoc. Prof.

Study Record Dates

• First Submitted: April 28, 2025
• First Posted: May 15, 2025
• Study Start: June 7, 2021
• Primary Completion: December 19, 2022
• Study Completion: December 19, 2022
• Last Updated: May 15, 2025

Record last verified: 2025-05

Locations

TU (1)