Evaluation of the Effect of Smoking on Pyroptosis in Periodontal Pathogenesis

This study investigates the impact of smoking on pyroptosis and inflammatory signaling pathways in systemically healthy individuals with various periodontal conditions. The research compares smokers and non-smokers across three clinical categories, resulting in six study groups: Healthy Control, Gingivitis, and Periodontitis. The study specifically evaluates the levels of IL-1beta, IL-18, and NLRP3 through ELISA, and the gene expression of TLR1, TLR4, and TLR6 through RT-PCR, to understand how cigarette smoke modulates innate immune responses in the absence of other systemic diseases.

Study Overview

• Status: Active, not recruiting
• Conditions: Periodontitis; Periodontal Diseases; Gingivitis; Smoking ( Cigarette)

Detailed Description

The aim of this research is to clarify the molecular link between tobacco use and pyroptotic cell death pathways in the pathogenesis of periodontal disease. To eliminate systemic bias, only systemically healthy participants are included. The study design consists of 6 distinct groups based on a 2x3 matrix (Smoking Status: Yes/No × Periodontal Status: Healthy/Gingivitis/Periodontitis).

Protein levels of the NLRP3 inflammasome and pro-inflammatory cytokines (IL-1beta, IL-18) are measured in periodontal tissues using the ELISA method. Furthermore, the mRNA expressions of Toll-like receptors (TLR1, TLR4, and TLR6) are analyzed via RT-PCR. These findings will help determine how smoking influences tissue destruction mechanisms specifically through the pyroptosis pathway in different stages of gum disease.

• Study Type: Observational
• Enrollment (Actual): 113

Contacts and Locations

Study Locations

• Turkey (Türkiye)
  • Isparta
    • Isparta, Isparta, Turkey (Türkiye), 32000
      • Suleyman Demirel University, Faculty of Dentistry, Department of Periodontology

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

The study population consists of individuals who applied to the Department of Periodontology at the Faculty of Dentistry. Participants include systemically healthy volunteers diagnosed with different periodontal conditions (healthy, gingivitis, or periodontitis) and categorized by their smoking status.

Description

Inclusion Criteria:

• Individuals who voluntarily agreed to participate in the study,
• Smokers (gingivitis and stage 3/4 generalized periodontitis) and non-smokers (gingivitis and stage 3/4 generalized periodontitis) who applied to the Periodontology Clinical of Suleyman Demirel University Faculty of Dentistry,
• Individuals without any systemic diseases and without using any medications that affect periodontal health were included.

Exclusion Criteria:

• Individuals who refuse to participate in the study or who cannot comply with the study protocol,
• Individuals participating in another clinical trial concurrently, Individuals who have received periodontal treatment within the last 6 months,
• Individuals who have used medications (antibiotics, anti-inflammatory drugs, etc.) that may affect periodontal parameters within the last 3 months,
• Individuals with systemic diseases that may affect the periodontium,
• Individuals with systemic diseases presenting with oral symptoms,
• Individuals with malignant disease,
• Pregnant or lactating women,
• Individuals with psychological or physical disabilities,
• Individuals with substance abuse,
• Individuals with severe malocclusion, undergoing orthodontic treatment, using appliances, or using removable dentures,
• Individuals with acute dental infection or pain (cavities, abscesses, etc.).

Study Plan

How is the study designed?

Number of groups / cohorts

6

Cohorts and Interventions

Group / Cohort
Healthy Non-Smoker; Systemically healty individulas with clinically healthy periodontal tissues who have never smoked or have not smoked for least 5 years.
Gingivitis Non-Smoker; Systemically healty individulas diagnosed with gingivitis who are non-smokers.
Periodontitis Non-Smoker; Systemically healty individulas diagnosed with Stage III or IV periodontitis who are non-smokers.
Healthy Smoker; Systemically healty individulas with clinically healthy periodontal tissues who have been smoking least 10 cigarettes per day for at least 1 year.
Gingivitis Smoker; Systemically healty individulas with gingivitis who have been smoking least 10 cigarettes per day for at least 1 year.
Periodontitis Smoker; Systemically healty individulas diagnosed with Stage III or IV periodontitis who who have been smoking least 10 cigarettes per day for at least 1 year.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Relative mRNA expression levels of TLR1,TLR4 and TLR6	Metric:Fold change (2^-ΔΔCt method). Measurement of gene expression levels in gingival tissues using the RT-PCR method.	Baseline (At the clinical examination)
Protein concerntrations of NLRP3, IL-18 and IL-1β	Metric: Picograms per milliliter (pg/mL). Evaluation of protein concentrations in gingival tissue samples using the ELISA method.	Baseline (At the time of clinical examinat

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Periodontal Inflamed Surface Area (PISA)	Metric: square millimeters (mm²). Calculation of the total periodontal inflamed surface area to quantify the inflammatory burden.	Baseline (At the time of clinical examination)
Periodontal Clinical Measurements (PD and CAL)	Metric: millimeters (mm). Measurements of Probing Depth (PD) and Clinical Attachment Level (CAL) using a periodontal probe.	Baseline (At the time of clinical examination)
Periodontal Indices (PI and GI)	Metric: 0-3 numerical scale. Assessment of Plaque Index (PI) and Gingival Index (GI) to evaluate oral hygiene and gingival inflammation.	Baseline (At the time of clinical examination)

Collaborators and Investigators

• Sponsor: Suleyman Demirel University

Study record dates

Study Major Dates

• Study Start (Actual): October 15, 2024
• Primary Completion (Actual): November 18, 2025
• Study Completion (Estimated): June 30, 2026

Study Registration Dates

• First Submitted: April 11, 2026
• First Submitted That Met QC Criteria: April 28, 2026
• First Posted (Actual): May 1, 2026

Study Record Updates

• Last Update Posted (Actual): May 1, 2026
• Last Update Submitted That Met QC Criteria: April 28, 2026
• Last Verified: April 1, 2026

More Information

Terms related to this study

• Keywords: pyroptosis; Cigarette Smoking; NLRP3 Inflammasome; Interleukin-18 (IL-18); TLR4; Interleukin-1 beta ( IL-1β); TLR1; TLR6
• Additional Relevant MeSH Terms: Mouth Diseases; Stomatognathic Diseases; Infections; Gingival Diseases; Behavior; Smoking; Tobacco Smoking; Tobacco Use; Periodontitis; Periodontal Diseases; Gingivitis; Cigarette Smoking
• Other Study ID Numbers: 79-38

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No