Single-cell and Spatial Transcriptomic Profiling of Gingival Tissues in Aggressive and Chronic Periodontitis: Deciphering Cellular Heterogeneity and Inflammatory Microenvironment Features

The goal of this observational study is to investigate cellular heterogeneity and inflammatory microenvironment features in gingival tissues from patients with aggressive periodontitis compared to chronic periodontitis and healthy controls, using single-cell and spatial transcriptomics combined with whole-genome sequencing.

The main question(s) it aims to answer are:

What are the differences in cellular composition, mesenchymal stem cell subpopulations, and gene expression profiles between aggressive periodontitis, chronic periodontitis, and healthy gingival tissues? Which key molecular markers, signaling pathways, and spatially resolved microenvironment patterns distinguish aggressive periodontitis from the more common chronic form? Are there disease-specific genetic variants associated with aggressive periodontitis identified through whole-genome sequencing of peripheral blood DNA?

Participants (already scheduled for tooth extraction or periodontal surgery as part of routine clinical care) will undergo:

Standard clinical periodontal examination and CBCT imaging Collection of a small gingival tissue sample (~1×3×1 mm) during the planned extraction/surgery under local anesthesia Collection of 5 mL peripheral blood for DNA extraction No additional intervention, treatment, or follow-up visit is required beyond routine dental care.

Last updated: December 23, 2025

Study Overview

• Status: Recruiting
• Conditions: Aggressive Periodontitis; Periodontis

• Study Type: Observational
• Enrollment (Estimated): 9

Contacts and Locations

• Study Contact: Name: Li-li Zhou; Phone Number: +86 18329193003; Email: sophiazhou04@163.com

Study Locations

• China
  • Zhejiang
    • Hangzhou, Zhejiang, China
      • Recruiting
      • 2nd Affiliated Hospital, School of Medicine, Zhejiang University, China
      • Contact: Li Li Zhou, doctor; Phone Number: 18329193003; Email: sophiazhou04@163.com

Participation Criteria

Eligibility Criteria

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

• Sampling Method: Non-Probability Sample

Study Population

This cross-sectional observational study will recruit 9 adults (n=3 per group) from the Department of Periodontology, Second Affiliated Hospital, Zhejiang University School of Medicine. All participants are already scheduled for routine tooth extraction or periodontal surgery, allowing incidental collection of a small gingival tissue sample (~1×3×1 mm) and 5 mL peripheral blood during standard care. No extra procedures are added.

Aggressive Periodontitis (AP) Group (n=3) Age 20-35 years

• 30% vertical bone loss at ≥2 sites on radiographs Stage III/IV, Grade C (2018 classification) With or without family history Minimal local factors (negligible calculus/trauma) Chronic Periodontitis (CP) Group (n=3) Age 30-40 years
• 3 sites with probing depth ≥5 mm Stage I/II, Grade A/B (2018 classification) No family history Disease driven by plaque/calculus Healthy Controls (n=3) Age 20-40 years No significant bone loss on radiographs Clinically healthy periodontium (no gingivitis/periodontitis)

Description

Inclusion Criteria:

Aggressive Periodontitis Patients:

1. Aged 20-35 years
2. Radiographic evidence of at least 30% vertical bone loss at ≥2 tooth sites
3. Can be classified as Stage III/IV, Grade C periodontitis according to the 2018 new classification of periodontitis
4. With or without family history of periodontitis
5. No significant local contributing factors (e.g., calculus, occlusal trauma, etc.)

Chronic Periodontitis Patients:

1. Aged 30-40 years
2. ≥3 sites with probing depth ≥5 mm
3. Can be classified as Stage I/II, Grade A/B periodontitis according to the 2018 new classification of periodontitis
4. No significant family history
5. Disease predominantly driven by local factors (dental plaque, calculus, etc.)

Healthy Controls:

1. Aged 20-40 years
2. Radiographic examination shows no significant alveolar bone loss
3. Clinically cannot be diagnosed with gingivitis or periodontitis

Exclusion Criteria:

1. Use of antibiotics within the past 3 months
2. Smoking >10 cigarettes per day
3. Uncontrolled severe diabetes (HbA1c level ≥6.7%)
4. Planning pregnancy, currently pregnant, or breastfeeding
5. Alcohol or drug addiction
6. Long-term use of immunosuppressive agents
7. Infectious diseases such as hepatitis or AIDS/HIV
8. History of malignant tumor, radiotherapy, or chemotherapy within the past 5 years

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort
control group
aggressive periodontitis group
chronic periodontitis group

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Landscape of Cellular Heterogeneity in Periodontal Tissues	Single-cell RNA sequencing (scRNA-seq) will be used to evaluate the cellular heterogeneity of gingival tissues from patients with aggressive periodontitis (AP group), chronic periodontitis (CP group), and healthy controls, with a particular focus on mesenchymal stem cells. This includes identification of cell subpopulations and quantification of these subpopulations based on gene expression profiles.	1 year
Identification of Key Molecular Markers and Signaling Pathways	To discover differentially expressed molecular markers and signaling pathways between the AP group, CP group, and healthy group. By integrating single-cell RNA sequencing and spatial transcriptomics data, the analysis will identify potential therapeutic targets.	1 year

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Spatial Transcriptomic Expression Profiles	Evaluation of the spatial distribution patterns of gene expression in gingival tissues from the aggressive periodontitis (AP) group, chronic periodontitis (CP) group, and healthy controls, as well as treatment-induced spatial changes. Spatial transcriptomics technology will be used to map cell-cell interactions and differences in the microenvironment.	1 year
Genetic Variants Identified by Whole-Genome Sequencing	Through whole-genome resequencing (WGS) of peripheral blood DNA, identification of genetic susceptibility variants associated with aggressive periodontitis and chronic periodontitis, with a particular focus on variants related to familial aggregation, genetic predisposition, disease progression, and treatment response.	1 year

Collaborators and Investigators

• Sponsor: Second Affiliated Hospital, School of Medicine, Zhejiang University

Study record dates

Study Major Dates

• Study Start (Actual): December 26, 2025
• Primary Completion (Estimated): December 26, 2026
• Study Completion (Estimated): December 30, 2026

Study Registration Dates

• First Submitted: March 17, 2026
• First Submitted That Met QC Criteria: March 17, 2026
• First Posted (Actual): March 23, 2026

Study Record Updates

• Last Update Posted (Actual): March 23, 2026
• Last Update Submitted That Met QC Criteria: March 17, 2026
• Last Verified: December 1, 2025

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Periodontal Diseases; Mouth Diseases; Stomatognathic Diseases; Periodontitis; Aggressive Periodontitis
• Other Study ID Numbers: 2025-1735

Drug and device information, study documents

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