Periodontal Inflammation Is Associated With Disruption of Gingival Circadian Clock Gene and Protein Expression in Individuals With Comparable Chronotype Profiles
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 入组人数
- 60
- 试验地点
- 1
- 主要终点
- Relative mRNA Expression Levels of Circadian Clock Genes in Gingival Tissue
研究概览
简要总结
This observational study aims to investigate whether periodontal inflammation is associated with alterations in the expression of circadian clock-related genes and proteins in gingival tissues. Circadian rhythms regulate many biological processes, including immune responses and inflammation. Although experimental studies suggest a link between circadian disruption and periodontal disease, human data under controlled chronotype conditions are limited.
A total of 60 systemically healthy, non-smoking individuals aged 22-45 years with comparable sleep patterns (intermediate chronotype and 6-9 hours of sleep) were included. Participants were classified as periodontally healthy, gingivitis, or stage III grade B periodontitis according to established diagnostic criteria. Gingival tissue samples were collected during clinically indicated procedures within a standardized morning time window (09:00-11:00).
Gene expression levels of circadian clock components (CLOCK, BMAL1, PER1-3, CRY1-2, Rev-Erb-β, ROR-α) and inflammatory mediators (IL-1β, IL-6, TNF-α, NF-κB, IFN-γ, RANKL, OPG) were analyzed using RT-qPCR, Western blot, and ELISA techniques. Associations between molecular findings and clinical periodontal parameters were evaluated.
The study seeks to clarify whether periodontal disease itself may disrupt local circadian regulatory mechanisms in gingival tissues.
详细描述
Circadian rhythms are generated through transcriptional-translational feedback loops involving core clock genes such as CLOCK, BMAL1, PER1-3, CRY1-2, ROR-α, and REV-ERB-β. These molecular oscillators regulate immune function, inflammatory signaling, and bone metabolism. Experimental evidence suggests that circadian dysregulation may aggravate periodontal inflammation and alveolar bone loss; however, comprehensive human data under controlled chronotype conditions remain limited.
This single-center, observational case-control study includes 60 systemically healthy, non-smoking individuals (30 males, 30 females) aged 22-45 years. Chronotype was determined using the Munich Chronotype Questionnaire, and only individuals with intermediate chronotype and self-reported sleep duration between 6 and 9 hours were included to minimize circadian variability.
Participants were classified into three groups (n=20 per group): periodontally healthy, gingivitis, and stage III grade B periodontitis according to the 2017 World Workshop criteria. Comprehensive periodontal examination included plaque index, gingival index, bleeding on probing, probing depth, and clinical attachment loss measurements. Gingival tissue biopsies were obtained during clinically indicated procedures and collected between 09:00 and 11:00 a.m. to standardize circadian timing. Samples were stored at -80°C until molecular analysis.
Total RNA was extracted from gingival tissues, and gene expression was quantified using RT-qPCR with normalization to β-actin and analysis via the 2^-ΔΔCT method. Protein expression of circadian clock components was assessed by Western blot, and inflammatory cytokine levels (IL-1β, IL-6) were quantified by ELISA. Correlation analyses were performed to evaluate associations between circadian gene expression, inflammatory mediators, and clinical periodontal parameters.
The primary objective is to determine whether periodontal inflammation is associated with disruption of gingival circadian clock gene and protein expression in individuals with comparable chronotype profiles.
研究设计
- 研究类型
- Observational
- 观察模型
- Case Control
- 时间视角
- Cross Sectional
入排标准
- 年龄范围
- 25 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Adults aged 18 years or older
- •Systemically healthy individuals
- •Presence of at least 20 natural teeth
- •Individuals classified as periodontally healthy, gingivitis, or Stage III Grade B periodontitis according to the 2018 classification of periodontal diseases
- •Willingness to provide written informed consent
排除标准
- •Presence of any systemic disease affecting periodontal status (e.g., diabetes mellitus, autoimmune diseases, cardiovascular diseases)
- •Use of antibiotics or anti-inflammatory medications within the previous 3 months
- •Periodontal therapy within the last 6 months
- •Current smokers or individuals who quit smoking within the past 5 years
- •Pregnancy or lactation
- •Use of medications known to influence immune or inflammatory responses
- •History of systemic conditions that may affect wound healing
研究组 & 干预措施
Periodontally Healthy
Systemically healthy individuals with clinically healthy periodontal tissues, no attachment loss, and no radiographic bone loss.
干预措施: Gingival Tissue Biopsy (Procedure)
Gingivitis
Systemically healthy individuals presenting with gingival inflammation without clinical attachment loss or radiographic bone loss.
干预措施: Gingival Tissue Biopsy (Procedure)
Stage III Grade B Periodontitis
Systemically healthy individuals diagnosed with Stage III Grade B periodontitis according to the 2018 classification of periodontal diseases.
干预措施: Gingival Tissue Biopsy (Procedure)
结局指标
主要结局
Relative mRNA Expression Levels of Circadian Clock Genes in Gingival Tissue
时间窗: At the time of gingival tissue collection (single time point)
Quantitative assessment of BMAL1, CLOCK, PER1, PER2, PER3, CRY1, CRY2, REV-ERBβ, and ROR-α mRNA expression levels in gingival tissue samples using real-time quantitative polymerase chain reaction (RT-qPCR). Expression levels will be calculated as relative fold changes normalized to housekeeping genes and compared among periodontally healthy, gingivitis, and Stage III Grade B periodontitis groups.
次要结局
- Pro-inflammatory Cytokine Expression Levels(At the time of tissue collection)
- NF-κB Expression Level(At the time of tissue collection)
- Bone Metabolism Markers(At the time of tissue collection)
研究者
Cuneyt Asim Aral
Professor, Department of Periodontology
Inonu University