Investigation of Circadian Clock Gene/Proteins in Individuals With and Without Disrupted Circadian Rhythm: A Prospective Cross-Sectional Clinical Study
试验速览
- 阶段
- 不适用
- 状态
- 尚未招募
- 入组人数
- 116
- 试验地点
- 1
- 主要终点
- GCF Circadian Rhythm Protein Levels
研究概览
简要总结
This prospective cross-sectional clinical study aims to investigate the relationship between circadian rhythm disruption and periodontal inflammation by evaluating circadian clock protein levels, inflammatory (IL-1beta) and anti-inflammatory (IL-10) cytokine levels in gingival crevicular fluid (GCF) of individuals with and without circadian rhythm disruption.
Participants aged 20-50 years will be classified into four groups based on their circadian rhythm status (disrupted/normal) and gingival health status (gingivitis/healthy). Clinical periodontal parameters including plaque index, gingival index, bleeding on probing, and probing depth will be assessed. Circadian rhythm status will be determined using validated questionnaires (Morningness-Eveningness Questionnaire and Munich Chronotype Questionnaire). Night-shift workers will represent the circadian rhythm disruption group.
GCF samples will be analyzed for circadian clock proteins (BMAL-1, CLOCK, PER-1, PER-2, PER-3, CRY-1, CRY-2, REV-ERB-beta, MTNR1B) and cytokines (IL-1beta, IL-10) using ELISA. Serum cortisol and melatonin levels will be measured for biochemical verification of circadian rhythm status.
Gingivitis groups will receive standard periodontal treatment and be re-evaluated at 2 weeks post-treatment. A total of 116 participants (29 per group) are planned for enrollment.
详细描述
Gingival tissue plays a critical role in maintaining oral health as a fundamental component of periodontal tissues. Recent evidence suggests that circadian rhythm mechanisms, beyond microbial factors, may significantly influence periodontal disease pathogenesis through modulation of host immune responses.
The circadian rhythm is an endogenous mechanism regulating biological processes in approximately 24-hour physiological and behavioral cycles, controlled centrally by the suprachiasmatic nucleus in the hypothalamus. At the molecular level, core clock genes (CLOCK, BMAL1, PER1-3, CRY1-2) operate through the Transcription-Translation Feedback Loop (TTFL). CLOCK and BMAL1 proteins form heterodimers in the cell nucleus, bind to E-box regions on DNA, and initiate transcription of PER and CRY genes. Subsequently, PER and CRY proteins synthesized in the cytoplasm return to the nucleus and inhibit the CLOCK-BMAL1 complex, creating approximately 24-hour rhythmic gene expression cycles. Regulatory nuclear receptors such as REV-ERBa and ROR also contribute to the fine-tuning of this mechanism.
Recent studies have demonstrated the presence of functional peripheral clock mechanisms in periodontal tissues. CLOCK, BMAL1, PER1-3, and CRY1-2 gene expression has been identified in gingival fibroblasts and periodontal ligament fibroblasts. Experimental studies have shown that circadian rhythm disruption can increase inflammatory processes, enhance macrophage activation, elevate inflammatory cytokine production, and accelerate alveolar bone loss in periodontitis models.
Despite these findings, human studies evaluating the relationship between inflammatory cytokines and circadian rhythm proteins in gingival crevicular fluid (GCF) remain limited. This study addresses this gap by evaluating gingival health status using clinical periodontal parameters in individuals with and without circadian rhythm disruption.
Study Design:
研究设计
- 研究类型
- Observational
- 观察模型
- Case Control
- 时间视角
- Prospective
入排标准
- 年龄范围
- 20 Years 至 50 Years（Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Aged between 20 and 50 years
- •Systemically healthy
- •Diagnosed as periodontally healthy or with gingivitis based on clinical periodontal examination
- •Having at least 20 natural teeth in the oral cavity
- •Willing to provide written informed consent
- •Agreeing to participate in circadian rhythm assessment and completing the Morningness-Eveningness Questionnaire (MEQ) and Munich Chronotype Questionnaire (MCTQ)
- •Sleep duration of 6-9 hours per day
- •For periodontally healthy: PD <=3 mm, BOP <10%, CAL=0, no radiographic bone loss
- •For gingivitis: PD <=3 mm, 10%<=BOP<=30%, CAL=0, no radiographic bone loss
- •For circadian rhythm disruption group: being a night-shift worker
排除标准
- •Use of antioxidant or melatonin supplements
- •Presence of any systemic disease
- •Use of antibiotics, anti-inflammatory drugs, or immunosuppressants within the past 3 months
- •Having received periodontal treatment within the past 6 months
- •Pregnancy or lactation
- •Use of tobacco or tobacco products
- •Presence of acute oral infection, abscess, or advanced periodontal disease
- •Currently undergoing orthodontic treatment or having fixed orthodontic appliances
- •Inability to comply with study procedures or refusal to provide informed consent
- •Self-reported sleep duration less than 6 hours or greater than 9 hours per day
研究组 & 干预措施
Normal Circadian / Periodontally Healthy
Individuals with normal circadian rhythm and periodontal health (control group). Inclusion: PD <=3 mm, BOP <10%, CAL=0, no radiographic bone loss. n=29
Normal Circadian / Gingivitis
Individuals with normal circadian rhythm and gingivitis. Inclusion: PD <=3 mm, 10%<=BOP<=30%, CAL=0, no radiographic bone loss. These participants will receive standard periodontal treatment and be re-evaluated at 14 days post-treatment. n=29
干预措施: Standard Periodontal Treatment (Other)
Disrupted Circadian / Periodontally Healthy
Night-shift workers with circadian rhythm disruption and periodontal health. Circadian rhythm status verified by MEQ and MCTQ questionnaires. Inclusion: PD <=3 mm, BOP <10%, CAL=0, no radiographic bone loss. n=29
Disrupted Circadian / Gingivitis
Night-shift workers with circadian rhythm disruption and gingivitis. Circadian rhythm status verified by MEQ and MCTQ questionnaires. Inclusion: PD <=3 mm, 10%<=BOP<=30%, CAL=0, no radiographic bone loss. These participants will receive standard periodontal treatment and be re-evaluated at 14 days post-treatment. n=29
干预措施: Standard Periodontal Treatment (Other)
结局指标
主要结局
GCF Circadian Rhythm Protein Levels
时间窗: Baseline (T0) and 14 days post-treatment (T1) for gingivitis groups
Concentrations of circadian clock proteins (BMAL-1, CLOCK, PER-1, PER-2, PER-3, CRY-1, CRY-2, REV-ERB-beta, MTNR1B) measured in gingival crevicular fluid (GCF) using ELISA. GCF collected with Periopaper strips placed in the gingival sulcus for 30 seconds. Volumes quantified using a Periotron device.
GCF Inflammatory Cytokine Levels
时间窗: Baseline (T0) and 14 days post-treatment (T1) for gingivitis groups
Interleukin-1 beta (IL-1beta, pro-inflammatory) and Interleukin-10 (IL-10, anti-inflammatory) cytokine concentrations in gingival crevicular fluid measured by ELISA.
次要结局
未报告次要终点
研究者
Cuneyt Asim Aral
Professor of Periodontology, Inonu University Faculty of Dentistry
Inonu University