Circadian Clock Proteins in Gingival Crevicular Fluid of Individuals With and Without Circadian Rhythm Disruption

NCT07592052 | Not Yet Recruiting

• 2 other identifiers: IUCREC.2026/9750Will be applied
• Study Type: observational
• Target: 116
• Locations: 1 country
• Sites: 1

Brief Summary

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.

Conditions

Gingivitis; Circadian Rhythm Disruption; Periodontal Inflammation; Circadian Rhythm Sleep Disorder, Shift Work Type

Keywords

Circadian rhythm; Periodontal health; Gingivitis; Gingival crevicular fluid; Clock genes; BMAL1; Night shift work; IL-1beta; IL-10; Chronotype

Outcome Measures

Primary Outcomes (2)

• GCF Circadian Rhythm Protein Levels

  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.

  Baseline (T0) and 14 days post-treatment (T1) for gingivitis groups

• GCF Inflammatory Cytokine Levels

  Interleukin-1 beta (IL-1beta, pro-inflammatory) and Interleukin-10 (IL-10, anti-inflammatory) cytokine concentrations in gingival crevicular fluid measured by ELISA.

  Baseline (T0) and 14 days post-treatment (T1) for gingivitis groups

Study Arms (4)

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

Other: Standard Periodontal Treatment

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

Other: Standard Periodontal Treatment

Interventions

Standard Periodontal Treatment OTHER

Standard periodontal treatment including detailed oral hygiene instruction and professional dental surface cleaning (scaling and polishing). Applied to gingivitis groups (Group 2 and Group 4) at baseline, with clinical and biochemical re-evaluation at 14 days post-treatment.

Disrupted Circadian / Gingivitis; Normal Circadian / Gingivitis

Eligibility Criteria

• Age: 20 Years - 50 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)
• Sampling Method: Non-Probability Sample

Study Population

Systemically healthy volunteers aged 20-50 years presenting to Inonu University Faculty of Dentistry, Department of Periodontology. The study population includes individuals with normal circadian rhythm and night-shift workers with circadian rhythm disruption. Participants are classified as periodontally healthy or with gingivitis based on clinical periodontal examination. Circadian rhythm status is verified using the Morningness-Eveningness Questionnaire (MEQ) and Munich Chronotype Questionnaire (MCTQ).

You may qualify if:

• 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

You may not qualify if:

• 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

Sponsors & Collaborators

• Inonu University: lead
• University of Minnesota: collaborator
• Harvard University: collaborator
• University of Turku: collaborator

Study Sites (1)

Inönü University Faculty of Dentistry, Department of Periodontology

Malatya, Malatya, 44210, Turkey (Türkiye)

Location

Related Publications (7)

• Wang D, Wei Y, Xiang Q, Yang H, Shan Y. Association of disrupted circadian rhythms with self-reported periodontal diseases: Insights from 94,305 UK Biobank participants. J Periodontol. 2026 Mar;97(3):540-551. doi: 10.1002/jper.11388. Epub 2025 Aug 21.

  PMID: 40838345 BACKGROUND

• Ma X, Chen X, Duan Z, Wu Y, Shu J, Wu P, Zhao Y, Wang X, Wang Y. Circadian rhythm disruption exacerbates the progression of macrophage dysfunction and alveolar bone loss in periodontitis. Int Immunopharmacol. 2023 Mar;116:109796. doi: 10.1016/j.intimp.2023.109796. Epub 2023 Feb 1.

  PMID: 36731157 BACKGROUND

• Wang Y, Li R, Ye Q, Fei D, Zhang X, Huang J, Liu T, Wang J, Wang Q. Circadian disruption by simulated shift work aggravates periodontitis via orchestrating BMAL1 and GSDMD-mediated pyroptosis. Int J Oral Sci. 2025 Feb 25;17(1):14. doi: 10.1038/s41368-024-00331-x.

  PMID: 40000642 BACKGROUND

• Zheng C, Li Y, Zhang Y, Geng T, Zhou Z, Jin B, Wang L. Circadian rhythm disruption affects cellular senescence through the BMAL1/CRY2/PER1 signaling pathway in periodontitis. J Mol Histol. 2026 Apr 16;57(3):141. doi: 10.1007/s10735-026-10794-3.

  PMID: 41989670 BACKGROUND

• Liu X, Cao N, Liu X, Deng Y, Xin Y, Fu R, Xin X, Hou Y, Yu W. Circadian Rhythm Disorders Aggravate Periodontitis by Modulating BMAL1. Int J Mol Sci. 2022 Dec 26;24(1):374. doi: 10.3390/ijms24010374.

  PMID: 36613816 BACKGROUND

• Janjic K, Kurzmann C, Moritz A, Agis H. Expression of circadian core clock genes in fibroblasts of human gingiva and periodontal ligament is modulated by L-Mimosine and hypoxia in monolayer and spheroid cultures. Arch Oral Biol. 2017 Jul;79:95-99. doi: 10.1016/j.archoralbio.2017.03.007. Epub 2017 Mar 14.

  PMID: 28350992 BACKGROUND

• Papagerakis S, Zheng L, Schnell S, Sartor MA, Somers E, Marder W, McAlpin B, Kim D, McHugh J, Papagerakis P. The circadian clock in oral health and diseases. J Dent Res. 2014 Jan;93(1):27-35. doi: 10.1177/0022034513505768. Epub 2013 Sep 24.

  PMID: 24065634 BACKGROUND

Biospecimen

Retention: SAMPLES WITHOUT DNA

Gingival crevicular fluid (GCF) collected using sterile Periopaper strips, unstimulated whole saliva, and serum samples obtained from venous blood. All samples are stored at -80°C until analysis by ELISA for circadian rhythm proteins and inflammatory cytokines.

MeSH Terms

Conditions

Gingivitis; Sleep Disorders, Circadian Rhythm

Condition Hierarchy (Ancestors)

Infections; Gingival Diseases; Periodontal Diseases; Mouth Diseases; Stomatognathic Diseases; Chronobiology Disorders; Nervous System Diseases; Dyssomnias; Sleep Wake Disorders; Occupational Diseases; Mental Disorders

Study Officials

• Cuneyt A Aral, Professor, DDS, PhD

  Inonu University

  STUDY CHAIR

Central Study Contacts

Cüneyt A Aral, Chair, Professor Dr, DDS, PhD

CONTACT

+905424577657; cuneytasimaral@gmail.com

Kübra Aral, Assoc. Professor, DDS, PhD

CONTACT

+905330578801; drkubraaral@gmail.com

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Professor of Periodontology, Inonu University Faculty of Dentistry

Study Record Dates

• First Submitted: May 11, 2026
• First Posted: May 18, 2026
• Study Start (Estimated): August 1, 2026
• Primary Completion (Estimated): January 1, 2027
• Study Completion (Estimated): March 1, 2027
• Last Updated: May 18, 2026

Record last verified: 2026-05

Locations

TU (1)