Hippo Signaling Pathway Proteins in Gingival Crevicular Fluid of Individuals With Periodontal Disease

NCT07759466 | Completed

• 2 other identifiers: IUCREC.2024/07TDH-2024-3461
• Study Type: observational
• Target: 75
• Locations: 1 country
• Sites: 1

Brief Summary

Periodontal disease is a chronic inflammatory condition of the tooth-supporting tissues in which the host inflammatory response drives the destruction of alveolar bone and periodontal attachment. The Hippo signaling pathway is a conserved regulatory network that controls cell proliferation, apoptosis, and bone homeostasis, but its role in human periodontal disease remains unknown. This observational, case-control study compared the levels of Hippo signaling pathway proteins (YAP, TAZ, MST1, TEAD, LATS1, MAP4K4), the anti-apoptotic protein BCL-2, and the bone metabolism marker alkaline phosphatase (ALP) in gingival crevicular fluid across 75 systemically healthy participants grouped as periodontally healthy, gingivitis, or stage 3 grade B periodontitis. The aim was to determine whether these proteins are differentially expressed across periodontal health and disease states and whether they correlate with markers of apoptosis and bone metabolism.

Conditions

Periodontal Disease; Gingivitis and Periodontal Diseases; Biomarker Discovery and Validation; Pathways

Keywords

Hippo signaling pathway; Periodontitis; Gingivitis; Periodontal disease; Gingival crevicular fluid; Yes-associated protein (YAP); transcriptional co-activator with PDZ-binding motif (TAZ); mammalian sterile 20-like kinase 1 (MST1); TEA domain family members (TEAD); large tumor suppressor kinase 1 (LATS1); mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4); BCL-2; Alkaline phosphatase; Apoptosis; Bone metabolism; Periodontal inflammation

Outcome Measures

Primary Outcomes (6)

• Gingival crevicular fluid level of Yes-associated protein (YAP)

  The level of Yes-associated protein (YAP) in gingival crevicular fluid was quantified by enzyme-linked immunosorbent assay and compared among periodontally healthy, gingivitis, and stage 3 grade B periodontitis groups. YAP was reported both as concentration (ng/mL) and as total amount per 30-second sampling interval.

  At the time of gingival crevicular fluid sampling (cross-sectional; single time point)

• Gingival crevicular fluid level of transcriptional co-activator with PDZ-binding motif (TAZ)

  The level of transcriptional co-activator with PDZ-binding motif (TAZ) in gingival crevicular fluid was quantified by enzyme-linked immunosorbent assay and compared among periodontally healthy, gingivitis, and stage 3 grade B periodontitis groups. TAZ was reported both as concentration (ng/mL) and as total amount per 30-second sampling interval.

  At the time of gingival crevicular fluid sampling (cross-sectional; single time point)

• Gingival crevicular fluid level of mammalian sterile 20-like kinase 1 (MST1)

  The level of mammalian sterile 20-like kinase 1 (MST1) in gingival crevicular fluid was quantified by enzyme-linked immunosorbent assay and compared among periodontally healthy, gingivitis, and stage 3 grade B periodontitis groups. MST1 was reported both as concentration (ng/mL) and as total amount per 30-second sampling interval.

  At the time of gingival crevicular fluid sampling (cross-sectional; single time point)

• Gingival crevicular fluid level of TEA domain family member (TEAD)

  The level of TEA domain family member (TEAD) in gingival crevicular fluid was quantified by enzyme-linked immunosorbent assay and compared among periodontally healthy, gingivitis, and stage 3 grade B periodontitis groups. TEAD was reported both as concentration (ng/L) and as total amount per 30-second sampling interval.

  At the time of gingival crevicular fluid sampling (cross-sectional; single time point)

• Gingival crevicular fluid level of large tumor suppressor kinase 1 (LATS1)

  The level of large tumor suppressor kinase 1 (LATS1) in gingival crevicular fluid was quantified by enzyme-linked immunosorbent assay and compared among periodontally healthy, gingivitis, and stage 3 grade B periodontitis groups. LATS1 was reported both as concentration (pg/mL) and as total amount per 30-second sampling interval.

  At the time of gingival crevicular fluid sampling (cross-sectional; single time point)

• Gingival crevicular fluid level of mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4)

  The level of mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4) in gingival crevicular fluid was quantified by enzyme-linked immunosorbent assay and compared among periodontally healthy, gingivitis, and stage 3 grade B periodontitis groups. MAP4K4 was reported both as concentration (ng/mL) and as total amount per 30-second sampling interval.

  At the time of gingival crevicular fluid sampling (cross-sectional; single time point)

Secondary Outcomes (2)

• Gingival crevicular fluid level of B-cell lymphoma 2 (BCL-2)

  At the time of gingival crevicular fluid sampling (cross-sectional; single time point)

• Gingival crevicular fluid level of alkaline phosphatase (ALP)

  At the time of gingival crevicular fluid sampling (cross-sectional; single time point)

Other Outcomes (6)

• Plaque Index (PI)

  At the time of periodontal examination (cross-sectional; single time point)

• Gingival Index (GI)

  At the time of periodontal examination (cross-sectional; single time point)

• Probing Depth (PD)

  At the time of periodontal examination (cross-sectional; single time point)

Study Arms (3)

Periodontally healthy

Systemically healthy adults with clinically healthy periodontium, defined by probing depth of 3 mm or less, bleeding on probing below 10 percent, no clinical attachment loss, and no radiographic bone loss. Gingival crevicular fluid was collected from non-inflamed sites.

Diagnostic Test: ELISA Biomarker Analysis

Gingivitis

Systemically healthy adults with gingival inflammation without attachment loss, defined by probing depth of 3 mm or less, generalized bleeding on probing above 30 percent, no clinical attachment loss, and no radiographic bone loss. Gingival crevicular fluid was collected from inflamed sites.

Diagnostic Test: ELISA Biomarker Analysis

Stage 3 grade B periodontitis

Systemically healthy adults with stage 3 grade B periodontitis, defined by probing depth of 6 mm or more, clinical attachment loss of 5 mm or more, bleeding on probing of 30 percent or more, and radiographic alveolar bone loss extending to the middle or apical third of the root. Gingival crevicular fluid was collected from diseased sites.

Diagnostic Test: ELISA Biomarker Analysis

Interventions

ELISA Biomarker Analysis DIAGNOSTIC_TEST

Gingival crevicular fluid samples were collected from each participant using standardized paper strips placed into the periodontal pocket for a fixed interval, and fluid volume was measured electronically. Samples were pooled per participant and stored until analysis. The levels of Hippo signaling pathway proteins (YAP, TAZ, MST1, TEAD, LATS1, MAP4K4), the anti-apoptotic protein BCL-2, and the bone metabolism marker alkaline phosphatase were quantified by enzyme-linked immunosorbent assay according to the manufacturers' instructions. Standard curves were generated individually for each biomarker, and results were expressed both as total amounts and as concentrations to account for differences in fluid volume between groups.

Gingivitis; Periodontally healthy; Stage 3 grade B periodontitis

Eligibility Criteria

• Age: 18 Years - 65 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

The study population consisted of 75 systemically healthy adults aged 18 to 65 years who presented to the Department of Periodontology, Faculty of Dentistry, Inönü University, between January and May 2024. Participants were allocated into three equally sized groups according to the 2017 World Workshop classification: periodontally healthy individuals, patients with gingivitis, and patients with stage 3 grade B periodontitis. Eligible participants had at least 20 teeth excluding third molars and no systemic disease, no antibiotic use or periodontal treatment within the preceding six months, no smoking or alcohol use, and no pregnancy or lactation. Gingival crevicular fluid samples were collected from group-specific sites for quantification of Hippo signaling pathway proteins, BCL-2, and alkaline phosphatase.

You may qualify if:

• Systemically healthy individuals aged 18-65 years
• Non-smokers or smoking cessation more than 5 years ago
• Healthy group: No history of periodontal disease, ≥20 teeth, probing depth (PD) ≤3 mm at all sites, bleeding on probing (BOP) \<10%, radiographic CEJ-to-alveolar bone crest distance ≤3 mm in ≥95% of teeth
• Gingivitis group: No history of periodontal disease, ≥20 teeth, PD ≤3 mm at all sites, BOP ≥30%, radiographic CEJ-to-alveolar bone crest distance ≤3 mm in ≥95% of teeth
• Periodontitis group: ≥15 teeth, \>30% of teeth affected by periodontal disease, PD ≥6 mm, clinical attachment loss (CAL) ≥5 mm, vertical bone loss ≥3 mm, Class II or III furcation involvement, radiographic alveolar bone loss extending to the middle third or beyond (≥33%), bone loss-to-age ratio between 0.25 and 1.00

You may not qualify if:

• Periodontal treatment or antibiotic use within the past 6 months
• Fewer than 20 teeth (excluding third molars)
• Presence of any systemic disease
• Current smoking or cessation within the past 5 years
• Use of immunosuppressive medication
• Alcohol consumption
• Regular medication use
• Pregnancy or lactation
• Need for antibiotic prophylaxis prior to dental procedures
• Prosthetic restorations on the teeth selected for sampling

Sponsors & Collaborators

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

Study Sites (1)

Inonu University Faculty of Dentistry

Malatya, 44210, Turkey (Türkiye)

Location

Related Publications (10)

• Zheng Y, Pan D. The Hippo Signaling Pathway in Development and Disease. Dev Cell. 2019 Aug 5;50(3):264-282. doi: 10.1016/j.devcel.2019.06.003.

  PMID: 31386861 BACKGROUND

• Qian S, Wei Z, Yang W, Huang J, Yang Y, Wang J. The role of BCL-2 family proteins in regulating apoptosis and cancer therapy. Front Oncol. 2022 Oct 12;12:985363. doi: 10.3389/fonc.2022.985363. eCollection 2022.

  PMID: 36313628 BACKGROUND

• Kegelman CD, Mason DE, Dawahare JH, Horan DJ, Vigil GD, Howard SS, Robling AG, Bellido TM, Boerckel JD. Skeletal cell YAP and TAZ combinatorially promote bone development. FASEB J. 2018 May;32(5):2706-2721. doi: 10.1096/fj.201700872R. Epub 2018 Jan 10.

  PMID: 29401582 BACKGROUND

• Jia L, Gu W, Zhang Y, Jiang B, Qiao X, Wen Y. Activated Yes-Associated Protein Accelerates Cell Cycle, Inhibits Apoptosis, and Delays Senescence in Human Periodontal Ligament Stem Cells. Int J Med Sci. 2018 Jul 30;15(11):1241-1250. doi: 10.7150/ijms.25115. eCollection 2018.

  PMID: 30123063 BACKGROUND

• Hao Y, Chun A, Cheung K, Rashidi B, Yang X. Tumor suppressor LATS1 is a negative regulator of oncogene YAP. J Biol Chem. 2008 Feb 29;283(9):5496-509. doi: 10.1074/jbc.M709037200. Epub 2007 Dec 24.

  PMID: 18158288 BACKGROUND

• Gamonal J, Bascones A, Acevedo A, Blanco E, Silva A. Apoptosis in chronic adult periodontitis analyzed by in situ DNA breaks, electron microscopy, and immunohistochemistry. J Periodontol. 2001 Apr;72(4):517-25. doi: 10.1902/jop.2001.72.4.517.

  PMID: 11338305 BACKGROUND

• Fu M, Hu Y, Lan T, Guan KL, Luo T, Luo M. The Hippo signalling pathway and its implications in human health and diseases. Signal Transduct Target Ther. 2022 Nov 8;7(1):376. doi: 10.1038/s41392-022-01191-9.

  PMID: 36347846 BACKGROUND

• Dong T, Sun X, Jin H. Role of YAP1 gene in proliferation, osteogenic differentiation, and apoptosis of human periodontal ligament stem cells induced by TNF-alpha. J Periodontol. 2021 Aug;92(8):1192-1200. doi: 10.1002/JPER.20-0176. Epub 2020 Nov 3.

  PMID: 32997793 BACKGROUND

• Bulut S, Uslu H, Ozdemir BH, Bulut OE. Expression of caspase-3, p53 and Bcl-2 in generalized aggressive periodontitis. Head Face Med. 2006 Jun 20;2:17. doi: 10.1186/1746-160X-2-17.

  PMID: 16787530 BACKGROUND

• Abuhussein H, Bashutski JD, Dabiri D, Halubai S, Layher M, Klausner C, Makhoul H, Kapila Y. The role of factors associated with apoptosis in assessing periodontal disease status. J Periodontol. 2014 Aug;85(8):1086-95. doi: 10.1902/jop.2013.130095. Epub 2013 Dec 22.

  PMID: 24359166 BACKGROUND

Biospecimen

Retention: SAMPLES WITHOUT DNA

Biospecimen Description: Gingival crevicular fluid (GCF) collected from the sulcus/pocket of three single-rooted teeth per participant using standardized paper strips (Periopaper, Oraflow Inc., USA) with absorbed volume measured by Periotron 8000. All samples stored in phosphate-buffered saline at -80°C until ELISA analysis.

MeSH Terms

Conditions

Periodontal Diseases; Gingivitis; Periodontitis; Warts; Hereditary Sensory and Autonomic Neuropathies

Condition Hierarchy (Ancestors)

Mouth Diseases; Stomatognathic Diseases; Infections; Gingival Diseases; Papillomavirus Infections; DNA Virus Infections; Virus Diseases; Skin Diseases, Viral; Tumor Virus Infections; Skin Diseases, Infectious; Skin Diseases; Skin and Connective Tissue Diseases; Nervous System Malformations; Nervous System Diseases; Heredodegenerative Disorders, Nervous System; Neurodegenerative Diseases; Polyneuropathies; Peripheral Nervous System Diseases; Neuromuscular Diseases; Congenital Abnormalities; Congenital, Hereditary, and Neonatal Diseases and Abnormalities; Genetic Diseases, Inborn

Study Officials

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

  Inonu University

  STUDY CHAIR

Study Design

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

Study Record Dates

• First Submitted: August 7, 2026
• First Posted: August 12, 2026
• Study Start: January 1, 2024
• Primary Completion: March 1, 2025
• Study Completion: April 1, 2025
• Last Updated: August 12, 2026

Record last verified: 2026-08

Locations

TU (1)