Periodontal DiseaseBYHN-GCF-HIPPONCT07759466
Hippo Signaling Pathway Proteins in Gingival Crevicular Fluid of Individuals With Periodontal Disease
An observational study of Periodontal Disease and Gingivitis and Periodontal Diseases, run by Inonu University at 1 site in Turkey (Türkiye).
Trial 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. Read more about this study
- Who is it for?
- Ages 18 to 65 yearsHealthy volunteers can take part
- Conditions the study is for
- Periodontal Disease, Gingivitis and Periodontal Diseases, Biomarker Discovery and Validation
- What is being tested?
- ELISA Biomarker Analysis
- How many people
- 75 participants
- Where
- 1 siteTurkey (Türkiye)
- Study dates
- 1 Jan 2024to 1 Apr 2025
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 65 years
- All sexes
- Healthy volunteers accepted
- Adult, Older adult
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.
Who may be able to join
- 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
Who cannot join
- 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
Show all 10Show fewer
- Regular medication use
- Pregnancy or lactation
- Need for antibiotic prophylaxis prior to dental procedures
- Prosthetic restorations on the teeth selected for sampling
Trial locations and contacts
1 site in 1 country.
Turkey (Türkiye)1 site
Inonu University Faculty of Dentistry
Malatya, 44210
About this study
Official title: Hippo Signaling Pathway Proteins and Their Correlation With Markers of Apoptosis and Bone Metabolism in the Gingival Crevicular Fluid of Individuals With Periodontal Disease
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.
Read the detailed description
Background and rationale
Periodontal disease is a common infectious and inflammatory condition of the periodontium, presenting clinically as gingivitis or periodontitis. Gingivitis is a reversible inflammation confined to the gingiva, whereas periodontitis is a chronic, progressive disease characterized by irreversible loss of periodontal attachment and alveolar bone. Although microbial biofilms containing periodontal pathogens initiate the disease, its severity and progression are largely governed by the host immune-inflammatory response.
The Hippo signaling pathway is a highly conserved regulatory network that integrates mechanical and biochemical cues to control cellular polarity, differentiation, proliferation, apoptosis, immune responses, tissue homeostasis, and organ size. Its core components include 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), and mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4). Upstream kinases such as MST1/2 and MAP4K4 activate LATS1/2, which phosphorylate YAP and TAZ, leading to their cytoplasmic retention or degradation and preventing TEAD-mediated transcriptional activity in the nucleus.
Preclinical evidence has implicated this pathway in mineralized tissue homeostasis of the alveolar bone and periodontal complex. Inhibition of YAP has been reported to suppress proliferation and induce apoptosis of periodontal ligament stem cells through the BCL-2 family pathway, and YAP/TAZ-deficient animal models show enhanced resorption of alveolar bone and cementum together with reduced alkaline phosphatase expression. Despite these findings, no study to date has directly evaluated the role of Hippo pathway proteins in human periodontal disease. This study was therefore designed to address that gap.
Objective and hypothesis
The study aimed to examine changes in the Hippo signaling pathway proteins YAP, TAZ, MST1, TEAD, LATS1, and MAP4K4, together with the osteogenic marker ALP and the anti-apoptotic protein BCL-2, in gingival crevicular fluid across periodontal health and disease states. It was hypothesized that the gingival crevicular fluid levels of Hippo pathway-related proteins are altered in periodontal disease and correlate with markers of bone turnover and apoptosis.
Study population and grouping
Systemically healthy adults aged 18 to 65 years who presented to the Department of Periodontology were assessed for eligibility. Participants were allocated into three groups of equal size according to the 2017 World Workshop on the Classification of Periodontal Diseases: periodontally healthy individuals, patients with gingivitis, and patients with stage 3 grade B periodontitis. Group assignment was based on probing depth, bleeding on probing, clinical attachment level, and radiographic evidence of bone loss.
Clinical examination
For all participants, Plaque Index, Gingival Index, Probing Depth, Bleeding on Probing, and Clinical Attachment Level were recorded by a single calibrated periodontist at six sites per tooth. Panoramic radiographs were used to assess alveolar bone loss.
Gingival crevicular fluid sampling
Gingival crevicular fluid samples were collected one day after periodontal examination. Participants were instructed not to eat, drink, or brush their teeth for at least two hours before sampling. Sampling sites were isolated with cotton rolls, supragingival plaque was removed, and tooth surfaces were gently dried. Paper strips were placed into the pocket and left in position for a fixed interval, and samples contaminated with blood or saliva were discarded. Fluid volume was measured electronically, and samples from each participant were pooled and stored at low temperature until analysis. Sampling sites within each group were defined according to group-specific clinical criteria.
Laboratory analysis
The levels of YAP, TAZ, MST1, LATS1, MAP4K4, TEAD, ALP, and BCL-2 were determined using commercially available enzyme-linked immunosorbent assay kits 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 gingival crevicular fluid volume between health and disease states.
Analytical approach
Both total amounts and concentrations of the target proteins were compared among the three groups. Correlation analyses were performed to evaluate the relationships among the measured proteins and between these proteins and clinical periodontal parameters. Reporting protein levels in two complementary ways was intended to distinguish true biological differences from those attributable to changes in fluid volume associated with inflammation.
- Conditions
- Periodontal Disease, Gingivitis and Periodontal Diseases, Biomarker Discovery and Validation, Pathways
- Being tested
- ELISA Biomarker Analysis
- 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
- Medical subject headings
- Periodontal Diseases, Gingivitis, Periodontitis, Warts, Hereditary Sensory and Autonomic Neuropathies
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
ELISA Biomarker AnalysisDiagnostic 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.
| Study group | Role | What they receive |
|---|---|---|
| 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. Receives: 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. Receives: 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. Receives: Diagnostic Test: ELISA Biomarker Analysis |
What is being measured
The primary outcome is the main question the study is designed to answer.
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.
Measured: Baseline
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.
Measured: Baseline
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.
Measured: Baseline
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.
Measured: Baseline
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.
Measured: Baseline
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.
Measured: Baseline
Secondary outcomes (2)
Gingival crevicular fluid level of B-cell lymphoma 2 (BCL-2)
The level of the anti-apoptotic protein B-cell lymphoma 2 (BCL-2) in gingival crevicular fluid was quantified by enzyme-linked immunosorbent assay and compared among periodontally healthy, gingivitis, and stage 3 grade B periodontitis groups. BCL-2 was reported both as concentration (ng/mL) and as total amount per 30-second sampling interval.
Measured: Baseline
Gingival crevicular fluid level of alkaline phosphatase (ALP)
The level of the bone metabolism marker alkaline phosphatase (ALP) in gingival crevicular fluid was quantified by enzyme-linked immunosorbent assay and compared among periodontally healthy, gingivitis, and stage 3 grade B periodontitis groups. ALP was reported both as concentration (ng/mL) and as total amount per 30-second sampling interval.
Measured: Baseline
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted12 Aug 2026
- Study start1 Jan 2024
- Primary completion1 Mar 2025
- Study completion1 Apr 2025
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (10)Papers the sponsor has linked to this study.
- Other trials for Periodontal DiseaseStudies of the same condition, open and under way.
- More trials run by Inonu UniversityThe sponsor's other registered studies.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Case-control
- Time perspective
- Cross-sectional
- Enrolment
- 75 participants (actual)
Sponsor and investigators
- Lead sponsor
- Inonu University
- Sponsor type
- University, hospital or other
- Collaborators
- Harvard University, University of Minnesota
- Responsible party
- Principal investigator, Cuneyt Asim Aral, Inonu University
- Investigators
- Cüneyt A Aral, Chair, Professor, DDS, PhD, Study chair, Inonu University
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT07759466
- Sponsor's protocol ID
- IUCREC.2024/07
- Other IDs
- TDH-2024-3461
- Acronym
- BYHN-GCF-HIPPO
- Status last verified
- Aug 2026
- First submitted
- 7 Aug 2026
- Last updated
- 13 Aug 2026
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- Undecided
Publications
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.
PubMed 31386861Qian 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.
PubMed 36313628Kegelman 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.
PubMed 29401582Jia 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.
PubMed 30123063Hao 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.
PubMed 18158288Gamonal 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.
PubMed 11338305Fu 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.
PubMed 36347846Dong 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.
PubMed 32997793Bulut 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.
PubMed 16787530Abuhussein 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.
PubMed 24359166
Study at a glance
From the registry record, last updated by the sponsor on 13 Aug 2026. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Hippo Signaling Pathway Proteins and Their Correlation With Markers of Apoptosis and Bone Metabolism in the Gingival Crevicular Fluid of Individuals With Periodontal Disease
- ClinicalTrials.gov ID
- NCT07759466
- Acronym / sponsor ID
- BYHN-GCF-HIPPO / IUCREC.2024/07
- 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)
- Study type
- Observational
- Recruitment status
- Completed
- Enrolment
- 75 participants (actual)
- Lead sponsor
- Inonu UniversityUniversity, hospital or other
- Collaborators
- Harvard University, University of Minnesota
- Study start
- 1 Jan 2024 (actual)
- Primary completion
- 1 Mar 2025 (actual)
- Study completion
- 1 Apr 2025 (actual)
- First posted
- 12 Aug 2026 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
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.
Detailed description
Background and rationale
Periodontal disease is a common infectious and inflammatory condition of the periodontium, presenting clinically as gingivitis or periodontitis. Gingivitis is a reversible inflammation confined to the gingiva, whereas periodontitis is a chronic, progressive disease characterized by irreversible loss of periodontal attachment and alveolar bone. Although microbial biofilms containing periodontal pathogens initiate the disease, its severity and progression are largely governed by the host immune-inflammatory response.
The Hippo signaling pathway is a highly conserved regulatory network that integrates mechanical and biochemical cues to control cellular polarity, differentiation, proliferation, apoptosis, immune responses, tissue homeostasis, and organ size. Its core components include 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), and mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4). Upstream kinases such as MST1/2 and MAP4K4 activate LATS1/2, which phosphorylate YAP and TAZ, leading to their cytoplasmic retention or degradation and preventing TEAD-mediated transcriptional activity in the nucleus.
Preclinical evidence has implicated this pathway in mineralized tissue homeostasis of the alveolar bone and periodontal complex. Inhibition of YAP has been reported to suppress proliferation and induce apoptosis of periodontal ligament stem cells through the BCL-2 family pathway, and YAP/TAZ-deficient animal models show enhanced resorption of alveolar bone and cementum together with reduced alkaline phosphatase expression. Despite these findings, no study to date has directly evaluated the role of Hippo pathway proteins in human periodontal disease. This study was therefore designed to address that gap.
Objective and hypothesis
The study aimed to examine changes in the Hippo signaling pathway proteins YAP, TAZ, MST1, TEAD, LATS1, and MAP4K4, together with the osteogenic marker ALP and the anti-apoptotic protein BCL-2, in gingival crevicular fluid across periodontal health and disease states. It was hypothesized that the gingival crevicular fluid levels of Hippo pathway-related proteins are altered in periodontal disease and correlate with markers of bone turnover and apoptosis.
Study population and grouping
Systemically healthy adults aged 18 to 65 years who presented to the Department of Periodontology were assessed for eligibility. Participants were allocated into three groups of equal size according to the 2017 World Workshop on the Classification of Periodontal Diseases: periodontally healthy individuals, patients with gingivitis, and patients with stage 3 grade B periodontitis. Group assignment was based on probing depth, bleeding on probing, clinical attachment level, and radiographic evidence of bone loss.
Clinical examination
For all participants, Plaque Index, Gingival Index, Probing Depth, Bleeding on Probing, and Clinical Attachment Level were recorded by a single calibrated periodontist at six sites per tooth. Panoramic radiographs were used to assess alveolar bone loss.
Gingival crevicular fluid sampling
Gingival crevicular fluid samples were collected one day after periodontal examination. Participants were instructed not to eat, drink, or brush their teeth for at least two hours before sampling. Sampling sites were isolated with cotton rolls, supragingival plaque was removed, and tooth surfaces were gently dried. Paper strips were placed into the pocket and left in position for a fixed interval, and samples contaminated with blood or saliva were discarded. Fluid volume was measured electronically, and samples from each participant were pooled and stored at low temperature until analysis. Sampling sites within each group were defined according to group-specific clinical criteria.
Laboratory analysis
The levels of YAP, TAZ, MST1, LATS1, MAP4K4, TEAD, ALP, and BCL-2 were determined using commercially available enzyme-linked immunosorbent assay kits 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 gingival crevicular fluid volume between health and disease states.
Analytical approach
Both total amounts and concentrations of the target proteins were compared among the three groups. Correlation analyses were performed to evaluate the relationships among the measured proteins and between these proteins and clinical periodontal parameters. Reporting protein levels in two complementary ways was intended to distinguish true biological differences from those attributable to changes in fluid volume associated with inflammation.
Study design
- Observational model
- Case-control
- Time perspective
- Cross-sectional
- Number of arms
- 3
| Arm | Type | Receives |
|---|---|---|
| Periodontally healthy | – | Diagnostic Test: ELISA Biomarker AnalysisSystemically 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. |
| Gingivitis | – | Diagnostic Test: ELISA Biomarker AnalysisSystemically 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. |
| Stage 3 grade B periodontitis | – | Diagnostic Test: ELISA Biomarker AnalysisSystemically 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. |
Who may qualify
- Ages
- 18 to 65 years
- Sex
- All
- Healthy volunteers
- Accepted
- Age groups
- Adult, Older adult
- 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.
Key inclusion criteria (5 listed)
- 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
Key exclusion criteria (10 listed)
- 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
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| ELISA Biomarker AnalysisGingival 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. | Diagnostic test | Gingivitis, Periodontally healthy, Stage 3 grade B periodontitis |
Primary outcome
Gingival crevicular fluid level of Yes-associated protein (YAP)
Time frame: Baseline
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.
Gingival crevicular fluid level of transcriptional co-activator with PDZ-binding motif (TAZ)
Time frame: Baseline
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.
Gingival crevicular fluid level of mammalian sterile 20-like kinase 1 (MST1)
Time frame: Baseline
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.
Gingival crevicular fluid level of TEA domain family member (TEAD)
Time frame: Baseline
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.
Gingival crevicular fluid level of large tumor suppressor kinase 1 (LATS1)
Time frame: Baseline
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.
Gingival crevicular fluid level of mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4)
Time frame: Baseline
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.
2 secondary outcomes
Gingival crevicular fluid level of B-cell lymphoma 2 (BCL-2)
Time frame: Baseline
Gingival crevicular fluid level of alkaline phosphatase (ALP)
Time frame: Baseline
Results and safety
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 13 Aug 2026.
- Background: PubMed 31386861Zheng 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.
- Background: PubMed 36313628Qian 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.
- Background: PubMed 29401582Kegelman 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.
- Background: PubMed 30123063Jia 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.
- Background: PubMed 18158288Hao 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.
- Background: PubMed 11338305Gamonal 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.
- Background: PubMed 36347846Fu 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.
- Background: PubMed 32997793Dong 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.
- Background: PubMed 16787530Bulut 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.
- Background: PubMed 24359166Abuhussein 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.
- Individual participant dataSharing planned: Undecided
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Cüneyt A Aral, Chair, Professor, DDS, PhD study chair
Inonu University
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Periodontal Disease and Gingivitis and Periodontal Diseases, run by Inonu University at 1 site in Turkey (Türkiye).
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 5 inclusion and 10 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: finished, as of the sponsor's last update on 13 Aug 2026. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Turkey (Türkiye). Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Gingival crevicular fluid level of Yes-associated protein (YAP) (time frame: Baseline). The study lists 6 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT07759466, which also lists 10 linked publications.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 13 Aug 2026.