Analysis of Genetic Variations in Patients With Peri-implantitis

March 22, 2022 updated by: Egas Moniz - Cooperativa de Ensino Superior, CRL

Association Between IL-1 and IL-1Ra Gene Polymorphisms With Peri-implantitis

The primary objective of this study is to evaluate interleukin-1(IL-1) gene group polymorphisms in patients with peri-implantitis and to compare them with patients with peri-implant health (control group), taking into account smoking status, gender, age and history of periodontitis.The aim of this investigation is also to look at the levels of the inflammatory response markers IL-1 beta, prostaglandin E2 (PGE2) and tumor necrosis factor alpha (TNF alpha) in the peri-implant crevicular fluid of patients with healthy oral implants in comparison with individuals with peri-implantitis, under consideration of the patients´ individual IL-1 genotype.

The main hypothesis is that individuals carrying the polymorphism in the IL-1 gene cluster are susceptible to develop peri-implantitis through altered IL-1 and IL-1 receptor antagonist (IL-1Ra) production. A second hypothesis is that both in healthy individuals and especially pronounced in patients having peri-implantitis, an IL-1-positive genotype will result in higher levels of inflammatory cytokines (IL-1 beta, PGE2 and TNF alpha) than an IL-1- negative genotype.

Patients included in the study will be recruited from the Dental Clinic Egas Moniz implant maintenance program and will only be used for this study. All possible candidates will receive a questionnaire and if the patient's medical history is in accordance with study inclusion criteria, and if they agree to participate, informed consent will be signed. All clinical data will be collected by the same examiner.

Genes IL-1A and IL-1B control the production of the proinflammatory proteins, IL-1 alpha and IL-1 beta, respectively. IL-1RN gene controls the synthesis of the IL-1Ra, which impedes the function of IL-1alpha and IL-1beta by competing for receptor binding. Polymorphisms of the following genes will be analyzed: IL-1A-889, IL-1B + 3954, IL-1B-511 and IL-1 RN from patients with peri-implantitis and peri-implant health. For the investigation of genetic polymorphisms, it will be collected a sample of cells from the jugal mucosa with the aid of a swab.

For the biochemical analysis of the inflammatory mediators IL-1 beta, TNF alpha and PGE2 a peri-implant crevicular fluid collection will be performed by inserting paper strips into peri-implant sulcus or pocket, in situations of peri-implantitis from the most affected location, while in situations of peri-implant health from the mesio-buccal location.

Study Overview

Status

Recruiting

Conditions

Intervention / Treatment

Detailed Description

The study will be conducted according to the principles of the Helsinki Declaration (reviewed in 2013). Appropriate measures will be taken for the protection of patient data. The study was approved by Instituto Universitário Egas Moniz (IUEM) Scientific Council on 3 July 2019 and the approval of the Ethics Committee of Egas Moniz was received on 24 July 2019. Authorization was also obtained by the Clinical Direction of Egas Moniz Dental Clinic to carry out research work at the Clinic of the University.

The study will consist of two groups, the first consisting of patients with peri-implantitis (bleeding and / or suppuration on probing, probing depth (PD) equal or greater than 6mm and bone loss equal to or greater than 3mm) and the second group (control group) formed by patients with peri-implant health (peri implant mucosa without inflammatory signs and absence of peri-implant bone loss). The definition of peri-implantitis and peri-implant health will be in accordance with the new classification of periodontal and peri-implant diseases (American Academy of Periodontology (AAP) and European Federation of Periodontology (EFP) 2017)

A peri-implant probing examination (6 locations with a CP12 graduated periodontal probe) and evaluation of plaque accumulation and gingival indexes (4 locations) will be performed as usual at implant maintenance appointments. The presence of bleeding on probing or pus will also be evaluated. An implant control radiograph will be taken in cases where there is a probing depth more than 6mm. All clinical data will be collected by the same examiner. Prior to the study, a training and calibration exercise will be performed for PD measurements using patients not included in the study.

The periodontal condition of the patients will be also evaluated. To consider periodontal health, patients should have the following clinical characteristics: up to 10% of locations with bleeding on probing, absence of erythema and edema, absence of patient symptoms, absence of clinical attachment loss and bone loss and PD < 3mm (AAP and EFP 2017). A patient is a periodontitis case if: interdental clinical attachment loss is detectable at least 2 non-adjacent teeth, or buccal or oral clinical attachment loss >3mm with pocketing >3mm is detectable at least 2 teeth. The classification of periodontitis will be done on a multi-dimensional staging and grading system.

In the case of a peri-implant health situation, plaque control measures will be taken, while in cases of peri-implantitis the patient will be referred for a specific treatment appointment. In situations of patients with more than one implant affected by peri-implantitis, the one with the greatest bone loss will be chosen for the study.

The genetic analysis will be done at the IUEM Molecular Biology Laboratory with the DNA extraction technique followed by polymerase chain reaction followed by restriction fragment length polymorphism.

For the biochemical analysis the paper strips will be inserted into the peri-implant sulcus/ pocket until resistance is felt, waiting 30 seconds before removal. Visually contaminated samples (blood) will be discarded. Subsequently paper strips will be placed on Periotron® 8000 (Periotron Ide-interstate, NY, USA) for quantification of the collected peri-implant crevicular fluid volume. The strips of paper will then be placed in eppendorf tubes with filter and frozen at -80ºC until shipment to the Universidad Complutense de Madrid. At this university, the concentration of inflammatory mediators IL-1 beta and TNF alpha will be analyzed using multiplexed fluorescent sphere immunoassays using the Luminex® 100/200™ system and PGE2 by enzyme-linked immunosorbent assay (ELISA).

The prevalence of IL-1 gene group polymorphisms in patients with peri-implantitis and healthy ones will be compared by using appropriate inferential statistics methodologies (bivariate analysis/association tests), taking into account several sociodemographic and clinical variables: smoking status, gender, age and history of periodontitis. Based on a medium effect size (w=0.3), with 80% power and a 5% alpha level, the sample size for this study was calculated as n=88, thus, n= 44 for each group (periimplantitis and peri-implant health). Further, a multivariable regression logistic model will be constructed in order to evaluate the effect of those covariables and quantify the risk, through determination of the correspondent adjusted Odds Ratio (OR), towards the presence of peri-implantitis. Additionally, the levels of the inflammatory response markers IL-1 beta, PGE2 and TNF-alpha, present in the peri-implant crevicular fluid, will be compared between the group of patients with peri-implantitis and the patients with healthy oral implants, taking into account the above mentioned variables, by using appropriate inferential statistics methodologies (parametric/non parametric tests). All inferential analysis will be performed at a 5% significance level.

Study Type

Observational

Enrollment (Anticipated)

88

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Contact

Study Locations

  • Portugal
    • Monte De Caparica
      • Almada, Monte De Caparica, Portugal, 2829-511
        • Recruiting
        • Instituto Universitário Egas Moniz

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Ages Eligible for Study

  • ADULT
  • OLDER_ADULT
  • CHILD

Accepts Healthy Volunteers

Yes

Genders Eligible for Study

All

Sampling Method

Non-Probability Sample

Study Population

Patients included in the study will be recruited from the Dental Clinic Egas Moniz implant maintenance program and will only be used for this study. All possible candidates will receive a questionnaire and if the patient's medical history is in accordance with study inclusion criteria, and if they agree to participate, informed consent will be signed.

Description

Inclusion Criteria:

  • patients with Caucasian parents and grandparents
  • patients with dental implants placed at the Dental Clinic Egas Moniz or out
  • patients with peri-implantitis diagnosis
  • implants in function for at least 1 year
  • patients included in a periodontal / peri-implant support program
  • patients who agree to participate in the study and sign informed consent

Exclusion Criteria:

  • patients with any systemic condition that affects the immune system
  • pregnant women
  • patients who have taken chronic antibiotics or anti-inflammatory drugs in the last 6 months
  • patients who have undergone peri-implant treatment in the area to be evaluated

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

Cohorts and Interventions

Group / Cohort
Intervention / Treatment
Peri-implant health
Includes patients with peri implant mucosa without inflammatory signs and absence of peri-implant bone loss.
Genetic: Analysis of genetic polymorphisms
For the investigation of genetic polymorphisms, it will be collected a sample of cells from the jugal mucosa with the aid of a swab. The genetic analysis will be done with the DNA extraction technique followed by PCR-RFLP techniques (polymerase chain reaction followed by restriction fragment length polymorphism).
Other: Biochemical analysis
For the biochemical analysis of the inflammatory mediators IL-1beta, TNF-alpha and PGE2 a peri-implant crevicular fluid collection will be performed by inserting paper strips into peri-implant sulcus or pocket, in situations of peri-implantitis from the most affected location, while in situations of peri-implant health from the mesio-buccal location.
Peri-implantitis
Includes patients with bleeding and / or suppuration on probing, probing depth equal or greater than 6mm and bone loss equal to or greater than 3mm.
Genetic: Analysis of genetic polymorphisms
For the investigation of genetic polymorphisms, it will be collected a sample of cells from the jugal mucosa with the aid of a swab. The genetic analysis will be done with the DNA extraction technique followed by PCR-RFLP techniques (polymerase chain reaction followed by restriction fragment length polymorphism).
Other: Biochemical analysis
For the biochemical analysis of the inflammatory mediators IL-1beta, TNF-alpha and PGE2 a peri-implant crevicular fluid collection will be performed by inserting paper strips into peri-implant sulcus or pocket, in situations of peri-implantitis from the most affected location, while in situations of peri-implant health from the mesio-buccal location.

What is the study measuring?

Primary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Frequency of Participants with polymorphism in the IL-1 gene cluster
Time Frame: 1 month after sample collection
In the IL-1A-889 gene, two possible alleles are expected to be found, allele 1 (83 base pairs (bp) + 16 bp), allele 2 - allele with nucleotide sequence changed (99 bp). In the IL-1B + 3954 gene, two possible alleles are expected to be found, allele 1 (12 bp + 85 bp + 97 bp), allele 2 (12 bp + 182 bp). In the IL-1B-511 gene, two possible alleles are expected to be found, allele 1 (190bp + 114bp), allele 2 (304 bp). In relation to the polymorphism of the IL-1RN gene, 5 possible alleles are expected to be found according to the repeated base pair sequences: allele 1 (410 bp), allele 2 (240 bp), allele 3 (500 bp), allele 4 (325 bp), allele 5 (595 bp).
1 month after sample collection

Secondary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Concentration of inflammatory response markers IL-1 beta, PGE2 and TNF-alpha
Time Frame: 1 month after sample collection
The paper strips with peri-implant crevicular fluid (PICF) will be placed on Periotron® 8000 (Periotron Ide-interstate, NY, USA) for quantification of the collected PICF volume.The concentration of inflammatory mediators IL-1beta and TNF-alpha will be analyzed using multiplexed fluorescent sphere immunoassays using the Luminex® 100/200™ system and PGE2 by ELISA (enzyme-linked immunosorbent assay).
1 month after sample collection

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Sponsor

Egas Moniz - Cooperativa de Ensino Superior, CRL

Collaborators

Oral Reconstruction Foudation

Investigators

  • Principal Investigator: Ricardo Alves, Phd, Egas Moniz

Publications and helpful links

The person responsible for entering information about the study voluntarily provides these publications. These may be about anything related to the study.

General Publications

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (ACTUAL)

March 9, 2021

Primary Completion (ANTICIPATED)

July 29, 2022

Study Completion (ANTICIPATED)

July 29, 2022

Study Registration Dates

First Submitted

March 18, 2021

First Submitted That Met QC Criteria

March 18, 2021

First Posted (ACTUAL)

March 19, 2021

Study Record Updates

Last Update Posted (ACTUAL)

March 23, 2022

Last Update Submitted That Met QC Criteria

March 22, 2022

Last Verified

March 1, 2022

More Information

Terms related to this study

Keywords

Additional Relevant MeSH Terms

Other Study ID Numbers

  • PHDJMC

Plan for Individual participant data (IPD)

Plan to Share Individual Participant Data (IPD)?

NO

IPD Plan Description

The collections of DNA samples were collected only for the purpose of analyzing the polymorphisms of the IL-1 gene group. Participants authorized their collection only for this purpose. For ethical and confidentiality reasons, these samples may not be accessible to other researchers. DNA samples will be eliminated after conducting the study.

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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