The Role of Anti-inflammatory Cytokines and Antimicrobial Peptide LL-37 Biomarkers in the Treatment of Periodontal Disease.

NCT04404335 | Completed

• 1 other identifier: LL-37PERIOURJC
• Study Type: observational
• Target: 60
• Locations: 1 country
• Sites: 1

Brief Summary

Periodontal disease is an infectious pathology that consists of the destruction of the gums and supporting structures around the teeth. There are several mechanisms involved in this process. In healthy conditions there is a balance between the bacteria present in the gums and the defence mechanisms of the host to fight the disease. One of the substances involved in favouring this protective mechanism is a peptide called LL-37. The aim of this study is to find out more about the rol of this peptide during this process by using a non invasive method of analysis consisting of analysing some samples of the fluid existing between the gums and the teeth. This clinical study will take place at the University Clinic at the University Rey Juan Carlos, Alcorcon (Madrid), Spain. Patients that attend the clinic suffering from gum disease will be offered the possibility to participate in this study. A full questionnaire indicating the risks and benefits of participating in the study as well as the management of confidential personal data will be discussed with the participants in full detail. Full consent will be sought before commencing the study. Firstly, all participants will receive a through periodontal (gum) examination to assess if they suffer from periodontal disease and to what extent (severity). Based on this preliminary data, patients will be classified in different categories. Patients will be divided in two groups:

• 30 Healthy patients (no signs of periodontitis)
• 30 Periodontal patients (subdivided in 2 groups of 30, depending of the severity of their disease:
• 15 patients with Stage I-II periodontitis (mild/moderate)
• 15 patients with Stage III-IV periodontitis (severe). The intervention will consist on taking samples of crevicular fluid (the fluid found in the space between the gum and the root of the tooth) to quantify the presence of LL-37 as well as IL-4, IL-6, IL-10, which seems to be associated with the defence mechanisms (immunity) of the host against periodontal disease. These samples will be analysed in the laboratory by using some specific procedure called ELISA. All patients from the test group will receive a basic course of periodontal treatment (root scale and polish), which is intended to remove the bacteria causing the disease from the space between the gums and the roots (periodontal pocket). Samples of crevicular fluid will be obtained before commencing the treatment. After 4-6 weeks, a new periodontal examination will take place to assess if there is an improvement of the condition and also to quantify the levels of LL-37, IL-4, IL-6, IL-10. The findings of this study will help to understand the role of this peptide in the defence mechanism of the host against periodontal disease. This could ultimately serve to develop new therapies that could include the use of this peptide in addition to the routine periodontal treatment prescribed to improve the healing conditions of the patient against periodontal disease.

Conditions

Periodontal Diseases; Periodontitis

Keywords

Antimicrobial peptides; Defensines; LL-37; Periodontal disease; Crevicular fluid; IL-4; IL-6; IL-10

Outcome Measures

Primary Outcomes (1)

• LL-37 in crevicular fluid in healthy/periodontal patients.

  Crevicular fluid samples will also be obtained following the manufacturers protocol using perio papers (Periopaper, Applied Biosystems, Inc, Foster City, CA). Samples will be stored at -80 C at the laboratory of Departamento de Epidemiología y Salud Pública, Edificio Departamental I, Fundación Clínica Universitaria, URJC (Alcorcon). Once all samples are obtained, appropriate transport will be arranged to the Department of Natural Sciences, Biomedical Sciences, Hendon Campus, Middlesex University of London. There ELISA analysis of all samples will be performed by using a specific ELISA kit commercially available for human LL-37 (HyCult Biotechnology, Uden, the Netherlands).

  At baseline for healthy (control) and periodontal (test) groups and at 4-6 weeks after treatment for test group

Secondary Outcomes (5)

• Periodontal parameters

  At baseline for healthy (control) and periodontal (test) groups and at 4-6 weeks after treatment for test group

• Probing depth (PD)

  At baseline for healthy (control) and periodontal (test) groups and at 4-6 weeks after treatment for test group

• Clinical attachment level (CAL)

  At baseline for healthy (control) and periodontal (test) groups and at 4-6 weeks after treatment for test group

• Plaque index (PI)

  At baseline for healthy (control) and periodontal (test) groups and at 4-6 weeks after treatment for test group

• Blood on probing (BOP)

  At baseline for healthy (control) and periodontal (test) groups and at 4-6 weeks after treatment for test group

Study Arms (2)

Healthy (control group)

This group will include a total of 30 patients free of periodontitis Samples of crevicular fluid to quantity levels of antimicrobial peptide LL-37 will be taken at baseline ELISA analysis of all samples will be performed by using a specific ELISA kit commercially available for human LL-37 and also IL-4, IL-6 and IL-10

Diagnostic Test: Crevicular fluid analysis by using ELISA testing.

Periodontitis (test group)

30 Periodontal patients (subdivided in 2 groups of 15, depending of the severity of their disease): * 15 patients with Stage I-II periodontitis (mild/moderate) * 15 patients with Stage III-IV periodontitis (severe). Two sets of samples of crevicular fluid will be obtained before and after receiving a course of routine basic periodontal treatment (root scaling and planing). A reevaluation visit to re-assess clinical periodontal parameters will take place at 4-6 weeks after treatment. New samples of crevicular fluid will then be taken at this stage to compare the level of LL-37 before and after the treatment. ELISA analysis of all samples will be performed by using a specific ELISA kit commercially available for human LL-37 (HyCult Biotechnology, Uden, the Netherlands).

Diagnostic Test: Crevicular fluid analysis by using ELISA testing.

Interventions

Crevicular fluid analysis by using ELISA testing. DIAGNOSTIC_TEST

Samples of crevicular fluid (fluid existing in the space between the gums and the roots of the teeth) will be obtained from healthy and periodontal patients. In periodontal patients, samples will be taken before and after receiving a routine course of periodontal treatment (root scale and polish) to assess the levels of a peptide called LL-37 and inflammatory cytokines (IL-4, IL-6, IL-10): Samples will then be analysed using laboratory procedures (ELISA)

Healthy (control group); Periodontitis (test group)

Eligibility Criteria

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

Study Population

This clinical study will take place at the University Clinic at the University Rey Juan Carlos, Alcorcon (Madrid), Spain. Patients that attend the clinic suffering from gum disease will be offered the possibility to participate in this study. A full questionnaire indicating the risks and benefits of participating in the study as well as the management of confidential personal data will be discussed with the participants in full detail. Full consent will be sought before commencing the study. Patients will be divided in two groups: * 30 Healthy patients (no signs of periodontitis) * 30 Periodontal patients (subdivided in 2 groups of 15, depending of the severity of their disease: * 15 patients with Stage I-II periodontitis (mild/moderate) * 15 patients with Stage III-IV periodontitis (severe).

Sponsors & Collaborators

• Universidad Rey Juan Carlos: lead

Study Sites (1)

Clinica Universitaria Universidad Rey Juan Carlos

Alcorcón, Madrid, 28923, Spain

Location

Related Publications (11)

• Davidopoulou S, Diza E, Sakellari D, Menexes G, Kalfas S. Salivary concentration of free LL-37 in edentulism, chronic periodontitis and healthy periodontium. Arch Oral Biol. 2013 Aug;58(8):930-4. doi: 10.1016/j.archoralbio.2013.01.003. Epub 2013 Feb 8.

  PMID: 23778112 BACKGROUND

• Turkoglu O, Emingil G, Kutukculer N, Atilla G. Gingival crevicular fluid levels of cathelicidin LL-37 and interleukin-18 in patients with chronic periodontitis. J Periodontol. 2009 Jun;80(6):969-76. doi: 10.1902/jop.2009.080532.

  PMID: 19485828 BACKGROUND

• Putsep K, Carlsson G, Boman HG, Andersson M. Deficiency of antibacterial peptides in patients with morbus Kostmann: an observation study. Lancet. 2002 Oct 12;360(9340):1144-9. doi: 10.1016/S0140-6736(02)11201-3.

  PMID: 12387964 BACKGROUND

• de Haar SF, Hiemstra PS, van Steenbergen MT, Everts V, Beertsen W. Role of polymorphonuclear leukocyte-derived serine proteinases in defense against Actinobacillus actinomycetemcomitans. Infect Immun. 2006 Sep;74(9):5284-91. doi: 10.1128/IAI.02016-05.

  PMID: 16926422 BACKGROUND

• Barros SP, Williams R, Offenbacher S, Morelli T. Gingival crevicular fluid as a source of biomarkers for periodontitis. Periodontol 2000. 2016 Feb;70(1):53-64. doi: 10.1111/prd.12107.

  PMID: 26662482 BACKGROUND

• Kinney JS, Morelli T, Oh M, Braun TM, Ramseier CA, Sugai JV, Giannobile WV. Crevicular fluid biomarkers and periodontal disease progression. J Clin Periodontol. 2014 Feb;41(2):113-120. doi: 10.1111/jcpe.12194. Epub 2013 Dec 12.

  PMID: 24303954 BACKGROUND

• McCrudden MT, Orr DF, Yu Y, Coulter WA, Manning G, Irwin CR, Lundy FT. LL-37 in periodontal health and disease and its susceptibility to degradation by proteinases present in gingival crevicular fluid. J Clin Periodontol. 2013 Oct;40(10):933-41. doi: 10.1111/jcpe.12141. Epub 2013 Aug 18.

  PMID: 23952216 BACKGROUND

• Greer A, Zenobia C, Darveau RP. Defensins and LL-37: a review of function in the gingival epithelium. Periodontol 2000. 2013 Oct;63(1):67-79. doi: 10.1111/prd.12028.

  PMID: 23931055 BACKGROUND

• Mallapragada S, Wadhwa A, Agrawal P. Antimicrobial peptides: The miraculous biological molecules. J Indian Soc Periodontol. 2017 Nov-Dec;21(6):434-438. doi: 10.4103/jisp.jisp_325_16.

  PMID: 29551860 BACKGROUND

• Khurshid Z, Naseem M, Yahya I Asiri F, Mali M, Sannam Khan R, Sahibzada HA, Zafar MS, Faraz Moin S, Khan E. Significance and Diagnostic Role of Antimicrobial Cathelicidins (LL-37) Peptides in Oral Health. Biomolecules. 2017 Dec 5;7(4):80. doi: 10.3390/biom7040080.

  PMID: 29206168 BACKGROUND

• Madruga D, Garcia MM, Martino L, Hassan H, Elayat G, Ghali L, Ceballos L. Positive correlational shift between crevicular antimicrobial peptide LL-37, pain and periodontal status following non-surgical periodontal therapy. A pilot study. BMC Oral Health. 2023 May 28;23(1):335. doi: 10.1186/s12903-023-03023-w.

  PMID: 37246231 DERIVED

Biospecimen

Retention: SAMPLES WITHOUT DNA

Crevicular fluid

MeSH Terms

Conditions

Periodontal Diseases; Periodontitis

Condition Hierarchy (Ancestors)

Mouth Diseases; Stomatognathic Diseases

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: PROSPECTIVE
• Target Duration: 2 Months
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Associate Professor

Study Record Dates

• First Submitted: May 17, 2020
• First Posted: May 27, 2020
• Study Start: May 17, 2021
• Primary Completion: June 30, 2022
• Study Completion: July 15, 2022
• Last Updated: September 30, 2022

Record last verified: 2022-09

Data Sharing

• IPD Sharing: Will not share

Locations

ES (1)