Free Amino-acid Composition of Peri-implant and Periodontal Conditions

NCT ID: NCT07187323

Last Updated: 2025-09-23

Basic Information

• Recruitment Status: COMPLETED
• Total Enrollment: 120 participants
• Study Classification: OBSERVATIONAL
• Study Start Date: 2024-08-02
• Study Completion Date: 2025-08-20

Brief Summary

Peri-implant diseases, encompassing peri-implant mucositis and peri-implantitis, are biologically driven inflammatory conditions that threaten the long-term success of dental implants. The microbial signatures and their metabolic products have prompted growing interest in salivary biomarkers, particularly amino acids (AAs), which may reflect both microbial activity and host response.However, despite increasing evidence of periodontal disease, the role of salivary AAs in peri-implant conditions remains underexplored.The aim of this study is to comprehensively evaluate and compare the salivary free amino acid profiles among individuals with peri-implantitis, peri-implant mucositis, and healthy peri-implant mucosa, as well as those with periodontitis, gingivitis, and a healthy periodontium, and seeks to investigate whether distinct salivary amino acid signatures are linked to different stages and types of peri-implant and periodontal inflammation, and to evaluate their potential as non-invasive biomarkers for disease differentiation, activity, and severity

Detailed Description

The study included one-hundred and thirty-two patients who applied to the Department of Periodontology at Istanbul Medipol University, Faculty of Dentistry. Four of the one-hundred and thirty-two patients declined to participate in the trial. Two patients were eliminated because they had recently taken antibiotics, six patients were discarded because they had recently received periodontal therapy, and six patients were excluded because they had systemic diseases, resulting in a final sample size of one-hundred and twenty patients .Quantitative analysis of amino acids in saliva was carried out using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with a Thermo Scientific TSQ Quantum Access MAX system.

Conditions

Peri-Implant Health; Peri Implant Mucositis; Peri Implantitis

Study Design

• Observational Model Type: OTHER
• Study Time Perspective: CROSS_SECTIONAL

Study Groups

Saliva collection

Saliva and serum sampling Saliva were collected to analyze the selected markers as unstimulated samples during the early hours of the day.

The saliva was centrifuged and then transferred into Eppendorf tubes. Venous puncture was performed after saliva collection and 10 mL of blood samples were collected by qualified staff from each participant. Saliva and serum were then stored at -80 °C until analysis.

Saliva sampling

Intervention Type: OTHER

Unstimulated, whole saliva was collected from all participants across the six study groups following a detailed explanation of the procedure. Initially, individuals were asked to rinse their mouths with filtered water and remain seated in a relaxed position for five minutes. Then, they were instructed to passively drool into a sterile collection tube over a 10-minute period.31 The collected saliva samples were then centrifuged at 3000 ×g for 10 minutes to remove debris and prepare them for storage at -80°C .

Amino Acid Analysis

Quantitative analysis of amino acids in saliva was carried out using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with a Thermo Scientific TSQ Quantum Access MAX system (Thermo Scientific, Schaumburg, IL, USA).

Amino Acid Analysis

Intervention Type: OTHER

Saliva samples were deproteinized by mixing with 6% sulfosalicylic acid in a 1:1 ratio, followed by incubation at room temperature for 5 minutes. The mixtures were then centrifuged at 15,000 rpm for 5 minutes. A 10 µL aliquot of the resulting supernatant was diluted with 800 µL of 2 mM tridecafluoroheptanoic acid containing 0.375 M glucosaminic acid and internal standards. The prepared solutions were transferred into autosampler vials for analysis. Calibration standards were generated by combining known concentrations of amino acid standards.

Interventions

Saliva sampling

Unstimulated, whole saliva was collected from all participants across the six study groups following a detailed explanation of the procedure. Initially, individuals were asked to rinse their mouths with filtered water and remain seated in a relaxed position for five minutes. Then, they were instructed to passively drool into a sterile collection tube over a 10-minute period.31 The collected saliva samples were then centrifuged at 3000 ×g for 10 minutes to remove debris and prepare them for storage at -80°C .

Intervention Type: OTHER

Amino Acid Analysis

Saliva samples were deproteinized by mixing with 6% sulfosalicylic acid in a 1:1 ratio, followed by incubation at room temperature for 5 minutes. The mixtures were then centrifuged at 15,000 rpm for 5 minutes. A 10 µL aliquot of the resulting supernatant was diluted with 800 µL of 2 mM tridecafluoroheptanoic acid containing 0.375 M glucosaminic acid and internal standards. The prepared solutions were transferred into autosampler vials for analysis. Calibration standards were generated by combining known concentrations of amino acid standards.

Intervention Type: OTHER

Eligibility Criteria

Inclusion Criteria

• be between 18 and 65 years of age
• have a minimum of 20 natural teeth, excluding third molars (for non-implant-treated groups)
• be edentulous patients treated with implants loaded at least 1 year prior (for implant-treated groups)
• be in a systemically healthy condition.

Exclusion Criteria

• current tobacco use
• chronic use of any systemic medication
• use of antibiotics, anti-inflammatory drugs (including corticosteroids and NSAIDs), immunosuppressants, beta-blockers, calcium channel blockers, anticoagulants, or hormonal contraceptives within the 3 months prior to the study
• receipt of periodontal therapy within the past 6 months
• pregnancy or breastfeeding
• presence of orthodontic appliances a DMFT index (Decayed, Missing, and Filled Teeth per tooth) greater than 2

• Minimum Eligible Age: 20 Years
• Maximum Eligible Age: 65 Years
• Eligible Sex: ALL
• Accepts Healthy Volunteers: Yes

Sponsors

Lokman Hekim University

OTHER_GOV

Sponsor Role: collaborator

Ankara University

OTHER

Sponsor Role: collaborator

Ankara Medipol University

OTHER

Sponsor Role: collaborator

Istanbul Medipol University Hospital

OTHER

Sponsor Role: lead

Responsible Party

Nur Balci, DDs, PhD

Assoc Prof Dr

Responsibility Role: PRINCIPAL_INVESTIGATOR

Locations

Istanbul Medipol University, School of Dentistry

Istanbul, Istanbul, Turkey (Türkiye)

Countries

Turkey (Türkiye)

Other Identifiers

Istanbul Medipol University

Identifier Type: OTHER

Identifier Source: secondary_id

645

Identifier Type: -

Identifier Source: org_study_id