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EVALUATION OF SALIVA AND SERUM HEME OXYGENASE, ARYLESTERASE AND NUCLEAR FACTOR ERYTHROID 2-RELATED FACTOR 2 LEVELS IN PATIENTS WITH STAGE III PERIODONTITIS

EVALUATION OF SALIVA AND SERUM HEME OXYGENASE, ARYLESTERASE AND NUCLEAR FACTOR ERYTHROID 2-RELATED FACTOR 2 LEVELS IN PATIENTS WITH STAGE III PERIODONTITIS

Status
Completed
Phases
Unknown
Study type
Observational
Source
ClinicalTrials.gov
Registry ID
NCT06986044
Enrollment
74
Registered
2025-05-22
Start date
2023-05-01
Completion date
2024-09-15
Last updated
2025-05-22

For informational purposes only — not medical advice. Sourced from public registries and may not reflect the latest updates. Terms

Conditions

Periodontal Health, Periodontitis Stage III

Keywords

total antioxidan status, total oxidan status, oxidative stress index, arylesterase, heme oxygenase, nuclear factor erythroid 2 related factor 2, saliva, serum, stage III periodontitis

Brief summary

Periodontal disease is a chronic and progressive inflammatory disease in which the hard and soft tissues that support the teeth are damaged. It is caused by the interaction between harmful bacteria and the body's immune responses, and most periodontal tissues are damaged by the body's abnormal response to these microorganisms and their products. When bacteria enter the body, immune cells (neutrophils) produce reactive oxygen species (ROS) in a process called the respiratory burst. These ROS damage cells, causing tissue destruction through a variety of mechanisms, including DNA damage, fat oxidation and protein damage. Studies have shown that neutrophils from individuals with periodontal disease produce more ROS than neutrophils from healthy individuals. High amounts of ROS lead to oxidative damage to gum tissue, periodontal ligament and alveolar bone. Oxidative stress occurs when antioxidants in the body are insufficient or when high levels of ROS are present. Therefore, disruption of the balance between oxidant and antioxidant activities is considered an important cause of oxidative damage in periodontal tissues. Parameters such as total antioxidant status (TAS), total oxidant status (TOS) and oxidative stress index (OSI) are used to determine oxidative stress. Furthermore, some enzymes such as arylesterase (ARE), heme oxygenase (HO) and nuclear factor erythroid 2-related factor 2 (NRF-2) are involved in defense mechanisms against oxidative stress. Many recent studies have shown a strong association between oxidative stress and periodontal disease.

Detailed description

This study was divided into 2 groups, 37 systemically and periodontally health individuals and 37 systemically healthy individuals with stage III grade B periodontitis were included. Clinical periodontal parameters plaque index (PI), gingival index (GI), probing pocket depth (SCD), bleeding on probing (BOP) and clinical attachment level (CAL) were recorded and saliva and serum samples were collected. ELISA method was used for analyses of TOS, TAS, OSI, ARE, HO-1, NRF-2 levels.

Interventions

DIAGNOSTIC_TESTSerum Oxidative Stress Index

Serum Oxidative Stress Index

DIAGNOSTIC_TESTSalivary Oxidative Stress Index

Salivary Oxidative Stress Index

DIAGNOSTIC_TESTSerum Arylesterase

Serum Arylesterase

DIAGNOSTIC_TESTSalivary Arylesterase

Salivary Arylesterase

DIAGNOSTIC_TESTSerum Total Oxidant Status

Serum total oxidant status

DIAGNOSTIC_TESTSalivary Total Oxidant Status

Salivary Total Oxidant Status

DIAGNOSTIC_TESTSerum Total Antioxidant Status

Serum Total Antioxidant Status

DIAGNOSTIC_TESTSalivary Total Antioxidant Status

Salivary Total Antioxidant Status

DIAGNOSTIC_TESTSerum Heme Oxygenase

Serum Heme Oxygenase

DIAGNOSTIC_TESTSalivary Heme Oxygenase

Salivary Heme Oxygenase

DIAGNOSTIC_TESTSerum Nuclear Factor Erythroid 2 Related Factor 2

Serum Nuclear Factor Erythroid 2 Related Factor 2

DIAGNOSTIC_TESTSalivary Nuclear Factor Erythroid 2 Related Factor 2

Salivary Nuclear Factor Erythroid 2 Related Factor 2

DIAGNOSTIC_TESTGingival Index

Gingival Index

DIAGNOSTIC_TESTPlaque Index

Plaque Index

DIAGNOSTIC_TESTBleeding on Probing Index

Bleeding on Probing Index

DIAGNOSTIC_TESTClinical Attachment Loss

Clinical Attachment Loss

DIAGNOSTIC_TESTPocket Deep

Pocket Deep

Sponsors

Saglik Bilimleri Universitesi
Lead SponsorOTHER

Study design

Observational model
CASE_CONTROL
Time perspective
CROSS_SECTIONAL

Eligibility

Sex/Gender
ALL
Age
18 Years to 65 Years
Healthy volunteers
Yes

Inclusion criteria

* systemically healthy * between the ages of 18-65

Exclusion criteria

* Patients with systemic diseases and/or conditions that may affect periodontal tissues (cardiovascular diseases, immune system diseases, rheumatoid arthritis, diabetes mellitus, chemotherapy/radiotherapy). * taking any medication that may have an effect on periodontal tissues, * less than 20 natural teeth excluding third molars * used antibiotics, anti-inflammatory or immunosuppressive drugs * periodontal treatment in the last 6 months * smokers * pregnant or lactating women.

Design outcomes

Primary

MeasureTime frameDescription
serum and saliva HO-1(ng/ml) levels6 monthsThe concentration of HO-1 was measured using an ELISA kit (BT Lab.) Wells were pre-coated with human HO-1-specific antibodies. Following the addition of samples, human HO-1 proteins bound to the immobilized antibodies. A biotinylated anti-HO-1 antibody and subsequently streptavidin-HRP conjugate were introduced. After removal of unbound components via washing, a substrate solution was added to initiate color development, which was proportional to the HO-1 concentration. The reaction was stopped using an acidic solution, and absorbance was recorded at 450 nm. Compared to healthy subjects, HO-1 is considered to be less expressed in periodontitis patients, potentially compromising antioxidant and anti-inflammatory defenses
serum and saliva TAS (mmol Trolox Eq/L) levels6 monthsTAS concentrations were evaluated utilizing an innovative automated colorimetric assay, originally developed by Erel. The outcomes were reported as millimoles of Trolox equivalent per liter (mmol Trolox Eq/L). Total antioxidant levels are significantly lower in periodontitis patients compared to healthy individuals, suggesting increased oxidative stress associated with periodontal inflammation.
serum and saliva TOS (µmol H₂O₂ Eq/L) levels6 monthsTOS concentrations were determined through a distinct automated colorimetric technique, also described by Erel. Results were expressed as micromoles of hydrogen peroxide equivalent per liter (µmol H₂O₂ Eq/L). Compared to healthy controls, periodontitis patients show higher total oxidant levels, which may contribute to tissue destruction and disease progression.
serum and saliva OSI levels6 monthsTAS values (initially in mmol Trolox Eq/L) were converted to µmol Trolox Eq/L. OSI was then computed using the following formula: OSI = \[(TOS, µmol/L) / (TAS, µmol Trolox Eq/L)\] × 100. The oxidative stress index is significantly higher in periodontitis patients compared to healthy individuals, reflecting an imbalance between oxidants and antioxidants.
serum and saliva ARE (U/L) levels6 monthsArylesterase activity was assessed using commercially available assay kits (Rel Assay Diagnostics, Turkey). For the measurement of arylesterase activity, phenylacetate was used as the substrate. The enzymatic activity was calculated based on the molar extinction coefficient of the phenol formed (1,310 M-¹cm-¹). One unit of arylesterase activity was defined as the amount of enzyme required to hydrolyze 1 µmol of phenol per minute under the assay conditions and was also expressed as U/L. Arylesterase activity is significantly decreased in periodontitis patients compared to healthy controls, suggesting impaired antioxidant enzyme function.
serum and saliva NRF-2 (ng/ml) levels6 monthsThe level of NRF-2 was quantified using an enzyme-linked immunosorbent assay (ELISA) kit (BT lab.). In this assay, wells were pre-coated with human NRF-2-specific antibodies. Samples containing NRF2 were added to the wells, allowing target binding. Subsequently, a biotin-conjugated anti-NRF-2 antibody was applied, followed by streptavidin-horseradish peroxidase (HRP) conjugate. After washing to remove unbound components, a substrate solution was added, producing a colorimetric reaction proportional to the amount of NRF-2 present. The reaction was terminated by adding an acidic stop solution, and absorbance was measured at 450 nm. It is suggested that the reduced expression of NRF-2 in periodontitis patients, relative to healthy controls, contributes to increased oxidative stress and tissue damage.

Secondary

MeasureTime frameDescription
Plaque Index (PI)6 monthsThe presence of plaque and the Silness and Löe plaque index (PI) were obtained from 4 surfaces of the teeth after drying the teeth and index values between 0-3 were given. Plaque accumulation, as reflected by higher plaque index scores, is more prominent in periodontitis patients than in healthy controls.
Bleeding on Probing Index (BoP)6 monthsBleeding in probing was evaluated as (+) if there was bleeding in the relevant area within 30 seconds after applying pressure to the gingival sulcus with the periodontal probe in 4 regions of all teeth with the weight of the probe itself, and as (-) if there was no bleeding. Compared to healthy controls, periodontitis patients demonstrate a higher incidence of bleeding on probing, indicating active inflammation and compromised periodontal tissue integrity.
Clinical Attachment Loss (CAL) (millimeter)6 monthsThe distance between the cementoenamel junction of the tooth and the base of the periodontal pocket is calibrated millimetrically, with the clinical attachment level on 6 surfaces of all teeth. using a Williams periodontal probe. Clinical attachment level is significantly increased in periodontitis patients compared to healthy individuals, indicating the extent of periodontal tissue destruction.
Pocket Depth (PD) (millimeter)6 monthsProbing pocket depth was performed on 6 surfaces of all teeth using a millimetrically calibrated Williams periodontal probe. The distance of the free gingival margin to the pocket bottom was measured in millimeters. The probe was held parallel to the long axis and the force applied to the probe was released when resistance was encountered during the measurement. Probing pocket depth is significantly greater in periodontitis patients compared to healthy individuals, reflecting deeper periodontal pockets due to tissue destruction.
Gingival Index (GI)6 monthsGingival Index (GI) (Löe ve Silness) : The presence of gingival inflammation was determined on 4 surfaces of all teeth using the GI score defined by Löe and Silness. The periodontal probe was moved over the tooth and index values between 0-3 were determined for each tooth. Compared to healthy individuals, a high gingival index score is frequently observed in periodontitis patients, indicating greater gingival inflammation.

Countries

Turkey (Türkiye)

Outcome results

None listed

Source: ClinicalTrials.gov · Data processed: Feb 4, 2026