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© Borgis - New Medicine 4/2019, s. 145-151 | DOI: 10.25121/NewMed.2019.23.4.145
Paula Piekoszewska-Ziętek1, Emilia Raćkowska2, Natalia Korytowska2, *Dorota Olczak-Kowalczyk1
Salivary antioxidant status and oral health in children and adolescents
Status antyoksydacyjny a stan zdrowia jamy ustnej u dzieci i młodzieży
1Department of Paediatric Dentistry, Medical University of Warsaw, Poland
Head of Department: Professor Dorota Olczak-Kowalczyk, MD, PhD
2Department of Bioanalysis and Drug Analysis, Medical University of Warsaw, Poland
Head of Department: Professor Piotr Wroczyński, MD, PhD
Streszczenie
Wstęp. Patogeneza chorób jamy ustnej może być związana ze stresem oksydacyjnym. Układ antyoksydacyjny śliny jest jednym z kluczowych mechanizmów obronnych śliny skierowanych przeciwko patogenom, a także czynnikiem ochronnym jamy ustnej.
Cel pracy. Analiza związku między stanem zdrowia jamy ustnej (poziomem higieny, stanem dziąseł i zębów), wiekiem i płcią a parametrami potencjału antyoksydacyjnego u dzieci i młodzieży z uzębieniem stałym.
Materiał i metody. Badaniem objęto 87 pacjentów. Dokonano oceny wskaźników PUWZ/PUWP i białych plam próchnicowych (ang. white spot lesions ? WSL), poziomu higieny jamy ustnej oraz stanu dziąseł. Od wszystkich pacjentów pobrano próbki śliny. Oznaczono wydzielanie śliny spoczynkowej (niestymulowanej) oraz dokonano badania próbek śliny pod kątem całkowitego potencjału antyoksydacyjnego (ang. total antioxidant capacity ? TAC) oraz aktywności antyoksydacyjnej FRAP.
Wyniki. U pacjentów z próchnicą, aktywnym procesem próchniczym, białymi plamami próchniczymi, niskim poziomem higieny jamy ustnej i stanem zapalnym dziąseł odnotowano niższe parametry potencjału antyoksydacyjnego, jednak różnice te nie były statystycznie istotne. U pacjentów z niskimi wartościami wydzielania śliny spoczynkowej stwierdzono istotnie wyższe parametry stresu oksydacyjnego. Analiza korelacji rang Spearmana nie wykazała związku między wartościami TAC i FRAP a płcią pacjenta, ale wykazano dodatnią korelację między wartościami TAC/FRAP a wiekiem pacjentów.
Wnioski. Parametry potencjału antyoksydacyjnego śliny różnią się w zależności od stanu zdrowia jamy ustnej. Potencjał antyoksydacyjny śliny jest skorelowany z wiekiem pacjenta.
Summary
Introduction. The pathogenesis of oral diseases may be associated with oxidative stress. Salivary antioxidant system constitutes one of the key salivary defence mechanisms against pathogens and a protective factor for oral cavity.
Aim. To investigate the relationship between oral health (hygiene level, gingival and dental health), age and gender and antioxidant capacity parameters in children and adolescents with permanent dentition.
Material and methods. A total of 87 patients were examined. DMFT/DMFS and white spot lesions (WSL), oral hygiene level and gingival health were assessed. Salivary samples were collected from all participants. Unstimulated salivary flow was calculated and salivary samples were assayed for total antioxidant capacity (TAC) and ferric reducing antioxidant power (FRAP).
Results. Antioxidant capacity parameters were lower in patients with caries, active caries, white spot lesions, poor oral hygiene and gingivitis, but the differences were not statistically significant. Oxidative stress parameters were significantly higher in low unstimulated salivary flow. Spearman’s rank correlation analysis revealed no relationship between TAC or FRAP values and patients’ gender, but there was a positive correlation between TAC/FRAP and patients’ age.
Conclusions. Salivary antioxidant capacity parameters differ in certain oral conditions. There is a correlation between salivary antioxidant capacity parameters and patients’ age.
Introduction
Oxidative stress is defined as an imbalance between excessively reactive oxygen species (ROS) and antioxidant mechanisms (enzymatic and non-enzymatic). ROS, such as the superoxide radical, hydrogen peroxide and hydroxyl radical, are products of cellular metabolism (1). This imbalance may be a result of infection, inflammation or disease (2). Several studies indicate that the pathogenesis of various systemic inflammatory diseases, including oral ones, can be associated with oxidative stress. Many of these include periodontitis and mucosal lesions, but also dental caries (3-6). Salivary antioxidant system is one of the key salivary defence mechanisms against pathogens and a protective factor for oral mucosa (4). There are multiple different antioxidant compounds in human saliva. The salivary antioxidant system is made of enzymes (for example: peroxidase, super oxide dismutase, glutathione peroxidase, catalase), as well as small molecules such as uric acid or ascorbic acid (7, 8). According to Battino el al., these elements have additive effects rather than act alone. For this reason, measuring individual antioxidants separately can be confusing and lead to misinterpretation. Studies should focus on the whole antioxidant status, which can be expressed, for example, as total antioxidant capacity (TAC) or ferric reducing antioxidant power (FRAP) (9-11).
Aim
The aim of the study was to investigate the relationship between oral health (hygiene level, gingival and dental health), age and gender and antioxidant capacity para-meters (TAC, FRAP) in children and adolescents with permanent dentition.
Material and methods
The study conformed to the STROBE guidelines (STrengthening the Reporting of OBservational studies in Epidemiology).
Study population
The study was conducted in 87 patients aged 11-16 years with permanent dentition. The participants were recruited from the Department of Paediatric Dentistry, Medical University of Warsaw. The research was conducted in full accordance with the World Medical Association Declaration of Helsinki. The Bioethical Commission of Warsaw Medical University (No. KB/194/2015) approved the study and informed written consent was obtained from all parents. The inclusion criteria were as follows: no systemic diseases, no chronic pharmacotherapy, no developmental defects of tooth hard tissues, no previous orthodontic treatment. Patients who did not meet the above mentioned criteria were excluded from the study.
Clinical examination
Clinical examinations were conducted in a dental office by one examiner using standardised WHO (World Health Organization) criteria. Caries index, expressed as mean DMFT/DMFS and white spot lesion (WSL) score, was evaluated (12). Oral hygiene level was assessed using ODI-S (the Debris Index), part of Oral Hygiene Index by Greene and Vermillion (13) and Approximal Plaque Index (API%) index by Lange et al. (14). Gingival health was assessed based on the Gingival Index (GI) by SILNESS and Löe (15), on four surfaces of permanent teeth 16, 12, 24, 36, 32, 44.
Sample collection for salivary analysis
Saliva samples were collected from all participants in the morning (8.00-10.00 am). The participants were fasted. Salivette collection tube (Sarstedt, Nümbrecht, Germany) was used. The swab was placed in the mouth and allowed to absorb resting saliva for 5 min. Immediately after collection, the tube was stored at +4°C and centrifuged within 2 hours. The centrifugation to recover saliva from the swab was performed at 1000 x g at +4°C for 15 min. Unstimulated salivary flow rate was calculated by dividing the patient’s salivary volume by collection time. The saliva samples were frozen and stored at -80°C until the analysis.
Total antioxidant capacity of saliva
TAC was assessed using a modified method developed by Erel (10). The assay was based on the principle of reduction of 2,2’-azino-bis (3-ethylbenz-thiazoline-6-sulfonic acid) cation radical (ABTS •+) (whose solution is blue-green) to ABTS (colorless solution) by antioxidants present in the sample. The reaction changed the colour of the reaction mixture. The green-blue colour disappeared. The reaction was measured spectrophotometrically at 660 nm. The change in absorbance was linked with the level of antioxidants in the sample. Calibration of the results was performed using Trolox. The results of TAC were expressed in mmol Trolox Equiv./L.
Ferric reducing antioxidant power
FRAP was determined using a method developed by Benzie and Strain (11). The method is based on the ability of antioxidants to reduce the complex of ferric tripyridyltriazine (Fe3+-TPTZ) to Fe2+-TPTZ at low pH (3.6). As a result of reaction, the yellow colour of Fe3+-TPTZ changed during the reduction to Fe2+-TPTZ, developing an intense blue colour with a maximum absorbance at 593 nm. The reaction was monitored spectrophotometrically. The change in absorbance was directly proportional to the level of antioxidants with a reduction potential in the sample. FRAP value of a sample was determined using calibration curve method. Calibration was performed using aqueous solutions of known Fe2+ (iron (II) sulphate heptahydrate (FeSO4 x 7H2O)) concentration. The results are expressed in mmol/L.
Statistical analysis
The analyses were performed using Statistica 12.0 software and R package. For quantitative variables, descriptive statistics characterising their variability were calculated. For binomial variables, the proportions (%) with division into groups were calculated. Comparison of quantitative variables between two groups was performed using the Mann-Whitney U test and the chi-square test, while the comparison of three or more groups was performed using the ANOVA test. In the case of categorical variables (including binomial variables), chi-square test was used to compare the groups. Spearman’s rank correlation coefficient was calculated. In all analyses, the statistical significance level was set at 0.05. Missing data were excluded from the analysis.
Results
A total of 87 patients, including 46 (52.9%) boys and 41 (47.1%) girls, were examined. The mean age ± SD was 13 ± 2 years. Caries (DMFT > 0) was present in 81 patients (93.1%), active caries (DT > 0) in 47 patients (54.3%), and white spot lesions (WSL > 0) in 41 patients (47.1%). The results for the whole group and those depending on sex are presented in table 1.
Tab. 1. Caries prevalence in the study group
ParametersTotalBoysGirlsp
Incidence [%]
DMFTa > 0 93.293.692.70.868
DTb > 0 54.357.451.20.562
WSLc > 0 47.141.353.60.251
Mean ± SD
DMFTa 6.30 ± 4.986.30 ± 5.096.37 ± 4.920.948
DTb 2.09 ± 2.812.30 ± 3.021.85 ± 2.560.458
MTd 0.06 ± 0.280.02 ± 0.150.10 ± 0.370.181
FTe 4.18 ± 3.783.98 ± 3.784.41 ± 3.810.599
DMFSf 8.93 ± 8.829.02 ± 9.738.83 ± 7.780.921
WSLc 3.47 ± 5.603.96 ± 6.302.93 ± 4.710.395
aDecayed, Missing, Filled Teeth
bDecayed Teeth
cWhite Spot Lesions
dMissing Teeth
eFilled Teeth
fDecayed, Missing, Filled Surfaces

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otrzymano: 2019-11-25
zaakceptowano do druku: 2019-12-16

Adres do korespondencji:
*Dorota Olczak-Kowalczyk
Zakład Stomatologii Dziecięcej Warszawski Uniwersytet Medyczny
ul. Binieckiego 6, 02-097 Warszawa
tel.: +48 (22) 116-64-24
dorota.olczak-kowalczyk@wum.edu.pl

New Medicine 4/2019
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