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Seyyedi, et al.: Salivary biomarkers and oral surgery
in biological samples could be an appropriate indicator of brushing their teeth 90 min before sample collection. All surgeries
antioxidant defence. [7] were performed by the same oral and maxillofacial surgeon with
Saliva is believed to be the first defence barrier against OS, 15 years of experience. Nerve block technique was implemented
using one carpule of 2% lidocaine with 1:100,000 epinephrine as
comprising crucial mechanisms such as uric acid, albumin, a local anaesthetic. Most surgeries lasted 30–60 min. Horizontal
ascorbic acid and glutathione. It has been established that and releasing incisions were made to have sufficient access.
[8]
saliva plays an essential role in the early stages of wound Next, the flap was carefully reflected and the alveolar bone was
healing following tooth extraction, by modulating the partially removed using a round bur in a surgical handpiece.
inflammatory mediators. Furthermore, a broad variety Following the tooth extraction, the socket was curetted in all
[9]
of hormonal, infectious, immunological and toxicological patients and irrigated with sterile saline solution and sutures were
biomarkers can be measured through saliva, consequently performed consequently. In addition to standard post‑operative
making it a diagnostic tool for numerous oral and systemic instructions, all patients were prescribed 500 mg amoxicillin for
diseases. In addition, collecting salivary samples is rather three days (in allergic cases, 500 mg erythromycin was prescribed
[10]
simple, low‑cost and non‑invasive in comparison to blood b.i.d. x4 days) and 200 mg ibuprofen t.i.d. x2 days. The patients
and biopsy samples, which proves to be highly significant. [10]
were advised to avoid consumption of any other drugs, and in
In previous studies, a higher level of OS biomarkers, including case of post‑operative complications, to only seek help from the
MDA and TAC, has been associated with periodontitis, conductors of the study.
[11]
dental caries, impacted wisdom tooth follicles as well as Saliva samples were collected on two separate occasions:
[13]
[12]
multiple oral diseases such as lichen planus and aphthous the initial samples were taken preoperatively, prior to local
[14]
stomatitis. [15] Refahee et al., [16] demonstrated a significant anaesthetic injection; and the second samples were taken
increase in serum OS biomarkers after extraction of premolars seven days after the operation, before the suture removal.
and canines of dogs. Few studies conducted on human subjects,
some with rather contrary results regarding MDA levels, have The subjects were instructed to rinse their mouths thoroughly with
reported an increase of serum OS markers after third molar deionised water and then were seated upright and asked to rest
extraction. [17,18] There is a scarce evidence on OS changes for 5 min before sample collection. To reduce possible circadian
in salivary samples. Therefore, extensive investigation is interference, all samples were collected between 10 and 12 a.m.
[19]
demanded for further confirmation of previous findings. This Unstimulated whole saliva was collected by asking the subjects to
study aimed to investigate the changes in OS biomarkers in lean forward and spit into sterile 5 mL test tubes (Falcon, Falcon
patients undergoing third molar surgery by the assessment of Co, China) every 60 s; the lids of the tubes were tightly closed
[20]
salivary MDA and TAC levels. with parafilm (Jinhua Hisure Scientific Co., Ltd, Zhejiang, China)
and labelled accordingly. The collected tubes were immediately
SubjectS and MethodS coded and sent to the biochemistry laboratory of Urmia University
This study included patients (120 males and 90 females) who of Medical Sciences to be centrifuged at 2000 rpm for 10 min to
remove debris. The supernatants were transferred to microtubes,
were referred to the Department of Oral Surgery of Urmia coded and stored at −20°C until testing.
University of Medical Sciences (in Urmia, Iran) for surgical
extraction of an impacted or partially impacted third molar The MDA measurement method was based on its reaction with
between 2014 and 2016. Subjects of the study were generally thiobarbituric acid (TBA), (Merck AG, Darmstadt, Germany),
healthy individuals aged 18 years or older. The exclusion criteria to form MDA‑TBA; absorption measurement was obtained by
included history of systemic diseases affecting periodontal spectrophotometry at 532 nm wavelength and was compared
health (e.g., diabetes mellitus, immunocompromised status, with a standard curve. To determine the TAC level in salivary
[4]
cardiovascular diseases or rheumatologic disorders), chronic samples, ferric reducing acid antioxidant power assay was
oral mucosal diseases (e.g., oral lichen planus, pemphigus performed, based on the reduction of Fe3+ to Fe2+ with
vulgaris and chronic aphthous stomatitis), history of drug 2,4,6‑Tris (2‑pyridyl)‑S‑triazine using a spectrophotometer at
allergies, smoking or alcohol use, recent history of antioxidant 539 nm, compared against the FeSO standard curve. The
[21]
4
medication consumption (e.g., vitamin A) and pregnant or salivary MDA and TAC concentrations were reported in µmol/ml.
post‑menopausal females. The present study was designed Sample size was calculated using Sample1 Software
in accordance with the guidelines issued and approved (Stoccu Inc. RaoSoft Co, Georgia, USA) with confidence
by the Ethics Committee of Urmia University of Medical interval of 95% and statistical power of 80%. For statistical
Sciences (code: IR. UMSU. REC.1395.137. Date: 2016/07/19). analysis, the results were presented as mean ± standard
All participants were informed about the objectives of the deviation (SD) for quantitative variables. The paired t‑test
study and were requested to fill in a written consent form. The or Kruskal–Wallis H‑test was used to assess the change in
study protocols were conducted according to the Declaration quantitative parameters for statistical analysis. P ≤ 0.05 was
of Helsinki for medical research.
considered statistically significant. The statistical software
Panoramic radiographs were examined preoperatively. All SPSS version 23.0 for Windows (IBM, Armonk, New York)
participants were instructed to abstain from eating, drinking and was used for the statistical analysis.

Annals of Maxillofacial Surgery ¦ Volume 14 ¦ Issue 2 ¦ July-December 2024 197