Periodontitis Clinical Trial
Official title:
Levels of Netrin 1 and Its Receptor Unc5b in Gingival Crevicular Fluid, Saliva and Serum in Periodontal Disease and Health
This study aimed to investigate gingival crevicular fluid (GCF), saliva and serum Netrin 1 and Unc5b levels in periodontal health and disease. A total of 60 individuals, 20 patients with periodontitis, 20 with gingivitis and 20 periodontally healthy individuals were included. Whole-mouth and site-specific clinical periodontal parameters including probing depth, clinical attachment level, bleeding on probing, gingival index, plaque index and papillar bleeding index were recorded. GCF, salivary and serum Netrin 1 and Unc5b levels were measured by enzyme-linked immunosorbent assay. Statistical analysis was performed by using non-parametric tests.
A total of 60 individuals, comprising 20 patients with Periodontitis (P Group), 20 patients
with gingivitis (G Group), and 20 periodontally healthy controls (H Group) were involved in
the study. The patients were diagnosed according to the new classification scheme. Stage and
extent of periodontitis were also determined.
The clinical periodontal criteria for the diagnosis of the groups were as follows: the
periodontitis group had interdental clinical attachment loss (CAL) ≥5 mm at site of greatest
loss and periodontal probing depth (PD) ≥ 6 mm in one or more sites of the 2 non-adjacent
teeth in at least two quadrants of the mouth. Accordingly, Stage III generalized
periodontitis patients were included in the study. Patients were diagnosed as gingivitis with
bleeding on probing ≥ 50 %, PD ≤3 mm, and no radiographic bone loss or CAL. Periodontally
healthy individuals had no recorded history of periodontal problems, with PD ≤3 mm, no
radiographic bone loss, good oral hygiene, no gingival inflammation.
Inclusion criteria were as follows: (1) aged >18 years, (2) having at least 16 natural teeth
(excluding third molar),(3) nonsmokers with no history of smoking, (4) not having any
diagnosed medical illness or drug intake that could affect the periodontal condition. The
patients who had (1) taken antibiotics, nonsteroidal antiinflammatory or any other drugs
within the past 3 months,(2) received nonsurgical or surgical periodontal treatment, (3) a
restorative and endodontic therapy requirement, (4) a removable partial denture and/or having
orthodontic therapy were excluded from the study. Current pregnancy or lactation and having
serum C reactive protein (CRP) > 3mg/L were also the exclusion criteria.
All individuals were examined at baseline and four weeks after non-surgical periodontal
treatment including, whole mouth probing depth (PD), CAL, presence of bleeding on probing
(BOP), papillar bleeding index (PBI), gingival index (GI) and plaque index (PI) except the
third molars. PD and CAL were measured at six sites per tooth using a manual periodontal
probe.
The non-surgical periodontal treatment for periodontitis group included supra- and
subgingival scaling, root planing and oral hygiene instructions. Subgingival scaling and root
planing were performed under local anesthesia in two sessions within the 48-72 h. Gingivitis
group had supra-and subgingival scaling, polishing and oral hygiene instructions. The
treatment of gingivitis and periodontitis patients were performed by a periodontist (BM)
using hand and ultrasonic instruments. All measurements were performed by the same calibrated
examiner (SG).
Saliva sampling
Whole unstimulated saliva samples were collected. Each participant was asked first to rinse
the mouth completely with tap water for 1 minutes, wait for 10 minutes, and then expectorate
into sterile polypropylene tube for 5 minutes.
Gingival crevicular fluid (GCF) sampling
GCF samples were obtained from two nonadjacent interproximal sites in one single- rooted and
one multi-rooted teeth by standardized filter paper strips. GCF was sampled from two sites
with GI < 1, PD ≤ 3 mm and without BOP in the healthy group; two sites with GI ≥ 2, PD ≤ 3 mm
and positive BOP (visible signs of inflammation) in the gingivitis group; and two deepest
pocket (≥ 5 mm) with GI ≥ 2 and positive BOP in periodontitis group.
Serum sampling
Serum samples were taken following saliva and GCF sampling before the periodontal treatment.
Six milliliters of venous blood were obtained by a standard venipuncture method and the serum
was separated from blood by centrifugation at 1,500 g for 20 minutes.
Measurement of Netrin 1 and Unc5b levels in GCF, Saliva and Serum Samples
Netrin 1 and Unc5b levels in GCF, saliva and serum samples were measured by the enzyme-linked
immunosorbent assay (ELISA) using commercial kits according to the manufacturer's guidelines.
Statistical Analysis
All data analyses were performed using a statistical software package. Comparisons of
clinical and biochemical parameters between the study groups were performed using the
Kruskal- Wallis with Mann Whitney U test with Bonferroni correction method. The intragroup
comparisons (at baseline and first month) were performed using Wilcoxon test for paired
samples. Associations among levels of the GCF, saliva and serum biomarkers and clinical
parameters were also examined using the Spearman rank correlation analysis.
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