Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT05680441 |
| Other study ID # |
CANSU35 |
| Secondary ID |
|
| Status |
Completed |
| Phase |
|
| First received |
|
| Last updated |
|
| Start date |
September 1, 2022 |
| Est. completion date |
December 16, 2022 |
Study information
| Verified date |
January 2023 |
| Source |
Dokuz Eylul University |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
The study aimed to investigate gingival crevicular fluid (GCF) levels of possible novel
biomarkers Annexin-A1 (ANX A1), Carbonic anhydrase- 1 (CA I), and Elongation Factor-1 Gamma
(EF1-Ɣ) in health along with different periodontal diseases.
In total, 80 systemically healthy individuals were included in this study; 20 with
periodontitis stage 3 grade B , 20 with periodontitis stage 3 grade C (P-Stage III/C), 19
with gingivitis, and 21 with clinically healthy periodontium. Probing depth, clinical
attachment level, plaque index, and papillary bleeding index were recorded. GCF ANX A1, CA I
and EF1-Ɣ levels were analyzed by enzyme-linked immunosorbent assay (ELISA). Receiver
operating characteristics curve was used for estimating the under the curve.
Description:
Diagnosis of periodontal diseases and conditions was arranged according to the radiographic
and clinical diagnostic criteria proposed by the 2017 World Workshop on Classifcation of
Periodontal and Peri-implant Diseases and Conditions; 20 patients with Stage III Grade B
generalized periodontitis , 20 patients with Stage III Grade C generalized periodontitis , 19
gingivitis patients and periodontally healthy volunteers were included in this study.
The clinical periodontal examination of the healthy and periodontitis subjects consisted of
plaque index (PI), probing pocket depth (PPD), clinical attachment level (CAL), and bleeding
on probing (BOP) recorded. CAL was calculated by adding GR to the PPD values. Averages for
full mouth PPD, CAL, and the percentage of sites with BOP were calculated for each subject. A
single calibrated examiner (VO) conducted a full mouth periodontal examination of all
participants. The measurements were performed using a Williams periodontal probe (Hu-Friedy,
Chicago, IL, USA). All measurements were performed full mouth and at 4 sites (mesio-buccal,
mid-buccal, distobuccal, and mid-lingual) for each tooth. Before clinical measurements,
intra-examiner calibration was performed by measuring PPD and CAL values twice on five
patients with one day interval resulting in intraclass correlation coefficients were 0.92 for
PD and 0.90 for CAL.
GCF samples were taken from the buccal aspects of two nonadjacent interproximal sites in
single-rooted teeth. In periodontitis groups, GCF was sampled from two deepest pockets of
single-rooted teeth. Samples were collected from the sites with visible signs of inflammation
in gingivitis group and without BOP in the healthy group. The selected areas were carefully
cleared of supragingival plaque using sterile curettes, isolated with cotton rolls, and
slightly air-dried to avoid contamination.
Standardized filter paper strips (PerioPaper, Proflow, Amityville, NY) were used for GCF
sampling. Sterile paper strips were gently inserted into the gingival sulcus or pocket until
mild resistance was felt and left there for 30 seconds. Mechanical irritation was avoided and
strips visually contaminated with blood were discarded. A precalibrated electronic device
measured the absorbed fluid volume (Periotron 8010, Oraflow, Amityville, NY). The readings
were converted to an actual volume (microliter, μL) by reference to the standard curve. The
paper strips were individually placed into a sterile polypropylene tube and stored at -80◦C
for further analysis.
Annexin-A1 levels were studied with commercially available kits using the ELISA method
(Bioscience SRB, catalog no: 201-12-3158). The optical density was measured
spectrophotometrically at a wavelength of 450 nm (Tecan). The assay ranges for the Annexin-A1
kit were 0.20-20 ng/mL, sensitivity 0.2 ng/mL, and the intra- and interassay coefficients of
variance (CV%) were<10%. The results were presented as ng.
Carbonic anhydrase 1 levels were studied with commercially available kits using the ELISA
method (Bioscience SRB, catalog no: SRB-T-88927). The optical density was measured
spectrophotometrically at a wavelength of 450 nm (Tecan). The assay ranges for the CA I kit
were 3.12-200ng/mL, sensitivity <1.875ng/mL, and the intra- and interassay coefficients of
variance (CV%) were<10%. The results were presented as ng.
EEF1G levels were studied with commercially available kits using the ELISA method (Bioscience
SRB, catalog no: 201-12-3732). The optical density was measured spectrophotometrically at a
wavelength of 450 nm (Tecan). The assay ranges for the EF1- Ɣ kit were 31.25-2000pg/mL,
sensitivity <12.4pg/mL, and the intra- and interassay coefficients of variance (CV%)
were<10%. The results were presented as ng.
Statistical analysis was performed using non-parametrical techniques. Comparisons between the
study groups were performed using the Kruskal-Wallis test. When there were significant
differences (p < 0.05), post-hoc 2-group comparisons were assessed with Bonferroni-corrected
Mann-Whitney U tests, and p-values < 0.05 was considered significant. All data analysis was
performed using a statistical package (Abacus Concepts Inc., Berkeley, CA, USA). The
diagnostic accuracy, sensitivity, specificity, positive predictive value (PPV), negative
predictive value (NPV), receiver operating characteristic (ROC) curve, and area under the ROC
curve (AUC) of the test dataset were assessed. P values < 0.05 were considered statistically
significant, and 95% confidence intervals (CIs) were calculated.
