Periodontal Health Clinical Trial
Official title:
Effect of Non-surgical Periodontal Treatment on GCF HIF-1α, VEGF and TNF-α Levels in Generalized Aggressive Periodontitis Patients.
Hypoxia-inducible angiogenic pathway involving hypoxia inducible factor-1 alpha (HIF-1α), vascular endothelial growth factor (VEGF) and tumour necrosis factor- alpha (TNF-α) may regulate the several biological processes related to inflammation. Generalized aggressive periodontitis (G-AgP) is a rare but highly destructive form of inflammatory periodontal disease. The present study aimed to assess the effect of non-surgical periodontal treatment on gingival crevicular fluid (GCF) HIF-1α, VEGF and TNF-α levels in G-AgP patients. 20 G-AgP and 20 periodontally healthy subjects were enrolled. At baseline, GCF samples were collected and whole mouth clinical periodontal parameters were recorded. G-AgP patients received non-surgical periodontal treatment. Clinical parameters and GCF cytokines were re-measured at 1 and 3 months after treatment. GCF HIF-1α, VEGF and TNF-α levels were analyzed by ELISA. Data were analyzed using appropriate statistical tests.
Study Population and Clinical Examination
A total of 40 individuals were recruited for the present study. Inclusion criteria included:
1) aged 25 to 45 years 2) non-smokers with no history of smoking 3) having at least 20
natural teeth. Exclusion criteria were as follows: 1) having any diagnosed medical disorders
such as diabetes mellitus, cardiovascular diseases, rheumatoid arthritis, immunological and
mucocutaneous diseases 2) usage of antibiotics, non-steroidal anti-inflammatory drugs and
immunosuppressive agents within the past 6 months 3) having any non-inflammatory destructive
periodontal disease 4) nonsurgical/surgical periodontal therapy received in the past year 5)
having a restorative and endodontic treatment requirement 6) having orthodontic appliances or
a removable partial denture 7) pregnant/ lactating/ postmenopausal females.
The whole mouth clinical periodontal examination included measurement of probing depth (PPD),
clinical attachment level (CAL), presence of bleeding on probing (BOP), gingival index (GI)
and plaque index (PI) at 6 sites per tooth, except the third molars. The presence and type of
the alveolar bone loss were assessed on the digital panoramic radiograph in each participant,
which was supplemented with periapical radiographs if necessary.
Periodontal status of each patient was evaluated by a single calibrated periodontists with a
manual probe. The diagnosis of G-AgP or periodontally health was determined according to the
International Classification of Periodontal Diseases in 1999. The patients in G-AgP group
(n=20) had a minimum three teeth apart from the first molars and incisors showing CAL ≥5 mm
and PPD ≥6 mm. Radiographic bone loss was ≥30 % of root length affecting ≥3 teeth other than
first molars and incisors. This generalized pattern of severe destruction was inconsistent
with the amount of microbial deposits. Patients had at least one other family member
presenting with or having a history of severe periodontal problems. Periodontally healthy
individuals (n=20) in the control group had no sites with PPD >3 mm and CAL >2 mm and also no
radiographic evidence of alveolar bone loss. BOP was <15% in the whole mouth.
Treatment
The recruited G-AgP patients received conventional quadrant scaling and root planning (SRP)
under local anaesthesia for 3 weeks. SRP was performed by the same periodontist (who was
different the investigator recorded periodontal parameters) using ultrasonic inserts and
manual periodontal curettes. Re-evaluations were performed at 1 and 3 months following the
completion of the SRP in the lower right quadrant. No periodontal intervention was carried
out in the periodontally healthy controls.
GCF sampling
GCF was sampled from two deepest pockets of single-rooted teeth at baseline in G-AgP group
immediately before the treatment of upper right quadrant. Sampling was repeated at the same
sites following the completion of the SRP in the lower right quadrant. GCF was sampled from
the buccal aspects of two nonadjacent interproximal sites in single-rooted teeth with.
Samples were collected from the sites without BOP in the periodontally healthy group.
Standardized filter paper strips 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 s. The absorbed fluid volume was measured with a precalibrated electronic device. The
paper strips were stored at −40◦C for further analysis.
Measurement of HIF-1α, VEGF and TNF-α Levels in GCF
Two paper strips were pooled, placed in 300 µL PBS-T (0.05%). HIF-1α, VEGF and TNF-α levels
in GCF samples were measured by the enzyme-linked immunosorbent assay using commercial kits
in line with the manufacturer's guidelines. GCF results were expressed as both total amounts
at two sites per sampling time.
Statistical Analysis
All statistical analyses were carried out with the standard statistical software package. For
the intra-group comparisons, if the data were not normally disturbed, Friedman test and the
Dunn test with the Bonferroni correction were used to analyze the change between baseline and
1 month and 3 months after treatment. For inter-group comparisons, Mann-Whitney U test for
normally and non-normally disturbed data. The Spearman's rank correlation test was used to
detect the correlations of biochemical parameters with clinical parameters and each others in
diseased group before and after treatment. All tests were performed at significance level of
P <0.05.
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