Effect of Periodontal Treatment on Salivary and Serum Biomarkers in Periodontitis
Condition: Periodontal Health; Periodontitis
MeSH terms: Periodontitis
Study type: Interventional

Summary: 
      The present study aimed to determine the effect of non-surgical periodontal treatment on
      serum and salivary SIRT-1, MMP-9, and T-SOD levels in periodontitis stage III grade B
      (P-III-B) and C (P-III-C) patients. 17 periodontally healthy, 16 P-III-B and 16 P-III-C
      subjects were enrolled. At baseline, serum and saliva samples were collected and the whole
      mouth clinical periodontal parameters were recorded. Periodontitis patients received
      non-surgical periodontal treatment. Clinical parameters were re-measured and samples were
      re-collected at 3 months after treatment. Serum and salivary SIRT-1, MMP-9, and T-SOD levels
      were analyzed by ELISA. Data were analyzed using appropriate statistical tests.
    

Detailed description: 
      Periodontitis is a dysbiotic chronic inflammatory disease that compromises the integrity of
      the tooth-supporting tissues. When periodontitis occurs, reactive oxygen species, which are
      overproduced mostly by hyperactive neutrophils, can not be balanced by the antioxidant
      defense system and cause tissue damage. Cytoprotective enzymes such as superoxide dismutase
      (SOD), catalase (Cat) and the regulatory pathways that influence them, play a critical role
      in the scavenging and detoxification of ROS. Sirtuin-1 (SIRT-1) is a nicotinamide adenine
      dinucleotide (NAD)dependent histone deacetylase. SIRT1 is known to deacetylate FOXO3a, which
      induces antioxidant responses via modulation in SOD2 and CAT. Moreover, SIRT1 overexpression
      downregulates the pro-inflammatory cytokines such as interleukin (IL)-1α, IL-6, IL-8 and
      tumor necrosis factor-α (TNF-α) synthesis which are associated with the onset and progression
      of the periodontal disease. MMP-9 is a host-derived proteolytic enzyme that leads to
      periodontal tissue breakdown and is activated by oxidative stress.

      This study aims to examine the effect of non-surgical periodontal treatment (NSPT) on saliva
      and serum SIRT-1, SOD and MMP-9. It is the first controlled clinical study investigating the
      effect of NSPT on salivary SIRT-1 levels in different periodontitis groups. The sample size
      was calculated based on a previous study investigating the level of salivary MMP-9 and the
      power of the test was 95%, alfa value: 0.05. The estimated sample size was 10 individuals for
      each group. Therefore, a total of 49 systemically healthy patients; 17 periodontally healthy,
      16 P-III-B, 16 P-III-C were included in this study. Periodontal examination was performed,
      and full medical and dental histories were collected by a single examiner at baseline and 3
      months after NSPT. 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 periodontitis or periodontal health was determined according
      to the 2017 World Workshop on Classification of Periodontal and Peri-Implant Diseases and
      Conditions. Periodontally healthy individuals (n=17) in the control group had no sites with
      PD >3 mm and CAL >2 mm and also no radiographic evidence of alveolar bone loss. BOP was <10%.

      The periodontitis stage III patients had a minimum of three teeth apart from the first molars
      and incisors showing CAL ≥5 mm and PD ≥6 mm and showed no>4 teeth loss because of
      periodontitis. Radiographic bone loss extending from coronal to middle third or beyond.
      Radiographic bone loss was determined from the tooth showing the most severe bone loss as a
      percentage of root length. If the values of bone loss %/age were between 0.25 and 1.0, the
      patients were assigned to grade B (n=16). If higher than 1.0, the patients were assigned to
      grade C (n=16).

      Treatment

      Patients in the periodontitis groups underwent quadrant-wise supra and subgingival mechanical
      scaling and root planing using ultrasonic scalers and manual instruments, after
      administration of local anesthesia. No periodontal intervention was carried out in the
      periodontally healthy controls.

      Saliva and Serum Sampling

      A total of 5 mL of unstimulated whole saliva was collected by passive drool method between
      9:00 and 10:00 am. The participants were advised to avoid food consumption for three hours
      before sample collection. The participants were seated upright, and saliva was collected over
      a period of 5 minutes with instructions to pool saliva in the floor of the mouth and
      passively drool it into a sterile glass beaker. Then saliva samples are immediately
      transferred to a 2 mL polypropylene tube and stored at -80°C. A total of 5 mL of blood was
      collected from the antecubital fossa by vene puncture method. Serum was isolated from the
      blood by centrifuging at 5000 rpm for 10 minutes followed by its rapid transfer to a sterile
      polypropylene tube and storage at -80°C.

      Biomarker Immunoassays Saliva and serum samples were thawed on ice. The saliva samples were
      centrifuged at 5.000 rpm for 15 minutes at room temperature, and supernatants were
      immediately used for assays. Serum and salivary samples of SIRT-1 , MMP-9, MIP-1α*, T-SOD
      were measured by ELISA using commercial kits.

      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 distributed, Wilcoxon-signed rank
      test and the Dunn test with the Bonferroni correction were used to analyze the change between
      baseline and 3 months after treatment. For inter-group comparisons, Mann-Whitney U test for
      normally and non-normally distributed data. The Spearman's rank correlation test was used to
      detect the correlations of biochemical parameters with clinical parameters and each other's
      in diseased group before and after treatment. All tests were performed at significance level
      of P <0.05.
    

Intervention name(s): Non-surgical periodontal treatment
Intervention type(s): Procedure
Intervention description(s): Non-surgical periodontal treatment involves the removal of microbial dental plaque and calculus by mechanical debridement using manual and ultrasonic instruments. Motivating and educating patients about oral hygiene procedures is another important part of the treatment. The aim is to stop the progression of the disease and to prevent the occurrence of active clinical inflammation.
Allocation: Non-Randomized
Intervention model: Parallel Assignment
Masking: None (Open Label)
Primary purpose: Treatment
Keywords: periodontitis; periodontal treatment