Brief resume of the intended work: Need for the study

Brief resume of the intended work:

Need for the study: New diagnostic methods are needed that can detect disease active periodontal sites and can accurately predict those destined to become active. Levels of the enzyme, Aspartate aminotransferase have been used to identify inflammatory lesions in the heart, liver and kidney for many years. A very strong association has been demonstrated between periodontal disease active sites and increased levels of Aspartate aminotransferase.

On the other hand Alkaline phosphatase has been found in various periodontal tissues and its level can be used as an indicator for bone formation.

The purpose of this study is to estimate the levels of the enzymes Aspartate aminotransferase and Alkaline phosphatase in saliva of patients suffering from Chronic Generalised Periodontitis, Chronic Generalised Gingivitis and to compare them with healthy individuals.

Review of literature:
Studies have suggested that the use of saliva as a periodontal diagnostic marker holds promise due to the numerous biochemical indicators present in it.¹
Saliva offers as an alternative biological fluid that can be analyzed for diagnostic purposes.²
The levels of Aspartate aminotransferase in Saliva is higher when related to Periodontal pockets, gingival bleeding and suppuration was the conclusion of a clinical trial.³
Saliva can be used as a diagnostic fluid due to the various salivary enzymes present in it.⁴
Saliva offers a basis for a patient specific biomarker assessment for periodontitis and other systemic diseases.⁵
A study has concluded that passive smoke exposure levels leads to increase in the levels of Aspartate aminotransferase.⁶


Objectives of the study:
1) To determine and evaluate the relationship between the enzyme, Aspartate aminotransferase levels in saliva of patients suffering from Chronic Generalised Periodontitis, Chronic Generalised Gingivitis and to compare them with healthy individuals using the Spectrophotometric Analysis.

2) To determine and evaluate the relationship between the enzyme, Alkaline Phosphatase levels in saliva of patients suffering from Chronic Generalised Periodontitis, Chronic Generalised Gingivitis and to compare them with healthy individuals using the Spectrophotometric Analysis.

Methodology:

Source of data:
The study will be conducted in 90 patients coming to the Department of Periodontics, Yenepoya Dental College, Mangalore.
Total Number of Subjects:

90 subjects will be examined in all (45 males and 45 females). Subjects will be classified into 3 groups of 30 (15 males and 15 females) patients each; Chronic Generalised Periodontitis, Chronic Generalised Gingivitis and healthy individuals.

Informed consent will be obtained from all subjects.
Clinical examination will include:

1. 
   Gingival Index by Loe and Silness (1963)
   
2. 
   Community Periodontal Index by W.H.O (1994)
   

Inclusion Criteria

• 
  Patients who on clinical examination are classified as suffering from Chronic Generalised Periodontitis.
  
• 
  Patients who are classified as Chronic Generalised Gingivitis on clinical examination.
  
• 
  Subjects who on clinical examination show no signs of gingivitis will serve as the control group.
  

Exclusion Criteria

• 
  Patients giving a history of any systemic diseases.
  
• 
  Patients who have undergone any periodontal therapy in the last 6 months.
  
• 
  Pregnant women.
  
• 
  Patients who have taken any Antibiotics or Anti-Inflammatory agents in the last 3 months.
  

Method of collection of sample:
Immediately after a single mouth rinse with 15ml of water to wash out exfoliated cells, about 1ml of unstimulated saliva will be collected in a sterile vial. It will be transported to the Biochemical Laboratory immediately.

Method of assessing Aspartate aminotransferase and Alkaline Phosphatase
- Method used is UV Kinetic method

- Spectrophotometric Analysis

- Sample is treated with a substrate and decrease in concentration of NADPH is measured at 340nm for the assessment of Aspartate aminotransferase.

• 
  Sample is treated with a substrate and increase in concentration of NADPH is measured at 405nm for the assessment of Alkaline Phosphatase.
  

Statistical Analysis:
Statistical Analysis will be done by the “ANOVA” (Analysis of Variance) method.
List of references:

1. 
   Kaufman E, Lamster IB: Analysis of saliva for periodontal diagnosis. A review. J Clin Periodontol 2000; 27: 453-465
   
2. 
   Kaufman E, Lamster IB: The Diagnostic Applications of Saliva- A Review. Crit Rev Oral Biol Med 2002; 13;2 : 197-212
   
3. 
   Cesco R de T, Ito IY, Albuquerque Junior RF: Levels of aspartate aminotransferase (AST) in saliva of patients with different periodontal conditions. J Clin Periodontol 2003; 30: 752-755
   
4. 
   Sinem E.S & Robert E.C: Analysis of host responses and risk for disease progression. Periodontology 2000 2004; 34: 57-83
   
5. 
   Mario Taba Jr, Janet Kinney, Amy S. Kim, William V. Giannobile: Diagnostic Biomarkers for Oral and Periodontal Diseases. Dent Clin N Am 2005; 49: 551-571
   
6. 
   Nishida N et al. Association between passive smoking and salivary markers related to periodontitis. J Clin Periodontol 2006; 33: 717-723