|Year : 2011 | Volume
| Issue : 2 | Page : 64-67
Estimation of gingival crevicular fluid aspartate aminotransferase levels in periodontal health and disease
Priti B Patil1, Basgauda Patil2
1 Department of Periodontology, Tatyasaheb Kore Dental College and Research Centre, New Pargaon, Dist Kolhapur, India
2 Department of Pedodontics and Preventive Dentistry, Dr. D. Y. Patil Dental College and Hospital, Pimpri, Pune, Maharashtra, India
|Date of Web Publication||21-Nov-2013|
Priti B Patil
Department of Periodontology, Tatyasaheb Kore Dental College and Research Centre, New Pargaon, 416137, Dist Kolhapur, Maharashtra
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Various enzymes have been assessed as biochemical markers and aspartate aminotransferase (AST) is one such marker that has received considerable attention recently. Analysis of gingival crevicular fluid (GCF) has been pursued as a means of identifying the sites undergoing active disease. A problem central to periodontology today is the inability to detect actively deteriorating sites and highly susceptible patients other than by longitudinal observations of attachment. Hence, AST levels from samples of GCF can be taken as an indication for active periodontal tissue destruction. Aim: To estimate the levels of AST in the GCF in periodontal health and disease. Materials and Methods: This study was an in vivo, case control, and clinico-biochemical assay. Eighty samples were selected which were divided into four groups of 20 patients each based on Russell's Periodontal Index. Statistical analysis: The values obtained for AST level in the different groups were subjected to Student's " t" test. Results: The mean of AST level showed an increase from Group I to Group IV. These values ran parallel with the values of clinical index, i.e. more severe the inflammation, higher the index score and higher was the AST level. Conclusions: It was concluded that as the severity of inflammation increases, there is a significant increase in the AST levels suggesting that there is a direct relationship between the AST levels in the GCF and periodontal destruction.
Keywords: Aspartate aminotransferase, biomarker, gingival crevicular fluid, inflammation, periodontitis
|How to cite this article:|
Patil PB, Patil B. Estimation of gingival crevicular fluid aspartate aminotransferase levels in periodontal health and disease. J Int Clin Dent Res Organ 2011;3:64-7
|How to cite this URL:|
Patil PB, Patil B. Estimation of gingival crevicular fluid aspartate aminotransferase levels in periodontal health and disease. J Int Clin Dent Res Organ [serial online] 2011 [cited 2019 Apr 18];3:64-7. Available from: http://www.jicdro.org/text.asp?2011/3/2/64/121864
| Introduction|| |
Periodontal disease is thought to progress inevitably from chronic gingivitis through periodontitis to eventually tooth loss. 
Periodontal disease activity refers specifically to the stage of the disease characterized by loss of supporting bone and connective tissue attachment. ,
Diagnostic methods used for periodontal disease are clinical, biochemical, microbiological, immunological, and genetic. ,,
Amongst all these, the biochemical markers have widely gained acceptance and importance.
The markers receiving the most attention in active periodontal lesions are aspartate aminotransferase (AST), collagenase, prostaglandin E2, Beta-glucoronidase, lactate dehydrogenase, arylsulfatase, and elastase. These enzymes are released from dead/dying cells, neutrophils, epithelial cells, and connective tissue at the affected sites. , AST, previously termed serum glutamic oxaloacetate transferase (SGOT), is one such marker of tissue destruction, suggested by its successful use as a diagnostic adjunct in human cardiac and hepatic tissue necrosis. ,,
Following tissue damage, AST is released from injured and dead cells into extracellular fluid and can be readily assayed in serum, tears, and gingival crevicular fluid (GCF). ,
Hence, in the present study, an attempt has been made to estimate the levels of AST in GCF in periodontal health and disease.
| Materials and Methods|| |
This study was an in vivo study conducted in accordance with the ethical standards on human experimentation and was approved by the ethical committee of S.D.M. College of Dental Sciences, Dharwad.
In this case control study, 80 subjects were recruited randomly. The subjects in this study were divided into four groups of 20 each on the basis of Russell's Periodontal Index. The inclusion criteria for the study were as follows:
- Patients of both sexes
- Age ranging from 20 to 45 years
- No history of any periodontal treatment in the past 6 months
- No history of antibiotic intake or oral antiseptic therapy for at least 4-6 weeks
The exclusion criteria for the study were as follows:
- Patients with history of cardiac/liver diseases or muscular dystrophy, as in such patients enzyme activity could be altered
- Pregnancy, immunocompromised patients, malignancy, and acute gingival diseases
- Any other systemic disorders that could alter the course of disease
All the subjects were informed about the study and a written consent was taken.
The subjects were then classified into four groups depending on Periodontal Index (PI) Score.
Group I represented the control group. It consisted of 20 patients (15 males and 5 females) without any signs of gingival inflammation with the PI Score ranging from zero to 0.2. Group II consisted of 20 patients (14 males and 6 females) with the PI Score ranging from 0.3 to 0.9. Group III consisted of 20 patients (9 males and 11 females) with the PI Score ranging from 1.0 to 2.0. Twenty patients (11 males and 9 females) formed Group IV, with the PI Score ranging from 2.1 to 5.0 and above.
The procedure was simple and direct. The patients were asked to gargle vigorously with a glass of sterile water to cleanse the tooth of loosely adherent debris. The test site was dried and isolated with cotton rolls. Volumetric micropipettes were placed extracrevicularly and a standardized volume of 1 μl was collected with the help of a plunger and measured within the micropipette. Multiple sites were selected to collect GCF.
The test sites that did not express any volume of fluid and the micropipettes contaminated with blood and saliva were not included in the study.
The GCF collected was immediately transferred to a test tube containing 0.1 ml of phosphate-buffer saline (pH 7, 0.05 M). The test tubes were immediately taken to the Department of Biochemistry for analysis. About three to four samples were collected per day, and all the samples were collected in the morning.
The Stangen SGOT (AST) kit is intended for the in vitro quantitative determination of SGOT (AST) activity. This kit is based on Reitman and Frankel's method.
The assay was carried out as per the manufacturer's instructions. For the enzyme assay, clean and dry test tubes were taken and were labeled as Blank (B) and Test (T) as shown below.
Buffered substrate (1) 0.5 ml 0.5 ml
Incubate at 37°C for 3 min
Phosphate-buffered saline containing GCF ------ 0.1 ml
Mix well and incubate at 37°C for 60 min
Dinitro phenyl hydrazine (DNPH) color reagent (2)
Mix well and allow to stand for 20 min
Distilled water 0.1 ml ------
Working sodium hydroxide 5.0 ml 5.0 ml
(obtained by diluting sodium hydroxide with distilled water in the ratio of 1:10)
Mix well, allow to stand at room temperature for 10 min, and measure the absorbance of Test (T) against Blank (B) on a spectrophotometer at 505 nm. This gives the optical density of the Test (T) at 505 nm.
To get the enzyme activity, a calibration curve was plotted by using pyruvate standard, and the absorbance (optical density) of Test (T) on the y-axis was read against the corresponding enzyme activity on the x-axis.
The entire procedure from the placement of micropipettes till the determination of enzyme activity took about 3 h.
The various findings in relation to the selected individuals were recorded using a proforma designed for this purpose.
The mean values obtained for AST level in the different groups were subjected to statistical analysis by applying Student's "t" test.
| Results|| |
It consisted of 20 patients, of whom 7 did not show any AST level in the GCF collected whereas the remaining patients showed AST level in the range of 2.50-5.00 IU/ml. The mean was 2.313 and standard deviation was 1.998.
It consisted of 20 patients. Only one patient did not show any AST level in the GCF collected, whereas the remaining patients showed AST levels in the range of 2.50-10.00 IU/ml. The mean and standard deviation were 5.375 and 2.871, respectively.
It consisted of 20 patients. All the 20 patients showed raised AST levels in the GCF collected, in the range of 5.00-20.00 IU/ml. The mean and standard deviation were 8.662 and 4.648, respectively.
It consisted of 20 patients. All the patients showed AST levels in the range of 5.00-37.50 IU/ml in the GCF collected. The mean was 10.00 and standard deviation was 7.906.
In the computed results [Table 1], when Group I was compared to groups II, III, and IV, it was found to be highly significant at <0.001 probability (P < 0.001). The probabilities were 0.0004, 0.000, and 0.00001, respectively.
When Group II was compared to groups III and IV, it was found to be significant at <0.05 probability (P < 0.05). The probabilities were 0.0105 and 0.0186, respectively.
When Group III was compared to Group IV, it was found to be nonsignificant (P > 0.05), the probability being 0.5182.
The mean of AST level showed an increase from Group I to Group IV. These values ran parallel with the values of clinical index, i.e. more severe the inflammation, higher the index score and higher was the AST level [Figure 1].
| Discussion|| |
Periodontitis in humans is an inflammatory disease, and microorganisms or their products and components are the driving force behind the observed tissue destruction. 
Evidence from a variety of sources suggests that human periodontal diseases may occur by a series of repeated episodic exacerbations. Periodontal disease is seen to proceed in bursts of activity that occur randomly at periodontal sites throughout the mouth. Following these destructive bursts, periodontal sites appear to remain quiescent for an undetermined period of time.  Some of the procedures traditionally used cannot distinguish between disease activity and inactivity at a given point of time.
Thus, a problem central to periodontology today is the inability to detect actively deteriorating sites and highly susceptible patients other than by longitudinal observations of attachment. There is a major need for objective tests that accurately detect deteriorating periodontal sites and distinguish them from healthy sites. A large amount of effort is being committed to solve this problem. 
The various diagnostic aids now elicited in the literature and currently in use are clinical, biochemical, microbiological, immunological, and genetic methods. ,, As periodontal diseases are characterized by destruction of the tooth-supporting tissues, quantitation of tissue breakdown products have been assessed by biochemical analysis and GCF has been pursued as a means of identifying sites undergoing active disease. 
Various enzymes have been assessed as biochemical markers and AST is one such marker that has received considerable attention recently.
In the present study, AST levels in GCF were compared using clinical index (Russell's Periodontal Index).  The results clearly showed that as the severity of clinical inflammation increases, AST level also increases.
In Group I, 7 out of 20 cases did not show any AST level in the GCF collected indicating that the periodontal tissues were in a healthy state and the enzyme AST could not be detected. The remaining patients did show a very slight amount of AST level. This could be due to the presence of subclinical inflammation. Since gingival tissues are directly exposed to external environment; hence, the presence of an inflammatory reaction is obvious. Even in clinically normal gingiva, small foci of inflammatory cells are found in the connective tissue near the base of the sulcus. Their presence is believed to be related to the penetration of antigenic substances via the sulcular epithelium. 
In Group II, all the patients showed a slight increase in the AST level suggesting that there was some amount of destruction going on, and hence, the enzyme could be detected in all but one individual. This is in agreement with the study reports of Chambers et al.  and Persson et al. 
Group III patients showed a significant increase in the AST level, which was due to the presence of disease activity. This increase is probably due to the presence of periodontopathic microorganisms that cause destruction of the periodontal tissues by activation of host cells and systems to produce and release enzymes and other molecules that destroy the periodontal tissues, and these activities are guided and regulated by the mediators of inflammation. 
In Group IV, i.e. severe inflammation with deep pockets, few patients showed very high levels of the AST enzyme, some showed a significant increase, while a few showed a slight increase. Patients who showed high and significant AST levels might be in an active state of the disease, whereas the others may be in a state of quiescence due to the chronicity of the lesion.
The results of this study show that as the severity of inflammation increases, the AST level in GCF also increases. So, from this study it can be deduced that AST is a potential biochemical marker of tissue destruction and can be used to measure active tissue destruction and thereby disease activity.
| Conclusion|| |
The study sample consisted of 80 patients who were divided into four groups of 20 patients each as per Russell's Periodontal Index. GCF was collected for biochemical analysis to estimate the AST levels.
From the results obtained, the following conclusions were arrived at:
- As the severity of inflammation increases, there is a significant increase in the AST levels suggesting that there is a direct relationship between the AST levels in the GCF and periodontal destruction.
- GCF in the destructive phase of periodontal disease possesses a greater amount of AST.
- AST may be used as a potential biochemical marker for the assessment of gingival inflammation and periodontal disease activity.
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