|Year : 2017 | Volume
| Issue : 1 | Page : 12-15
Estimation and comparison of salivary calcium levels in healthy controls and patients with generalized gingivitis and chronic periodontitis
Madhura Vijay Rane1, Girish Suragimath2, Siddhartha Varma2, Sameer Anil Zope2, SR Ashwinirani3
1 Intern, School of Dental Sciences, KIMSDU, Karad, Maharashtra, India
2 Department of Periodontology, School of Dental Sciences, KIMSDU, Karad, Maharashtra, India
3 Department of Oral Medicine and Radiology, School of Dental Sciences, KIMSDU, Karad, Maharashtra, India
|Date of Web Publication||2-Mar-2017|
Department of Periodontology, School of Dental Sciences, KIMSDU, Karad - 415 110, Maharashtra
Source of Support: None, Conflict of Interest: None
Aim: To evaluate salivary calcium levels in healthy subjects and patients with chronic gingivitis and Chronic Periodontitis.
Materials and Methods: One fifty subjects in the age range between 20 – 45 years were randomly selected and subjected to periodontal examination using gingival index, plaque index, oral Hygiene Index and clinical attachment loss. Following periodontal examination, subjects were divided in three groups of 50 patients each: Group A: healthy subjects, Group B: gingivitis patients, Group C: periodontitis patients. Saliva samples from the study subjects was collected and subjected to estimation of salivary calcium levels. The obtained results were subjected to statistical analysis. The significance of difference in means was tested by ANOVA test.
Results: The levels of salivary calcium increased as the disease progressed from healthy to gingivitis and periodontitis. There was statistically significant difference observed between healthy to gingivitis group and gingivitis to periodontitis group.
Conclusion: Salivary calcium levels can be used as a biomarker to assess the periodontal disease progression. Early diagnosis of periodontal disease by estimation of calcium levels in saliva can help in prevention of gingivitis or periodontitis by various therapeutic measures.
Keywords: Gingivitis, periodontitis, saliva, salivary calcium
|How to cite this article:|
Rane MV, Suragimath G, Varma S, Zope SA, Ashwinirani S R. Estimation and comparison of salivary calcium levels in healthy controls and patients with generalized gingivitis and chronic periodontitis. J Oral Res Rev 2017;9:12-5
|How to cite this URL:|
Rane MV, Suragimath G, Varma S, Zope SA, Ashwinirani S R. Estimation and comparison of salivary calcium levels in healthy controls and patients with generalized gingivitis and chronic periodontitis. J Oral Res Rev [serial online] 2017 [cited 2017 Sep 25];9:12-5. Available from: http://www.jorr.org/text.asp?2017/9/1/12/201404
| Introduction|| |
Periodontitis is defined as “an inflammatory disease of the supporting tissues of the teeth caused by specific microorganisms, or group of specific microorganisms, resulting in progressive destruction of the periodontal ligament and alveolar bone with increased probing depth, recession or both.”
Periodontal disease results from a complex interplay between subgingival biofilm and the host immune inflammatory events that develop in the gingival and periodontal tissues in response to the challenge presented by the bacteria. Plaque consists of different proteins derived from saliva and gingival crevicular fluid (GCF), in which a number of microorganisms are embedded to form a highly organized matrix. Plaque accumulation is the first step toward initiation of periodontal problems. The primary cause of gingival inflammation is bacterial-therapy, self-inflicted injuries, use of tobacco and others. Calculus is one of the major predisposing factors. It consists of mineralized bacterial plaque that forms on the surface of natural teeth and dental prosthesis. The soft plaque is hardened by the precipitation of mineral salts present in the saliva.
Saliva is one of the most complex, versatile, and important body fluids supplying large range of physiological needs. Saliva is an exocrine secretion consisting of approximately 99% water and organic and inorganic molecules. It contains a variety of electrolytes such as calcium, magnesium, potassium, sodium, chloride, bicarbonate, and phosphate. Among these factors, some of them can be used as biomarkers to assess the periodontal status. Biomarkers are measurable and quantifiable biological parameters that have an important impact on clinical situations. Some studies have assessed the relationship existing between periodontal disease and these ions. Among these components, elevated levels of calcium are one of the factors seen to be associated with periodontal diseases.
Salivary calcium plays a major role in the formation of supra and subgingival calculus in the presence of unmineralized dental plaque. The dental calculus formed act as niches for further plaque accumulation which are the causative agents of periodontal diseases. As the mineral content increases in saliva, the plaque mass becomes calcified to form calculus. This is difficult to remove through routine oral hygiene measures. It is hypothesized that high salivary calcium content in saliva would result in a more rapid rate of plaque mineralization leading to periodontal diseases.
There is a need to estimate the salivary calcium levels and compare them between the healthy controls and patients with generalized gingivitis and chronic periodontitis to determine whether calcium levels in saliva can be used as an early diagnostic marker and help in the prevention of periodontal diseases. The objective of the study is to estimate, compare, and correlate salivary calcium levels in healthy controls, generalized gingivitis, and chronic periodontitis patients.
| Materials and Methods|| |
A total of 150 patients reporting to the Department of Periodontology were enrolled for the study after due approval of the ethical committee.
The subjects in the study were explained about the objectives of the study, and informed consent was obtained before commencing the study.
The study subjects were divided equally into three groups according to the following parameters:
- Group A (50): Healthy controls with absence of gingival inflammation and bleeding on probing 
- Group B (50): Generalized gingivitis patients with gingival inflammation and bleeding on probing without clinical attachment loss (CAL)
- Group C (50): Chronic periodontitis patients who had bleeding on probing with CAL.
- Subjects who had received any periodontal treatment during the past 3 months
- Subjects who had taken any antibiotic therapy during the past 3 months
- Subjects who were suffering from any known systemic diseases or conditions
- Tobacco users in smoked and smokeless form
- Pregnant and lactating women
- Subjects with less than 20 teeth.
All the subjects were interviewed, and their general information, medical history, and periodontal parameters were recorded in a specially designed proforma. The enrolled subjects were examined by a single calibrated examiner under the supervision of a senior Periodontist and categorized as a healthy, generalized gingivitis or chronic periodontitis according to consensus of World Workshop for the classification of periodontal diseases (1999). Clinical parameters assessed:
- Gingival index (Loe and Sillness 1963)
- Plaque index (Turesky–Gilmore–Glickman modification of the Quigley-Hein Plaque Index, 1970)
- Oral Hygiene Index-Simplified (John C. Greene, 1960)
After a detailed dental examination, the subjects were rescheduled for the collection of saliva samples.
Saliva samples were collected from 10:00 am, 2 h after the last meal to standardize the collections according to the circadian rhythm. Two milliliter unstimulated whole saliva samples were collected following a brief rinsing of the mouth with distilled water. The subjects were refrained from talking and asked to drop down the head. They were then asked to let the saliva pool in their floor of the mouth to their maximum extent and then expectorate into the collecting vessel till the desired quantity was collected. The samples were then transferred into sterile eppendorf tube, and 2 drops of 1% sodium azide was added which served as an antibacterial agent. The saliva samples were then transported to the laboratory immediately using standard gel coolant packs to maintain the temperature between 2°C and 4°C.
The biochemist who analyzed the salivary samples was unaware of the periodontal status of the patients. The estimation of inorganic salivary calcium was carried out using Arsenazo reagent (Accucare Calcium Arsenazo III Lab Care Diagnostics, India Pvt. Ltd.). Biochemical assay of saliva samples was carried out to quantify the salivary calcium using a calcium test kit O-Cresolphthalein complex one) and end point assay (Span Cogent Diagnostic Ltd. India Erba EM 360 fully automated auto analyzer (Erba Diagnostic, Mannheim, Germany).
All the data collected was statistically analyzed using Statistical Package for the Social Sciences software version 19 (Armonk, New York: IBM. Corporation, USA).
The results were expressed in means and percentages; P ≤ 0.05 was considered statistically significant.
The significance of difference in means was tested by ANOVA test.
| Results|| |
The 150 enrolled subjects were equally divided into three groups of fifty patients. In Group A, 19 patients were male and 31 were female, in Group B, 28 were male and 22 were female whereas in Group C, 35 were male and 15 were female patients. Therefore, out of 150 patients 82 (54.67%) patients were males and 68 (45.33%) patients were females. The above data suggest that more number of males are affected by periodontitis than the females [Table 1].
The mean salivary calcium in group A was found to be 4.84 ± 1.07 mg/dl, in Group B is 5.43 ± 0.90 mg/dl and in Group C is 5.91 ± 1.46, which showed that as the periodontal disease progressed there was an increase in the calcium levels in saliva [Table 2]. The comparison of salivary calcium in Group A, B, and C revealed that there was a statistically significant difference observed between Group A and Group B: group A and Group C. There was no statistical difference observed between Group B and Group C [Figure 1].
| Discussion|| |
Saliva and GCF not only play a decisive role in preventing periodontal disease but also ironically in the induction of periodontal pathology., Salivary calcium has more affinity toward plaque. Therefore, it is an important factor not only with regard to periodontitis but also with regard to dental health.
Calcifying plaque increases plaque retention limiting oral hygiene and hence causing gingivitis. The continuous, apically growing, calcifying plaque may be sufficient enough to cause periodontitis, despite further efforts to improve oral hygiene. The mineralized plaque acts as niche where further plaque accumulation takes place, and due to constant irritation caused by the increasing bacterial load, the healthy gingiva starts to show signs of inflammation further leading to periodontitis.
In most of the previous studies stimulated saliva was used for analysis. but in the present study, we used unstimulated whole saliva for analysis as it predominantly bathes the oral cavity most of the time, and is a representative of pooled subgingival plaque samples.
The results of the present study revealed that the subjects in the periodontitis group had significantly higher levels of salivary calcium than gingivitis and healthy group. There was a linear increase in the salivary calcium levels as the disease progressed from healthy gingiva to periodontitis. Similar results were reported by several authors (Sewon and Karjalaine et al. and Fiyaz et al.,, Rajesh et al.).
Sewon and Karjalaine et al. suggested that higher calcium concentration of plaque is associated with low caries incidence. The periodontitis-affected group seemed to have more intact teeth than the controls. It supports the view that periodontitis-affected subjects may have a higher remineralization potential than individuals with no signs of periodontitis. This is partly due to the excess of salivary calcium. However, in contrast, the results obtained by Sewon and Makela, found that higher calcium level was related to good dental health, but there was no relation to periodontal bone destruction.
In the current study population, the prevalence of gingivitis and periodontitis was more in male subjects than female subjects. These results were similar to study conducted by Shaju et al. who reported more prevalence of periodontal disease in male subjects than female subjects in the rural population.
The calcium level of saliva may reflect fluctuations in both dietary calcium and general calcium turnover but the mechanisms are far from clear. In the present study, the effect of diet and age wise calcium changes was not considered. Larger sample size could be considered to substantiate the results.
Further studies with larger sample size can be undertaken to prove that salivary calcium levels can be used as a risk biomarker for periodontal disease.
| Conclusion|| |
It can be concluded that the levels of salivary calcium increased as the disease progressed from healthy to gingivitis and periodontitis. Salivary calcium levels can be used as a biomarker to assess the periodontal disease progression. Early diagnosis of periodontal disease by estimation of calcium levels in saliva can help in the prevention of gingivitis or periodontitis by various therapeutic measures.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Nepale MB, Varma S, Suragimath G, Abbayya K, Kale V. A prospective case control study to assess and compare the role of disclosing agent in improving the patient compliance in plaque control. J Oral Res Rev 2014;6:45-8.
Edgar WM. Saliva: Its secretion, composition and functions. Br Dent J 1992;172:305-12.
Nagar P, Vishwanath D, Prabhuji MV. Pentaxins and immunity. J Oral Res Rev 2014;6:30-3.
Miller CS, King CP Jr., Langub MC, Kryscio RJ, Thomas MV. Salivary biomarkers of existing periodontal disease: A cross-sectional study. J Am Dent Assoc 2006;137:322-9.
Loe H, Silness J. Periodontal disease in pregnancy. I. Prevalence and severity. Acta Odontol Scand 1963;21:533-51.
Armitage GC. Development of a classification system for periodontal diseases and conditions. Ann Periodontol 1999;4:1-6.
Schipper R, Loof A, de Groot J, Harthoorn L, Dransfield E, van Heerde W. SELDI-TOF-MS of saliva: Methodology and pre-treatment effects. J Chromatogr B Analyt Technol Biomed Life Sci 2007;847:45-53.
Mandel ID. The diagnostic uses of saliva. J Oral Pathol Med 1990;19:119-25.
Sewón L, Söderling E, Karjalainen S. Comparative study on mineralization-related intraoral parameters in periodontitis-affected and periodontitis-free adults. Scand J Dent Res 1990;98:305-12.
Sewón L, Mäkelä M. A study of the possible correlation of high salivary calcium levels with periodontal and dental conditions in young adults. Arch Oral Biol 1990;35 Suppl:211S-2S.
Cortelli SC, Feres M, Rodrigues AA, Aquino DR, Shibli JA, Cortelli JR. Detection of Actinobacillus actinomycetemcomitans
in unstimulated saliva of patients with chronic periodontitis. J Periodontol 2005;76:204-9.
Fiyaz M, Ramesh A, Ramalingam K, Thomas B, Shetty S, Prakash P. Association of salivary calcium, phosphate, pH and flow rate on oral health: A study on 90 subjects. J Indian Soc Periodontol 2013;17:454-60.
Rajesh KS, Zareena, Hegde S, Arun Kumar MS. Assessment of salivary calcium, phosphate, magnesium, pH, and flow rate in healthy subjects, periodontitis, and dental caries. Contemp Clin Dent 2015;6:461-5.
Shaju JP, Zade RM, Das M. Prevalence of periodontitis in the Indian population: A literature review. J Indian Soc Periodontol 2011;15:29-34.
[Table 1], [Table 2]