|Year : 2017 | Volume
| Issue : 2 | Page : 96-100
An attempt to correlate biochemical parameters in saliva with dental carries in children of two different age groups: A comparative study
Santosh Mahajan1, Bharat Suneja2, Paramjot Kaur3
1 Department of Biochemistry, Baba Jaswant Singh Dental College Hospital and Research Institute, Ludhiana, Punjab, India
2 Department of Pedodontics, Baba Jaswant Singh Dental College Hospital and Research Institute, Ludhiana, Punjab, India
3 Department of Oral and Maxillofacial Surgery, Baba Jaswant Singh Dental College Hospital and Research Institute, Ludhiana, Punjab, India
|Date of Web Publication||8-Jan-2018|
Dr. Santosh Mahajan
Department of Biochemistry, Baba Jaswant Singh Dental College Hospital and Research Institute, Sector 40, Chandigarh Road, Ludhiana, Punjab
Source of Support: None, Conflict of Interest: None
Aim: Dental caries is one of the most common chronic dental diseases of childhood. Saliva by virtue of its chemical composition provides the main host defence in the oral cavity and plays an essential role in maintaining the integrity of oral structure. The present study was an attempt to correlate the changes in biochemical parameters with dental caries in the saliva samples of children of two different age groups.
Materials and Methods: The level of calcium, phosphorous, α-amylase, and pH was measured in the saliva of caries-free (control group with decayed, missing, and filled teeth [DMFT] = 0) and caries-affected (test group with DMFT ≥5) children of 3–8 years and 9–14 years of age.
Results: Statistical analysis of the data thus obtained revealed no significant difference in the mean values of calcium, phosphorus, α-amylase, and pH in caries-affected children when compared to their age-matched control group except phosphorous which increased significantly in caries-affected children of 3–8 years of age. There was also a significant correlation between calcium and phosphorous levels (r = 0.192*, P = 0.04) irrespective of age group.
Conclusions: This study indicated a direct relationship between phosphorous level and dental caries in children of younger age group (3–8 years of age) and was attributed to their greater susceptibility to dental caries leading to their more DMFT status than children of 9–14 years of age. Greater susceptibility of primary enamel to demineralization is well documented in the literature, and it is because of its less mineralized, significantly softer and less elastic nature and higher organic content that dissolves faster in the acidic environment than the permanent enamel. Increased phosphorus in the saliva sample of caries-affected children might be due to the hydrolysis of organic phosphates of their enamel. Significant correlation between calcium and phosphorous supports the involvement of these two minerals in the formation of hydroxyapatite of tooth.
Keywords: Calcium, caries, children, decayed, missing and filled teeth, pH, phosphorous, saliva, α-amylase
|How to cite this article:|
Mahajan S, Suneja B, Kaur P. An attempt to correlate biochemical parameters in saliva with dental carries in children of two different age groups: A comparative study. Int J Oral Health Sci 2017;7:96-100
|How to cite this URL:|
Mahajan S, Suneja B, Kaur P. An attempt to correlate biochemical parameters in saliva with dental carries in children of two different age groups: A comparative study. Int J Oral Health Sci [serial online] 2017 [cited 2019 Oct 21];7:96-100. Available from: http://www.ijohsjournal.org/text.asp?2017/7/2/96/222403
| Introduction|| |
Dental caries (a multifactorial disease of the calcified tissues), though prevalent in both the sexes of every age group and all socioeconomic strata, is one of the most common dental disease of childhood. Despite the general decrease in dental caries in the past decade childhood caries remains a major problem in developing countries and in a few industrialized nations., As the children reach school age, they have an increasing incidence of carious lesions due to the change in dietary habits, in addition to oral hygiene, tooth structure, and composition of saliva, etc. The balance between protective and pathological factors determines the progression or reversal of dental caries. The disease starts with the fermentation of carbohydrates producing organic acids that decrease the pH of the oral cavity resulting in demineralization  and creating an environment favorable for the growth of Streptococcus mutans. Saliva provides the main host defence against these cariogenic challenges. By virtue of its chemical composition, it is capable of rehardening the buffer softened enamel surface, resulting in remineralization but at a slower rate than demineralization. Since the reparative capacity of saliva is related to its mineral constituents bathing the teeth, a considerable research attention has been directed to evaluate salivary calcium and phosphorous level in relation to dental caries. However, no conclusive evidence has been presented to support that the concentration of either is of any importance in caries prevention. Amylase, besides its function in digestion, is of considerable significance in dental health because of its intraoral action., However, its overall effect on caries is still questionable. In view of this, the present study was an attempt to estimate the level of calcium, phosphorous, α-amylase, and pH in the saliva samples of caries-free and caries-affected children of two different age groups and elucidate if any correlation exists between these parameters and dental caries.
| Materials and Methods|| |
The study sample was comprised of a total of 120 children from 3 to 8 years and 9–14 years of age attending the Pedodontics Outpatient Department in Baba Jaswant Singh Dental College and Research Institute, Ludhiana. This study was approved by the ethical committee of institute. Informed consent was obtained from the parents of children included in this study after explaining them the study protocol.
Screening of the children for their decayed, missing, and filled status of both primary and permanent teeth (decayed, missing, and filled teeth [DMFT]) was done using dental mirror and explorer following Cappeli and Mobley. Children were divided into four groups of 30 children in each group. Group I and group III comprised caries-free children with DMFT score 0 (control group) of 3–8 and 9–14 years of age, whereas group II and IV comprised children having DMFT score ≥5 (test group) from the respective age group.
To control the circadian rhythm, the saliva samples of the children were collected between 10:00 am and 11:30 am. The children were asked not to eat or drink 1 h before the collection of the sample. Stimulated saliva samples were collected in sterilized vials by churning 10 ml distilled water in the mouth for 2–3 min and were analyzed biochemically.
Each sample was analyzed for calcium by titrimetric method of Clark and Collip. Level of phosphorous and α-amylase was measured following the colorimetric methods of Kuttner and Lichenstein, and Bansal et al., respectively. The pH of all the samples was noted by pH meter. The data obtained were analyzed statistically.
Mean ± Standard deviation (SD) value for different parameters in the two age groups and the comparison of their levels in control with the test was done using Student's t-test. The correlations between different parameters were analyzed following nonparametric Spearman's correlation. The level of significance was established at P < 0.05.
| Results|| |
A total of 120 children (71 boys and 49 girls) were included in this study. Range and mean ± SD values of age, DMFT, calcium, phosphorous, α-amylase, and pH in different study groups have been shown in [Table 1]. Comparison of the mean values of Calcium, phosphorous, α-amylase, and pH of control group with age-matched test group is given in [Table 2]. Difference in the mean ± values of calcium, phosphorous, amylase, and pH between caries-affected children was not significant statistically when compared to their age-matched control group except phosphorous which increased significantly (P = 0.010) in caries-affected children of 3–8 years of age [Figure 1], [Figure 2], [Figure 3], [Figure 4]. Spearman's correlation revealed no significant correlation between different parameter studied except between calcium and phosphorous levels (r = 0.192, P = 0.04) [Table 3].
|Table 1: Range and mean±S.D values of age, decayed, missing, and filled teeth, calcium, phosphorous, α-amylase, and pH in different study groups (n=30 in each age group)|
Click here to view
|Table 2: Comparison of mean±standard error values of calcium, phosphorous, amylase, and pH between control and respective test group|
Click here to view
|Figure 1: Intergroup comparison of mean values of calcium, phosphorous, amylase, and pH between control and respective test group|
Click here to view
|Table 3: Correlations among decayed, missing, and filled teeth, salivary calcium, phosphorous, amylase, and pH irrespective of age group|
Click here to view
| Discussion|| |
The composition of saliva in relation to dental caries has been studied extensively showing positive, negative, or no correlation.,,,, Protective role of saliva against dental caries is a complex phenomenon involving its composition, tooth structure, hormonal status, external factors such as microbial flora oral hygiene. In the present study, no significant difference is found in the level of calcium, phosphorous, amylase, and pH between caries-free and caries-affected children except phosphorous which increased significantly in caries affected children of 3–8 years when its level was compared with their age-matched control group. The results are “partially” in accordance with Shahrabi et al. who could not establish any relationship between dental caries and the level of calcium, phosphorous, and alkaline phosphatase in children of 3–5 years age group. The results are also in line with Hubbel and Bunting  who did not find any difference in the concentration of calcium and phosphorous in caries-free and caries-affected children of 7–16 years. Bagherian and Asadikaram  did not find statistically significant difference in the level of calcium and phosphorus in children with and without early childhood caries. The results of the present study are suggestive of the fact that demineralization and remineralization might be taking place simultaneously, thereby maintaining the level of these minerals same in control and test group. However, as demineralization occurs at faster rate than remineralization, it leads to the formation of a cavity. It is further hypothesized that as saliva is a blood filtrate, unaltered calcium level in caries affected children might be due to the regulatory role of parathyroid hormone (PTH) maintaining its level in the caries-affected children to the level as found in caries-free children. Role of PTH in regulating serum calcium level is well documented. Significant increase in phosphorous levels in caries-affected children of 3-8 years indicated a direct relationship between phosphorus and dental caries in this age group and that might be because of their more susceptibility to dental caries than children of 9–14 years of age. It has been documented in the literature that the primary enamel is significantly softer and elastic, exhibit more diffusion coefficient  and showed a significantly greater susceptibility to demineralization in the acidic medium due to more organic content that dissolves faster than the permanent enamel. Increased phosphorus in the caries affected children of 3–8 years might have been derived from the hydrolysis of phosphoprotein and/or phospholipids (the organic content) of their enamel leading to their more DMFT status than 9–14-year-old children [Table 1] and [Figure 5]. Direct relationship between alkaline phosphatase, phosphorus, and dental caries has been established in children of 4–6-year-old by Gandhy and Damle. On the other hand, Watanbe et al. indicated that calcium concentration of resting saliva has no relationship with the DMFT status of children. Mean ± SD values of amylase in caries affected children was less as compared to their age-matched control groups. However, the decrease was not significant statistically. It is proposed that as amylase is purely of salivary origin, decrease in mean values of amylase in caries-affected children might be because of decrease in salivary flow rate hence caries development., Further, calcium acts as cofactor for α-amylase and the activity of α-amylase is strongly linked with the level of calcium. No significant decrease in the level of amylase in caries affected children is attributable to their unaltered calcium level. The present study revealed no significant difference in salivary pH in caries-free and caries-affected children. These findings find support from Mukherjee  who did not find any change in pH and phosphorus level in caries susceptible and calculus susceptible subjects aged 6–50 years. Strong correlation was noted between calcium and phosphorous with correlation coefficient of 0.192 and P = 0.040 irrespective of age group reflects that these two minerals are the integral constituents of tooth structure.
| Conclusions|| |
Significantly increased phosphorus in caries-affected children of 3–8 years might have been derived from the hydrolysis of more organic content in their enamel that dissolves faster leading to more DMFT status than the children of 9–14 years of age group.
Pitfalls and future recommendations
Saliva samples were taken only once. The future studies with multiples of saliva samples are needed to confirm the present results. The studies involving the organic composition of the primary and the permanent, and the role of PTH on salivary calcium should be undertaken to support the present study.
We are highly indebted to the children who participated willingly in the present study. We are also thankful to the technical staff of the Department of Pedodontics and the Department of Biochemistry for the successful completion of the study. Special thank goes to Master Raunak Mahajan (Son of Dr. Santosh Mahajan and a 7th standard student) for helping in the typing the manuscript.
Financial support and sponsorship
The work would not have been completed without the financial support by the institute.
Conflicts of interest
There are no conflflicts of interest.
| References|| |
McDonald RE, Avery DR, Stookey GK. Dental caries in child and adolescent. In: McDonald RE, Avery DR, Dean JA, editors. Dentistary for the Child and Adolescent. 8th
ed. St. Louis: Mosbey Company; 2004. p. 202-35.
Tsai AI, Chen CY, Li LA, Hsiang CL, Hsu KH. Risk indicators for early childhood caries in Taiwan. Community Dent Oral Epidemiol 2006;34:437-45.
Seow WK. Biological mechanisms of early childhood caries. Community Dent Oral Epidemiol 1998;26:8-27.
Shafer W, Hine MK, Levy BM. Textbook of Oral Pathology. 4th
ed. Philadelphia: W.B. Saunder's Company; 1993. p. 406-78.
Loesche WJ. Role of Streptococcus mutans
in human dental decay. Microbiol Rev 1986;50:353-80.
Dashper SG, Reynolds EC. Effects of organic acid anions on growth, glycolysis, and intracellular pH of oral streptococci. J Dent Res 2000;79:90-6.
Koulourides T. Dynamics of tooth surface-oral fluid equilibrium. Adv Oral Biol 1966;2:149-71.
Afonsky D. Saliva and its Relation to Oral Health: A Survey of the Literature. Birmingham: University of Alabama Press; 1961.
Edgar WM. Saliva: Its secretion, composition and functions. Br Dent J 1992;172:305-12.
Walsh LJ. Preventive dentistry for the general dental practitioner. Aust Dent J 2000;45:76-82.
Cappeli DP, Mobley CC. Prevention in Clinical Oral Health Care. 1st
ed. Philladelphia: Elsevier; 2008.
Dezan CC, Nicolau J, Souza DN, Walter LR. Flow rate, amylase activity, and protein and sialic acid concentrations of saliva from children aged 18, 30 and 42 months attending a baby clinic. Arch Oral Biol 2002;47:423-7.
Clark EP, Collip JB. Determination of serum calcium. J Biol Chem 1925;63:461.
Kuttner T, Lichenstein L. Determination of serum phosphorous. J Biol Chem 1930;86:671.
Bansal DD, Khardori R, Gupta MM. Estimation of amylase. Pract Biochem 1985;1:63-5.
Ashley FP. The relationship between diet, saliva, plaque and caries. J Dent Res 1972;51:1234.
Elizarova VM, Petrovich IuA. Ionized calcium in the saliva of children with multiple caries. Stomatologiia (Mosk) 1997;76:6-8.
Anderson P, Hector MP, Rampersad MA. Critical pH in resting and stimulated whole saliva in groups of children and adults. Int J Paediatr Dent 2001;11:266-73.
Prabhakar AR, Shubha AB, Mahantesh T. Estimation of calcium, phosphorous and amylase concentrations in stimulated whole saliva of children with different caries status. A comparative study. Malasian Dent J 2008;29:6-13.
Bagherian A, Asadikaram G. Comparison of some salivary characteristics between children with and without early childhood caries. Indian J Dent Res 2012;23:628-32.
] [Full text]
Lenander-Lumikari M, Loimaranta V. Saliva and dental caries. Adv Dent Res 2000;14:40-7.
Shahrabi M, Nikfarjam J, Alikhani A, Akhoundi N, Ashtiani M, Seraj B, et al.
A comparison of salivary calcium, phosphate, and alkaline phosphatase in children with severe, moderate caries, and caries free in Tehran's kindergartens. J Indian Soc Pedod Prev Dent 2008;26:74-7.
] [Full text]
Hubbel RB, Bunting RW. Calcium and phosphorous of saliva in relation to dental caries. J Nut 1932;5:599-605.
Vasudevan DM, Sreekumari S, Vaidyanathan K. Textbook of Biochemistry for Medical Students. 6th
ed. Vol 1: Jaypee Brothers Medical Publishers Pvt. Ltd.; 2011. p. 413-5.
Lussi A, Kohler N, Zero D, Schaffner M, Megert B. A comparison of the erosive potential of different beverages in primary and permanent teeth using an in vitro
model. Eur J Oral Sci 2000;108:110-4.
Lindén LA, Björkman S, Hattab F. The diffusion in vitro
of fluoride and chlorhexidine in the enamel of human deciduous and permanent teeth. Arch Oral Biol 1986;31:33-7.
Wang LJ, Tang R, Bonstein T, Bush P, Nancollas GH. Enamel demineralization in primary and permanent teeth. J Dent Res 2006;85:359-63.
Gandhy M, Damle SG. Relation of salivary inorganic phosphorus and alkaline phosphatase to the dental caries status in children. J Indian Soc Pedod Prev Dent 2003;21:135-8.
Watanabe Y, Mizoguchi H, Masamura K, Nagaya T. No relationship of salivary flow rate or secretory immunoglobulin A to dental caries in children. Environ Health Prev Med 1997;2:122-5.
Fiehn NE, Oram V, Moe D. Streptococci and activities of sucrases and alpha-amylases in supragingival dental plaque and saliva in three caries activity groups. Acta Odontol Scand 1986;44:1-9.
Scully C. Sjögren's syndrome: Clinical and laboratory features, immunopathogenesis, and management. Oral Surg Oral Med Oral Pathol 1986;62:510-23.
van der Reijden WA, van der Kwaak JS, Veerman EC, Nieuw Amerongen AV. Analysis of the concentration and output of whole salivary constituents in patients with Sjögren's syndrome. Eur J Oral Sci 1996;104:335-40.
Axelson P. Diagnosis and Risk Prediction of Dental Caries. Vol. 2. Chicago, Illianos: Illianos Quintessence Book; 2000.
Mukherjee S. The state of calcium phosphate in saliva of caries susceptible and calculus susceptible children and adults. J Pedod 1986;11:76-82.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2], [Table 3]