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Journal of Indian Society of Pedodontics and Preventive Dentistry Official publication of Indian Society of Pedodontics and Preventive Dentistry
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Year : 2008  |  Volume : 26  |  Issue : 2  |  Page : 74-77

A comparison of salivary calcium, phosphate, and alkaline phosphatase in children with severe, moderate caries, and caries free in Tehran's kindergartens

1 Department of Pedodontics, Dental School, Tehran University, Tehran, Iran
2 Department of Pathology, School of Medicine, Tehran University, Tehran, Iran

Correspondence Address:
B Seraj
Department of Pedodontics , Dental School, Tehran University, Tehran
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0970-4388.41621

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The most common dental disease in childhood is dental caries. This study was carried out to recognize the components of saliva which are protective factors in children to evaluate and predict caries susceptible and caries resistant individuals. Unstimulated whole saliva was obtained from 75 children aged 3-5 years. They divided into three groups: decayed missing and filled teeth (dmft) > 6 (severe caries), 1 < dmft < 6 (moderate caries), and dmft < 1 (caries free). Unstimulated whole saliva assayed by biochemical methods to determine salivary calcium, inorganic phosphate, and alkaline phosphatase. There was no significant changes in salivary calcium, alkaline to phosphate and alkaline phosphatase activity and their ratio with progress of caries ( P > 0.05 ). Although the results showed that salivary phosphate and alkaline phosphatase in caries free group and calcium in the group with severe caries were somewhat more than those in other groups. Despite of the results of the present study, the relationship between salivary components and caries rate in children remains controversial. So more studies are necessary to achieve some practical criteria for predicting dental caries, recognition of susceptible persons, and finally prevention of caries in children.

Keywords: Calcium, dental caries, phosphate and alkaline phosphatase, saliva

How to cite this article:
Shahrabi M, Nikfarjam J, Alikhani A, Akhoundi N, Ashtiani M, Seraj B. 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

How to cite this URL:
Shahrabi M, Nikfarjam J, Alikhani A, Akhoundi N, Ashtiani M, Seraj B. 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 [serial online] 2008 [cited 2022 Jan 28];26:74-7. Available from: https://www.jisppd.com/text.asp?2008/26/2/74/41621

   Introduction Top

Several investigators have reported variations in the calcium and phosphorous concentrations of dental plaque according to site of collection. [1],[2],[3],[4],[5],[6] These variations have generally followed the pattern of distribution of supragingival calculus and have been attributed to the relationship of the plaque to the orifices of the salivary glands.

In view of these observations, a direct relationship between the calcium and phosphorous concentrations of saliva and plaque would not be unexpected. However, Tatevassian reported no correlation between the composition of resting mixed saliva obtained on waking and the composition of aqueous plaque extract in 40 subjects aged 18-21 years. [7]

Much of the early evidence relating calcium and phosphorous levels in plaque to caries experience is also contradictory. [8]

Dawes and Jenkins suggested that the low susceptibility of lower incisor teeth to caries was related to the higher calcium and phosphorous concentration observed in plaque from this area, compared with plaque from other areas of the mouth. [9] In several later studies, with more standardized recording methods, when the total caries activity of the mouth was assessed and subjects were divided into those showing either high caries activity or low caries activity, significant inverse relationships were found between caries experience and plaque calcium or phosphorous levels. [10],[11],[12],[13]

In most studies, the Decay, Missing, Filling (DMF) index has been used as measure of caries experience, but caries progression is discontinous and hence a caries lesion or filling is an indication of past events and not necessarily of current caries activity. In the study reported by Luma, [14] the subjects were divided on the basis of DMF and caries increment over a one-year period. Low extracellular inorganic phosphouous was found in plaque from caries active individuals compared with low-caries group.

The purpose of the present investigation is to relate the calcium, phosphate, and alkaline phosphatase levels in saliva of several caries, moderate and caries free children selected from Tehran's kindergartens.

   Materials and Methods Top

Subjects were 75 children between 3 and 5 years which examined clinically by a flashlight, explorer, and mirror. The obtained information from the questionnaire was to eliminate the altered factors such as metabolic disease, Rickets, and deformans osteitis which cause increase the level of alkaline phosphatase activity of serum. [15]

To unify the cases, they were asked about their mother's occupation, residence in 2 years ago, cookies, and snack consumption between meals (at least four times in 24 h), breast milk or formula consumption at least for 2 years and brushing teeth once a day. To unify the fluoride consumption through the water tap, the information was obtained from Tehran water organization.

The clinical diagnosis of dental caries was based on to see an undermined cavity in fissures and the presence of pit on the smooth surfaces. [16] The children were divided into three groups: dmft > 6 severe caries (case), 1 < dmft < 6 moderate caries (control 1), and dmft < 1 caries free (control 2).

Unstimulated whole saliva was collected in tubes. After collecting 3 mL saliva the tubes' cap closed and delivered to the biochemistry Lab in <2 h.

The measurement of salivary calcium by methyl-thymol blue. In colorometric method, calcium and methyl-thymol blue makes a color that is proportional with the calcium ion in solution. [17]

The measurement of phosphate was done by Ultraviolet (UV) method which measured the optic density of phosphorous in complex of phosphorous and ammonium molibidate in 340 nm wave length. [18]

Alkaline phosphatase was measured by kinetic method. Alkaline phosphatase effects on 4-nitrophenyl phosphate and converts if to yellow 4-nitrophenol which could absorb the light. The absorption of light is proportional with alkaline phosphatase enzyme activity.

Statistical analysis

Summary statistical measures (mean and standard deviation) were calculated for continuous variables. For discrete variables, frequency distributions were studied.

Distribution normality assumptions of calcium, phosphorous, alkaline phosphatase, the proportions of calcium to alkaline, alkaline to phosphorous and calcium to phosphorous variables were assessed by the Kolmogorov-Smirnov test. For comparison of these variables between clinical characteristic, one-way analysis of variance (ANOVA) was used. In some comparisons when normality would not hold, non-parametric Kruskal-Wallis test were used. The effect of age was taken into account using the ANCOVA test. In our discussion, the correlations between every two variables were analyzed with Spearman's correlation. The level of statistical significance was established at P < 0.05.

The ANOVA, ANCOVA, and Kruskal-Wallis tests [Table 1] show that there is no differences (not significant) between calcium, phosphorous, alkaline phosphatase, the proportions of calcium to alkaline, alkaline to phosphorous, and calcium to phosphorous in three groups of children ( P value > 0.05).

[Figure 1] shows the mean of calcium, phosphorous, alkaline phosphatase, the proportions of calcium to alkaline, alkaline to phosphorous, and calcium to phosphorous in three groups of children [Table 2].

The result of Spearman's correlation test shows there is no statistical relationship between gender, order of birth, number of children in a family, age, mother's, and father's education in three groups ( P value > 0.05) [Figure 2],[Figure 3],[Figure 4 ].

   Discussion Top

Saliva has an important role to protect the oral health. [16],[19] The sufficient amount of saliva is necessary to protect the oral tissue. [19] The balance between demineralization and remineralization depends on the salivary calcium and phosphate concentration as well as the level of the salivary alkaline phosphatase. [15] This is an important point that the calcium and phosphate should be saturated in saliva to effect on demineralization and remineralization. Conflicting results have been obtained from investigations of calcium and phosphate contents of saliva and their relationship to dental caries. Karshan observed that salivary calcium concentration decreased with increasing caries activity. [20] Ashley found that the salivary calcium increased with decreasing caries activity. [21] Shannon and Feller also found the inverted relationship between parotid saliva and Decay, Missing, Filling Surface (DMFS).[22] On the other hand Turtola in 1978 showed the salivary calcium concentration increased with increasing caries activity. [23] Also Elizarova and Petrovich in 1977 investigated in 4- to 12-year-old children and announced that the calcium level of saliva is higher in children with multiple caries than in children with single caries. [24] This study demonstrates the mean of salivary calcium in children with severe caries in more than the two other groups, but this difference was not significant after statistical analysis which confirms the result of the study of Masamurak et al ., [25] Marray and Shaw,[26] Shafizadeh et al ., [27] Cornejo et al. , [ 28] and Gandhy and Damle [15] investigated the relationship between alkaline phosphatase and inorganic phosphorous with decay, filling, surface and they observed the level of alkaline phosphatase and inorganic phosphorous in rampant caries children was higher than the caries free children. [15],[28] Also Shafizadeh et al . demonstrated the same result. [27] However, Mandel and Karshon, and Ashley and Shaw reported low phosphate in children with high dental caries. [24],[27] Also in Shannons and Feller study, there was an inverted relationship between phosphate level and parotid saliva and DMFT. [22]

In this study, the mean of inorganic phosphate and alkaline phosphatase in caries free group was higher than the other groups, but this difference was not significant after statistical analysis. In Shaw study, [26] the calcium to phosphorous proportion was reported significantly higher in caries free group however in our study there was not any significant difference in calcium to phosphorous, alkaline to phosphorous, and calcium to alkaline proportions.

   Conclusion Top

This study demonstrates no significant difference between calcium, phosphate, alkaline phosphatase, and their proportions also there was no relationship between the other probable factors effect on caries process (such as order of birth, gender) in three groups. Since the proportion of the salivary contents is more important in susceptibility or resistance to dental caries, it is suggested to do more investigation in this field. Also by controlling the definite and probable factors effect on dental caries, the effect of inherent difference between people has been confirmed.

   References Top

1.Allen WI, Moore BW. Calcium content of plaque and saliva. Internat. ASS. For dent. Res. Preprinted abstracts, 35th general meeting, Abstract 108. 1957.  Back to cited text no. 1    
2.Dawes C, Jenkins GN. Some inorganic constituents of dental plaque and their relationship to early calculus formation and caries. Arch Oral Biol 1962;7:161-72.  Back to cited text no. 2  [PUBMED]  
3.Schoroeder HE. Inorganic content and histology of early dental calculus in man. Helv Odont Acta 1963;7:17-29.  Back to cited text no. 3    
4.Kaminisky FS, Kleinberg I. Comparison of calcium, phosphorus and ash levels of plaques of different ages on the labial and interproximal surfaces of the anterior teeth. Interant Ass For dent Res. Preprinted abstracts, 45the general meeting, abstract 206. 1967.  Back to cited text no. 4    
5.Zuniga MA, Lopez H, Sandham HJ, Bradley EL, Koulourides T. Calcium and phosphorus contents of dental plaques and microhardness changes of sample enamel in the human mouth. Ala J Med Sci 1973;10:3-10.  Back to cited text no. 5  [PUBMED]  
6.Ashley FP. Calcium and phosphorus levels in human dental plaque-variations according to site of collection. Arch Oral Biol 1975;20:167-70.  Back to cited text no. 6  [PUBMED]  
7.Tatevossian A. The relationship of composition of human resting saliva, obtained at waking, to the metabolic activity in saliva, DMFT index and the composition of and aqueous plaque extract. J Dent Res 1971;50:212.  Back to cited text no. 7    
8.Afonsky D. Saliva and its relation to oral health. Birmingham: University of Alabama Press; 1961.  Back to cited text no. 8    
9.Jenkins N. Physiology and biochemistry of the mouth, 4th ed. Oxford: Blask Well; 1978.  Back to cited text no. 9    
10.Kleinberg I, Chatterjee R, Castaldi CR, Chebib F. Relations between plaque ash, calcium and phosphorus levels and dental caries.   Back to cited text no. 10    
11.Ashley FP. The relationship between diet, saliva, plaque and caries. J Dent Res 1972;51:1234.   Back to cited text no. 11    
12.Ashley FP. Calcium and phosphorus concentration of dental plaque related to dental caries in 11- to 14-year-old male subjects. Caries Res 1975;9:351-62.  Back to cited text no. 12  [PUBMED]  
13.Scham Schula RG, Agus H, Kunzel M, Adkins BL, Barmes DE. The concentrations of selected major and trace minerals in human dental plaque. Arch Oral Biol 1977;22:34-5.   Back to cited text no. 13    
14.Luoma H. Liability of inorganic phosphate in dental plaque and saliva. Acta Dont Scand 1964;22:41.   Back to cited text no. 14    
15.Gandhy M, Damle G. Relation of salivary inorganic phosphorous and alkaline phosphatase to the dental caries status in children. J Indian Soc Pedod Prev Dent 2002;21:135-8.  Back to cited text no. 15    
16.Mahejabeen R, Sudha P, Kulkarni SS, Anegundi R. Dental prevalence among preschool children of Hubli: Dharwand city. J Indian Soc Pedod Prev Dent 2006;24:19-22.  Back to cited text no. 16    
17.Grindler E, King JD. Calcium, apparent calcium absorption and calcium balance in young and elderly subjects: Influence of protein intake. Am J Clin Pathol 1972;58:376.  Back to cited text no. 17    
18.Daly JA, Erthingshausen G. Direct method for determining inorganic phosphate in serum with the "contrifichem". Clin Chem 1972;18:263-5.  Back to cited text no. 18    
19.Jenknes GN. Edgar WM. The effect of daily gum-chewing on salivary flow rate in man. J Dent Res 1989;68:786-90.  Back to cited text no. 19    
20.Karshan M. Factors in saliva correlated with dental caries. J Dent Res 1939;18:395-407.  Back to cited text no. 20    
21.Ashley FP, Wilson RF. The relationship between calcium and human saliva and dental plaque. Archs Oral Boil 1978;23:69-73.  Back to cited text no. 21    
22.Shannon, Feller RP. Parotid saliva flow rate, calcium, phosphorous and magnesium concentration in relation to dental caries experience in children. Pediatr Dent 1979;1:16-20.   Back to cited text no. 22    
23.Turtola L. Dental caries and its prevention. Pro Finn Den Soc 1978;74:36-7.  Back to cited text no. 23    
24.Elizarora VM. Petrovich IU. Ionized calcium in the saliva of children with multiple caries. Stomatologia 1997;76:6-8.  Back to cited text no. 24    
25.Masumurak, Inaba R, Iwata H. Salivary calcium and total protein in relation to dental caries. Nippon Zasshi 1995;50:882-92.  Back to cited text no. 25    
26.Shaw L, Marray JJ, Burchell CK, Best JS. Calcium and phosphorous content of plaque and saliva in relation to dental caries. Caries Res 1983;17:543-8.  Back to cited text no. 26    
27.Afshar H, Seraj B, Shafizadeh N. The relationship between rampant caries and salivary situation of 4-5 year old children living in Tehran. J Islamic Dent Assoc Iran 2001;13:18-35.  Back to cited text no. 27    
28.Cornejo LS, Brunotto M, Hilas E. Salivary factors associated to the prevalence and increase of dental caries in rural schoolchildren. Rev Saude Publica 2008;42:19-25.  Back to cited text no. 28  [PUBMED]  [FULLTEXT]


  [Figure 1], [Figure 2], [Figure 3], [Figure 4 ]

  [Table 1], [Table 2]

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