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ORIGINAL ARTICLE
Year : 2008  |  Volume : 26  |  Issue : 4  |  Page : 158-161
 

Quantitative assessment of IgA levels in the unstimulated whole saliva of caries-free and caries-active children


Department of Pediatric Dentistry, Meenakshi Ammal Dental College, Alapakkam Main Road, Maduravoyal, Chennai, India

Correspondence Address:
M S Muthu
Department of Pediatric Dentistry, Meenakshi Ammal Dental College, Alapakkam Main Road, Maduravoyal, Chennai
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0970-4388.44031

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   Abstract 

Saliva is commonly referred to as the blood stream of the oral cavity. It has many functions, one of the major functions being protection of teeth against dental caries. There are many components in saliva, each one having a specific role in the prevention of dental caries. The composition of saliva varies from individual to individual and in the same individual it varies between the glands. The composition of whole saliva, especially when unstimulated, has gained much interest, because it is this which constantly bathes the teeth. The aim of this study was to determine the IgA levels in the unstimulated whole saliva of caries-free and caries-active children aged 3-6 years and to correlate its role in protection of the tooth against dental caries.


Keywords: Immunoturbidimetry, salivary IgA, unstimulated whole saliva


How to cite this article:
Shifa S, Muthu M S, Amarlal D, Prabhu V R. Quantitative assessment of IgA levels in the unstimulated whole saliva of caries-free and caries-active children. J Indian Soc Pedod Prev Dent 2008;26:158-61

How to cite this URL:
Shifa S, Muthu M S, Amarlal D, Prabhu V R. Quantitative assessment of IgA levels in the unstimulated whole saliva of caries-free and caries-active children. J Indian Soc Pedod Prev Dent [serial online] 2008 [cited 2019 Jul 22];26:158-61. Available from: http://www.jisppd.com/text.asp?2008/26/4/158/44031



   Introduction Top


Dental caries is the most common infectious disease of the oral cavity. [1] The United States surgeon general's report on oral health in America published in May 2000 reported dental caries as the single most common chronic childhood disease. [2] Though this disease is very common the mechanism of how some children remain caries free has been the topic of interest in the field of dental research for decades. Dental caries is a multifactorial disease and one of the major contributing factors is saliva. Salivary components, its flow, viscosity, buffering capacity, etc. play a major role in the prevention, initiation, and progression of the disease. [3] It helps in the prevention of the caries by its antibacterial effect. Many constituents of the saliva, both immunoglobulins and nonimmunoglobulins contribute to this antibacterial effect. [3] Salivary immunoglobulins include IgA, IgG, and IgM. Salivary IgA (sIgA) is a secretory immunoglobulin and is different from the serum IgA by having a dimeric structure compared to the monomeric structure of the latter. . Salivary IgA contributes 60% of the total immunoglobulin count in the saliva and helps in the antibacterial action of the saliva by neutralizing the bacterial toxins and enzymes, and preventing the adherence of the bacteria to the tooth surface by blockage of bacterial adhesions, reduction of hydrophobicity, and agglutination of the bacteria. [4] Previous investigations on the role of IgA in dental caries have reported contradictory results. Some authors reported higher levels of sIgA in caries-resistant individuals in relation to caries-susceptible ones, suggesting an effective protective function. [5],[6],[7],[8],[9],[10] Legler et al. [6] observed higher caries incidence in patients with IgA deficiency. On the other hand, other authors did not observe any correlation between caries activity and sIgA levels. [11],[12],[13]

The aim of this study was to assess the levels of sIgA in the unstimulated whole salivary samples of caries-free and caries-active children and to correlate IgA's quantitative role in prevention of dental caries.


   Materials and Methods Top


The study was conducted in the Department of Pediatric dentistry, Meenakshi Ammal Dental College, Meenakshi University, Chennai, India. Twenty children of both the sexes aged 3-6 years were selected depending on their caries experience. They were divided into two groups. Group I included ten children who had active carious lesions with DMFT scores of ≥5. Group II included ten children who were caries free, that is, their DMFT were '0'. The selection criteria for the study samples were:

  1. Children aged between 3-6 years.
  2. No reported positive medical history.
  3. No antibiotic and analgesic intake in past two-weeks.
  4. For the children in group I the DMFT scores had to be ≥5.


The unstimulated whole salivary samples were collected two hours after any oral or visual exposure to food stuffs. All the salivary samples were collected in sterile vials [Figure 1] between 10-11 am in order to prevent any bias in the concentration of the saliva due to the circadian rhythm. After obtaining informed consent from parents, the unstimulated whole salivary samples were collected by the method suggested by Colin Dawes. [14] Children were asked to pool the saliva in the floor of their oral cavity and spit into a sterile vial intermittently [Figure 2]. These samples were transported at -70 ░C. [15]

The samples were then tested for the levels of sIgA. The investigator who analyzed the sIgA level was blinded. The procedure by which the levels were analyzed was immunoturbidimetry with the help of SpinReact (SpinReact , SA, Gerona , Spain). The principle of the procedure is that IgA antibodies when mixed with salivary samples form insoluble complexes. These complexes cause an absorbance change in the spectrophotometer [Figure 3], dependent upon the IgA concentration of the patient sample. The reading in the spectrometer can be compared with a calibrator of known IgA concentration. The calibrator used in this study was human serum. The contents of SpinReact kit are two reagents R 1 and R 2 .

R 1 contains tris buffer 20 mmol/L, PEG 8000, sodium azide 0.95 g/L, pH 8.2. R 2 contains goat serum, antihuman IgA, sodium azide 0.95 g/L, pH 7.5.

The procedure is as follows: 950 Ál of R 1 was dispensed into the test tube to which 7 Ál of the sample was added. Both were mixed and the absorbance reading was taken as A 1. Later, 50 Ál of R 2 was added and the absorbance reading A 2 was taken after one minute . The values were substituted using the following formula:

A 2 - A 1 = ΔA

ΔA x factor = IgA level

Where, factor is (calibrator concentration)/(ΔA of the calibrator) equaling 1736. Thus, the quantity of IgA in each sample was calculated.


   Results Top


The sIgA level in salivary samples of group I and II are shown in [Table 1] and [Table 2], respectively. The respective mean values for group I and group II were 221.1 and 229.9. Thus, the mean value for group II was numerically greater than group I. The values of both the study groups were analyzed using the student's t -test. The P value was 0.76, which was >0.05. Therefore, the values were not statistically significant at 5% confidence interval.


   Discussion Top


The infectious nature of dental caries assumes the hypothesis that some form of host immunity can regulate caries activity. [15] If immunity can regulate caries activity then secretory IgA should be the antibody that provides protection because it has shown to do so in other body secretions. [16] This study was conducted to correlate the relationship between dental caries and sIgA in children with complete primary dentition. As there are not many studies pertaining to the primary dentition and salivary IgA levels, children between ages 3-6 years were chosen. Mandel and khurana found that the IgA and protein concentration decreased with increased salivary flow from the parotid and submaxillary salivary glands. [17] Stimulated saliva could have decreased the concentration of the IgA, hence unstimulated whole saliva was collected.

Saliva was collected by the method suggested by Colin Dawes as it was easy to obtain the child's cooperation. [14] In this method, the child was asked to pool the saliva in the floor of the oral cavity and asked to spit intermittently. All the samples were collected between 10-11 am. This time was two hours after any oral or visual exposure to food stuffs. This was done to prevent the effect of circadian rhythm on the salivary concentration. [18] The salivary IgA levels were determined using immunoturbidimetry. The principle of the immunoturbidimetry is that IgA antibodies when mixed with samples containing IgA form insoluble complexes. These complexes cause an absorbance change in the spectrophotometer, dependent upon the IgA concentration of the patient sample. The reading in the spectrometer can be noted and compared with a calibrator of known IgA concentration. The control used in our study was human serum.

Many similar studies were conducted in the past and contradictory results were obtained. Studies conducted by Lehner et al. , [6] Gregory et al. , [5] Cogulu et al. , [7] Tenovuo et al. , [8] Parkash et al. , [9] and Rose et al. [10] reported an increase in the level of salivary IgA with decrease in caries activity. Whereas studies conducted by deFarias et al. [19] and Bruno et al. [20] found an increase in IgA level with increase in caries activity. Our study results are in agreement with Koga et al. , [11] Camling et al. , [12] and Everhart et al. [13] who found no correlation between dental caries and IgA levels. However, these results are not directly comparable with the results of the study conducted by Koga et al. [11] and Camling et al. [12] as they had measured IgA specific to  Streptococcus mutans Scientific Name Search  ELISA method, whereas in our study, we have used immunoturbidimetry and measured whole salivary IgA levels.


   Conclusion Top


The IgA levels of the whole unstimulated saliva and its protective mechanism against dental caries are still unclear. Though there was an increase in the mean IgA value of the caries-resistant group, the results were not statistically significant. Our study did not show any correlation between sIgA levels and caries status. In future, an increase in sample size with analysis of the IgA specific to streptococcus mutans might give a clear correlation.


   Acknowledgments Top


We are grateful to Mrs. N.S. Jagadeeswari, Department of Biochemistry Dr. Raguram, Professor and Head, Department of Microbiology, Dr. Rajesh (Intern) and Dr. Vinodh (Intern) for their support during the study.

 
   References Top

1.Caufield PW, Griffen AL. Dental caries: An Infectious and transmissible disease. Pediatr Clin North Am 2000;47:1001-19.  Back to cited text no. 1    
2.US Department of Health and Human Services: Oral health in America: A report of surgeon general. Rockville, MD: US Department of Health and Human Services, National institute of Dental and craniofacial Research, National Institutes of Health; 2000.  Back to cited text no. 2    
3.Whelton H. The anatomy and physiology of salivary glands. In: Edgar WM, O'Mullane DM, editors. Saliva and oral health, 2 nd ed. British Dental Association; 1996. p. 2.  Back to cited text no. 3    
4.Newbrun E. Current concepts of caries etiology. In: Newbrun E, editor. Cariology. 3rd ed. Baltimore: Williams and Wilkins; 1978. p. 43.  Back to cited text no. 4    
5.Gregory RL, Hobbs LC, Kindle JC, VanTo T, Malmstrom HS. Immunodominant antigens of Streptococcus mutans in dental caries-resistant subjects. Hum Antibodies Hybridomas 1990;1:132-6.  Back to cited text no. 5    
6.Legler DW, McGhee JR, Lynch DP, Mesteck JF, Shaefer ME, Carson J, et al. Immunodeficiency disease and dental caries in man. Arch Oral Biol 1981;26:905-10.  Back to cited text no. 6    
7.Cogulu D, Sabah E, Kutukculer N, Ozkinay F. Evaluation of the relationship between caries indices and salivary secretory IgA, salivary pH, buffering capacity and flow rate in children with Down's syndrome. Arch Oral Biol 2006;51:23-8.   Back to cited text no. 7    
8.Tenovuo J, Jentsch H, Soukka T, Karhuvaara L. Antimicrobial factors of saliva in relation to dental caries and salivary levels of mutans streptococci. J Biol Buccale 1992;20:85-90.  Back to cited text no. 8    
9.Parkash H, Sharma A, Banerjee U, Sidhu SS, Sundaram KR. Humoral immune response to mutans streptococci associated with dental caries. Natl Med J India 1994;7:263-6.  Back to cited text no. 9    
10.Rose PT, Gregory RL, Gfell LE, Hughes CV. IgA antibodies to Streptococcus mutans in caries-resistant and -susceptible children. Pediatr Dent 1994;16:272-5.  Back to cited text no. 10    
11.Koga-Ito CY, Martins CA, Balducci I, Jorge AO. Correlation among mutans streptococci counts, dental caries, and IgA to Streptococcus mutans in saliva. Pesqui Odontol Bras 2004;18:350-5.  Back to cited text no. 11    
12.Camling E, Kohler B. Infection with the bacterium Streptococcus mutans and salivary IgA antibodies in mothers and their children.Arch Oral Biol 1987;32:817-23.  Back to cited text no. 12    
13.Everhart DL, Rothenberg K, Carter WH, Klapper B. The determination of antibody to streptococcus mutans serotypes in saliva for children ages three to seven years. J Dent Res 1978;57:631-5.  Back to cited text no. 13    
14.Dawes C. Factors influencing salivary flow rate and composition. Edgar WM, O'Mullane DM, editors. Saliva and oral health. 2nd ed. British Dental Association; 1996. P. 27.   Back to cited text no. 14    
15.Bolton RW, Hlava GL. Evaluation of salivary IgA antibodies to cariogenic microorganisms in children: Correlation with dental caries activity. J Dent Res 1982;61:1225-8.  Back to cited text no. 15    
16.Orstavik D, Brandzaeg P. Secretion of parotid IgA in relation to gingival inflammation and dental caries experience in man. Arch Oral Biol 1975;20:701-4.  Back to cited text no. 16    
17.Mandel ID, Khurana HS. The relation of human salivary IgA globulin and albumin to flow rate. Arch Oral Biol 1970;14:1433-5.  Back to cited text no. 17    
18.Lehtonen OP, Grεhn EM, Stεhlberg TH, Laitinen LA. Amount and avidity of salivary and serum antibodies against streptococcus mutans in two groups of human subjects with different dental caries susceptibility. Infect Immun 1984;43:308-13.  Back to cited text no. 18    
19.de Farias DG, Bezerra AC. Salivary antibodies, amylase and protein from children with early childhood caries. Clin Oral Investig 2003;7:154-7.   Back to cited text no. 19    
20.Bruno B, Pezzini A, Menegazzi M. Salivary levels of immunoglobulin and dental caries in children. Boll Soc Ital Biol Sper 1985;61:381-6.  Back to cited text no. 20    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2]


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