Estimation of salivary immunoglobulin A levels in gingivitis and chronic periodontitis patients before and after phase I periodontal therapy

Kalakonda Butchibabu; Mythili Swaminathan; Senthil Kumar; Pradeep Koppolu; Kotha Kiran; Tupili Muralikrishna

doi:10.4103/2277-8632.128486

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ORIGINAL ARTICLE

Year : 2014 | Volume : 3 | Issue : 5 | Page : 23-27

Estimation of salivary immunoglobulin A levels in gingivitis and chronic periodontitis patients before and after phase I periodontal therapy

Kalakonda Butchibabu¹, Mythili Swaminathan², Senthil Kumar², Pradeep Koppolu¹, Kotha Kiran³, Tupili Muralikrishna¹
¹ Department of Periodontics, Sri Sai College of Dental Surgery, Vikarabad, Andhra Pradesh, India
² Rajah Muthiah Dental College & Hospital, Annamalai University, Tamil Nadu, India
³ Triveni Institute of Dental Sciences, Hospital & Research Centre, Bilaspur, Chhattisgarh, India

Date of Web Publication

10-Mar-2014

Correspondence Address:
Kalakonda Butchibabu
Department of Periodontics, Sri Sai College of Dental Surgery, Vikarabad, Andhra Pradesh
India

Source of Support: None, Conflict of Interest: None

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DOI: 10.4103/2277-8632.128486

Abstract

Background: The purpose of this study was to estimate the levels of immunoglobulin A (IgA) in the saliva of healthy controls and patients with gingivitis and chronic periodontitis.
Materials and Methods: The study population consisted of 45 male adult patients, of 20-50 age group who were grouped into three different groups viz. Group A (healthy subjects), Group B (Gingivitis), Group C (Periodontitis) based on clinical parameters. The clinical parameters including plaque index (PlI), gingival index (GI), probing pocket depth (PPD), Russell's periodontal index (PI), clinical attachment level (CAL), and serum IgA levels were recorded at baseline in Group A and at baseline and after 4 weeks in Group B and C. Levels of salivary IgA were measured using Enzyme-linked Immunosorbent Assays (ELISA) technique after collecting 5 ml of unstimulated pooled saliva from patients in two different samples.
Results: There was a significant reduction in salivary IgA levels, PlI, GI and PPD after phase I periodontal therapy in Group B and C; whereas a decrease in Russell's PI and gain in the CAL after phase I periodontal therapy were not statistically significant in Group C (periodontitis) patients. Salivary IgA levels, PlI, GI, and Russell's PI were significantly lower in the Group A (control) patients compared with Group B (gingivitis) and Group C (periodontitis) patients.
Conclusion: Our study showed increased levels of salivary IgA in patients with periodontal disease compared with the healthy controls. The less severe the periodontal involvement, the more dependable was the reduction in IgA concentration following phase I periodontal therapy.

Keywords: Gingivitis, periodontitis, salivary immunoglobulin A

How to cite this article:
Butchibabu K, Swaminathan M, Kumar S, Koppolu P, Kiran K, Muralikrishna T. Estimation of salivary immunoglobulin A levels in gingivitis and chronic periodontitis patients before and after phase I periodontal therapy. J NTR Univ Health Sci 2014;3, Suppl S1:23-7

How to cite this URL:
Butchibabu K, Swaminathan M, Kumar S, Koppolu P, Kiran K, Muralikrishna T. Estimation of salivary immunoglobulin A levels in gingivitis and chronic periodontitis patients before and after phase I periodontal therapy. J NTR Univ Health Sci [serial online] 2014 [cited 2023 Mar 27];3, Suppl S1:23-7. Available from: https://www.jdrntruhs.org/text.asp?2014/3/5/23/128486

Introduction

The host response to infectious and non-infectious substances is physiological function of the immune system. Specifically, microbial substances including lipids, proteins and polysaccharides trigger immune responses that either eliminate the pathogens or non-self-substances that results in pathological consequences in the host. ^[1]

Haffajee and Socransky ^[2] proposed new models to suggest that chronic periodontitis progresses as bursts of activity, which occurs with high frequency over finite periods of a patient's life. This hypothesis initiated intense research to identify host response markers in Gingival crevicular fluid and saliva, which ultimately with proven sensitivity and specificity could be used as a diagnostic test for progressive periodontal disease. ^[3] Salivary variables cannot give site specific information but may be hopeful candidates for the whole mouth follow-up. ^[4]

Salivary IgA is considered as the principal line of defense in the oral cavity against invading microorganisms and plays a vital part in host bacterial interactions. ^[5],[6] Since saliva can be collected more effortlessly and less invasively compared with blood; we have considered the estimation of salivary IgA in this study.

Though many studies exist relating IgA to periodontal disease, studies evaluating the role of salivary IgA as a potential diagnostic marker in periodontal diseases does not exist. Hence our study was designed to evaluate and compare the levels of salivary IgA in healthy and diseased patients before and after phase I periodontal therapy and to assess the diagnostic potential of salivary IgA.

Materials and Methods

This study was conducted in the Department of Periodontics in association with the Department of Biochemistry. A total of 45 male adult patients, of 20-50 age groups, were randomly selected from the out-patient of the Department of Periodontics. The following inclusion and exclusion criteria were considered.

Inclusion criteria

Non-Smoking, healthy individuals with no signs and/or symptoms of systemic disease
Individuals who have not undergone professional oral prophylaxis during the last 1 year
Patients who have not received any antibiotic therapy 6 months prior to the commencement of the study
Patients who were compliant and willing to return after the phase I therapy.

Exclusion criteria

Patients who are suffering from any systemic diseases (e.g. Diabetes mellitus, connective tissue disorders like rheumatoid arthritis)
Patients who are immunocompromised (e.g. Human immunodeficiency virus positive, primary and secondary immunodeficiency disorders, etc.)
Patients with a history of upper respiratory diseases of recent occurrence (within 4 weeks), allergic disorders or autoimmune disorders and infected caries lesions
Patients with salivary gland hypofunction
Patients undergoing chemotherapy.

Study design

Study design included the following steps:

Clinical examination
Collection of saliva
Phase I periodontal therapy
Biochemical investigation

Clinical examination

The following clinical parameters were assessed:

PlI - (Silness and Loe-1964)
GI - (Loe and Silness-1963)
PPD (from the gingival margin to the base of the gingival sulcus or the pocket) was measured to the nearest mm using a William's periodontal probe
PI - (Russell-1956)
CAL - (Ramfjord-1967).

Based on the clinical parameters assessed above the patients were grouped as follows:

Group A: Control group

Fifteen systemically healthy individuals with no signs of gingival and periodontal disease.

Group B: Gingivitis group

Fifteen patients with the presence of bleeding on probing. PPD not exceeding 4 mm and without any attachment or bone loss.

Group C: Chronic periodontitis group

Fifteen patients with at least 16 natural teeth excluding third molars probing depth of ≥5 mm with attachment loss and radiographic evidence of alveolar bone loss.

Scaling and root planning (SRP), occlusal adjustment and splinting procedures were performed as required for Group B and C patients.

Collection of saliva

From the patients chosen above for the study two samples of 5 ml of unstimulated pooled saliva were collected for IgA assay. First sample was collected soon after preliminary examination and second sample after a period of 4 weeks following phase I therapy. Subjects were asked to accumulate saliva in the floor of the mouth and then expectorate in to sterile plastic cup using a draining method. Samples were frozen and kept at −70°C until analyzed.

Biochemical investigation

The samples that have been frozen at −70°C were then thawed and centrifuged for 15 min at 12000g at 4°C to remove the mucin and debris. Concentration of IgA was evaluated using ELISA. Levels of salivary IgA were estimated following the guidelines of the commercial kit provided by Bethyl Laboratories Inc., Montgomery, TX, USA catalog no E80-102. The ELISA protocol was performed in the Department of Biochemistry.

Statistical analysis

Following the biochemical analysis of salivary samples, the results were obtained for all the groups and were subjected to statistical analysis. Statistical package for social sciences package version 14 was used for the statistical evaluation. Mean values in the diseased group before and after treatment were compared by using the Student's paired 't' test, whereas the mean values between the diseased groups were compared using the Student's independent 't' test. The mean values in all the study groups before and after treatment were tested using one-way ANOVA and Tukey - Honestly significant difference(HSD) procedure. Pearson's correlation coefficient analysis was used to find out the relationship between salivary IgA and the selected variables.

Results

For each subject, the mean of the values of the clinical indices for attachment level, PPD, plaque and bleeding along with biochemical parameter like salivary IgA were assessed for 45 patients (20-50 years). Summary statistics for these means across all subjects are shown in [Table 1], [Table 2], [Table 3] and [Figure 1] and [Figure 2].

Table 1: Comparison of parameters for group A (control) with group B (gingivitis) and group C (periodontitis) patients before and after treatment

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Table 2: Comparison of mean values of the variables (before treatment) between different study groups

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Table 3: Comparison of mean values of the variables (after treatment) between different study groups

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Figure 1: Comparison of clinical parameters for Group A (control) with Group B (gingivitis) and Group C (periodontitis) patients before and after treatment

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Figure 2: Comparison of the mean salivary IgA levels for Group A (control) with Group B (gingivitis) and Group C (periodontitis) patients before and after treatment

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The mean value of salivary IgA (mg/dl) in the Group A individuals is 21.25 ± 1.68 while in Group B and C before treatment, the values were 22.3 ± 1.91 and 40.74 ± 5.48 respectively. After treatment, the values were 22.3 ± 1.91 and 39.18 ± 5.69 respectively in Group B and C and change before and after treatment was statistically significant in both the groups (P < 0.05) [Table 1], [Figure 2].

At baseline, the mean PlI in the Groups A, B, and C was 0.29 ± 0.15, 1.52 ± 0.29, and 1.61 ± 0.14 respectively. After treatment, the values were 0.07 ± 0.03 and 0.19 ± 0.27 respectively in Group B and C. There was a significant improvement in both groups and was found to be statistically highly significant (P < 0.001) [Table 1], [Figure 1].

Periodontal pockets vary in their location and depth; hence changes in the mean PPD for the entire mouth provide realistic information. Accordingly, the mean PPD for the entire mouth at the beginning of study for Groups A, B, and C were 2.51 ± 0.20 mm, 2.73 ± 0.11 mm, and 5.48 ± 0.50 mm respectively. After treatment, the mean PPD in Group B and C are 2.51 ± 0.08 mm and 4.96 ± 0.45 mm respectively. The effect of treatment and change in the mean PPD was found to be statistically significant (P < 0.001) in both groups [Table 1], [Figure 1].

The mean GI for Groups B and C patients before treatment was 1.43 ± 0.42, 1.77 ± 0.09, which had decreased to 0.65 ± 0.36 and 1.12 ± 0.13 after the treatment. Thus, there was a mean reduction of 0.78 ± 0.18, which was statistically significant (P < 0.001) [Table 1], [Figure 1].

The mean PI (Russell's) for Group B and C patients before treatment was 1.58 ± 0.31 and 5.84 ± 0.32, which had decreased to 1.09 ± 0.32 and 5.74 ± 0.28 after the treatment. However, the mean reduction was statistically significant in Group B (P < 0.001), but not statistically significant in Group C (P = 0.125) [Table 1], [Figure 1].

Discussion

Secretory immunoglobulin A (sIgA) constitutes the predominant immunoglobulin isotype in secretions including saliva. It is considered to be the principal line of defense of the host against pathogens, which inhabit or invade the surfaces inundated by external secretions. ^[5],[6] The foremost function of sIgA antibodies seems to be to frontier microbial adherence as well as permeations of foreign antigens into the mucosa. ^[7]

Naon et al.^[8] and Gronbald ^[9] also physiological stimulation has led to a decrease in salivary IgA concentration as salivary IgA is produced locally depending on selective transport and as such is not amplified due to release from local sites during stimulation. Present study included merely male patients as there could be an alteration in the rate of salivary flow in both genders. Bergdahl ^[10] reported differences in salivary flow between genders. Both unstimulated and stimulated salivary flow rates were significantly greater in men than in women. Patients selected for the present study were between 20 and 50 age group, excluding those above 50 years of age. Challacombe et al.^[11] and Miletic et al.^[12] demonstrated a significant reduction in the concentration and secretion rates of salivary IgA in the elderly people (60-80 age group) than in the young.

Henskens et al.^[13] and Hagewald et al.^[14] estimated levels of salivary IgA before and after SRP and found significant alteration in concentration. The patients enrolled in the study were reassessed 4 weeks after phase I periodontal therapy and the clinical parameters as well as the levels of salivary IgA were evaluated as evaluation of clinical changes occurring in the periodontal tissues following non-surgical therapy should be performed not earlier than 4 weeks Greenstein. ^[15] However, Henskens et al.^[13] and Darby et al. ^[16] reviewed the patients 6 weeks after the last clinical intervention. Levels of salivary IgA in gingivitis and periodontitis patients were found to be significantly higher when related to healthy controls in the present study.

Similarly, Demetriou et al.^[17] and Guven et al.^[18] also reported elevated levels of salivary IgA in periodontal disease. The higher levels of salivary IgA could be ascribed to the local production of IgA from the immunocytes of salivary glands, gingiva and also contribution from serum due to the antigenic stimulation by periodontopathogenic bacteria. Present study revealed a significant decrease in salivary IgA levels following phase I periodontal therapy in both gingivitis and periodontitis patients, which was in accordance with the studies of Reiff. ^[19]

Conclusion

Our study showed increased levels of salivary IgA in patients with periodontal disease compared with healthy controls. The less severe the periodontal involvement, the more consistent was the reduction in IgA concentration following phase I periodontal therapy. Further, longitudinal studies with a larger study population, with definitive treatment procedures like periodontal surgery and advanced immunological techniques investigating the concentration, specificity and avidity of antibodies directed to periodontal bacterial components must be carried out if relationships to risk, disease activity or prognosis are to be established.

References

1.	Snyderman R. The role of the immune response in the development of periodontal disease. Int Dent J 1973;23:310-6.
2.	Haffajee AD, Socransky SS. Attachment level changes in destructive periodontal diseases. J Clin Periodontol 1986;13:461-75.
3.	Greenstein G, Caton J. Periodontal disease activity: A critical assessment. J Periodontol 1990;61:543-52.
4.	Wilton JM, Curtis MA, Gillett IR, Griffiths GS, Maiden MF, Sterne JA, et al. Detection of high-risk groups and individuals for periodontal diseases: Laboratory markers from analysis of saliva. J Clin Periodontol 1989;16:475-83.
5.	Brandtzaeg P. Immunology of inflammatory periodontal lesions. Int Dent J 1973;23:438-54.
6.	Marcotte H, Lavoie MC. Oral microbial ecology and the role of salivary immunoglobulin A. Microbiol Mol Biol Rev 1998;62:71-109.
7.	Kerr MA. The structure and function of human IgA. Biochem J 1990;271:285-96.
8.	Ben-Aryeh H, Naon H, Szargel R, Horowitz G, Gutman D. The concentration of salivary IgA in whole and parotid saliva and the effect of stimulation. Int J Oral Maxillofac Surg 1986;15:81-4.
9.	Grönblad EA. Concentration of immunoglobulins in human whole saliva: Effect of physiological stimulation. Acta Odontol Scand 1982;40:87-95.
10.	Bergdahl M. Salivary flow and oral complaints in adult dental patients. Community Dent Oral Epidemiol 2000;28:59-66.
11.	Challacombe SJ, Percival RS, Marsh PD. Age-related changes in immunoglobulin isotypes in whole and parotid saliva and serum in healthy individuals. Oral Microbiol Immunol 1995;10:202-7.
12.	Miletic ID, Schiffman SS, Miletic VD, Sattely-Miller EA. Salivary IgA secretion rate in young and elderly persons. Physiol Behav 1996;60:243-8.
13.	Henskens YM, van der Weijden FA, van den Keijbus PA, Veerman EC, Timmerman MF, van der Velden U, et al. Effect of periodontal treatment on the protein composition of whole and parotid saliva. J Periodontol 1996;67:205-12.
14.	Hägewald SJ, Fishel DL, Christan CE, Bernimoulin JP, Kage A. Salivary IgA in response to periodontal treatment. Eur J Oral Sci 2003;111:203-8.
15.	Greenstein G. Periodontal response to mechanical non-surgical therapy: A review. J Periodontol 1992;63:118-30.
16.	Darby IB, Mooney J, Kinane DF. Changes in subgingival microflora and humoral immune response following periodontal therapy. J Clin Periodontol 2001;28:796-805.
17.	Demetriou N, Drikos G, Bambionitakis A. Relation between gingival fluid and mixed and parotid salivary IgA. J Periodontol 1978;49:64-6.
18.	Güven O, De Visscher JG. Salivary IgA in periodontal disease. J Periodontol 1982;53:334-5.
19.	Reiff RL. Serum and salivary IgG and IgA response to initial preparation therapy. J Periodontol 1984;55:299-305.

Figures

[Figure 1], [Figure 2]

Tables

[Table 1], [Table 2], [Table 3]

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