ABSTRACT
Introduction:
Dental caries is a multifactorial disease caused by combined effect of host, environment, and microorganism. Mutans Streptococci (MS) is the main microorganism in causing and progression of dental caries. Advances in science and technology led to attempt in managing dental caries by targeting its initiation and delaying its progress. Immunoglobulin Y has proven efficacy in reducing the colonisation of S. Mutans by affecting the adhesion.
Aim:
Thus the present study was undertaken to evaluate the effect of passive immunisation on S.Mutans levels in caries active children aged 8-11 yrs of age.
Materials and methods:
Total of 50 children were randomly divided into IgY mouthspray (Test Group) and 0.9% Saline (Control group). Saliva sample were collected and subjected to microbial examination using MSB agar at the end of 30 days of study period.
Results:
Within the limitations, it can be concluded that IgY mouthspray significantly reduces the adhesion and growth of the causative agent S.Mutans in caries active children.
Conclusion:
Use of passive immunisation significantly reduces the bacterial overload and thus decreasing the dental caries in high risk children.
KEYWORDS: Early childhood caries, IgY, immunoglobulin, passive immunity
INTRODUCTION
Dental caries has now become a chronic disease of childhood. It poses a major public health concern for a large section of society, along with being an economic burden. It is widespread, multifactorial, and infectious in nature, affecting children of every age group with varying severity.[1]
Keyes described the etiology as a triad of substrate, host, microorganism, and time, which was later modified by Newburn. A group of acidogenic streptococcal special is found to be mainly responsible for initiation of dental caries. This is known to increase the colony of bacteria and create a virulent biofilm to produce glucan by transferase enzyme.[2]
Caries to its widespread effect on nutrition, mastication, overall growth, and self-esteem, it becomes imperative to focus on preventive measures rather than therapeutic measures. Various modalities have been tried to reduce the adhesion and multiplication of microorganisms thereby preventing and delaying the development and progression of dental caries. Passive immunization using immunoglobulin IgY has been widely explored to prevent caries as it has been proven to interfere with the adhesion of S. mutans to enamel.
Chewable tablets of IgY has been investigated by a few researchers, however other forms of delivery has been suggested to enhance their effectiveness.[3] Thus, the study aimed to assess the effect of passive immunization on S.mutans in caries active children.
METHODOLOGY
A total of fifty children aged 8-11 were selected from residential schools in Bhubaneswar, Odisha. Institutional ethical clearance was obtained for the study. Written informed consent was obtained from the parents and head of the institution before the study intervention.
The children were randomly divided into two groups by closed envelop method each consisting of twenty-five children per group with dmft/DMFT score 3-6. Group 1 (test group) IgY mouth spray and group 2(control group) 0.9% saline mouth spray.
The children were supervised throughout the process and were advised not to drink or eat anything 1 hour prior. Resting unstimulated saliva was collected from each child in vials with baseline characteristics. Complete oral prophylaxis was performed to ensure uniformity, and children continued their daily oral hygiene routine and diet pattern during the study period. The DMFT/dmft scores were recorded by the investigator according to the WHO criteria.
Group 1 children were given IgY mouth spray and Group 2 children were given 0.9% Saline mouth spray. To avoid bias, the dispensing containers were identical and color-coded. The children were advised to take the spray every day twice after meals for 30 days, under the supervision of trained personnel at the school.
At the end of the study period, the saliva samples were collected in the vials and sent for microbiological analysis using mitis-salivarius-bacitracin (MSB) agar media for growth cultures. Results thus obtained were subjected to statistical analysis.
Microbiological assessment
Mitis-salivarius-bacitracin (MSB) agar media containing 10% sucrose and 1% potassium telluride were used to evaluate the growth count of s.mutans. The MSB growth plates were incubated for 24 hours in a 5% carbon dioxide jar at a temperature of 37°C. Colony were then quantified and expressed as CFU/ML (colony-forming –units- per milliliters)
RESULTS
The obtained data was tabulated and subjected to statistical analysis using SPSS version 22.0. The mean levels of S.mutans (CFU/ml) among the groups before and after the test intervention were assessed using Mann-Whitney Test. To compare the mean S.mutans level in each group before and after intervention, Wilcoxon signed-rank test was used. To compare the mean values of S.mutans in IgY and control group in pre- and post-intervention, based on age of children, the Krusal-Wallis test was performed. The level of significance was set at P ≤ 0.05.
Table 1 presents the distribution of the children based on age and gender among the test and control group. Table 2 shows the comparison of the mean S.mutans level between IgY (Test group) and Control group at pre- and postintervention period. The mean value of S. mutans showed significant reduction from 3.87 ± 3.1 to 1.87 ± 4.2 as compared to mean value S.mutans in control group from 3.56 ± 4.2 to 2.56 ± 4.2.
Table 1.
Age and gender distribution between IgY and Control study group
| Age | 8 | 3 (20.0) | 1 (6.7) | 0.25 |
|---|---|---|---|---|
| 9 | 2 (13.3) | 1 (6.7) | ||
| 10 | 3 (20.0) | 6 (40.0) | ||
| 11 | 7 (46.7) | 7 (46.7) | ||
| Gender | Males | 6 (40.0) | 6 (40.0) | 1.00 |
| Females | 9 (60.0) | 9 (60.0) |
Table 2.
Comparison of mean S. mutans levels (CFU/ml) between IgY and control group at pre and post-intervention period
| Mean S. mutans (CFU/ml) between IgY and 0.9% Saline group | ||||
|---|---|---|---|---|
|
| ||||
| Time | Group | n | Mean±S.D | P |
| Pre-intervention | IgY (Test ) | 15 | 3.87±3.1 | 0.44 |
| 0.9% Saline (control) | 15 | 3.56±4.2 | ||
| Post - intervention | IgY (Test) | 15 | 1.87±2.1 | 0.001* |
| 0.9% Saline (control) | 15 | 2.56±4.2 | ||
Comparing the mean S. mutans levels between the pre and post-intervention periods in IgY (Test Group) and control groups by Wilcoxon Signed Rank test revealed a significant reduction within the test group, however, statistically insignificant reduction was observed in the control group [Table 3].
Table 3.
Comparison of mean S. Mutans levels between pre – and post-intervention period in IgY (Test Group) and control groups by Wilcoxon Signed Rank test
| Mean S. Mutans (CFU/ml) between IgY and 0.9% Saline group pre and post –intervention time | |||||
|---|---|---|---|---|---|
|
| |||||
| Group | Time | n | Mean±SD | Mean difference | P |
| IgY (Test) | Pre | 15 | 3.87±3.1 | 2.09 | 0.001* |
| Post | 15 | 1.87±2.1 | |||
| 0.9% Saline (control) | Pre | 15 | 3.56±4.2 | 1.02 | 0.07 |
| Post | 15 | 2.56±4.2 | |||
DISCUSSION
In the present era with advances in science and technology, the focus remains on preventive measures than the treatment and surgical model of disease. Various researchers in cariology has proven that saliva serves as a potent agent in caries prevention and thus can be targeted, modulated, and utilized for prevention. Saliva is known to contain varying levels of immunoglobulin and enzymes such as amylase, lactoferrin, lactoperoxidase, and histantins. IgY if enhanced in the salivary concentration has shown an inhibitory effect on adhesion and growth of S.mutans and biofilm.[4,5]
The transfer of active humoral antibodies as ready-made antibodies from one individual to another is termed Passive Immunity. In recent caries research, antigen-specific IgY has gained special importance owing to its global availability and prior usage in the medical and research field for many decades. It has already been in use for treating infection in the alimentary canal.[6]
In the present study, results obtained are similar to research conducted by Ramachandra and Gopinath in 2022 and Mary and Mathew in 2020, to evaluate the effect of IgY on S.mutans in caries active children using microbiological assays.[1,7] However in the present study, the delivery agent used was in the form of mouth spray, which ensured ease of use and uniform quantity. This explained the numerically and statistically significant reduction in the levels of S.mutans in just one month period. Studies reported by Batchiar et al. and Zhou et al. showed reduction in S.mutans count but with different delivery system and periods of intervention in humans and animals.[8,9]
In an in-vitro study, Carlander et al.[10] studied the residual effect of the IgY and concluded that anti-S.mutans Immunoglobulin Y binds to the pathogen and thereby preventing its adherence and biofilm adhesion and growth. It also affects the aciduric property of S.mutans thereby inhibiting the key step of dental caries formation. This explained the greater reduction in S.mutans in the IgY test group in comparison to the isotonic saline control group.
IgY is routinely present in regular diet and derived mainly from poultry egg. This also presents minimal systemic risks and carries less chances of the development of resistance, as it doesn’t pass through the system in this intact form. However, concerns have been raised about allergic reactions from egg-derived components thus needing the preparation in water-soluble form. Also, the risk is minimal when administered in oral form.[11]
CONCLUSION
The present study presents with limitations of small sample size and shorter period. However, within the limitations, it can fairly be concluded that IgY mouth spray significantly reduces the adhesion and growth of the causative agent S.mutans in caries active children. However, larger sample size and longer duration study are needed for results to be generalized as a preventive protocol for dental caries by passive immunotherapeutic.
Conflicts of interest
There are no conflicts of interest.
Funding Statement
Nil.
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