Estimation of Levels of Salivary Cytokines Interleukin-6 and Tumor Necrosis Factor-α in Children with Mild Intellectual Disability After Total Oral Rehabilitation- An In Vivo Study

Children between the age group of 3-6 years of either gender with mild intellectual disability having Intelligence Quotient in the range of 55-69 as specified by DSM-5

• classification given by American Psychiatric Association with early childhood caries will be considered for the study.
• Salivary samples will be collected from all the children for baseline data.
• Total oral rehabilitation in children (including oral prophylaxis, pit and fissure sealants, fluoride application, atraumatic restorative treatment, pulp therapy, extractions) followed by diet counselling and reinforcement of oral hygiene instructions for the parents/care givers will be carried out.
• Post treatment salivary samples will be collected from the children, 1 month and 3 months after total oral rehabilitation, diet counselling and reinforcement of oral hygiene instructions.
• Diet counselling

1.Interview will be conducted and diet diary will be introduced. Parent will be asked to maintain the diary and the amount, consistency and frequency of food consumed for the 6 consecutive days will be analyzed

2.Diet modification will be  done based on the   child’s preference which includes:

a)Educate the parents about the nutritional value of the food

b) Educating parents about the role of sugar in the decay process

c)Emphasize the role of  diet sugar rich foods like cookies, candies, sugar syrups etc  in causing dental caries and limit their usage to mealtime only

d)Emphasize the importance of  unrefined foods like vegetable and fruits  and their role in maintaining the overall nutritional  and oral  health status of the child

e) Daily menu can be customized according to child’s needs, likes and preferences

3.Recall visits:

a) To reinforce the above measures

b) To motivate the parents and child so as to maintain a healthy balanced diet

4.Oral hygiene instructions to parents which include: a) Analysis of the brushing technique

b) providing knowledge about the importance of brushing twice a day

c)teaching proper tooth brushing techniques, usage of fluoridated toothpastes, mouthwashes, floss and other at home practices to maintain oral hygiene.

1)Collection of salivary samples-

• The participating children will be asked to refrain from eating and drinking for at least 1 hour prior to saliva collection.
• The participating children will be made to sit quietly with the head bent down (Coachman position) and the mouth open to allow the saliva to drip passively from the lower lip into the sterile plastic container.   For uncooperative children, saliva collection will be done by micro-pipetting method, using needleless syringes to collect unstimulated saliva or by cotton swab method.
• 2 ml of unstimulated saliva samples will be collected from the participating children early in the morning (9-11am) in a sterile container, which will be placed in a storage box at a temperature of 40 Celsius. 12
• Saliva samples will be transported to Central Research Laboratory KSHEMA. Each sample must be frozen, thawed and centrifuged at least once in order to separate the mucins by centrifugation and stored in an ultra-cold storage unit at -800 Celsius until further usage.12
• The samples will then be subjected to solid phase enzyme-linked immunosorbent      assay (ELISA) for salivary interleukin-6 & Tumor Necrosis Factor-α level

estimation. 12

 

2. Evaluation of salivary interleukin-6

 

• The Salivary ELISA kit will be used for salivary interleukin-6 evaluation as per manufacturer’s instructions .

    

3)Evaluation of salivary Tumor Necrosis Factor-α will be done by:

• Salivary Tumor Necrosis Factor-α levels will be measured with a solid phase sandwich enzyme linked immunosorbent assay (ELISA) as per manufacturer’s instructions.

**8)  METHOD OF STATISTICAL ANALYSIS:

• Data will be statistically analyzed using Shapiro Wilk test and paired t test/Wilcoxon Signed Rank Test.
• p value <0.05 will be considered statistically significant.
• Statistical software SPSS version 20 will be used to analyse the data.**