Difference between Oral Health Status, Dental Caries Experience and Salivary Biomarkers in TYPE–1 Diabetic children and non diabetic children.
- Conditions
- Type 1 diabetes mellitus without complications,
- Registration Number
- CTRI/2025/05/086675
- Lead Sponsor
- Dr Chetan Patil
- Brief Summary
**TITLE:**
**COMPARISON OF ORAL HEALTH STATUS, DENTAL CARIESEXPERIENCE AND SALIVARY BIOMARKERS IN TYPE 1 DIABETIC CHILDREN AND NON DIABETICCHILDREN**
**INTRODUCTION:**
**Type 1 Diabetes mellitus (T1DM), also known as autoimmunediabetes, is a chronic disease characterized by insulin deficiency due topancreatic B-cell loss and leads to hyperglycemia. Type 1 Diabetes is usually present in individuals without a family history.Only 10-15% of the patients have a first or second-degree relative with thedisease. However, the lifetime risk for developing Type 1 Diabetes Mellitus issignificantly increased in relatives of patients, as about 6% of children, 5%of siblings and 50% of monozygotic twins present the disease compared to 0.4%prevalence of the general population. People with diabetes in 1,000,000 are approximately 74,194.7 in India.According to 2021 census report, the 0-14 age group is 25.31% of the totalpopulation of India, among which, Type 1 diabetic children (0 to 14 years) in1,000,000 are approximately 126.4 in India. In India, new cases of type 1diabetes (0-14 years) in 1,000,000 are 19.2. Higher prevalence of dental diseases like dental caries, xerostomia, gingivalinflammation are reported for children with Type 1 diabetes when compared tosystemically healthy children. The present study attempted to compare the oral health status, dental cariesexperience, salivary flow rate, salivary pH, salivary glucose level, salivarysialic acid and total salivary antioxidant levels in Type 1 Diabetic Childrenand Non Diabetic Children. Reason for selecting parameters:**
- **Healthy flow of saliva is essential for maintenance of both oral and general health. It provides a cleansing effect. Saliva may constitute a first line of defense against free radical mediated oxidative stress. Salivary flow is significantly reduced in type 1 diabetic children.**
- **Serum sialic acid levels are found to be raised in storage diseases, cardiovascular diseases and cancers. Free sialic acid levels are raised in type 1 diabetes.**
- **Harmful effects of oxygen are due to formation of Reactive Oxygen Species in diseased states. To protect us against the oxidizing action of free radicals, every individual has an army of antioxidants which fight against free radicals. Thus in any diseased state, free radicals produced are quite higher. Type 1 Diabetes is one such diseased state in which free radicals are increased significantly.**
- **The chronic hyperglycemia of diabetes is associated with long-term damage, dysfunction of different organs and impaired salivary gland functions leading to changes in saliva composition. In type 1 diabetes, salivary glucose levels get elevated. Increasing the level of glucose in saliva affects the activity of microorganisms.**
- **The Salivary Flow Rate, Salivary pH, Salivary Glucose, Salivary Sialic Acid, Salivary Total Anti-Oxidants Levels may be altered in Type 1 Diabetic patients and due to which, poor oral hygiene and increased dental caries experience may be seen.**
**Since diabetes has an influence on oral health, it isimportant for the dentist to be aware of newer advances in the field ofdiabetes and to recognize specific oral problems related to diabetes. Thus, thedentist becomes an important part of the healthcare team for the patients withdiabetes. This study will help us to prevent deterioration of oral healthstatus in Type 1 Diabetic Children by taking appropriate measures to caries andperiodontal diseases. Many researchers have tried to compare dental caries experience or gingivalhealth status in Type 1 Diabetic Children and Non Diabetic Children but thereis still a gap of knowledge to associate changes in salivary composition orsalivary biomarkers to increased oral health problems in Type 1 Diabetic Children.Also, the association of diabetes and dental caries has received much lessattention and the results have been controversial. Most studies arecross-sectional and show either higher, similar or lower caries prevalenceamong diabetic than among the controls. This study includes evaluation of bothchanges in salivary composition or salivary biomarkers and deteriorated oralhealth of Type 1 Diabetic Children.**
**OBJECTIVES:**
1. **To evaluate Oral Health Status, Dental Caries Experience, Salivary Flow Rate, Salivary pH, Salivary Glucose, Salivary Sialic Acid and Salivary Total Antioxidant Capacity in Type 1 Diabetic children and Non Diabetic children.**
2. **To compare Oral Health Status, Dental Caries Experience, Salivary Flow Rate, Salivary pH, Salivary Glucose, Salivary Sialic Acid and Salivary Total Antioxidants Capacity in Type 1 Diabetic children with Non Diabetic children.**
**RESEARCH QUESTION: Is there a difference between OralHealth Status, Dental Caries Experience, Salivary Biomarkers such as SalivaryFlow Rate, Salivary pH, Salivary Glucose, Salivary Sialic Acid, Salivary TotalAntioxidant Capacity in Type 1 Diabetic children and Non Diabetic children?**
**RESEARCH HYPOTHESIS:**
- **Null hypothesis: There is no significant difference in Oral Health Status, Dental Caries Experience, Salivary Biomarkers such as Salivary Flow Rate, Salivary pH, Salivary Glucose, Salivary Sialic Acid, Salivary Total Antioxidant Capacity in Type 1 Diabetic children when compared to Non Diabetic children.**
- **Alternative hypothesis: There is a significant difference in Oral Health Status, Dental Caries Experience, Salivary Biomarkers such as Salivary Flow Rate, Salivary pH, Salivary Glucose, Salivary Sialic Acid, Salivary Total Antioxidant Capacity in Type 1 Diabetic children when compared to Non Diabetic children.**
**METHODOLOGY:**
- **STUDY TYPE: Analytical Study**
- **STUDY DESIGN: Case Control Study**
- **SAMPLE SIZE: 50 Case (Type 1 Diabetic children) and 50 Control (Non Diabetic children)**
**Group 1 (Study Group): 50 Case (Type 1 Diabetic children)(5 to 12 years) Group 2 (Control Group): 50 Control (Non Diabetic children) (5 to 12 years)**
**INCLUSION CRITERIA FOR CASE:**
- **Type 1 diabetic diagnosed children.**
- **Age group of 5 to 12 years.**
- **Male and female children.**
- **Children whose guardians are providing informed consent.**
**EXCLUSION CRITERIA FOR CASE:**
- **Presence of other systemic disease/s.**
- **Patient with cognitive impairment.**
**INCLUSION CRITERIA FOR CONTROL:**
- **Non diabetic children.**
- **Age group of 5 to 12 years.**
- **Male and female children.**
- **Children whose guardians are providing informed consent.**
**EXCLUSION CRITERIA FOR CONTROL:**
- **Presence of other systemic disease/s.**
- **Patient with cognitive impairment.**
**MATERIALS TO BE USED:**
1. **Disposable gloves**
2. **Disposable Mask**
3. **Protective eyewear**
4. **Plane mouth mirror – no.5**
5. **Williams graduated probe – no.15/734**
6. **Explorer – no.23 shepherd’s hook**
7. **Modified Who oral health assessment form 4**
8. **Eppendorf tube for Saliva collection**
9. **Measuring cylinder for salivary flow rate measurement**
10. **pH meter**
11. **Test tubes**
12. **Pipettes**
13. **Graph papers**
14. **Glass Beakers**
15. **Distilled water**
**Chemicals for tests:**
- **Salivary glucose estimation:**
- **3,5-Dinitrosalicylic acid**
- **Sodium hydroxide**
- **Sodium potassium tartarate**
- **Standard glucose**
- **Estimation salivary total antioxidant capacity:**
- **Ammonium molybdate**
- **Sodium phosphate**
- **Sulfuric acid**
- **Ascorbic acid (vit C)**
- **Alpha – tocopherol (vit A)**
- **Salivary sialic acid estimation:**
- **Ninhydrin Reagent**
- **Glacial acetic acid**
- **Acetone**
- **Sulfuric acid**
**Study Conduct: Information Dissemination: Information brochures will be provided to theparents, explaining the disease and its correlation with oral health in simpleand local language.**
**Informed Consent Process: Written informed consent willbe obtained in the local language with a verbal explanation provided to thechild’s parents. The consent will be easy to understand and include informationabout clinical checkups for oral health status, dental caries index, and stepsinvolved in saliva collection. Children whose parents agree to sign theinformed consent will be included in the study.**
**A thorough examination will be performed that willinclude demographic data and a complete intra-oral examination. Oral health anddental health status will be examined using the following indices:**
**DENTAL CARIES EXPERIENCE:**
Using a **Plain Mouth Mirror no. 5** and **Explorer no.23**, dental caries will be recorded and scored.
**Primary Teeth (Scores) | Permanent Teeth (Scores) | Status**
- A: 0 | Sound
- B: 1 | Caries
- C: 2 | Filled with caries
- D: 3 | Filled, no caries
- E: 4 | Missing due to caries
- -: 5 | Missing for other reasons
- F: 6 | Fissure Sealant
- G: 7 | Fixed Dental Prosthesis
- -: 8 | Unerupted
- -: 9 | Not Recorded
**DMFT** (Decayed, Missing, Filled Teeth) is deriveddirectly from the data in the boxes:
- **D** component: Teeth coded 1 or 2 (B or C).
- **M** component: Teeth coded 4 or 5 (E).
- **F** component: Teeth coded 3 or 6 (D or F).
**DMFT Calculation:** DMFT = D + M + F.
---
**ORAL HEALTH STATUS:**
Using **Williams Graduated Probe no. 15/734**, oralhealth status will be recorded and scored.
**Gingival Bleeding Scores:**
- 0: Absence of bleeding
- 1: Presence of bleeding
- 9: Tooth Excluded
- X: Tooth not present
**Pocket Score:**
- 0: Absence of condition
- 1: Pocket 4-5 mm
- 2: Pocket 6 mm or more
- 9: Tooth excluded
- X: Tooth not present
---
**Collection of Saliva:**
Saliva will be collected in the **morning hours (8 am to 10am)**, considering circadian rhythms. The patient will be instructed not toeat or drink anything **1 hour before** collection.
- **Unstimulated saliva (5 ml)** will be collected in a **funnel** inserted into a beaker, allowing the patient to drool passively.
- After collection, the saliva will be transferred into **plastic containers**, labeled, sealed, and transported in a **biohazardous bag** to prevent contamination.
---
**A) Estimation of Salivary Flow Rate:**
The **unstimulated salivary flow rate (USF)** will becalculated using the formula:
**USF = Total volume of collected saliva / Time period forcollection of saliva.**
The saliva will be stored at **-80 C** in a deep freezerat the College of Biosciences and Technology, Loni.
---
**B) Salivary pH:**
1. Salivary pH will be measured using a **pH meter** (Thermo Orion).
2. **1 ml of saliva** will be transferred to a glass beaker.
3. The tip of the pH meter will be dipped into the saliva.
4. The **pH value** will be displayed on the meter.
**Interpretation of Results:**
- pH 0 to 6.9: Acidic
- pH 7: Neutral
- pH above 7: Basic
---
**C) Salivary Sialic Acid:**
1. **Sialic acid** will react with **ninhydrin** in an acidic medium to form a colored product, which can be measured **spectrophotometrically** at **470 nm**.
2. A precipitate will form after **ethanol treatment**. To this precipitate, 1.0 ml of distilled water, 1.0 ml of glacial acetic acid, and 1.0 ml of acidic ninhydrin reagent will be added.
3. The reaction mixture will be heated for **10 minutes** in a boiling water bath. After cooling, absorbance will be measured at **470 nm**.
4. A **standard sialic acid curve** will be generated for comparison.
---
**D) Salivary Total Antioxidant Capacity:**
1. An aliquot of **0.1 ml of saliva** will be combined with **1 ml of a reagent solution** containing **0.6 M sulfuric acid**, **28 mM sodium phosphate**, and **4 mM ammonium molybdate**.
2. The tube will be incubated at **95 C** for **90 minutes**.
3. After cooling, the absorbance will be measured at **695 nm**.
4. The assay is based on the reduction of **phosphate-Mo (VI)** to **phosphate Mo (V)** and the subsequent formation of a **bluish-green color** complex.
---
**E) Salivary Glucose Test:**
1. **DNSA Method** will be used to estimate reducing sugars.
2. A standard curve will be created using **standard solutions** and the test sample will be compared.
3. The **optical density (O.D.)** at **540 nm** will be measured.
4. The glucose concentration will be determined using the standard graph.
---
**Statistical Analysis:**
- Data will be analyzed using **SPSS software v26.0**.
- A significance level of **5%** will be considered.
- **Normality tests** (Kolmogorov-Smirnov) will be performed. Parametric tests will be applied to normally distributed data, and non-parametric tests will be applied to skewed data.
- Demographic characteristics will be presented using **descriptive statistics**.
- Comparisons will be made using **independent t-tests** or **Mann-Whitney tests** for salivary biomarkers and other measures.
- **Chi-square tests** will be used for comparing dental caries, gingival bleeding, and pocket scores.
---
**Amendment of Protocol:**
No changes will be made to the study procedures withoutmutual agreement from the investigator, physician, and ethical committee.
---
**Confidentiality:**
Patient identity will be kept confidential. Data will onlybe available to the investigator and regulatory authorities, with any breach ofconfidentiality reviewed by the investigator and ethical committee.
---
**Ethical Considerations:**
- Ethical approval will be obtained before commencing the study from the institutional ethical committee.
- Informed consent will be obtained from all participants and their guardians.
---
**Implications:**
This study aims to enhance the **oral health care** ofType 1 Diabetic children by raising awareness of the connection between **salivarybiomarkers** and **oral health**. Improved dental care will contribute tobetter overall health for these children.
---
**References:**
1. Katsarou A, Gudbjörnsdottir S, Rawshani A et al. Type 1 diabetes mellitus. Nat Rev Dis Primers. 2017 March.
2. Paschou SA, et al. On type 1 diabetes mellitus pathogenesis. Endocr Connect. 2018 Jan;7(1):R38-R46.
3. IDF Diabetes Atlas 2021.
4. Rai K, Hegde AM, et al. Dental caries and salivary alterations in Type I Diabetes. J Clin Pediatr Dent. 2011 Winter;36(2):181-4.
5. Hegde AM, Joshi S, et al. Evaluation of oral hygiene status in acute lymphoblastic leukemic (ALL) children. J Clin Pediatr Dent. 2011 Spring;35(3):319-23.
6. Joshi S, Hegde AM, et al. Evaluation of salivary sialic acid levels in acute lymphoblastic leukemic children. J Clin Pediatr Dent. 2013 Spring;37(3):309-13.
7. Kenzaburoh Yao, Toshihiko Ubuka, et al. Direct determination of bound sialic acids. Analytical Biochemistry. 1989.
8. Prieto P, Pineda M, et al. Spectrophotometric quantitation of antioxidant capacity. Anal Biochem. 1999 May 1;269(2):337-41.
- Detailed Description
Not available
Recruitment & Eligibility
- Status
- Not Yet Recruiting
- Sex
- All
- Target Recruitment
- 50
- Type 1 diabetic diagnosed children.
- Age group of 5 to 12 years.
- Male and female children.
- Children whose guardians are providing informed consent.
- Presence of other systemic disease/s.
- Patient with cognitive impairment.
Study & Design
- Study Type
- Observational
- Study Design
- Not specified
- Primary Outcome Measures
Name Time Method The Type 1 Diabetic patients and their parents are preoccupied with their life-threatening problems arising from the disease and often neglecting the basic preventive dental care. This study would help the patients to take proper care of their oral hygiene as neglection of this may add to existing complications. 1 year Relation between salivary biomarkers and oral health status in Type 1 Diabetic children will help us in preventive dental care to avoid further severe oral health problems. 1 year
- Secondary Outcome Measures
Name Time Method
Trial Locations
- Locations (1)
Department of Pediatric and Preventive Dentistry,RDC,Loni
🇮🇳Ahmadnagar, MAHARASHTRA, India
Department of Pediatric and Preventive Dentistry,RDC,Loni🇮🇳Ahmadnagar, MAHARASHTRA, IndiaDrChetan PatilPrincipal investigator9370188635cp397654@gmail.com