1. What data in particular will be shared?
   Response - Individual participant data that underlie the results reported in this article, after de-identification (text, tables, figures, and appendices).
   


2. What additional supporting information will be shared?
   Response -  Study Protocol
   Response - Informed Consent Form
   Response - Clinical Study Report
   
   
3. Who will be able to view these files?
   Response - Researchers whose proposed use of the data has been approved by an independent review committee identified for this purpose.
   


4. For what types of analyses will this data be available?
   Response - To achieve aims in the approved proposal.
   


5. By what mechanism will data be made available?
   Response - Proposals should be directed to [sudhayadav2788@gmail.com].
   


6. For how long will this data be available start date provided 27-12-2027 and end date provided 27-12-2030?
   Response - Beginning 3 months and ending 5 years following article publication.
   


7. Any URL or additional information regarding plan/policy for sharing IPD? 
   Additional Information - NIL

The primary objective of this study is to quantify the key inflammatory biomarkers in pulpal blood, including interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α), and establishing cut-off values that correlate with clinical diagnoses of pulp inflammation. This data will be used to develop a Point-of-Care (POC) diagnostic kit for assessing the inflammatory status of dental pulp.

 Methodology

Type of Study: 

This study is a prospective observational study which will be conducted at the Department of Conservative Dentistry & Endodontics, Maulana Azad Institute of Dental Sciences (MAIDS), and the Multidisciplinary Research Unit (MRU), Maulana Azad Medical College (MAMC), New Delhi.

Study Site: 

- Department of Conservative Dentistry & Endodontics, MAIDS: Responsible for collecting pulpal blood samples from patients diagnosed with symptomatic irreversible pulpitis

- Multidisciplinary Research Unit, Department of Biochemistry, MAMC: Responsible for biomarker analysis and data interpretation.

Sample Size: 

Based on previous studies, the expected area under the ROC curve is 0.85. To achieve a precision of ±0.05 at a 95% confidence level, a total sample size of 228 subjects  is required.

Eligibility Criteria:

Inclusion Criteria

The study will include patients aged 18 -55 years with symptomatic irreversible pulpitis, characterized by deep carious lesions and symptoms such as continuous moderate to severe pain, which may be spontaneous or provoked by stimuli. Pulpal health status will be confirmed through clinical examinations and pulp sensibility tests. 

Exclusion Criteria

Exclusion criteria for the study include non-responsive pulp in sensibility tests, non-restorable teeth, significant periodontal disease, or the presence of sinus tracts. Additionally, patients with systemic diseases, pregnant women, and those who have recently taken NSAIDs or antibiotics will be excluded. Radiographic exclusion criteria include periapical pathology, internal or external resorption, cracked teeth, and immature teeth with open apices.

Sample Collection (Phase I): 

Patients presenting with symptomatic irreversible pulpitis and healthy controls indicated for root canal treatment or vital pulp treatment will be enrolled.

Sample Collection: 

Local anesthesia will be administered and the tooth will be isolated using dental dam. After complete caries removal and subsequent pulp exposure, pulpal blood sample will be collected from exposed pulp. 100μL of blood from the exposed surface of the pulp will be collected with a micropipette and transferred to an Eppendorf cup. The samples will be transported using cool packs at -25 ºC to the Department of Biochemistry, where they will be centrifuged for 1.000 x g for 1 min and stored at -80ºC until later laboratory processing.

 The teeth will undergo root canal treatment following the collection of pulpal blood, following a standardized treatment procedure. To outline the process briefly, once the access cavity is prepared, the working length will be determined using an apex locator (Root ZX, J Morita, Corp, Tokyo, Japan) and confirmed through radiography (Kodak). Biomechanical preparation will be carried out using Protaper Gold rotary files (Dentsply Tulsa, Tulsa, OK, USA) in conjunction with a 5.25% sodium hypochlorite irrigation solution. Final irrigation will be performed using 17% EDTA, followed by a sterile saline solution. The canals will be dried using paper points and obturated with a warm vertical obturation technique.

Biomarker Analysis: 

Quantification of biomarkers will be done. The analysis will focus on inflammatory markers such as IL-6, IL-8, TNF-α, and additional markers identified through literature review. The concentrations of these biomarkers will be converted to absolute values using standard curves derived from known concentrations.

Data Analysis: 

Statistical analysis will be performed using SPSS software. Key metrics will include sensitivity, specificity, and the area under the ROC curve. The analysis will establish cut-off values for biomarkers, aiding in the differentiation between healthy and inflamed pulp tissues.

 Outcome Measures:

·         Quantification of key inflammatory bio-markers in pulpal blood, including but not limited to interleukin-6, 8 (IL-6, 8), and tumor necrosis factor-alpha (TNF-α).

·         Measure the sensitivity, specificity, Positive Predictive Value, Negative Predictive Value of these inflammatory biomarkers

·         Establishment of cut-off values for each inflammatory bio-marker individually and in combination.

Based on the data obtained in Phase I, a Point of care diagnostic kit will be developed.