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 -  Statistical Analysis Plan
   Response - Informed Consent Form
   
   
3. Who will be able to view these files?
   Response - Researchers who provide a methodologically sound proposal.
   


4. For what types of analyses will this data be available?
   Response - For individual participant data meta-analysis.
   


5. By what mechanism will data be made available?
   Response - To gain access, data requestors will need to sign a data access agreement. Data are available for 5 years at a third party website (aarzoodogra2000@gmail.com).
   


6. For how long will this data be available start date provided 06-10-2025 and end date provided 09-07-2026?
   Response - Immediately following publication. No end date.
   


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

The study protocol will be explained to the parent or guardian. A written informed consent will be obtained prior to initiating the treatment. A total of eighty primary teeth requiring pulpectomy will be selected from children aged 4 to 8 years and isolated using a rubber dam following the administration of local anesthesia. A sterile round bur #41 will be used to create an initial access opening. Subsequently, a sterile round bur #46, mounted on a high-speed handpiece, will be employed to further access the pulp chamber and eliminate any dentinal ledges obstructing a direct pathway to the canal orifices. On gaining the access, a sterile paper point will be inserted to obtain root canal content which will then be immediately transferred to a test tube containing transport media. The tube will be tightly sealed for the further transfer to microbiological laboratory. This will constitute the “pre-irrigant culture”. 

After collecting the “pre-irrigant culture” the teeth will be randomly and equally distributed amongst the following four groups on the basis of sequentially numbered opaque sealed technique (SNOSE).

GROUP I- (n=20) Teeth to be irrigated with Triton New All-in-one irrigating solution

GROUP II- (n=20) Teeth to be irrigated with Sodium hypochlorite (3%) irrigating solution

GROUP III- (n=20) Teeth to be irrigated with Ethylenediaminetetraacetic acid (17%) irrigating solution

GROUP IV- (n=20) Teeth to be irrigated with Normal saline (control)

The endodontic therapy including root canal irrigation will be double blinded.

On the same day, the working length determination and biomechanical preparation will be performed using files three sizes larger than the initial instrument. Throughout the instrumentation, the root canal will be intermittently irrigated with 2ml of the designated irrigant after each step: 

GROUP I: During biomechanical preparation, canal will be irrigated with Triton New All-in-one irrigating solution using a sterile syringe with side vented needle that does not bind to the canal walls. The canal will then be dried with a sterile paper point. Following this, another sample will be taken for culture in an identical manner to pre-irrigant culture method. This will constitute the “post-irrigant culture”. The samples will be transferred to a test tube and sent to the microbiology department for analysis.

GROUP II: During biomechanical preparation, canal will be irrigated with 3% Sodium hypochlorite irrigating solution using a sterile syringe with side vented needle that does not bind to the canal walls. The canal will then be dried with a sterile paper point. Following this, another sample will be taken for culture in an identical manner to pre-irrigant culture method. This will constitute the “post-irrigant culture”. The samples will be transferred to a test tube and sent to the microbiology department for analysis.

GROUP III: During biomechanical preparation canal will be irrigated with 17% EDTA irrigating solution using a sterile syringe with side vented needle that does not bind to the canal walls. The canal will then be dried with a sterile paper point. Following this, another sample will be taken for culture in an identical manner to pre-irrigant culture method. This will constitute the “post-irrigant culture”. The samples will be transferred to a test tube and sent to the microbiology department for analysis.

GROUP IV: During biomechanical preparation canal will be irrigated with Normal saline (control group) using a sterile syringe with side vented needle that does not bind to the canal walls. The canal was then dried with a sterile paper point. Following this, another sample will be taken for culture in an identical manner to pre-irrigant culture method. This will constitute the “post-irrigant culture”. The samples will be transferred to a test tube and sent to the microbiology department for analysis.

Microbiological Study: The screw-capped test tube will be vigorously shaken to ensure uniform dispersion of the sample inside. The inoculum will be obtained from the inoculated Thioglycollate Broth (HIMEDIA, Mumbai, India), and a calibrated loop with a 3.26 mm diameter, containing 0.004 mL of broth, will be used for inoculation. The sample will be streaked onto two sheep blood agar plates—one for aerobic and the other for anaerobic bacterial culture.

For the pre-irrigation sample, the aerobic culture plate will be incubated for 24 hours, while the anaerobic culture plate will be incubated for 48 hours in an anaerobic chamber with a Gas-Pak System (HIMEDIA, Mumbai, India). bacterial count will be calculated. Similarly, for the post-irrigation sample, both aerobic and anaerobic culture plates will be incubated, and bacterial colonies will be enumerated accordingly.

COLONY FORMING UNITS (CFU) – 

CFU =  n×d

    s

where  c= concentration, CFU/ml

 n= number of colonies

 d= dilution

            s= volume transferred to plate