The proposed study shall be conducted in the Department of Conservative Dentistry & Endodontics, at Maulana Azad Institute of Dental Sciences, New Delhi on patients referred to the department for the management of their symptomatic carious permanent molar teeth.

A written informed consent (ANNEXURE II) will be obtained from all patients prior to procedure. A systematic approach of history taking, clinical and radiographic examination and pulp sensibility assessment (cold and EPT) will be done to formulate an appropriate diagnosis.

A.    Preparation of operative site

Local anaesthesia will be induced by the administration of 2% lignocaine with 1:80,000 epinephrine.  Calculus and debris will be removed from the tooth surface and subsequently isolated with a rubber dam. The tooth surface will be disinfected with gauze soaked in 5% sodium hypochlorite (NaOCl) prior to caries excavation.

Teeth that will meet the inclusion criteria will be treated with partial pulpotomy as per standardised protocol.

B.    Collection of pulpal blood sample

The cavity will be prepared using a sterile high-speed round bur under water coolant, whereas caries excavation will be performed using a slow-speed round bur. Haemorrhage on entering the pulp chamber confirmed the clinical diagnosis of a vital pulp. 100μL of blood from the exposed surface of the pulp was collected with a micropipette.

In addition, pulpal blood samples will also be collected from 10 non-carious teeth with a diagnosis of normal pulp and periapical tissue that required intentional root canal treatment for prosthetic reasons will also be  recruited in the study as controls for quantification of IL-8 in the healthy pulp.

C.    Transportation and Storage of blood sample

The pulpal blood samples will be immediately transferred from the micropipette to the Eppendorf tube

The samples will be stored at -80 ºC at the institute until it is transported to biochemistry laboratory for analysis. IL-8 assay will be done using ELISA.

A.    Partial Pulpotomy procedure

Approximately 2-3 mm of pulpal tissue underneath the exposure site will be amputated using a sterile high-speed round bur. The pulp wound will be flushed with 5ml of 2.5%-3% sodium hypochlorite and the bleeding will be controlled with a sterile cotton pellet moistened with 2.5%-3% sodium hypochlorite for 5 minutes and will be repeated if required up to 15 minutes. Time to achieve haemostasis will be recorded using a stopwatch by another investigator.

If the bleeding cannot be controlled within 15 minutes, the treated tooth will be considered a failure and excluded from the study and further treated by full pulpotomy or root canal treatment.

Biodentine will be mixed according to manufacturers’ instructions and then placed in a 2-3 mm thick layer above the pulp tissue using an amalgam carrier and gently packed using a condenser. After 12 min of waiting for initial setting, a 2-3 mm layer of RMGIC will be placed over the Biodentine and light-cured for 20 seconds. The remaining cavity will be selectively etched with 37% phosphoric acid for 30 seconds and rinsed. The bonding agent will be applied to the cavity with a microbrush brush, agitated for 20 seconds, and light-cured. The cavity will be restored with posterior composite resin and an immediate postoperative radiograph will be taken as a baseline data for further future evaluation.

B.    Follow up assessment

Patients will be called up for clinical (visual inspection, mobility testing, and pulp testing) and radiographic examination after 3 months and 6 months.

Clinical success criteria consists of :

·       Absence of spontaneous pain and pain during mastication,

·       Absence of swelling, sinus tract and fistula

·       Negative response to apical palpation and axial percussion.

Radiographic success criteria consists of:

·       No periapical or furcation changes.

·       Absence of internal and/or external root resorption.