This prospective clinical study aims to evaluate the association between the levels of selected pulpal biomarkers and the clinical success of vital pulp therapy in permanent teeth. Patients undergoing vital pulp therapy will be monitored over time, and biomarker expression levels will be analyzed in relation to treatment outcomes. The study intends to provide insight into the predictive value of pulp tissue biomarkers for long-term success in vital pulp procedures.

The aim of this study is to identify the biomarkers that influence the success of vital pulp therapy (VPT), a treatment that offers several advantages over conventional root canal therapy. The study also aims to evaluate the success of VPT in teeth diagnosed with irreversible pulpitis. The success of VPT largely depends on the accurate assessment of pulpal status.

This prospective clinical study will be conducted on 50 patients who apply to the Department of Endodontics, Faculty of Dentistry, Atatürk University, and meet the inclusion criteria. Informed consent will be obtained from all participants. A specially designed clinical assessment form will be completed for each patient to record clinical findings prior to the procedure.

Local anesthesia will be administered using 1.8 mL of 4% articaine with 1:100,000 epinephrine. Following anesthesia, the teeth will be isolated using a rubber dam. Caries removal will be performed under an operating microscope (OPMI) using a diamond bur and aerator. All infected dentin will be removed selectively until hard dentin is reached.

Pulpal bleeding will be controlled by applying a cotton pellet moistened with sterile saline into the cavity for one minute. Subsequently, pulpal fluid and blood samples will be collected using 5 mm-long sterile paper points to ensure equal sample volume per case. Each sample (one per patient) will be transferred into separate sterile Eppendorf tubes containing 500 µL of phosphate-buffered saline (PBS) and stored at -80°C until biochemical analysis. Total bleeding time will be recorded using a chronometer.

The first tube will be used for the analysis of mediators including RANTES (CCL5), MCP-1, TNF-α, TGF-β, IL-1β, TIMP-1, MMP-12, and MMP-1. The second tube will be used to measure MDA, SDA, TAS, and TOS levels.

After sample collection, the exposed pulp will be disinfected with a cotton pellet soaked in 2.5% sodium hypochlorite for 30 seconds. The cavity will then be rinsed with sterile saline and dried. The exposed pulp will be capped with Mineral Trioxide Aggregate (MTA) according to the manufacturer's instructions, and permanent restoration of the tooth will be completed.

Patients will be recalled for follow-up evaluations at 3, 6, and 12 months, during which pulpal vitality will be assessed. A correlation will be established between biomarker levels and clinical success of the vital pulp therapy.

Inclusion Criteria:

• Permanent teeth diagnosed with reversible pulpitis
• Patients diagnosed with irreversible pulpitis but without apical radiolucency
• Teeth showing a normal response to cold testing (carbon dioxide snow)
• Teeth with irreversible pulpitis demonstrating prolonged response to cold testing
• Teeth with no sensitivity to percussion or chewing
• In teeth with exposed pulp tissue, bleeding time less than 5 minutes
• In irreversible pulpitis cases, bleeding time less than 10 minutes
• No widening of the periodontal ligament space (periapical index [PAI] ≤ 2)
• Radiographic evidence of deep dentinal caries

Exclusion Criteria:

• Teeth with a negative response to cold testing (carbon dioxide snow)
• Presence of apical radiolucency (PAI > 2)
• Condensed apical periodontitis
• Internal or external root resorption
• History of dental trauma
• Longitudinal root fracture
• Presence of periodontal-endodontic lesions on the day of treatment
• Functional loss (e.g., Grade 3 tooth mobility)
• Swelling associated with the treated tooth
• Teeth that cannot be treated under rubber dam isolation
• Teeth from which less than 2.5 mL of pulpal blood can be collected
• Immunocompromised individuals
• Pregnant women at the time of treatment
• Use of antibiotics, bisphosphonates, or statins within 4 weeks prior to treatment

Participants were blinded to their group assignment, but investigators were aware of the clinical group classification.