To Assess the Cell Viability of Triple Antibiotic Paste under Confocal Microscope: An In Vitro Study

Background and aim: Regenerative endodontic procedures (REPs) rely on the viability of stem cells from the apical papilla (SCAPs) to promote tissue repair and regeneration in immature teeth. Triple antibiotic paste (TAP), a widely used intracanal medicament, has shown effectiveness in eradicating root canal infections. However, its cytotoxic effects on SCAPs remain underexplored. This study aimed to assess the viability of SCAPs exposed to varying concentrations of TAP using confocal laser scanning microscopy (CLSM). Materials and methods: Twelve healthy immature premolars, extracted for orthodontic reasons, were used to isolate SCAPs. The pulp was collected, and SCAPs were cultured in α-MEM medium supplemented with fetal bovine serum (FBS), penicillin, streptomycin, and Fungisome. TAP was prepared by mixing metronidazole, ciprofloxacin, and minocycline in a 1:1:1 ratio. SCAPs were exposed to three TAP concentrations: 10 μg/mL, 25 μg/mL, and 50 μg/mL for 1, 3, and 7 days. A control group was maintained without TAP exposure. Cell viability was assessed using CLSM with the Live/Dead BacLight Kit. Results: The results showed that cell viability was concentration dependent. At 10 μg/mL, SCAPs exhibited 95% viability, while at 25 μg/mL, viability decreased to 80%, and at 50 μg/mL, only 60% of SCAPs remained viable. The control group demonstrated 100% viability, indicating no cytotoxicity without TAP exposure. Conclusion: TAP demonstrates a concentration-dependent cytotoxic effect on SCAPs, with higher concentrations reducing cell viability significantly. The findings suggest that TAP at a concentration of 10 μg/mL may be a safer choice for clinical use in regenerative endodontics, balancing antimicrobial efficacy and stem cell preservation. Clinical significance: This study highlights the importance of selecting appropriate concentrations of TAP in REPs to ensure SCAP viability. Using lower concentrations, such as 10 μg/mL, may optimize antimicrobial action while minimizing cytotoxic effects, promoting successful tissue regeneration in immature teeth.

Abstract licence: CC BY-NC

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