Introduction: The purpose of this study was to determine the antibacterial effect and bioactivity of triple antibiotic paste (TAP), calcium hydroxide (Ca[OH]₂), and calcium hypochlorite (Ca[OCl]₂).

Methods: Root canals were infected with 3-week-old Enterococcus faecalis biofilm and then medicated for 7 days with TAP, Ca(OH)₂, or Ca(OCl)₂ (n = 10/group). Untreated and uninfected canals were used as positive and negative controls. The antibacterial effect was determined using colony-forming units and a Live/Dead bacterial viability kit. Dental pulp stem cells were seeded on medicated dentin surfaces for 7 days. Sodium thiosulfate and various concentrations of ascorbic acid (1%, 5%, and 10%) were also used to neutralize the samples treated with Ca(OCl)₂ before cell seeding (n = 3 in triplicate). Cell viability and morphology were evaluated using a viability assay and Live/Dead cell analysis. Alkaline phosphatase (ALP) activity was also measured to determine the cells' mineralization activity.

Results: All medicaments decreased the initial bacterial load (P < .05). The highest bacterial reduction in the main canal and dentinal tubules was observed in the Ca(OCl)₂ group (P < .05). TAP- or Ca(OH)₂-treated dentin surface improved cell viability and ALP activity compared with the untreated dentin surface (P < .05), whereas Ca(OCl)₂ decreased cell viability and ALP activity (P < .05). Ten percent ascorbic acid neutralized the effect of Ca(OCl)₂ on the treated dentin surface, showing higher cell viability (P < .05) and similar ALP activity with the untreated dentin surface and the other groups (P > .05).

Conclusions: Ca(OCl)₂ medication improved root canal disinfection against E. faecalis biofilm compared with TAP and Ca(OH)₂. The adverse effects caused by Ca(OCl)₂ on cell viability and mineralization activity can be neutralized with 10% ascorbic acid.