Achieving thorough disinfection is regarded as one of the pillars in endodontics. Although calcium hydroxide (CH) is one of the routinely used intracanal medicament in endodontics; alternative approaches are gaining popularity to mitigate endodontic pathology. However, CH has to be tested for its dissociation which is a rate-limiting attribute essential for its therapeutic action. The dissociation of CH into OH- and Ca2+ depends on the vehicle used to prepare the paste. This in-vitro study evaluated the use of ozonized olive oil in facilitating calcium ion release and change in pH when combined with CH. Fifty single rooted extracted human mandibular premolars were instrumented with NiTi rotary files (40/6). The teeth were divided into two groups (n = 25 per group) on the basis of vehicle: olive oil (CH + olive oil) and ozonized olive oil (CH + ozonized olive oil) groups. Both olive and ozonized olive oil vehicles allowed the diffusion of ions. However, pastes prepared with ozonized oil showed more ion diffusion, with marked calcium ion release after 15 days and alkalinity was maintained for complete period of 15 days, depicting better support for CH action. The change in calcium ion release and alkalinity were statistically significant in ozonized oil vehicle compared to olive oil vehicle. The present in-vitro study supports the use of ozonized olive oil as a vehicle to be used with CH as an intracanal medicament, considering its anti-microbial potential and sustainable release of calcium ions. The study was approved by the Institutional Ethical Committee of Manubhai Patel Dental College (approval No. MPDC_130/CONS-25/17) on June 4, 2018.