The objective of the study was to compare the frequency of contamination on used with unused (brand new) healing abutment after sterilization using Phloxine B dye and to determine the contamination on different sites of healing abutment using Phloxine B dye. A total of 60 healing abutment (Zimmer and Biohorizon) divided into equal test (unused new HA) and control groups (used HA) were evaluated for contamination after sterilization. Data was analyzed using SPSS version 22. Chi-square was used to compare the frequency of contamination between two groups. Effect modifiers like site of healing abutment and type of healing abutment were controlled through stratification. The level of significance was kept at p value ¿ 0.01. Frequency of contamination was significantly higher in test group compared to control group (p- value ¿ 0.001). Significant difference was also observed for frequency of contamination among different sites of healing abutment (p- value ¿ 0.001). The most frequently contaminated site was screw thread and screw driver engagement site, however type of healing abutment and contamination frequency showed no significant difference (p- value ¿ 0.01).