Objective: The estimation of time of death or the determination of the postmortem interval (PMI) is one of the most important issues in forensic medicine and odontology. However, evaluation of bone and dental hard tissues in PMI could be challenging due to the lack of objective methods with high accuracy. In this respect, micro-CT analysis which has not been used for postmortem evaluation would be beneficial in evaluating hard tissues such as bones and teeth. Therefore, the aim of the present study was to determine the alterations in the hard dental tissue, mineral density of enamel, and the surface abrasion of hard dental tissues of rats in the PMI period with a relatively novel method, micro-CT. Methods: The present study included 60 female Wistar rats which were divided into six study groups. The rats were sacrificed at the baseline and were left into nature putrefaction process. The study groups were created based on the PMI period as week—0, week—1, week—2, week—4, week—8, and week—12, which included 10 rats in each group. All hemi-mandibles were collected in the determined timelines and the micro-CT analysis was carried out on each group. Mineral density of enamel and the surface abrasion of hard dental tissues were determined. Results: The enamel and cement thickness remained the same in the examined PMI periods. Mineral density of the enamel tissues were also similar until the 8th week but the decrease was significant at 12th week (2.313 gHAp cm3). Surface abrasion of the dental tissues on weeks 4, 8, and 12 were 0.006, 0.024, and 0.024 mm, respectively. Conclusion: The present study indicated that surface abrasion and enamel mineral density evaluation via micro-CT can be considered as objective and precise parameters in PMI evaluation in forensic medicine and odontology.