Head and neck cancer patients are prone to dental caries after radiotherapy. An ex-vivo study was conducted to assess the feasibility of optical coherence tomography (OCT) to detect tooth demineralization due to caries in irradiated teeth. Thirty-nine human molar teeth were subjected to caries lesion induction through irradiation (Group 1), pH cycling (Group 2-1), and both (Group 2-2). The OCT signal attenuation coefficient, µR was assessed and validated against microhardness test and scanning electron microscope (SEM). The µR for Group 1 increased from 10 Gy to 40 Gy, and subsequently decreased after irradiated to 50 Gy and 60 Gy due to damaged enamel microstructure. In Group 2-1, the µR decreased with duration of pH cycling from day 1 to day 14 due to the increase of porosity in enamel layer. However, the µR showed decreasing trend from day 14 to day 28 of pH cycling, resulted from mineral deposition in the enamel layer. Although no significant difference was found in the µR between Group 2-1 and 2-2, SEM of Group 2-2 demonstrated visually higher porosity and larger gaps between microstructures. Irradiation may accelerate caries damage to tooth microstructure by increasing its porosity and brittleness, but larger sample size may be needed to further prove the effect. OCT could potentially be used for early detection of tooth demineralization in vivo based on the measurable µR changes for all groups which are shown negatively correlated with microhardness value (p