We report the use of optical coherence tomography (OCT) to detect and quantify demineralization process induced by S. mutans biofilm in third molars human teeth. Artificial lesions were induced by a S. mutans microbiological culture and the samples (N = 50) were divided into groups according to the demineralization time: 3, 5, 7, 9, and 11 days. The OCT system was implemented using a light source delivering an average power of 96 μW in the sample arm, and spectral characteristics allowing 23 μm of axial resolution. The images were produced with lateral scans step of 10 μm and analyzed individually. As a result of the evaluation of theses images, lesion depth was calculated as function of demineralization time. The depth of the lesion in the root dentine increased from 70 μm to 230 μm (corrected by the enamel refraction index, 1.62 @ 856 nm), depending of exposure time. The lesion depth in root dentine was correlated to demineralization time, showing that it follows a geometrical progression like a bacteria growth law.