New techniques for tissue engineering (TE) are rapidly emerging. The basic concept of autologous TE is to isolate cells from small biopsy specimens, and to expand these cells in culture for subsequent seeding onto biodegradable scaffolds. Nano-crystalline diamond (NCD) films have attracted the attention of researchers from a variety of different areas in recent years, due to their unique and exceptional properties. In this approach, human Dental Stem Cells (hDSC) were characterized by flow cytometry and grown on diamond films with hydrogen (H) and oxygen (O)-terminated surfaces for 28 days, and then removed by lysis and washing with distilled water. Energy Dispersive Spectroscopy (EDS) analysis were performed, showing that the regions with O-terminated surfaces contained much higher levels of deposited calcium, oxygen and phosphorus. These results suggest that the extra cellular matrix (ECM) was considerably more developed in the O-terminated regions, as compared with the H-terminated regions. In addition, optical microscopy of hDSC cultured on the diamond substrate with H and O-terminated surfaces, before washing with distilled water, showed preferential directions of the cells arrangement, where orthogonal lines suggest that the cells appeared to be following the O-terminated regions or hydrophilic surface. These findings suggest that oxygen-terminated diamond surface treated on biodegradable scaffold can be useful for mineralized dental tissue formation.