In this work, nitrogen doped diamond like carbon (DLC) films were fabricated on silicon wafers by varying CH4–N2 flow rates in an ECR-MPCVD system. The morphology, roughness and physicochemical nature of as-deposited films were characterized using SEM, AFM and XPS. With the increase of CH4 flow rate from 5 to 20 sccm, the coverage and surface roughness of DLC film have improved, while sp2/sp3 ratio decreases. Subsequently, the in vitro osteogenesis of N doped DLC films with 20 sccm CH4 was evaluated in terms of their apatite forming ability by soaking the samples in simulated body fluid (SBF) at 37 °C. The results show a very fast formation of crystalline apatite coatings on DLC films in 28 days, compared to reported more than 12 weeks. Moreover, the water contact angle (70°), the hemolysis rate (3.63%), the relative growth rate of HEK293T cell (101.2 ∼ 127.96%) and the cytotoxicity level (0) prove its promising potential for biomedical application.