Metallic porous materials are designed to allow the ingrowth of living tissue inside the pores and to improve the mechanical anchorage of the implant. In the present study, the production of highly porous Ti-6Al-4V parts by powder metallurgical technology and subsequently uses it in invitro bone tissue engineering is described. A space-holder method using carbamide to produce parts with porosities between 35 and 70% was used. Mechanical properties of parts were determined by compressive test. The porous parts are characterized by using scanning electron microscopy. Furthermore, study was demonstrated to investigate the effects of 3 different porosities of on proliferation, differentiation and cell-matrix interaction of mouse osteoblast-like cells, MC-3T3. Results showed the cell proliferation was significantly (p < 0.05) higher on 70% porous Ti-6Al-4V. However, synthesis of different types of extra cellular matrix proteins was also more abundant on 70% porous Ti-6Al-4V than 35 and 50% porous Ti-6Al-4V disk except some specific proteins. An increase in alkaline phosphate activity was significantly (p < 0.05) higher on 70 and 50% porous Ti-6Al-4V disk after 12 days of MC-3T3 cells incubation. The results indicated that porosity (nearly 70%) of porous Ti6Al-4V topography affects proliferation and differentiation of osteoblast like MC-3T3 cells. The results showed that this novel process is promise to fabricate porous biomaterials for bone implants.