Due to the increasing number of orthopedic implantation surgery and advancements in biomaterial manufacturing, chemistry and topography, there is an increasing need of reliable and rapid methods for the preclinical investigation of osseointegration and bone ingrowth. Implant surface composition and topography increase osteogenicity, osteoinductivity, osteoconductivity and osseointegration of a prosthesis. Among the biomaterials used to manufacture an orthopedic prosthesis, titanium alloy (Ti-6Al-4V) is the most used. Type I collagen (COLL I) induces cell function, adhesion, differentiation and bone extracellular matrix component secretion and it is reported to improve osseointegration if immobilized on the alloy surface. The aim of the present study was to evaluate the feasibility of an alternative ex vivo model, developed by culturing rabbit cortical bone segments with Ti-6Al-4V alloy cylinders (Ti-POR), fabricated through the process of electron beam melting (EBM), to evaluate osseointegration. In addition, a comparison was made with Ti-POR coated with COLL I (Ti-POR-COLL) to evaluate osseointegration in terms of bone-to-implant contact (BIC) and new bone formation (nBAr/TAr) at 30, 60 and 90 d of culture. After 30 and 60 d of culture, BIC and nBAr/TAr resulted significantly higher in Ti-POR-COLL implants than in Ti-POR. No differences have been found at 90 d of culture. With the developed model it was possible to distinguish the biomaterial properties and behavior. This study defined and confirmed for the first time the validity of the alternative ex vivo method to evaluate osseointegration and that COLL I improves osseointegration and bone growth of Ti-6Al-4V fabricated through EBM.