A new highly controlled powder sintering technique was used for the fabrication of a porous Ti6Al4V scaffold. The platelet-rich plasma (PRP) was prepared using whole blood. The PRP was used as a cell carrier to inject bone marrow mesenchymal stem cells (MSC) into the pores of the Ti6Al4V scaffold in the presence of calcium chloride and thrombin, and then the composite construct of porous Ti6Al4V loaded with PRP gel and MSC was obtained. The bare Ti6Al4V scaffold and the Ti6Al4V scaffold loaded with MSC were used as controls. The characteristics and mechanical properties of the scaffold, and the biological properties of the constructs were evaluated by a series of in vitro and in vivo experiments. The results show that the sintered porous Ti6Al4V has good biocompatibility, and high porosity and large pore size, which can provide sufficient space and sufficient mechanical support for the growth of cells and bones without an obvious stress shielding effect. However, Ti6Al4V/MSC/PRP showed a significantly higher cell proliferation rate, faster bone growth speed, more bone ingrowth, and higher interfacial strength. Therefore, the porous Ti6Al4V scaffolds incorporated with MSC and PRP may be more effective at enhancing bone regeneration, and is expected to be used for bone defect repair.