Standard X-ray micro-computed tomography is a technique that allows a good visualization of the structure of mineralized tissues and biomaterials, but it fails to finely discern soft tissues. Here, we used X-ray synchrotron radiation pseudo-holotomography to visualize, at three-dimensional (3D) level, microvascular networks for the first time with no need for contrast agents, and to extract quantitative structural data in a bone-engineered construct implanted for 24 weeks in a mouse. When compared to standard histology, pseudo-holotomography allowed a previously unavailable 3D resolution of the vessels, which in turn appeared more clearly visible. Thus, pseudo-holotomography is an innovative technique that offers a promising powerful tool to investigate angio- and microvasculogenesis in advanced biomedical research areas such as regenerative medicine and antiangiogenic cancer therapies.