Due to the limited supply of and numerous potential complications associated with current bone grafting materials, a tremendous clinical need exists for alternative biologically active implant materials capable of promoting bone regeneration in orthopaedic applications. Recent advances in tissue engineering technology have enabled the coating of biologically inactive materials, such as titanium fiber meshes, with a biologically active bone-like extracellular matrix produced by mesenchymal stem cells during in vitro culture. The resulting constructs can then be implanted as acellular scaffolds or as transplantation vehicles for mesenchymal stem cell populations to guide bone tissue regeneration in vivo. Such a novel tissue engineering strategy marks a paradigmatic shift in drug delivery approaches from delivering bioactive factors from a scaffold to generating constructs that contain biological signaling moieties produced by cells under engineered conditions in vitro. This chapter provides a brief introduction to general bone tissue engineering strategies and an overview of the seminal work from our laboratory in the application of mesenchymal stem cells in the in vitro generation of biologically active bone-like extracellular matrix constructs for bone tissue engineering.