This thesis reports recent innovations in stem cells, 3D printing, and layered biomaterial for the regeneration of bone, cementum, PDL, and bone-cementum-PDL complex. First, cell-homing was developed to circumvent the ethical issues associated with utilizing embryonic stem cells and reduce the risk of immunological rejection related to foreign cell transplantation. Secondly, 3D printing produced innovative scaffolds with customized biomimetic architectures and bioactive components more easily and reliably. Third, gene therapy could improve periodontal regeneration by carefully targeting the regulation of the defect of the microenvironment. Finally, combinational techniques for healing the bone-PDL-cementum system have been devised by establishing multiphasic components and cells to restore periodontal compartments synchronously, biomimetically, and entirely. The alternative techniques for fabricating fibre guiding scaffolds, such as Electrospun Fibers, Additive Manufacturing, the Soft Lithography method, and their periodontal regeneration capacity in animal models, have been reported.