The periodontium is a complex structure that is comprised of soft tissue components the gingiva and periodontal ligament, as well as hard tissue components in the form of alveolar bone and cementum. Human periodontal ligament cells (HPDLCs) and their extracellular matrix are regarded as essential components to achieve successful periodontal regeneration when treating periodontal lesions. In the present study, cell sheet technology was utilized to fabricate periodontal ligament cell sheets. These cell sheets were subsequently decellularized to isolate and preserve the extracellular component structural and functional characteristics, and were evaluated in vitro and in vivo. In all experiments that were undertaken in this study, melt electro-spun polycaprolactone (PCL) scaffolds were used as a carrier for the cell sheets, in order to support their fragile nature during the processes of decellularization. The combined cell sheet-PCL scaffold structure is referred to as a decellularized cell sheet construct.