Tissue-engineered grafts using a scaffold can be used in treating periodontal disease; however, previous scaffolds for the cultivations of the periodontal ligament cells have been structurally incompatible with the morphological requirements of human periodontal tissue. Here, we describe a self-organized honeycomb-patterned film (honeycomb film) that acted as an appropriate scaffold for periodontal tissue regeneration. The honeycomb films were prepared from biodegradable poly(ε-caprolactone) with highly regular three-dimensional micropatterned surface topography by casting a polymer solution of water-immiscible solvent under humid conditions. To evaluate its performance in activating the proliferation and organizing of cells, we have demonstrated specific behaviors of the cultured periodontal ligament cells on the self-organized honeycomb structures in vitro. Fibroblast-like cells derived from the periodontal ligament of extracted human molar teeth were cultivated on three types of honeycomb films with 5-, 10-, and 15-µm pore sizes for 4 h to 42 d. Morphological observation of the cultured tissues at 4—72 h revealed that the pseudopodiums of cell bodies were attached to the pillars in the honeycomb structure. A certain number of cells shifted their cell bodies into the honeycomb structural lumen through the oscula of 10- and 15-µm pores. After 28 and 42 d, the cells were observed to have formed multiple layers; further, each cell had penetrated through the 10- and 15-µm pores in the honeycomb film. The morphological examination of the honeycomb film along with the pillar structures revealed that the scaffold was clusteringly arrayed with interconnected structures, remarkably enhanced proliferation, and extension of the cultured cells. We consider that the film can be applied in periodontal therapy for use as a scaffold for periodontal tissue regeneration.
