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Abstract. There are many trials to regenerate one‘s defected organs or tissues by tissue engineered techniques. To date, synthetic polymers are frequently used as base materials. However, such polymers have unexpected side effects, such as cytotoxicity and genotoxicity. To solve these problems, biomaterials derived from natural resources are addressed as alternatives. We introduce the application of natural resource-derived biomaterials, such as equine bone powder and chitosan, with nanomaterials and 3D printing techniques. At first, equine bone powder and chitosan were solely used for tissue regeneration. In this research, their biocompatibilities were evaluated well. Next, their application with nanotechnologies were investigated. The application of nanomaterials, such as graphene, and electrospun nanofiber with natural resource-derived biomaterials resulted in enhanced tissue regenerations. Finally, we tried to apply natural resources into 3D printed tissue reconstruction. Under appropriate characterization, natural resource-derived biomaterials were successfully printed. Their application is expected to fulfill the limitation of conventional synthetic polymer-based therapies.