Nature is full of many interesting things to work with, but many natural resources are also protected. In this view the recycling of aquaculture and fishery residues may lead to the manufacture of new devices and the isolation of new molecules with potential application in medicine. The aim of the present study was to explore the possibility to transform the cuttlefish bone into an hydroxyapatite scaffold suitable for bone tissue engineering application. The mixture of different lamellar porous structure of cuttlefish bone from the species Sepia Officinalis was selected and characterized, according to morphology (including porosity, surface development, surface characteristics) and mechanical properties. The material was transformed into suitable scaffold for bone tissue regeneration, trying to totally or partially convert calcium carbonate (aragonite) into calcium phosphate (hydroxyapatite HA) using hydrothermal transformation. The studies on cell attachment and proliferation (by MTT assay at different experimental times), cell morphology with Scanning Electron Microscopy (SEM), alkaline phosphatase (ALP) and osteocalcin (OC) activities and expressions by mouse osteoblast-like MC3T3-E1 cells on HA were investigated at different experimental times in cultures, in comparison with those observed on titanium specimens used as a control (ET and ST). Cell proliferation was less in HA transformed cuttlefish bone scaffolds than in ET and ST specimens. In contrast, good performance for osteoblasts differentiation was observed on HA transformed cuttlefish bone scaffolds, similar to those observed onto titanium scaffolds.