PCL-coated hydroxyapatite scaffold derived from cuttlefish bone: In vitro cell culture studies

• Hydroxyapatite/poly(ε-caprolactone) scaffold with interconnected pores was prepared
• Cytotoxicity test showed that the scaffold was not cytotoxic towards MC3T3-E1 cells
• The scaffold supported the attachment, proliferation and differentiation of cells
• A 3D cell colonization was confirmed using the fluorescence microscopy
• The scaffold might be a promising candidate for bone tissue engineering

In the present study, we examined the potential of using highly porous poly(ε-caprolactone) (PCL)-coated hydroxyapatite (HAp) scaffold derived from cuttlefish bone for bone tissue engineering applications. The cell culture studies were performed in vitro with preosteoblastic MC3T3-E1 cells in static culture conditions. Comparisons were made with uncoated HAp scaffold. The attachment and spreading of preosteoblasts on scaffolds were observed by Live/Dead staining Kit. The cells grown on the HAp/PCL composite scaffold exhibited greater spreading than cells grown on the HAp scaffold. DNA quantification and scanning electron microscopy (SEM) confirmed a good proliferation of cells on the scaffolds. DNA content on the HAp/PCL scaffold was significantly higher compared to porous HAp scaffolds. The amount of collagen synthesis was determined using a hydroxyproline assay. The osteoblastic differentiation of the cells was evaluated by determining alkaline phosphatase (ALP) activity and collagen type I secretion. Furthermore, cell spreading and cell proliferation within scaffolds were observed using a fluorescence microscope.