Fabrication and in vitro cytocompatibility evaluation of porous bone scaffold based on cuttlefish bone-derived nano-carbonated hydroxyapatite reinforced with polyethylene oxide/chitosan fibrous structure

Abstract

A novel porous bone scaffold based on nano-carbonated hydroxyapatite reinforced with fibrous-like structured polyethylene oxide/chitosan network (nCHA/PEO/CS) was introduced and fabricated via freeze-drying. Prior to this, the nCHA was synthesized through a hydrothermal reaction based on cuttlefish bone (CFB, Sepia officinalis). The raw cuttlefish bone (raw-CFB) was first decomposed to obtain cuttlefish bone-derived calcium oxide (CaO-CFB) by calcination at 1000 °C, which was used for synthesizing nCHA. The chemical composition analysis showed that the nCHA formed AB-type CHA with a high carbonate content of 7.38 wt%, which is in the range of carbonate content in native bone (2–9 wt%). The Ca/P molar ratio of nCHA was 1.712, very close to the Ca/P of biological apatite of 1.71. Morphological analysis revealed that nCHA consists of nanosized particles, potentially offering a large surface area to volume to promote ion exchange and cell interaction. The excellent physicochemical and morphological properties of nCHA proposed suitability as a bone scaffold precursor combined with PEO and CS. The nCHA/PEO/CS scaffolds were freeze-dried with varying PEO/CS concentrations. Physicochemical analysis indicated that increasing the PEO/CS concentration decreased the crystallinity of the scaffold, causing it to be lower than the nCHA crystallinity, which may be beneficial for cell growth. Morphological analysis revealed that the scaffold structure comprised nCHA cross-linked within a fibrous-like structured PEO/CS network, which appropriately mimics the fibrous structure of extracellular matrix (ECM) in natural bone. However, the nCHA/PEO/CS-11 scaffold formed more appropriate pores with suitable porosity for cell development, blood vessel formation, and nutrient perfusion. The nCHA/PEO/CS-11 scaffold also demonstrated sufficient compressive strength and good swelling behavior, which may favor bone regeneration. The nCHA/PEO/CS-11 scaffold demonstrated high cytocompatibility and facilitated the adherence of MC3T3E1 cells on the scaffold surface. The nCHA/PEO/CS-11 scaffold also promoted cell osteogenic differentiation. Owing to its desirable and suitable characteristics, the nCHA/PEO/CS-11 scaffold is promising in bone tissue engineering.