Development of hydroxyapatite from eggshell waste and a chitosan-based composite: In vitro behavior of human osteoblast-like cell (Saos-2) cultures

Hydroxyapatite was successfully synthesized using eggshell waste as a raw material. Eggshell waste and orthophosphoric acid were co-precipitated for 2 h at an ambient temperature. The pH of the solution was adjusted to 10 using ammonium hydroxide. Then, 10-30 wt% of hydroxyapatite was loaded into the chitosan film. Scanning electron microscopy and energy dispersive analyses confirmed the morphological properties and dispersion. A thermogravimetric analysis showed a significant enhancement in the thermal stability of the composite. The existence of hydroxyapatite resulted in a higher thermal stability. Furthermore, atomic force microscopy was used to investigate the roughness of the surface. With the addition of hydroxyapatite, the roughness significantly increased. The swelling behavior of the composite was observed in phosphate buffer saline solution. The hydroxyapatite offered the inferiority on the swelling behavior. A preliminary investigation on the in vitro behavior of Saos-2 was also performed. The composite presented good cytotoxicity, and thus, excellent properties as a tissue engineering material.