Enzyme-mediated in situ preparation of biocompatible hydrogel composites from chitosan derivative and biphasic calcium phosphate nanoparticles for bone regeneration

Thi Phuong Nguyen, Bach Hai Phuong Doan, Dinh Vu Dang, Cuu Khoa Nguyen and Ngoc Quyen Tran

Abstract

Injectable chitosan-based hydrogels have been widely studied toward biomedical applications because of their potential performance in drug/cell delivery and tissue regeneration. In this study we introduce tetronic–grafted chitosan containing tyramine moieties which have been utilized for in situ enzyme-mediated hydrogel preparation. The hydrogel can be used to load nanoparticles (NPs) of biphasic calcium phosphate (BCP), mixture of hydroxyapatite (HAp) and tricalcium phosphate (TCP), forming injectable biocomposites. The grafted copolymers were well-characterized by ¹H NMR. BCP nanoparticles were prepared by precipitation method under ultrasonic irradiation and then characterized by using x-ray powder diffraction (XRD) and scanning electron microscopy (SEM). The suspension of the copolymer and BCP nanoparticles rapidly formed hydrogel biocomposite within a few seconds of the presence of horseradish peroxidase (HRP) and hydrogen peroxide (H₂O₂). The compressive stress failure of the wet hydrogel was at 591 ± 20 KPa with the composite 10 wt% BCP loading. In vitro study using mesenchymal stem cells showed that the composites were biocompatible and cells are well-attached on the surfaces