Bone-like apatite formation in biocompatible phosphate-crosslinked bacterial cellulose-based hydrogels for bone tissue engineering applications

Abstract

Addressing major bone injuries is a challenge in bone regeneration, necessitating innovative 3D hydrogel-based therapeutic approaches to enhance scaffold properties for better bioactivity. Bacterial cellulose (BC) is an excellent scaffold for bone tissue engineering due to its biocompatibility, high porosity, substantial surface area, and remarkable mechanical strength. However, its practical application is limited due to a lack of inherent osteogenic activity and biomineralization ability. In this study, we synthesized bone-like apatite in biocompatible BC hydrogel by introducing phosphate groups. Hydrogels were prepared using fibrous BC, acrylamide (AM), and bis [2-methacryloyloxy] ethyl phosphate (BMEP) as a crosslinker through free radical polymerization (P-BC-PAM). P-BC-PAM hydrogels exhibited outstanding compressive mechanical properties, highly interconnected porous structures, good swelling, and biodegradable properties. BMEP content significantly influenced the physicochemical and biological properties of the hydrogels. Increasing BMEP content enhanced the fibrous structure, porosity from 85.1 % to 89.5 %, and compressive mechanical strength. The optimized hydrogel (2.0P-BC-PAM) displayed maximum compressive stress, toughness, and elastic modulus at 75 % strain: 221 ± 0.08 kPa, 24,674.2 ± 978 kPa, and 11 ± 0.47 kPa, respectively. P-BC-PAM hydrogels underwent biomineralization in simulated body fluid (SBF) for 14 days, forming bone-like apatite with a Ca/P ratio of 1.75, similar to hydroxyapatite. Confirmed by Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and field-emission scanning electron microscopy (FE-SEM), this suggests their potential as scaffolds for bone tissue engineering. MC3T3-E1 osteoblast cells effectively attached and proliferated on P-BC-PAM. In summary, this study contributes insights into developing phosphate-functionalized BC-based hydrogels with potential applications in bone tissue engineering.

Keywords: Bacterial cellulose; Biocompatible; Biomineralization; Bone tissue engineering; Phosphate functionality.