In-vitro and in-vivo evaluation for the bio-natural Alginate/nano-Hydroxyapatite (Alg/n-HA) injectable hydrogel for critical size bone substitution

Currently, bio-natural injectable hydrogels are receiving a lot of attention due to their ability to control, adjust, and adapt to random bone defects, in addition, to their ability to mimic the composition of natural bones. From such a viewpoint, this study goal is to prepare and characterize the injectable hydrogels paste based on the natural alginate (Alg) derived from brown sea algae as a polysaccharide polymer, which coupled with nano biogenic-hydroxyapatite (n-HA) prepared from eggshells and enriched with valuable trace elements. The viscosity and mechanical properties of the paste were investigated. As well as the in-vitro study in terms of water absorption and biodegradability in the PBS, biocompatibility and the capability of the injectable Alginate/n-Hydroxyapatite (Alg/n-HA) to regenerate bone for the most suitable injectable form. The injectable hydrogel (BP -B sample) was chosen for the study as it had an appropriate setting time for injecting (13 mins), and suitable compressive strength reached 6.3 MPa. The in vivo study was also carried out including a post-surgery follow-up test of the newly formed bone (NB) in the defect area after 10 and 20 weeks using different techniques such as (SEM/EDX) and histological analysis, the density of the newly formed bone by Dual x-ray absorptiometry (DEXA), blood biochemistry and the radiology test. The results proved that the injectable hydrogels Alginate/n-Hydroxyapatite (Alg/n-HA) had an appreciated biodegradability and bioactivity, which allow the progress of angiogenesis, endochondral ossification, and osteogenesis throughout the defect area, which positively impacts the healing time and ensures the full restoration for the well-mature bone tissue that similar to the natural bone.