Evaluation of bioactive scaffold magnesium-containing hydroxyapatite from biomass resources for supporting osteogenesis, angiogenesis, andin vivobone regeneration

Translating artificial bone research into clinical applications requires numerous preclinical studies to evaluate the bioactivity of scaffolds in facilitating cell adhesion, proliferation, osteoblastic differentiation, and angiogenic differentiation. Additionally, developing materials with dual bioactivity to stimulate bone regeneration and vascularization remains a significant challenge. Previous research has successfully extracted hydroxyapatite (HA) from biomass sand lobster shells (HA-SLS;Panulirus homarus), which contain Mg²⁺ions that have the potential to accelerate osteoblast differentiation for bone regeneration compared with commercial HA (HA-C). The novelty of this research is the significant preclinical evaluation results of the novel HA-SLS bioactive ceramic scaffold in facilitating the adhesion and proliferation of pre-osteoblast MC3T3E1 cells, promoting human umbilical vein endothelial cells for angiogenesis, and accelerating femoral bone defect regeneration in white rats (Rattus norvegicus). The results showed that the HA-SLS scaffold could form an apatite layer in simulated body fluid, stimulate the attachment and proliferation of MC3T3E1 cells, and increase the levels of proangiogenesis biomarkers. Histology analysis of thein vivobone tissue defect expressed in mallory and hematoxylin-eosin (HE) staining showed that the percentage of collagen fibers and the number of osteoblast cells forming bone matrix after being implantation for 6 weeks in the HA-SLS group were 29.6% and 20.83% higher than those of HA-C, respectively. These results were confirmed by x-ray radiographic images, which also showed a faster reduction in defect depth in the HA-SLS group compared than in the HA-C group. These findings indicate the promising dual bioactivity of the developed HA-SLS bioceramics. However, several limitations of this study need to be addressed in future studies, including mechanical properties, long-term safety, and large-animal studies prior to translation to clinical application.