Lithium-incorporated deproteinized bovine bone substitute improves osteogenesis in critical-sized bone defect repair

This study aimed to explore the surface modification of deproteinized bovine bone using lithium-ion and evaluate its efficacy on osteogenesis improvement and critical-sized bone defect repair. Hydrothermal treatment was performed to produce lithium-incorporated deproteinized bovine bone. In vitro study, human osteosarcoma cell MG63 (MG63) was cultured with the bone substitute to evaluate the cell viability and then calcium deposition was measured to analyze the osteogenesis. In vivo studies, male adult goats were chosen to build critical-sized bone defect model and randomly divided into three groups. The goats were treated with autogenous cancellous bone, lithium-incorporated deproteinized bovine bone, and deproteinized bovine bone. Animals were evaluated using radiological analysis including X-ray, computed tomography, and Micro-CT; histological methods involving hematoxylin-eosin dyeing, Masson dyeing, and immunofluorescence detection at 4 and 12 weeks after surgery were carried out. According to the results, lithium-incorporated deproteinized bovine bone produced nano-structured surface layer. The lithium-incorporated deproteinized bovine bone could promote the osteoblast proliferation and increase the calcium deposition. In vivo studies, radiographic results revealed that lithium-incorporated deproteinized bovine bone scaffolds provided better performance in terms of mean gray values of X films, mean pixel values of computed tomography films, and bone volume and trabecular thickness of micro-computed tomography pictures when compared with the deproteinized bovine bone group. In addition, histological analysis showed that the lithium-incorporated deproteinized bovine bone group also significantly achieved larger new bone formation area. At the same time, when the expression of osteogenic factors in vivo was evaluated, runt-related transcription factor 2 (Runx2) and collagen type one (Col-1) were expressed more in lithium-incorporated deproteinized bovine bone group than those in deproteinized bovine bone group. However, the bone defect repair effect using autograft is still a little better than that of lithium-incorporated deproteinized bovine bone substitute based on our results. In conclusion, surface lithium-incorporated deproteinized bovine bone achieved improvement of osteogenesis effect and could enhance the new bone formation in critical-sized bone defects.