Abstract
Distraction osteogenesis (DO) is the gold standard to treat large bone defects, but long consolidation period is a major limitation. Innovative efforts to promote osteogenesis are needed. Porcine brain extract (PBE) was reported to enhance the proliferation and differentiation of multiple primary cells. In this study, we aimed to develop a method for collecting PBE and investigate its effects on osteogenic differentiation of rat bone marrow derived mesenchymal stem cells (rBMSCs) and bone consolidation in a rat DO model. The PBE was collected from neonatal brain tissues of porcine fetus and was used to treat rBMSCs. Following PBE treatment (700 ng/ml), osteogenic differentiation was assessed. Further, we locally injected PBE (7 μg/ml, 100μl) or PBS (100μl) into the gap in a Sprague-Dawley (SD) male rat DO model every three days till termination. X-rays, micro-computed tomography, mechanical testing, histology and immunohischemistry examinations were used to exam the quality of the regenerates. The alkaline phosphatase, calcium deposits, and steogenic markers in the PBE treated rBMSCs were significantly increased. In the rat model, new bone properties of bone volume/total tissue volume and mechanical strength were higher in the PBE treated group. Histological analysis also confirmed more mineralized bone after PBE treatment. The current study reports a standard protocol for PBE collection and demonstrated its positive effects on osteogenic differentiation and bone consolidation in DO. Since the PBE is readily available and very cost effective, PBE may be a potential new bio-source to promote bone formation in patients undergo DO treatment.
Highlights
Distraction osteogenesis (DO), an effective tool for repairing bone defects and correcting osseous deformities, has been used frequently in clinical practice [1]
Fresh porcine brain extract (PBE), PBE kept in frozen for 2, 4, and 6 weeks were used for testing the effects on osteogenesis of rat bone marrow derived mesenchymal stem cells (rBMSCs), and no difference on the effects of rBMSCs osteogenesis was found among the various preparation of PBE
To evaluate the effects of PBE on osteogenesis of rBMSCs, Alkaline phosphatase (ALP) and Alizarin Red S staining were performed at day 3 and day 7, respectively
Summary
Distraction osteogenesis (DO), an effective tool for repairing bone defects and correcting osseous deformities, has been used frequently in clinical practice [1]. In the report of Burgess et al [8], some factors from neural tissues were found to be potent mitogens for mesoderm-derived cells, for vascular endothelial cells and some ectoderm-derived cells. Owing to their ability to stimulate fibroblasts proliferation, these growth factors have been named brain fibroblast growth factors (FGFs), which stimulate bone consolidation [9,10,11]. We introduced a method of collecting porcine brain extract (PBE) in this study, and investigated the effect of PBE on osteogenic differentiation of rat bone marrow derived mesenchymal stem cells (rBMSCs) and bone consolidation in a rat DO model
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