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Effect of triple-primed mesenchymal stem cells on bone regeneration

Abstract

There is evidence that priming mesenchymal stem cells (MSCs) increase their efficacy for regenerative medicine procedures. Among the several priming methods, MSCs overexpressing growth factors, culturing MSCs on modified surfaces and MSCs treated with photobiomodulation (PBM) seem to stimulate tissue formation by MSCs. In keeping with this, the aim of this project is to evaluate the effect of cell therapy based on local injection of triple-primed MSCs (1) genetically edited to overexpress BMP-9, (2) cultured on microtopography substrate and (3) PBM laser irradiated on bone regeneration. For these, an immortalized bone marrow derived MSCs lineage will be CRISPR/Cas9 edited to overexpress BMP-9, cultured on tissue culture plastic with microtopography until confluence, enzymatically released and injected into rat calvaria defects (5´104 cells/50 mL of PBS) created 2 weeks earlier. PBM will be applied immediately, 48 and 96 h following cell injection (808 nm; 40 mW; 1,42 W/cm2; 4 J/cm2; 3 s; 0,12 J/point; 0,028 cm2) in 4 points (1 in the centre and 4 around the defect). Bone formation will be evaluated by in vivo microtomography immediately, at 2 and 4 weeks after cell injection and by histological analysis after 4 weeks. The results of this study would contribute to launch new treatments based on the concepts of cell therapy to promote bone regeneration under challenging sites with applications in distinct areas such as Oral and Maxillofacial Surgery, Periodontology, Implantology and Orthopedia. (AU)

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