Hypoxia induces osteogenesis in rabbit adipose‐derived stem cells overexpressing bone morphogenic protein‐2

Abstract

Objective Hypoxic culture potentiates mesenchymal stem cells ( MSC s) to survive and secrete various growth factors. Genetically modified stem cells overexpressing bone morphogenic protein‐2 ( BMP ‐2) demonstrate strong osteogenic ability. Hence, we investigated the coeffect of hypoxic culture conditions and BMP ‐2 overexpression on the osteogenic ability of rabbit adipose‐derived stem cells (r ASC s) in vitro . Materials and Methods Rabbit adipose‐derived stem cells with or without adenoviral‐ BMP ‐2 transduction were cultured in hypoxic (1%) and normoxic (21%) conditions. Cell viability, attachment, and proliferation were compared. Real‐time PCR amplification of osteogenic and angiogenic genes including alkaline phosphatase ( ALP ), osteocalcin ( OCN ), HIF ‐1α, and vascular endothelial growth factor ( VEGF ) was performed. Moreover, ALP activity, immunofluorescent staining of OCN , and mineralization assay by alizarin red S quantification and von K ossa staining were conducted. Results Cells under hypoxic conditions attached better within 12 h and proliferated faster. While BMP ‐2 overexpression and hypoxic condition separately elevated the transcription of key osteogenic and angiogenic genes, a cooperative effect was observed to enhance the upregulation of osteogenic as well as angiogenic genes. Identical changes were observed in ALP activity, immunofluorescent staining of OCN , and mineralization assay. Conclusions Hypoxic culture can enhance the osteogenic ability of BMP ‐2 gene‐modified r ASC s, which provides a strategy to improve the osteogenesis of r ASC s for in vivo bone regeneration.

Hypoxia induces osteogenesis in rabbit adipose‐derived stem cells overexpressing bone morphogenic protein‐2

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