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Effect Of Physiological Oxygen Levels On Osteogenic Differentiation Of Adipose-Derived Stem Cells, Suchit Sahai

Theses and Dissertations

Regenerative medicine presents exciting strategies for healing critical-size bone defects through the implantation of cells and biocompatible scaffolds. Most in vitro studies are performed in atmospheric oxygen conditions (~20%), which do not accurately mimic the CSD microenvironment. Due to damage to the vasculature at CSDs, oxygen levels fall into the hypoxic range (<5%), which can impact viability, proliferation and differentiation of the cells employed for bone regeneration. Understanding the cellular responses to hypoxia has grown primarily from study of individual molecular factors. The master regulator of adaptive responses to low oxygen availability is the nuclear factor, hypoxia-inducible factor-1. HIF-1 is a heterodimeric protein, stable below 6% O2 condition in the nucleus and has been shown to play a role in angiogenic-osteogenic coupling. We have developed a responsive, fluorescent, hypoxia detection system and determined whether HIF activity can be tracked in both 2-D and 3-D cultures. Adipose-derived stem cells (ASCs) were selected due to their broad utilization in tissue engineering strategies and characterized the influence of HIF signaling on its phenotype. The work done identified that of hypoxia impaired osteogenic differentiation of ASCs in both 2-D and 3-D cultures and HIF-1 did not mediate this effect. Ischemic preconditioning (IPC) strategy and varied protocols used represents a clinically feasible manipulation of cell preparation to help the survival of implanted ASCs and accelerate osteogenic differentiation at physiological oxygen levels.