Prolyl hydroxylase inhibitors increase neoangiogenesis and callus formation following femur fracture in mice

J Orthop Res. 2009 Oct;27(10):1298-305. doi: 10.1002/jor.20886.

Abstract

Skeletal trauma and impaired skeletal healing is commonly associated with diminished vascularity. Hypoxia inducible factor alpha (HIF-1) is a key transcription factor responsible for activating angiogenic factors during development and tissue repair. Small molecule inhibitors of the prolyl hydroxylase enzyme (PHD), the key enzyme responsible for degrading HIF-1, have been shown to activate HIF-1, and are effective in inducing angiogenesis. Here we examined the effects of several commercially available PHD inhibitors on bone marrow mesenchymal stromal cells (MSCs) in vitro and in a stabilized fracture model in vivo. Three PHD inhibitors [Desferrioxamine (DFO), L-mimosine (L-mim), and Dimethyloxalylglycine (DMOG)] effectively activated a HIF-1 target reporter, induced expression of vascular endothelial growth factor (VEGF) mRNA in vitro, and increased capillary sprouting in a functional angiogenesis assay. DFO and DMOG were applied by direct injection at the fracture site in a stabilized murine femur fracture model. PHD inhibition increased the vascularity at 14 days and increased callus size as assessed by microCT at 28 days. These results suggest that HIF activation is a viable approach to increase vascularity and bone formation following skeletal trauma.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids, Dicarboxylic / pharmacology*
  • Animals
  • Bony Callus / drug effects*
  • Bony Callus / metabolism
  • Bony Callus / physiopathology
  • Cell Proliferation / drug effects
  • Deferoxamine / pharmacology*
  • Endothelium, Vascular / cytology
  • Femoral Fractures / metabolism
  • Femoral Fractures / physiopathology*
  • Hypoxia-Inducible Factor 1 / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mimosine / pharmacology*
  • Models, Animal
  • Neovascularization, Physiologic / drug effects*
  • Neovascularization, Physiologic / physiology
  • Procollagen-Proline Dioxygenase / antagonists & inhibitors*
  • Procollagen-Proline Dioxygenase / metabolism
  • RNA, Messenger / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Amino Acids, Dicarboxylic
  • Hypoxia-Inducible Factor 1
  • RNA, Messenger
  • Vascular Endothelial Growth Factor A
  • Mimosine
  • Procollagen-Proline Dioxygenase
  • Deferoxamine
  • oxalylglycine