Background
The diabetic phenotype of wound healing is in part characterized by impaired neovascularization
and deficient endothelial progenitor cell (EPC) recruitment. Angiopoietin-1 (Ang-1)
is a potent mobilizer of EPCs from the bone marrow (BM). A suggested mechanism for
EPC mobilization from the BM is mediated by matrix metalloproteinase 9 (MMP-9) and
stem cell factor (SCF). Taken together, we hypothesized that overexpression of Ang-1
in diabetic wounds will recruit EPCs and improve neovascularization and wound healing.
Methods
An endothelial lineage BM-labeled murine model of diabetes was developed to track
BM-derived EPCs. FVBN mice were lethally irradiated and then reconstituted with BM
from syngeneic Tie2/LacZ donor mice. Diabetes was induced with streptozotocin. Dorsal
wounds in BM-transplanted mice were treated with Ad-Ang-1, Ad-GFP, or phosphate-buffered
saline. At day 7 after injury, wounds were harvested and analyzed. A similar experiment
was conducted in EPC mobilization deficient MMP-9 –/– mice to determine whether the
effects of Ang-1 were EPC-dependent.
Results
Overexpression of Ang-1 resulted in greatly improved re-epithelialization, neovascularization,
and EPC recruitment in diabetic BM-transplanted wounds at day 7. Ang-1 treatment resulted
in increased serum levels of proMMP-9 and SCF but had no effect on vascular endothelial
growth factor levels. According to our FACS results, peripheral blood EPC (CD34+/Cd133+/Flk1+) counts at day 3 after wounding showed impaired EPC mobilization in MMP-9 –/– mice
compared with those of wild-type controls. EPC mobilization was rescued by SCF administration,
validating this model for EPC-mobilization–deficient mechanistic studies. In MMP-9
–/– mice, Ad-Ang-1 accelerated re-epithelialization in a similar manner, but had no
effect on neovascularization.
Conclusion
Our results show that Ang-1 administration results in improved neovascularization
which is dependent on EPC recruitment and has direct effects on wound re-epithelialization.
These data may represent a novel strategy to correct the phenotype of impaired diabetic
neovascularization and may improve diabetic wound healing.
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Article info
Publication history
Accepted:
June 27,
2015
Footnotes
Swathi Balaji, PhD, and Nate Han, BS, contributed equally to this work.
Timothy M. Crombleholme, MD, and Sundeep G. Keswani, MD, are co-senior authors.
This study is supported by NIH-NIDDK awards R01-DK072446 and R01-DK074055 (T.M.C.) and NIH-NIGMS awards K08 GM098831 and 1R01GM111808 (S.G.K).
The work described here was conducted in Cincinnati, OH.
Abstract Control Number: ASC20151363.
Identification
Copyright
© 2015 Elsevier Inc. Published by Elsevier Inc. All rights reserved.
