Deep vein thrombosis resolution is not accelerated with increased neovascularization.
BACKGROUND
Deep venous thrombosis (DVT) resolution involves fibrinolysis, neovascularization, and fibrosis. We hypothesized that promoting neovascularization would accelerate DVT resolution.


METHODS
A rat model of stasis DVT was produced with proximal ligation of the inferior vena cava (IVC) and all visible tributaries. One microg of interferon inducible protein (IP-10; angiostatic chemokine), basic fibroblast growth factor (bFGF; pro-angiogenic cytokine), epithelial neutrophil activating protein (ENA-78; pro-angiogenic chemokine), or saline solution control was injected into the IVC after ligation, and then via tail vein injection daily until sacrifice at either 4 or 8 days. Peripheral blood counts were measured, and thrombus weight was recorded at sacrifice. Laser Doppler in vivo imaging was used to estimate post-thrombotic IVC blood flow. Immunohistologic assessment of the thrombosed IVC for polymorphonuclear neutrophils (PMNs), monocytes (ED-1), and laminin (neovascular channels) was performed or the thrombus was separated from the IVC and assayed for keratinocyte cytokine (KC), monocyte chemotactic protein-1 (MCP-1), bFGF with enzyme-linked immunosorbent assay (ELISA), and total collagen with a direct colorimetric assay.


RESULTS
Peripheral blood and intrathrombus PMNs and monocytes were not significantly different in the treated or control rats. There were no differences in any measure at 4 days. At 8 days, thrombus neovascularity, but not weight or collagen content, was increased in rats treated with bFGF or ENA-78 compared with control rats (17.6 +/- 0.93, 16.2 +/- 0.97 vs 13.2 +/- 0.79; channels/5 high-power fields (hpf; n = 6-10; P <.05). Post DVT IVC blood flow was significantly increased in bFGF-treated rats but not in rats treated with IP-10 or ENA-78, as compared with control rats. Rats treated with ENA-78 had increased intrathrombus bFGF compared with control rats (85 +/- 27 pg/mg protein vs 20 +/- 6 pg/mg protein; n = 6; P <.05), but other mediators were not significantly different in treated rats compared with control rats.


CONCLUSIONS
Pro-angiogenic compounds increase thrombus neovascularization, but this does not correlate with smaller or less fibrotic DVT. Mechanisms other than neovascularization may be more important to hasten DVT dissolution. Clinical relevance Improved therapy for deep venous thrombosis (DVT) will ideally increase the rate of thrombus dissolution and eliminate the bleeding risks of anticoagulation. This study evaluated promoting DVT neovascularization with angiogenic chemokines, and, while successful by experimental measures, this did not translate into smaller DVT. Solely promoting thrombus neovascularization will not likely speed resolution.
Varma
Manu R
MR
Department of Surgery, University of Michigan Medical School, Ann Arbor, USA.
Moaveni
Daria M
DM
Dewyer
Nicholas A
NA
Varga
Andrea J
AJ
Deatrick
K Barry
KB
Kunkel
Steven L
SL
Upchurch
Gilbert R
GR
Wakefield
Thomas W
TW
Henke
Peter K
PK
D000818:Animals; D054357:Chemokine CXCL10; D054365:Chemokine CXCL5; D019743:Chemokines, CXC / Q000502:physiology*; D004195:Disease Models, Animal; D016222:Fibroblast Growth Factor 2 / Q000502:physiology*; D016209:Interleukin-8 / Q000031:analogs & derivatives / Q000502:physiology*; D008297:Male; D018919:Neovascularization, Physiologic; D051381:Rats; D017207:Rats, Sprague-Dawley; D014463:Ultrasonography; D014682:Vena Cava, Inferior / Q000000981:diagnostic imaging / Q000473:pathology / Q000503:physiopathology*; D020246:Venous Thrombosis / Q000000981:diagnostic imaging / Q000473:pathology / Q000628:therapy*
C486263:CXCL5 protein, human; D054357:Chemokine CXCL10; D054365:Chemokine CXCL5; D019743:Chemokines, CXC; D016209:Interleukin-8; D016222:Fibroblast Growth Factor 2