Neovascularization during venous thrombosis organization: a preliminary study.
OBJECTIVE
Thrombus organization after venous thromboembolism leading to recanalization occurs at a variable rate. The angiogenic chemokine interleukin-8 (IL-8) has been found in thrombus months after thrombus initiation. We hypothesize that thrombus organization involves neovascularization and leukocyte influx and that IL-8 administered at thrombus induction will promote thrombus organization.


METHODS
A group of rats underwent inferior vena caval occlusive thrombosis. At thrombus induction and every 24 hours, the rats were administered IL-8 (1 microgram) or serum albumin. The rats were killed at either day 4, day 8, or day 12, and, at death, colloidal carbon was perfused via the heart. The inferior vena cava was isolated, measured, weighed, and formalin fixed. The sections were stained with anti-polymorphonuclear leukocyte antibody, the endothelial marker factor VIII-related antigen, and with hematoxylin and eosin. Thrombus neovascularization (colloidal carbon) with morphometric analysis was normalized to the total thrombus area. In addition, the rats underwent perfusion with fluorescein isothiocyanate dextran (molecular weight, 150,000) at death to correlate with colloidal carbon perfusion, and thrombus fluorescence was determined.


RESULTS
Thrombus cellularity initially involved neutrophils, followed by monocytes. Significantly more neutrophils, monocytes, and cells that were defined as spindle shaped (fibroblasts and endothelial cells) were noted in the animals treated with IL-8. Neovascularization was significantly increased at day 4 in the animals treated with IL-8 versus the animals treated with serum albumin and was corroborated with a significant increase in thrombus fluorescein isothiocyanate dextran fluorescence at day 4 in the rats treated with IL-8. Colloidal carbon perfusion was noted within vascular channels without extravasation and colocalized with factor VIII-related antigen.


CONCLUSIONS
This study shows that thrombus organization involves neovascularization and that IL-8 augments thrombus organization.
Wakefield
T W
TW
Jobst Vascular Surgery Laboratory, Section of Vascular Surgery, Department of Surgery, Division of Pulmonary and Critical Care Medicine, Ann Arbor, Michigan, USA.
Linn
M J
MJ
Henke
P K
PK
Kadell
A M
AM
Wilke
C A
CA
Wrobleski
S K
SK
Sarkar
M
M
Burdick
M D
MD
Myers
D D
DD
Strieter
R M
RM
D000818:Animals; D002244:Carbon; D004338:Drug Combinations; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D016209:Interleukin-8 / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D009389:Neovascularization, Pathologic; D011205:Povidone; D051381:Rats; D012709:Serum Albumin / Q000494:pharmacology; D014682:Vena Cava, Inferior; D020246:Venous Thrombosis / Q000473:pathology*
D004338:Drug Combinations; D005456:Fluorescent Dyes; D016209:Interleukin-8; D012709:Serum Albumin; C078073:carbon colloid; D002244:Carbon; D011205:Povidone; D016650:Fluorescein-5-isothiocyanate