Vein wall re-endothelialization after deep vein thrombosis is improved with low-molecular-weight heparin

Daria Moaveni, Erin M. Lynch, Cathy Luke, Vikram Sood, Gilbert R. Upchurch, Thomas W. Wakefield, Peter K. Henke

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Objective: Vein wall endothelial turnover after stasis deep vein thrombosis (DVT) has not been well characterized. The purpose of this study was to quantify re-endothelialization after DVT and determine if low-molecular-weight heparin (LMWH) therapy affects this process. Methods: Stasis DVT was generated in the rat by inferior vena cava ligation, with harvest at 1, 4, and 14 days. Immunohistologic quantification of vascular smooth muscle cells and luminal endothelialization was estimated by positive staining for α-smooth muscle actin and von Willebrand factor, respectively. In separate experiments, rats were treated either before or after DVT with subcutaneous LMWH (3 mg/kg daily) until harvesting at 4 and 14 days. The inferior vena cava was processed for histologic analysis or was processed for organ culture after the thrombus was gently removed. The vein wall was stimulated in vitro with interleukin-1β (1 ng/mL), and the supernatant was processed at 48 hours for nitric oxide. Cells were processed by real-time polymerase chain reaction for endothelial nitric oxide synthase, inducible nitric oxide synthase, cyclooxygenase-1 and -2, and thrombomodulin at 4 and 14 days, and collagen I and III at 14 days. Comparisons were done with analysis of variance or t test. A P < .05 was significant. Results: Thrombus size peaked at 4 days, whereas luminal re-endothelialization increased over time (1 day, 11% ± 2%; 4 days, 23% ± 4%; 14 days, 64% ± 7% (+) von Willebrand factor staining; P < .01, n = 3 to 4, compared with non-DVT control). Similarly, vascular smooth muscle cell staining was lowest at day 1 and gradually returned to baseline by 14 days. Both before and after DVT, LMWH significantly increased luminal re-endothelialization, without a difference in thrombus size at 4 days, but no significant difference was noted at 14 days despite smaller thrombi with LMWH treatment. Pretreatment with LMWH was associated with increased vascular smooth muscle cell area and recovery of certain inducible endothelial specific genes. No significant difference in nitric oxide levels in the supernatant was found at 4 days. At 14 days, type III collagen was significantly elevated with LMWH treatment. Conclusions: Venous re-endothelialization occurs progressively as the DVT resolves and can be accelerated with LMWH treatment, although this effect appears limited to the early time frame. These findings may have clinical relevance for LMWH timing and treatment compared with mechanical forms of therapy.

Original languageEnglish (US)
Pages (from-to)616-624
Number of pages9
JournalJournal of Vascular Surgery
Volume47
Issue number3
DOIs
StatePublished - Mar 2008
Externally publishedYes

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Low Molecular Weight Heparin
Venous Thrombosis
Veins
Thrombosis
Vascular Smooth Muscle
Smooth Muscle Myocytes
von Willebrand Factor
Inferior Vena Cava
Staining and Labeling
Nitric Oxide
Thrombomodulin
Cyclooxygenase 1
Collagen Type III
Nitric Oxide Synthase Type III
Organ Culture Techniques
Nitric Oxide Synthase Type II
Cyclooxygenase 2
Interleukin-1
Ligation
Smooth Muscle

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Surgery

Cite this

Vein wall re-endothelialization after deep vein thrombosis is improved with low-molecular-weight heparin. / Moaveni, Daria; Lynch, Erin M.; Luke, Cathy; Sood, Vikram; Upchurch, Gilbert R.; Wakefield, Thomas W.; Henke, Peter K.

In: Journal of Vascular Surgery, Vol. 47, No. 3, 03.2008, p. 616-624.

Research output: Contribution to journalArticle

Moaveni, Daria ; Lynch, Erin M. ; Luke, Cathy ; Sood, Vikram ; Upchurch, Gilbert R. ; Wakefield, Thomas W. ; Henke, Peter K. / Vein wall re-endothelialization after deep vein thrombosis is improved with low-molecular-weight heparin. In: Journal of Vascular Surgery. 2008 ; Vol. 47, No. 3. pp. 616-624.
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abstract = "Objective: Vein wall endothelial turnover after stasis deep vein thrombosis (DVT) has not been well characterized. The purpose of this study was to quantify re-endothelialization after DVT and determine if low-molecular-weight heparin (LMWH) therapy affects this process. Methods: Stasis DVT was generated in the rat by inferior vena cava ligation, with harvest at 1, 4, and 14 days. Immunohistologic quantification of vascular smooth muscle cells and luminal endothelialization was estimated by positive staining for α-smooth muscle actin and von Willebrand factor, respectively. In separate experiments, rats were treated either before or after DVT with subcutaneous LMWH (3 mg/kg daily) until harvesting at 4 and 14 days. The inferior vena cava was processed for histologic analysis or was processed for organ culture after the thrombus was gently removed. The vein wall was stimulated in vitro with interleukin-1β (1 ng/mL), and the supernatant was processed at 48 hours for nitric oxide. Cells were processed by real-time polymerase chain reaction for endothelial nitric oxide synthase, inducible nitric oxide synthase, cyclooxygenase-1 and -2, and thrombomodulin at 4 and 14 days, and collagen I and III at 14 days. Comparisons were done with analysis of variance or t test. A P < .05 was significant. Results: Thrombus size peaked at 4 days, whereas luminal re-endothelialization increased over time (1 day, 11{\%} ± 2{\%}; 4 days, 23{\%} ± 4{\%}; 14 days, 64{\%} ± 7{\%} (+) von Willebrand factor staining; P < .01, n = 3 to 4, compared with non-DVT control). Similarly, vascular smooth muscle cell staining was lowest at day 1 and gradually returned to baseline by 14 days. Both before and after DVT, LMWH significantly increased luminal re-endothelialization, without a difference in thrombus size at 4 days, but no significant difference was noted at 14 days despite smaller thrombi with LMWH treatment. Pretreatment with LMWH was associated with increased vascular smooth muscle cell area and recovery of certain inducible endothelial specific genes. No significant difference in nitric oxide levels in the supernatant was found at 4 days. At 14 days, type III collagen was significantly elevated with LMWH treatment. Conclusions: Venous re-endothelialization occurs progressively as the DVT resolves and can be accelerated with LMWH treatment, although this effect appears limited to the early time frame. These findings may have clinical relevance for LMWH timing and treatment compared with mechanical forms of therapy.",
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AU - Upchurch, Gilbert R.

AU - Wakefield, Thomas W.

AU - Henke, Peter K.

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N2 - Objective: Vein wall endothelial turnover after stasis deep vein thrombosis (DVT) has not been well characterized. The purpose of this study was to quantify re-endothelialization after DVT and determine if low-molecular-weight heparin (LMWH) therapy affects this process. Methods: Stasis DVT was generated in the rat by inferior vena cava ligation, with harvest at 1, 4, and 14 days. Immunohistologic quantification of vascular smooth muscle cells and luminal endothelialization was estimated by positive staining for α-smooth muscle actin and von Willebrand factor, respectively. In separate experiments, rats were treated either before or after DVT with subcutaneous LMWH (3 mg/kg daily) until harvesting at 4 and 14 days. The inferior vena cava was processed for histologic analysis or was processed for organ culture after the thrombus was gently removed. The vein wall was stimulated in vitro with interleukin-1β (1 ng/mL), and the supernatant was processed at 48 hours for nitric oxide. Cells were processed by real-time polymerase chain reaction for endothelial nitric oxide synthase, inducible nitric oxide synthase, cyclooxygenase-1 and -2, and thrombomodulin at 4 and 14 days, and collagen I and III at 14 days. Comparisons were done with analysis of variance or t test. A P < .05 was significant. Results: Thrombus size peaked at 4 days, whereas luminal re-endothelialization increased over time (1 day, 11% ± 2%; 4 days, 23% ± 4%; 14 days, 64% ± 7% (+) von Willebrand factor staining; P < .01, n = 3 to 4, compared with non-DVT control). Similarly, vascular smooth muscle cell staining was lowest at day 1 and gradually returned to baseline by 14 days. Both before and after DVT, LMWH significantly increased luminal re-endothelialization, without a difference in thrombus size at 4 days, but no significant difference was noted at 14 days despite smaller thrombi with LMWH treatment. Pretreatment with LMWH was associated with increased vascular smooth muscle cell area and recovery of certain inducible endothelial specific genes. No significant difference in nitric oxide levels in the supernatant was found at 4 days. At 14 days, type III collagen was significantly elevated with LMWH treatment. Conclusions: Venous re-endothelialization occurs progressively as the DVT resolves and can be accelerated with LMWH treatment, although this effect appears limited to the early time frame. These findings may have clinical relevance for LMWH timing and treatment compared with mechanical forms of therapy.

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