Human saphenous vein organ culture under controlled hemodynamic conditions

Ayumi Aurea Miyakawa, Luis Alberto Oliveira Dallan, Silvia Lacchini, Thaiz Ferraz Borin, Jose Eduardo Krieger

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

INTRODUCTION: Saphenous vein grafting is still widely used to revascularize ischemic myocardium. The effectiveness of this procedure is limited by neointima formation and accelerated atherosclerosis, which frequently leads to graft occlusion. A better understanding of this process is important to clarify the mechanisms of vein graft disease and to aid in the formulation of strategies for prevention and/or therapeutics. OBJECTIVE: To develop an ex vivo flow system that allows for controlled hemodynamics in order to mimic arterial and venous conditions. METHODS: Human saphenous veins were cultured either under venous (flow: 5 ml/min) or arterial hemodynamic conditions (flow: 50 ml/min, pressure: 80 mmHg) for 1-, 2- and 4-day periods. Cell viability, cell density and apoptosis were compared before and after these intervals using MTT, Hoeschst 33258 stain, and TUNEL assays, respectively. RESULTS: Fresh excised tissue segments were well preserved prior to the study. Hoechst 33258 and MTT stains showed progressive losses in cell density and cell viability in veins cultured under arterial hemodynamic conditions from 1 to 4 days, while no alterations were observed in veins cultured under venous conditions. Although the cell density from 1-day cultured veins under arterial conditions was similar to that of freshly excised veins, the TUNEL assay indicated that most of these cells were undergoing apoptosis. CONCLUSION: The results observed resemble the events taking place during early in vivo arterial-vein grafting and provide evidence that an ex vivo perfusion system may be useful for the identification of new therapeutic targets that ameliorate vein graft remodeling and increase graft patency over time.

Original languageEnglish (US)
Pages (from-to)683-688
Number of pages6
JournalClinics
Volume63
Issue number5
DOIs
StatePublished - Oct 28 2008

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Organ Culture Techniques
Saphenous Vein
Veins
Hemodynamics
Transplants
Cell Count
In Situ Nick-End Labeling
Cell Survival
Coloring Agents
Apoptosis
Bisbenzimidazole
Neointima
Atherosclerosis
Myocardium
Perfusion
Pressure
Therapeutics

Keywords

  • Ex vivo organ culture
  • Saphenous vein graft
  • Vascular biology

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Human saphenous vein organ culture under controlled hemodynamic conditions. / Miyakawa, Ayumi Aurea; Dallan, Luis Alberto Oliveira; Lacchini, Silvia; Borin, Thaiz Ferraz; Krieger, Jose Eduardo.

In: Clinics, Vol. 63, No. 5, 28.10.2008, p. 683-688.

Research output: Contribution to journalArticle

Miyakawa, AA, Dallan, LAO, Lacchini, S, Borin, TF & Krieger, JE 2008, 'Human saphenous vein organ culture under controlled hemodynamic conditions', Clinics, vol. 63, no. 5, pp. 683-688. https://doi.org/10.1590/S1807-59322008000500018
Miyakawa, Ayumi Aurea ; Dallan, Luis Alberto Oliveira ; Lacchini, Silvia ; Borin, Thaiz Ferraz ; Krieger, Jose Eduardo. / Human saphenous vein organ culture under controlled hemodynamic conditions. In: Clinics. 2008 ; Vol. 63, No. 5. pp. 683-688.
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N2 - INTRODUCTION: Saphenous vein grafting is still widely used to revascularize ischemic myocardium. The effectiveness of this procedure is limited by neointima formation and accelerated atherosclerosis, which frequently leads to graft occlusion. A better understanding of this process is important to clarify the mechanisms of vein graft disease and to aid in the formulation of strategies for prevention and/or therapeutics. OBJECTIVE: To develop an ex vivo flow system that allows for controlled hemodynamics in order to mimic arterial and venous conditions. METHODS: Human saphenous veins were cultured either under venous (flow: 5 ml/min) or arterial hemodynamic conditions (flow: 50 ml/min, pressure: 80 mmHg) for 1-, 2- and 4-day periods. Cell viability, cell density and apoptosis were compared before and after these intervals using MTT, Hoeschst 33258 stain, and TUNEL assays, respectively. RESULTS: Fresh excised tissue segments were well preserved prior to the study. Hoechst 33258 and MTT stains showed progressive losses in cell density and cell viability in veins cultured under arterial hemodynamic conditions from 1 to 4 days, while no alterations were observed in veins cultured under venous conditions. Although the cell density from 1-day cultured veins under arterial conditions was similar to that of freshly excised veins, the TUNEL assay indicated that most of these cells were undergoing apoptosis. CONCLUSION: The results observed resemble the events taking place during early in vivo arterial-vein grafting and provide evidence that an ex vivo perfusion system may be useful for the identification of new therapeutic targets that ameliorate vein graft remodeling and increase graft patency over time.

AB - INTRODUCTION: Saphenous vein grafting is still widely used to revascularize ischemic myocardium. The effectiveness of this procedure is limited by neointima formation and accelerated atherosclerosis, which frequently leads to graft occlusion. A better understanding of this process is important to clarify the mechanisms of vein graft disease and to aid in the formulation of strategies for prevention and/or therapeutics. OBJECTIVE: To develop an ex vivo flow system that allows for controlled hemodynamics in order to mimic arterial and venous conditions. METHODS: Human saphenous veins were cultured either under venous (flow: 5 ml/min) or arterial hemodynamic conditions (flow: 50 ml/min, pressure: 80 mmHg) for 1-, 2- and 4-day periods. Cell viability, cell density and apoptosis were compared before and after these intervals using MTT, Hoeschst 33258 stain, and TUNEL assays, respectively. RESULTS: Fresh excised tissue segments were well preserved prior to the study. Hoechst 33258 and MTT stains showed progressive losses in cell density and cell viability in veins cultured under arterial hemodynamic conditions from 1 to 4 days, while no alterations were observed in veins cultured under venous conditions. Although the cell density from 1-day cultured veins under arterial conditions was similar to that of freshly excised veins, the TUNEL assay indicated that most of these cells were undergoing apoptosis. CONCLUSION: The results observed resemble the events taking place during early in vivo arterial-vein grafting and provide evidence that an ex vivo perfusion system may be useful for the identification of new therapeutic targets that ameliorate vein graft remodeling and increase graft patency over time.

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