Limits...
Human saphenous vein organ culture under controlled hemodynamic conditions.

Miyakawa AA, Dallan LA, Lacchini S, Borin TF, Krieger JE - Clinics (Sao Paulo) (2008)

Bottom Line: Cell viability, cell density and apoptosis were compared before and after these intervals using MTT, Hoeschst 33258 stain, and TUNEL assays, respectively.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.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.

View Article: PubMed Central - PubMed

Affiliation: Heart Institute (InCor) and Department of Medicine-LIM 13, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil. ayumi.miyakawa@incor.usp.br

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.

Show MeSH

Related in: MedlinePlus

Morphological and integrity analyses of fresh excised human saphenous vein. HE stain (A) and VVG stain (B) demonstrating the tissue structure. Hoechst 33258-labeled cell nuclei (C) and TUNEL assay (D) showing tissue cellular density with no presence of apoptotic processes. 100X magnification
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2664728&req=5

f2-18-0106: Morphological and integrity analyses of fresh excised human saphenous vein. HE stain (A) and VVG stain (B) demonstrating the tissue structure. Hoechst 33258-labeled cell nuclei (C) and TUNEL assay (D) showing tissue cellular density with no presence of apoptotic processes. 100X magnification

Mentions: The human saphenous vein segments were cultured under venous and arterial hemodynamic conditions for 1, 2 and 4 days. Before and after these periods, analysis of cell viability, cell density, and apoptosis were performed. In the fresh excised segment of human saphenous vein, tissue morphology and integrity were verified by HE and VVG staining, and also by Hoechst 33258 nuclei stain and TUNEL assay (Figure 2). Furthermore, there was no evidence for apoptotic process indicating that the vessels used were well preserved (Figure 2).


Human saphenous vein organ culture under controlled hemodynamic conditions.

Miyakawa AA, Dallan LA, Lacchini S, Borin TF, Krieger JE - Clinics (Sao Paulo) (2008)

Morphological and integrity analyses of fresh excised human saphenous vein. HE stain (A) and VVG stain (B) demonstrating the tissue structure. Hoechst 33258-labeled cell nuclei (C) and TUNEL assay (D) showing tissue cellular density with no presence of apoptotic processes. 100X magnification
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2664728&req=5

f2-18-0106: Morphological and integrity analyses of fresh excised human saphenous vein. HE stain (A) and VVG stain (B) demonstrating the tissue structure. Hoechst 33258-labeled cell nuclei (C) and TUNEL assay (D) showing tissue cellular density with no presence of apoptotic processes. 100X magnification
Mentions: The human saphenous vein segments were cultured under venous and arterial hemodynamic conditions for 1, 2 and 4 days. Before and after these periods, analysis of cell viability, cell density, and apoptosis were performed. In the fresh excised segment of human saphenous vein, tissue morphology and integrity were verified by HE and VVG staining, and also by Hoechst 33258 nuclei stain and TUNEL assay (Figure 2). Furthermore, there was no evidence for apoptotic process indicating that the vessels used were well preserved (Figure 2).

Bottom Line: Cell viability, cell density and apoptosis were compared before and after these intervals using MTT, Hoeschst 33258 stain, and TUNEL assays, respectively.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.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.

View Article: PubMed Central - PubMed

Affiliation: Heart Institute (InCor) and Department of Medicine-LIM 13, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil. ayumi.miyakawa@incor.usp.br

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.

Show MeSH
Related in: MedlinePlus