Effects of various monomers and micro-structure of polyhydroxyalkanoates on the behavior of endothelial progenitor cells and endothelial cells for vascular tissue engineering

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  1. Topical Collection on Bio-Based Polymers

Abstract

Cardiovascular diseases are a leading cause of mortality in the world today. Vascular tissue engineering is an important and attractive research issue for the repair and regeneration of blood vessels. Two bio-based polymers, poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), which both belong to the polyhydroxyalkanoate (PHA) family, were used in this study. The aim of this study is to assess the potential application of PHB and PHBV to serve as a scaffold that is seeded with human umbilical vein endothelial cells (HUVECs) or endothelial progenitor cells (EPCs) for vascular tissue engineering. PHA films with various surface characteristics were prepared by solution-casting (surface roughness) and electrospinning (mesh-like structure). First, the mechanical and physical properties of various types of PHA films were analyzed. Then, the PHAs films were examined for cytotoxicity, biocompatibility and proliferation ability using cell lines (3 T3 and L929) and primary cells (HUVECs and EPCs). The cell morphology cultured on the PHA films was observed by fluorescence microscope and scanning electron microscopy. In addition, cultured EPCs on various types of PHA films were analyzed for whether the cells maintained the abilities of Ac-LDL uptake and UEA-1 lectin binding and exhibited specific gene expressions, including VEGFR-2, vWF, CD31, CD34 and CD133. Importantly, the cell retention rate and anti-coagulation ability of HUVECs or EPCs cultured on the various types of PHA films were also evaluated at the indicated time points. Our results showed that PHA films that were prepared using electrospinning methods (Ele-PHB and Ele-PHBV) had good mechanical and physical properties. HUVECs and EPCs can attach and grow on Ele-PHB and Ele-PHBV films without showing cytotoxicity. After a one-week culture, expanded HUVECs or EPCs maintained the correct cell morphologies and exhibited correct cell functions, such as high cell attachment rate and anti-coagulation ability. Taken together, Ele-PHB and Ele-PHBV films were ideal bio-based polymers to combine with HUVECs or EPCs for vascular tissue engineering.

Keywords

Polyhydroxyalkanoates Casting Electrospinning Endothelial progenitor cells Vascular tissue engineering 

Abbreviations

A540

The absorbance at 540 nm

Ac-LDL

Acetylated low-density lipoprotein

Cast-PHB

Solvent-cast PHB

Cast-PHBV

Solvent-cast PHBV

DAPI

4′,6-Diamidine-2′-phenylindole dihydrochloride

DMEM

Dulbecco’s modified Eagle’s medium

DMF

N,N-dimethyl formamide

EC

Endothelial cells

ECGS

Endothelial cell growth supplement

ECM

Extracellular matrix

Ele-PHB

Electrospun PHB

ElePHBV

Electrospun PHBV

EPC

Endothelial progenitor cell

F-actin

Filamentous actin

FBS

Fetal bovine serum

GAPDH

Glyceraldehyde 3-phosphate dehydrogenase

GPC

Gel permeation chromatography

HUVEC

Human umbilical vein endothelial cell

HV

Hydroxyvalerate

LSS

Laminar shear stress

MNC

Mononuclear cell

Mw

The weight-averaged molecular weight

PCR

Polymerase chain reaction

PDI

The polydispersity indice

PHA

Polyhydroxyalkanoate

PHB

Poly(3-hydroxybutyrate)

PHBV

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)

SEM

Scanning electron microscopy

TCPS

Tissue culture polystyrene

UCB

umbilical cord blood

UEA-1 Lectin

FITC-conjugated Lectin from Ulex europaeus

vWF

von Willebrand factor

Notes

Acknowledgements

This work was supported by the Ministry of Science and Technology, Taiwan, Republic of China [MOST 104-2628-E-155-002-MY3].

Compliance with ethical standards

Conflict of interest

The authors indicate no potential conflicts of interest.

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Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Chao-Ling Yao
    • 1
    • 2
  • Jian-Haw Chen
    • 1
  • Cheng-Hung Lee
    • 2
    • 3
    • 4
  1. 1.Department of Chemical Engineering and Materials ScienceYuan Ze UniversityTaoyuan CityTaiwan
  2. 2.Graduate School of Biotechnology and BioengineeringYuan Ze UniversityTaoyuan CityTaiwan
  3. 3.Department of General SurgeryBuddhist Dalin Tzu Chi HospitalChia-YiTaiwan
  4. 4.School of MedicineTzu Chi UniversityHualienTaiwan

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