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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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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.

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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

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Acknowledgements

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

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Authors and Affiliations

  1. Department of Chemical Engineering and Materials Science, Yuan Ze University, No. 135, Yuan-Tung road, Chung-Li District, Taoyuan City, 32003, Taiwan

    Chao-Ling Yao & Jian-Haw Chen

  2. Graduate School of Biotechnology and Bioengineering, Yuan Ze University, Chung-Li, Taoyuan City, 32003, Taiwan

    Chao-Ling Yao & Cheng-Hung Lee

  3. Department of General Surgery, Buddhist Dalin Tzu Chi Hospital, Chia-Yi, 62247, Taiwan

    Cheng-Hung Lee

  4. School of Medicine, Tzu Chi University, Hualien, 97002, Taiwan

    Cheng-Hung Lee

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Yao, CL., Chen, JH. & Lee, CH. Effects of various monomers and micro-structure of polyhydroxyalkanoates on the behavior of endothelial progenitor cells and endothelial cells for vascular tissue engineering. J Polym Res 25, 187 (2018). https://doi.org/10.1007/s10965-017-1341-1

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