Annals of Biomedical Engineering

, Volume 45, Issue 2, pp 360–377 | Cite as

Optimizing Photo-Encapsulation Viability of Heart Valve Cell Types in 3D Printable Composite Hydrogels

  • Laura Hockaday Kang
  • Patrick A. Armstrong
  • Lauren Julia Lee
  • Bin Duan
  • Kevin Heeyong Kang
  • Jonathan Talbot Butcher
The Pursuit of Engineering the Ideal Heart Valve Replacement or Repair

Abstract

Photocrosslinking hydrogel technologies are attractive for the biofabrication of cardiovascular soft tissues, but 3D printing success is dependent on multiple variables. In this study we systematically test variables associated with photocrosslinking hydrogels (photoinitiator type, photoinitiator concentration, and light intensity) for their effects on encapsulated cells in an extrusion 3D printable mixture of methacrylated gelatin/poly-ethylene glycol diacrylate/alginate (MEGEL/PEGDA3350/alginate). The fabrication conditions that produced desired hydrogel mechanical properties were compared against those that optimize aortic valve or mesenchymal stem cell viability. In the 3D hydrogel culture environment and fabrication setting studied, Irgacure can increase hydrogel stiffness with a lower proportional decrease in encapsulated cell viability compared to VA086. Human adipose derived mesenchymal stem cells (HADMSC) survived increasing photoinitiator concentrations in photo-encapsulation conditions better than aortic valve interstitial cells (HAVIC) and aortic valve sinus smooth muscle cells (HASSMC). Within the range of photo-encapsulation fabrication conditions tested with MEGEL/PEGDA/alginate (0.25–1.0% w/v VA086, 0.025–0.1% w/v Irgacure 2959, and 365 nm light intensity 2–136 mW/cm2), the highest viabilities achieved were 95, 93, and 93% live for HASSMC, HAVIC, and HADMSC respectively. These results identify parameter combinations that optimize cell viability during 3D printing for multiple cell types. These results also indicate that general oxidative stress is higher in photocrosslinking conditions that induce lower cell viability. However, suppressing this increase in intracellular oxidative stress did not improve cell viability, which suggests that other stress mechanisms also contribute.

Keywords

Extrusion bioprinting Oxidative stress Mesenchymal stem cells Photo-polymerization Bio-ink Biofabrication 

Abbreviations

CTR

CellTracker™ Red CMTPX

DCF

5-(and-6)-Chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate, acetyl ester CM-H2DCFDA

E5to15

Compressive modulus calculated from 5 to 15% strain

ECM

Extracellular matrix

HADMSC

Human adipose derived mesenchymal stem cells

HASSMC

Human aortic valve sinus smooth muscle cells

HAVIC

Human aortic valve interstitial cells

HBSS

Hank’s balanced salt solution

HLED

High powered light emitting diode

Irgacure 2959

2-Hydroxy-1(4-(hydroxyethox)pheny)-2-methyl-1-propanone

LED

Light emitting diode

MEGEL

Methacrylated gelatin

PBS

Phosphate buffered saline

PEG

Poly-ethylene glycol

PEGDA

Poly-ethylene glycol diacrylate

3D

Three dimensional

TEHV

Tissue engineered heart valves

2D

Two dimensional

UV

Ultraviolet

VA086

2,2′-Azobis[2-methyl-N-(2-hydroxyethyl)propionamide]

Supplementary material

10439_2016_1619_MOESM1_ESM.pdf (8.7 mb)
Supplementary material 1 (PDF 8896 kb)

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Copyright information

© Biomedical Engineering Society 2016

Authors and Affiliations

  • Laura Hockaday Kang
    • 1
  • Patrick A. Armstrong
    • 1
  • Lauren Julia Lee
    • 1
  • Bin Duan
    • 1
  • Kevin Heeyong Kang
    • 1
  • Jonathan Talbot Butcher
    • 1
    • 2
  1. 1.Nancy E. and Peter C. Meinig School of Biomedical EngineeringCornell UniversityIthacaUSA
  2. 2.Nancy E. and Peter C. Meinig School of Biomedical EngineeringCornell UniversityIthacaUSA

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