Optimizing Photo-Encapsulation Viability of Heart Valve Cell Types in 3D Printable Composite Hydrogels.
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.
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, 101 Weill Hall, Ithaca, NY, 14853, USA.
Laura Hockaday
LH
Kang
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, 101 Weill Hall, Ithaca, NY, 14853, USA.
Patrick A
PA
Armstrong
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, 101 Weill Hall, Ithaca, NY, 14853, USA.
Lauren Julia
LJ
Lee
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, 101 Weill Hall, Ithaca, NY, 14853, USA.
Bin
B
Duan
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, 101 Weill Hall, Ithaca, NY, 14853, USA.
Kevin Heeyong
KH
Kang
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, 101 Weill Hall, Ithaca, NY, 14853, USA. ajtbutcher@gmail.com.|Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, 304 Weill Hall, Ithaca, NY, 14850, USA. ajtbutcher@gmail.com.
Jonathan Talbot
JT
Butcher
D000464:Alginates / Q000737:chemistry / Q000494:pharmacology; D001021:Aortic Valve / Q000166:cytology / Q000378:metabolism*; D002470:Cell Survival; D002478:Cells, Cultured; D005780:Gelatin / Q000737:chemistry / Q000494:pharmacology; D020723:Glucuronic Acid / Q000737:chemistry / Q000494:pharmacology; D006603:Hexuronic Acids / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D020100:Hydrogels / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D055668:Photochemical Processes; D011092:Polyethylene Glycols / Q000737:chemistry / Q000494:pharmacology; D066330:Printing, Three-Dimensional; D054457:Tissue Scaffolds / Q000737:chemistry*
D000464:Alginates; D006603:Hexuronic Acids; D020100:Hydrogels; C437167:poly(ethylene glycol)diacrylate; D011092:Polyethylene Glycols; D020723:Glucuronic Acid; D005780:Gelatin
Bio-ink; Biofabrication; Extrusion bioprinting; Mesenchymal stem cells; Oxidative stress; Photo-polymerization