Journal
ACS BIOMATERIALS SCIENCE & ENGINEERING
Volume 5, Issue 9, Pages 4522-4530Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsbiomaterials.9b00257
Keywords
gradients; microscopy; consumption rate; engineered tissues; three-dimensional
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Funding
- National Institute of Allergy and Infectious Disease [U19AI11649]
- National Institute of Diabetes and Digestive and Kidney Diseases [F30DK108541]
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A major barrier to the creation of engineered organs is the limited diffusion of oxygen through biological tissues. Advances in biofabrication bring us increasingly closer to complex vascular networks capable of supplying oxygen to large cellularized scaffolds. However, technologies for monitoring oxygen levels in engineered tissues do not accommodate imaging depths of more than a few dozen micrometers. Here, we report the creation of fluorescent porphyrinhydrogel microparticles that can be used at depths of 2 mm into artificial tissues. By combining an oxygen-responsive porphyrin dye with a reference dye, the microparticles generate a ratiometric signal that is photostable, unaffected by attenuation from biological material, and responsive to physiological change in oxygen concentration. These microparticles can measure long-distance oxygen gradients within 3D, cellularized constructs and accurately report cellular oxygen consumption rates. Furthermore, they are compatible with a number of hydrogel polymerization chemistries and cell types, including primary human cells. We believe this technology will significantly advance efforts to visualize oxygen gradients in cellularized constructs and inform efforts to tissue engineer solid organs.
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