@article{f0b62f19de674a74b2a1896030945a68,
title = "Engineering a 3d-bioprinted model of human heart valve disease using nanoindentation-based biomechanics",
abstract = "In calcific aortic valve disease (CAVD), microcalcifications originating from nanoscale calcifying vesicles disrupt the aortic valve (AV) leaflets, which consist of three (biomechanically) distinct layers: the fibrosa, spongiosa, and ventricularis. CAVD has no pharmacotherapy and lacks in vitro models as a result of complex valvular biomechanical features surrounding resident mechanosensitive valvular interstitial cells (VICs). We measured layer-specific mechanical properties of the human AV and engineered a three-dimensional (3D)-bioprinted CAVD model that recapitulates leaflet layer biomechanics for the first time. Human AV leaflet layers were separated by microdissection, and nanoindentation determined layer-specific Young{\textquoteright}s moduli. Methacrylated gelatin (GelMA)/methacrylated hyaluronic acid (HAMA) hydrogels were tuned to duplicate layer-specific mechanical characteristics, followed by 3D-printing with encapsulated human VICs. Hydrogels were exposed to osteogenic media (OM) to induce microcalcification, and VIC pathogenesis was assessed by near infrared or immunofluorescence microscopy. Median Young{\textquoteright}s moduli of the AV layers were 37.1, 15.4, and 26.9 kPa (fibrosa/spongiosa/ ventricularis, respectively). The fibrosa and spongiosa Young{\textquoteright}s moduli matched the 3D 5% GelMa/1% HAMA UV-crosslinked hydrogels. OM stimulation of VIC-laden bioprinted hydrogels induced microcalcification without apoptosis. We report the first layer-specific measurements of human AV moduli and a novel 3D-bioprinted CAVD model that potentiates microcalcification by mimicking the native AV mechanical environment. This work sheds light on valvular mechanobiology and could facilitate high-throughput drug-screening in CAVD.",
keywords = "3D printing, Aortic valve, Bioprinting, Calcific aortic valve disease, Calcification, Mechanobiology, Microdissection, Nanoindentation",
author = "{van der Valk}, {Dewy C.} and {van der Ven}, {Casper F.T.} and Blaser, {Mark C.} and Grolman, {Joshua M.} and Wu, {Pin Jou} and Fenton, {Owen S.} and Lee, {Lang H.} and Tibbitt, {Mark W.} and Andresen, {Jason L.} and Wen, {Jennifer R.} and Ha, {Anna H.} and Fabrizio Buffolo and Mil, {Alain van} and Bouten, {Carlijn V.C.} and Body, {Simon C.} and Mooney, {David J.} and Sluijter, {Joost P.G.} and Masanori Aikawa and Jesper Hjortnaes and Robert Langer and Elena Aikawa",
note = "Funding Information: This research was funded by the National Institutes of Health (NIH) R01 grants R01HL114805, R01HL136431 and R01HL109889 (E.A.); the NIH/National Institute of Dental and Craniofacial Research (NIDCR) R01 grant DE013033 (D.M.); the Netherlands CardioVascular Research Initiative (CVON: The Dutch Heart Foundation, Dutch Federation of University Medical Centers, the Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Science) and Vrienden UMC Utrecht (C.V., J.S.); an unrestricted grant from CELLINK to Vrienden UMC Utrecht (C.V., J.S.); the Dutch Ministry of Education, Culture, and Science Gravitation Program grant 024.003.013 (C.V.C.B.); the Harvard Catalyst Advanced Microscopy Pilot grants (C.V., M.B., E.A.); and the NIH Ruth L. Kirschstein National Research Service Award F32HL122009 (M.W.T.). This work was conducted with support from Harvard Catalyst|The Harvard Clinical and Translational Science Center (National Center for Advancing Translational Sciences, National Institutes of Health Award UL1 TR001102) and financial contributions from Harvard University and its affiliated academic healthcare centers. This work was performed in part at Harvard University{\textquoteright}s Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Coordinated Infrastructure Network (NNCI), which is supported by the National Science Foundation under NSF award No. 1541959. We thank the Harvard Center for Biological Imaging for infrastructure and support. Funding Information: Funding: This research was funded by the National Institutes of Health (NIH) R01 grants R01HL114805, R01HL136431 and R01HL109889 (E.A.); the NIH/National Institute of Dental and Craniofacial Research (NIDCR) R01 grant DE013033 (D.M.); the Netherlands CardioVascular Research Initiative (CVON: The Dutch Heart Foundation, Dutch Federation of University Medical Centers, the Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Science) and Vrienden UMC Utrecht (C.V., J.S.); an unrestricted grant from CELLINK to Vrienden UMC Utrecht (C.V., J.S.); the Dutch Ministry of Education, Culture, and Science Gravitation Program grant 024.003.013 (C.V.C.B.); the Harvard Catalyst Advanced Microscopy Pilot grants (C.V., M.B., E.A.); and the NIH Ruth L. Kirschstein National Research Service Award F32HL122009 (M.W.T.). This work was conducted with support from Harvard Catalyst|The Harvard Clinical and Translational Science Center (National Center for Advancing Translational Sciences, National Institutes of Health Award UL1 TR001102) and financial contributions from Harvard University and its affiliated academic healthcare centers. This work was performed in part at Harvard University{\textquoteright}s Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Coordinated Infrastructure Network (NNCI), which is supported by the National Science Foundation under NSF award No. 1541959. We thank the Harvard Center for Biological Imaging for infrastructure and support. Publisher Copyright: {\textcopyright} 2018 by the authors. Licensee MDPI, Basel, Switzerland.",
year = "2018",
month = may,
day = "1",
doi = "10.3390/nano8050296",
language = "English",
volume = "8",
number = "5",
}