Engineering anisotropic biomimetic fibrocartilage microenvironment by bioprinting mesenchymal stem cells in nanoliter gel droplets.
Over the past decade, bioprinting has emerged as a promising patterning strategy to organize cells and extracellular components both in two and three dimensions (2D and 3D) to engineer functional tissue mimicking constructs. So far, tissue printing has neither been used for 3D patterning of mesenchymal stem cells (MSCs) in multiphase growth factor embedded 3D hydrogels nor been investigated phenotypically in terms of simultaneous differentiation into different cell types within the same micropatterned 3D tissue constructs. Accordingly, we demonstrated a biochemical gradient by bioprinting nanoliter droplets encapsulating human MSCs, bone morphogenetic protein 2 (BMP-2), and transforming growth factor β1 (TGF- β1), engineering an anisotropic biomimetic fibrocartilage microenvironment. Assessment of the model tissue construct displayed multiphasic anisotropy of the incorporated biochemical factors after patterning. Quantitative real time polymerase chain reaction (qRT-PCR) results suggested genomic expression patterns leading to simultaneous differentiation of MSC populations into osteogenic and chondrogenic phenotype within the multiphasic construct, evidenced by upregulation of osteogenesis and condrogenesis related genes during in vitro culture. Comprehensive phenotypic network and pathway analysis results, which were based on genomic expression data, indicated activation of differentiation related mechanisms, via signaling pathways, including TGF, BMP, and vascular endothelial growth factor.
Case Biomanufacturing and Microfabrication Laboratory, Mechanical and Aerospace Engineering Department, Department of Orthopaedics, Case Western Reserve University , Cleveland, Ohio 44106, United States.
Umut A
UA
Gurkan
Rami
R
El Assal
Simin E
SE
Yildiz
Yuree
Y
Sung
Alexander J
AJ
Trachtenberg
Winston P
WP
Kuo
Utkan
U
Demirci
D032701:Biomimetics / Q000379:methods*; D062028:Bioprinting / Q000379:methods*; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000235:genetics / Q000502:physiology*; D020219:Chondrogenesis / Q000235:genetics / Q000502:physiology; D051445:Fibrocartilage / Q000378:metabolism / Q000502:physiology*; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D020100:Hydrogels / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000378:metabolism / Q000502:physiology*; D010012:Osteogenesis / Q000235:genetics / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics; D023822:Tissue Engineering / Q000379:methods*; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
D055396:Bone Morphogenetic Protein 2; D020100:Hydrogels; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A