Probing mechanoregulation of neuronal differentiation by plasma lithography patterned elastomeric substrates.
Cells sense and interpret mechanical cues, including cell-cell and cell-substrate interactions, in the microenvironment to collectively regulate various physiological functions. Understanding the influences of these mechanical factors on cell behavior is critical for fundamental cell biology and for the development of novel strategies in regenerative medicine. Here, we demonstrate plasma lithography patterning on elastomeric substrates for elucidating the influences of mechanical cues on neuronal differentiation and neuritogenesis. The neuroblastoma cells form neuronal spheres on plasma-treated regions, which geometrically confine the cells over two weeks. The elastic modulus of the elastomer is controlled simultaneously by the crosslinker concentration. The cell-substrate mechanical interactions are also investigated by controlling the size of neuronal spheres with different cell seeding densities. These physical cues are shown to modulate with the formation of focal adhesions, neurite outgrowth, and the morphology of neuroblastoma. By systematic adjustment of these cues, along with computational biomechanical analysis, we demonstrate the interrelated mechanoregulatory effects of substrate elasticity and cell size. Taken together, our results reveal that the neuronal differentiation and neuritogenesis of neuroblastoma cells are collectively regulated via the cell-substrate mechanical interactions.
1] Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, Arizona 85721, USA [2] Centre for Analytical Instrumentation Development, The Korea Basic Science Institute, Deajeon305-806, Korea.
Ki-Hwan
KH
Nam
Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, Arizona 85721, USA.
Nima
N
Jamilpour
Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, Arizona 85721, USA.
Etienne
E
Mfoumou
The Key Laboratory for Complex Systems and Intelligence Science, The Institute of Automation, Chinese Academy of Sciences, Beijing, China.
Fei-Yue
FY
Wang
Department of Pharmacology and Toxicology, The University of Arizona, Tucson, Arizona. 85721, USA.
Donna D
DD
Zhang
Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, Arizona 85721, USA.
Pak Kin
PK
Wong
D002448:Cell Adhesion; D002450:Cell Communication; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D060833:Cellular Microenvironment; D003432:Cross-Linking Reagents / Q000737:chemistry; D055119:Elastic Modulus; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D040542:Mechanotransduction, Cellular; D055495:Neurogenesis; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D058626:Plasma Gases; D011327:Printing / Q000295:instrumentation / Q000379:methods*; D018874:Spheroids, Cellular / Q000378:metabolism* / Q000648:ultrastructure
D003432:Cross-Linking Reagents; D058626:Plasma Gases