Fabrication of hASCs-laden structures using extrusion-based cell printing supplemented with an electric field

Abstract

In this study, we proposed a hybrid cell-printing technique that combines a conventional extrusion-based cell-printing process with an electrohydrodynamic jet. The electric field stabilized the extruded struts of cell-embedding-hydrogel and reduced the damage to dispensed cells caused by the high wall shear stress in the dispensing nozzle. The new cell-printing process was optimized in terms of various processing parameters, applied electric field strength, nozzle movement speed, and distance between the nozzle tip and working stage. Using the optimal cell-embedding hydrogel composition (1×106cellsmL−1 in 4wt% alginate) and cell-printing process parameters (applied voltage, 1kV; nozzle movement speed, 12mms−1; distance, 0.7mm; current, 10.67±1.1nA), we achieved rapid and stable fabrication of a cell-laden structure without loss of cell viability or proliferation, the values of which were similar to those of the process without an electric field. Furthermore, by applying the same pneumatic pressure to fabricate cell-laden structures, considerably higher volume flow rate and cell viability at the same volume flow rate were achieved by the modified process compared with conventional extrusion-based cell-printing processes. To assess the feasibility of the method, the hydrogel containing human adipose stem cells (hASCs) and alginate (4wt%) was fabricated into a cell-laden porous structure in a layer-by-layer manner. The cell-laden structure exhibited reasonable initial hASC viability (87%), which was similar to that prior to processing of the cell-embedding-hydrogel. Statement of SignificanceThe extrusion-based cell-printing process has shortcomings, such as unstable flow and potential loss of cell viability. The unsteady flow can occur due to the high cell concentration, viscosity, and surface tension of bioinks. Also, cell viability post extrusion can be significantly reduced by damage of the cells due to the high wall shear stress in the extrusion nozzle. To overcome these limitations, we suggested an innovative cell-printing process that combines a conventional extrusion-based cellprinting process with an electric field. The electric field in the cell-printing process stabilized the extruded struts of bioink and dramatically reduced the damage to dispensed cells caused by the high wall shear stress in the dispensing nozzle.