Analysis of multiple types of human cells subsequent to bioprinting with electrospraying technology

Abstract

The aim of the present study was to investigate bioprinting with electrospraying technology using multiple types of human cell suspensions as bio-ink, in order to lay the initial foundations for the application of the bioprinting technology in tissue engineering. In the current study, six types of human cells were selected and cultured, including human fibroblasts, human adipose-derived stem cells (hADSCs), human periodontal ligament cells (HPDLCs), adult human retinal pigment epithelial cells (ARPE-19), human umbilical vascular endothelial cells (HUVECs) and human gastric epithelial cell line (GES-1). Each cell type was divided into two groups, the experimental and control group. All the experimental group cells were electrosprayed using an electrospraying printer (voltage, 15 kV; flow rate, 150 µl/min) and collected in a petri dish placed 15 cm away from the needle (needle diameter, 0.5 mm). Subsequently, cell viability was detected by flow cytometry with a Live/Dead Viability kit. In addition, the cell morphological characteristics were observed with a phase-contrast microscope after 6 h of culturing in order to obtain adherent cells, while cell proliferation was analyzed using a Cell Counting Kit-8 assay. The control groups, without printing, were subjected to the same procedures as the experimental groups. The results of the cell viability and proliferation assays indicated a statistically significant difference after printing between the experiments and control groups only for the hADSCs (P<0.05); by contrast, no significant difference was observed in cell viability and proliferation for the other five cell types (P>0.05). In addition, there were no observable differences between all experimental and the control groups at any examined time point in the terms of cell morphological characteristics. In conclusion, bioprinting based on electrospraying technology demonstrated no distinct negative effect on cell vitality, proliferation and morphology in the present study, and thus the application of this novel technology to cell printing may provide a promising method in tissue engineering.

3D Bioprinting Human Induced Pluripotent Stem Cell Constructs for In Situ Cell Proliferation and Successive Multilineage Differentiation

“…There are a number of different yet-to-be-tested bioprinting platforms that may be suitable for 3D iPSC deposition including inkjet-based, [20] laser-assisted, [21] and electrospray [22,23] bioprinting. While it will be important to determine the utility of these platforms, our work represents the first example of bioprinting iPSCs for ensuing culture and expansion within a 3D construct.…”

Electrospraying: Possibilities and Challenges of Engineering Carriers for Biomedical Applications—A Mini Review

“…BES is considered safe for cells due to the low current (usually in the nanoampere range) albeit the high voltage up to several kilovolts. Recently, a variety of cells have been electrosprayed, including fibroblasts, adipose-derived stem cells (ADSCs), periodontal ligament cells, retinal pigment epithelial cells, umbilical vascular endothelial cells, gastric epithelial cells (Xin et al, 2016), and bone marrow-derived mesenchymal stem cells (McCrea et al, 2018). The overall results demonstrated no distinct negative effect of the electrospraying process on cell vitality, morphology, and proliferation.…”

3D bioprinting using stem cells

“…Thus, 3D stem cell bioprinting approaches can have huge implications in regenerative medicine, for disease modeling and treatment of heart disease and heart failure, as well as for toxicological studies and personalized drug testing (19,25). In fact, engineered myocardium grafts are currently in preclinical studies and may one day serve as economical and efficient solutions to MI (7,26). 3D bioprinting also has potential in valvular disease, as the ability to accurately reconstruct native heart valves has enormous clinical implications, including surgical planning processes as well as regenerative medicine (22).…”

“…Heart tissue is thick and complex, and thus requires adequate vascularization and appropriate enervation to allow for functionality and biocompatibility (27). Thus, further study is needed in generating adequately vascularized heart tissue constructs of clinically-relevant thickness that can appropriately respond to electrical impulses and maintain a synchronous beating pattern, especially given that cardiomyocytes are metabolically active cells (26). This is especially a challenge given the hierarchical structure of native myocardium (13).…”