Abstract

Biocompatibility is very important for cell growth using 3D printers, but biocompatibility materials are very expensive. In this study, we investigated the possibility of cell culture by the surface modification of relatively low-cost industrial materials and an efficient three-dimensional (3D) scaffold made with an industrial ABS filament for cell proliferation, spheroid formation, and drug screening applications. We evaluated the adequate structure among two-layer square shape 3D scaffolds printed by fused deposition modeling with variable infill densities (10–50%). Based on the effects of these scaffolds on cell proliferation and spheroid formation, we conducted experiments using the industrial ABS 3D scaffold (IA3D) with 40% of infill density, which presented an external dimension of (XYZ) 7650 µm × 7647 µm × 210 µm, 29.8% porosity, and 225 homogenous micropores (251.6 µm × 245.9 µm × 210 µm). In the IA3D, spheroids of cancer HepG2 cells and keratinocytes HaCaT cells appeared after 2 and 3 days of culture, respectively, whereas no spheroids were formed in 2D culture. A gold nanoparticle-coated industrial ABS 3D scaffold (GIA3D) exhibited enhanced biocompatible properties including increased spheroid formation by HepG2 cells compared to IA3D (1.3-fold) and 2D (38-fold) cultures. Furthermore, the cancer cells exhibited increased resistance to drug treatments in GIA3D, with cell viabilities of 122.9% in industrial GIA3D, 40.2% in IA3D, and 55.2% in 2D cultures when treated with 100 µM of mitoxantrone. Our results show that the newly engineered IA3D is an innovative 3D scaffold with upgraded properties for cell proliferation, spheroid formation, and drug-screening applications.

Highlights

  • Scaffold-based three-dimensional (3D) cell cultures are important technologies with numerous applications in drug development, tissue engineering, and regenerative medicine [1,2]

  • Various polymers, such as polylactic acid (PLA), PCL, polyethylene glycol (PEG), and acrylonitrile butadiene styrene (ABS) have been used in 3D scaffold fabrication for cell culture and tissue engineering applications

  • We investigated a low-cost industrial ABS 3D scaffold (IA3D) printed by fused deposition modeling for cell proliferation, spheroid formation, and drug screening

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Summary

Introduction

Scaffold-based three-dimensional (3D) cell cultures are important technologies with numerous applications in drug development, tissue engineering, and regenerative medicine [1,2]. Three-dimensional models such as 3D scaffolds better mimic the in vivo conditions for cell studies, tissue organization, and drug screening applications, by comparison to conventional 2D models They can be considered as potent alternatives to animal testing. Apart from ABS-M30i, which is suitable for biotechnological applications, industrial ABS is known to be non-biodegradable and non-biocompatible [9] Various polymers, such as PLA, PCL, PEG, and ABS have been used in 3D scaffold fabrication for cell culture and tissue engineering applications. Our result is the first report of a gold nanoparticle-coated industrial ABS scaffold (GIA3D), which can be employed to stimulate cell viability and proliferation in vitro, as well as to mimic the in vivo conditions for drug testing

Fabrication and Characterization of IA3D
Cell Seeding in IA3D and GIA3D
Cell Morphology Cultured on Surface Coated with Au-NPs
Cytotoxicity of IA3D and GIA3D
Immunofluorescence Staining
Drug Screening in IA3D and GIA3D
Statistical Analysis
Properties of IA3D
Optimization of the Infill Density and Cell Seeding Density in IA3D
Improvement of Cell Viability in Surface-Modified GIA3D
Cytotoxicity of Drugs in HepG2 and HaCaT Cells Cultured on GIA3D
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