Abstract

3D printing is a rapidly evolving field for biological (bioprinting) and non-biological applications. Due to a high degree of freedom for geometrical parameters in 3D printing, prototype printing of bioreactors is a promising approach in the field of Tissue Engineering. The variety of printers, materials, printing parameters and device settings is difficult to overview both for beginners as well as for most professionals. In order to address this problem, we designed a guidance including test bodies to elucidate the real printing performance for a given printer system. Therefore, performance parameters such as accuracy or mechanical stability of the test bodies are systematically analysed. Moreover, post processing steps such as sterilisation or cleaning are considered in the test procedure. The guidance presented here is also applicable to optimise the printer settings for a given printer device. As proof of concept, we compared fused filament fabrication, stereolithography and selective laser sintering as the three most used printing methods. We determined fused filament fabrication printing as the most economical solution, while stereolithography is most accurate and features the highest surface quality. Finally, we tested the applicability of our guidance by identifying a printer solution to manufacture a complex bioreactor for a perfused tissue construct. Due to its design, the manufacture via subtractive mechanical methods would be 21-fold more expensive than additive manufacturing and therefore, would result in three times the number of parts to be assembled subsequently. Using this bioreactor we showed a successful 14-day-culture of a biofabricated collagen-based tissue construct containing human dermal fibroblasts as the stromal part and a perfusable central channel with human microvascular endothelial cells. Our study indicates how the full potential of biofabrication can be exploited, as most printed tissues exhibit individual shapes and require storage under physiological conditions, after the bioprinting process.

Highlights

  • Methodscomputer aided design (CAD) designComputer-aided design (CAD) of the geometries to be printed was performed with Solidworks Premium 2017 (Dassault Systèmes, France) and converted into STL-file with all test bodies arranged. (STL)-files.Machine-specific G-code-like files were generated from the STL-files for every printer using the individual company own software for the respective printer

  • To compare 3D printing methods and specific printers of the respective method, test bodies were designed to assess X, Y and Z resolution, printing of channels, angled overhangs and squared cups, which were used for measuring the leakage of the printed parts

  • The test bodies were designed in Solidworks software, containing the most digital information possible in the company’s own SLDPT-format (Fig 2A)

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Summary

Methods

CAD designComputer-aided design (CAD) of the geometries to be printed was performed with Solidworks Premium 2017 (Dassault Systèmes, France) and converted into STL-files.Machine-specific G-code-like files were generated from the STL-files for every printer using the individual company own software for the respective printer. For vaporised hydrogen peroxide (VH2O2) plasma sterilisation, the chamber of a plasma cleaner device (Pico LF PC 115656, Diener electronic GmbH & Co. KG, Germany) was preheated by an induced oxygen plasma with a generator power of 500 W for 12 min at a pressure of 0.3 mbar created by a gas flow of 12 standard cubic centimeters per minute (sccm). After the heating process, the foil-wrapped (Stericlin, VP Group, Germany) 3D printed parts and a metal vaporiser unit filled with 1.5 ml of 60% H2O2 solution (Thermo Fisher Scientific, USA) were placed in the plasma chamber. The atmosphere in the chamber was replaced by pure O2 gas to a pressure of 0.4 mbar before the start of the last plasma process at 300 W for 5 min. The chamber was flooded by pure oxygen gas for 120 s, before the atmosphere in the chamber was replaced by air

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