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1.
Biofabrication ; 16(1)2023 10 11.
Article in English | MEDLINE | ID: mdl-37769669

ABSTRACT

The outcome of three-dimensional (3D) bioprinting heavily depends, amongst others, on the interaction between the developed bioink, the printing process, and the printing equipment. However, if this interplay is ensured, bioprinting promises unmatched possibilities in the health care area. To pave the way for comparing newly developed biomaterials, clinical studies, and medical applications (i.e. printed organs, patient-specific tissues), there is a great need for standardization of manufacturing methods in order to enable technology transfers. Despite the importance of such standardization, there is currently a tremendous lack of empirical data that examines the reproducibility and robustness of production in more than one location at a time. In this work, we present data derived from a round robin test for extrusion-based 3D printing performance comprising 12 different academic laboratories throughout Germany and analyze the respective prints using automated image analysis (IA) in three independent academic groups. The fabrication of objects from polymer solutions was standardized as much as currently possible to allow studying the comparability of results from different laboratories. This study has led to the conclusion that current standardization conditions still leave room for the intervention of operators due to missing automation of the equipment. This affects significantly the reproducibility and comparability of bioprinting experiments in multiple laboratories. Nevertheless, automated IA proved to be a suitable methodology for quality assurance as three independently developed workflows achieved similar results. Moreover, the extracted data describing geometric features showed how the function of printers affects the quality of the printed object. A significant step toward standardization of the process was made as an infrastructure for distribution of material and methods, as well as for data transfer and storage was successfully established.


Subject(s)
Bioprinting , Humans , Bioprinting/methods , Reproducibility of Results , Tissue Scaffolds/chemistry , Biocompatible Materials , Printing, Three-Dimensional , Tissue Engineering/methods
2.
Polymers (Basel) ; 15(8)2023 Apr 09.
Article in English | MEDLINE | ID: mdl-37111976

ABSTRACT

Three-dimensional bioprinting and especially extrusion-based printing as a most frequently employed method in this field is constantly evolving as a discipline in regenerative medicine and tissue engineering. However, the lack of relevant standardized analytics does not yet allow an easy comparison and transfer of knowledge between laboratories regarding newly developed bioinks and printing processes. This work revolves around the establishment of a standardized method, which enables the comparability of printed structures by controlling for the extrusion rate based on the specific flow behavior of each bioink. Furthermore, printing performance was evaluated by image-processing tools to verify the printing accuracy for lines, circles, and angles. In addition, and complementary to the accuracy metrics, a dead/live staining of embedded cells was performed to investigate the effect of the process on cell viability. Two bioinks, based on alginate and gelatin methacryloyl, which differed in 1% (w/v) alginate content, were tested for printing performance. The automated image processing tool reduced the analytical time while increasing reproducibility and objectivity during the identification of printed objects. During evaluation of the processing effect of the mixing of cell viability, NIH 3T3 fibroblasts were stained and analyzed after the mixing procedure and after the extrusion process using a flow cytometer, which evaluated a high number of cells. It could be observed that the small increase in alginate content made little difference in the printing accuracy but had a considerable strong effect on cell viability after both processing steps.

3.
Front Bioeng Biotechnol ; 10: 831350, 2022.
Article in English | MEDLINE | ID: mdl-35309988

ABSTRACT

Bioprinting is increasingly regarded as a suitable additive manufacturing method in biopharmaceutical process development and formulation. In order to manage the leap from research to industrial application, higher levels of reproducibility and a standardized bioprinting process are prerequisites. This said, the concept of process analytical technologies, standard in the biopharmaceutical industry, is still at its very early steps. To date most extrusion-based printing processes are controlled over penumatic pressure and thus not adaptive to environmental or system related changes over several experimental runs. A constant set pressure applied over a number of runs, might lead to variations in flow rate and thus to unreliable printed constructs. With this in mind, the simple question arises whether a printing process based on a set flow rate could improve reproduciblity and transfer to different printing systems. The control and monitoring of flow rate aim to introduce the concept of PAT in the field of bioprinting. This study investigates the effect of different processing modes (set pressure vs. set flow rate) on printing reproducibility occurring during an extrusion-based printing process consisting of 6 experimental runs consisting of 3 printed samples each. Additionally, the influence of different filling levels of the ink containing cartridge during a printing process was determined. Different solutions based on a varying amount of alginate polymer and Kolliphor hydrogels in varying concentrations showed the need for individual setting of printing parameter. To investigate parameter transferability among different devices two different printers were used and the flow was monitored using a flow sensor attached to the printing unit. It could be demonstrated that a set flow rate controlled printing process improved accuracy and the filling level also affects the accuracy of printing, the magnitude of this effects varies as the cartridge level declined. The transferability between printed devices was eased by setting the printing parameters according to a set flow rate of each bioink disregarding the value of the set pressure. Finally, by a bioprinting porcess control based on a set flow rate, the coefficient of variance for printed objects could be reduced from 0.2 to 0.02 for 10% (w/v) alginate polymer solutions.

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