Journal Article

An optimized 3D-printed perfusion bioreactor for homogeneous cell seeding in bone substitute scaffolds for future chairside applications

Fechner, Carsten, Voges, Annika, Krause, Bernd Joachim, Siewert, Stefan, Khaimov, Valeria, Bergner, Carina, Engel, Nadja, Ott, Robert, Stenzel, Jan, Liese, Jan, Schmitz, Klaus-Peter, Frerich, Bernhard

2021

Overview

A clinical implementation of cell-based bone regeneration in combination with scaffold materials requires the development of efficient, controlled and reproducible seeding procedures and a tailor-made bioreactor design. A perfusion system for efficient, homogeneous, and rapid seeding with human adipogenic stem cells in bone substitute scaffolds was designed. Variants concerning medium inlet and outlet port geometry, i.e. cylindrical or conical diffuser, cell concentration, perfusion mode and perfusion rates were simulated in silico. Cell distribution during perfusion was monitored by dynamic [ 18 F]FDG micro-PET/CT and validated by laser scanning microscopy with three-dimensional image reconstruction. By iterative feedback of the in silico and in vitro experiments, the homogeneity of cell distribution throughout the scaffold was optimized with adjustment of flow rates, cell density and perfusion properties. Finally, a bioreactor with a conical diffusor geometry was developed, that allows a homogeneous cell seeding (hoover coefficient: 0.24) in less than 60 min with an oscillating perfusion mode. During this short period of time, the cells initially adhere within the entire scaffold and stay viable. After two weeks, the formation of several cell layers was observed, which was associated with an osteogenic differentiation process. This newly designed bioreactor may be considered as a prototype for chairside application.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 631/532 / 631/61 / 639/301 / 692/308 / Animals / Biomarkers / Bioreactors / Bone growth / Bone marrow / Bone Regeneration / Bone Substitutes / Cell Culture Techniques / Cell density / Cell Differentiation / Cells, Cultured / Confocal microscopy / Design / Equipment Design / Experiments / Flow rates / Geometry / Humanities and Social Sciences / Humans / Image processing / Immunohistochemistry / Maxillofacial surgery / Medical research / Mesenchymal Stem Cells - cytology / Mesenchymal Stem Cells - metabolism / multidisciplinary / Nuclear medicine / Osteogenesis / Perfusion / Plastic surgery / Positron emission tomography / Positron Emission Tomography Computed Tomography / Printing, Three-Dimensional / Regeneration / Science / Science (multidisciplinary) / Simulation / Stem cells / Tissue Engineering - methods / Tissue Scaffolds