title: A single short session of media perfusion induces osteogenesis in hBMSCs cultured in porous scaffolds, dependent on cell differentiation stage.
abstract: Perfusing culture media through porous cell-seeded scaffolds is now a common approach within many tissue engineering strategies. Human bone-marrow derived mesenchymal stem cells (hBMSC) are a clinically valuable source of osteoprogenitors that respond to mechanical stimuli. However, the optimal mechanical conditions for their osteogenic stimulation in vitro have not been defined. Whereas the effects of short durations of media fluid flow have been studied in monolayers of osteoblastic cells, in 3D culture continuous or repeated perfusion is usually applied. Here, we investigated whether a short, single perfusion session applied to hBMSCs cultured in 3D would enhance their osteogenesis in vitro. We cultured hBMSCs on gelatine-coated, porous polyurethane scaffolds with osteogenic supplements and stimulated them with a single 2-h session of unidirectional, steady, 2.5 mL/min media perfusion, at either early or late stages of culture in 3D. Some cells were pre-treated in monolayer with osteogenic supplements to advance cell differentiation, followed by 3D culture also with the osteogenic supplements. We report that this single, short session of media perfusion can markedly enhance the expression of bone-related transcription and growth factors, and matrix components, by hBMSCs but that it is more effective when cells reach the pre-osteoblast or osteoblast differentiation stage. These findings could aid in the optimization of 3D culture protocols for efficient bone tissue engineering. Biotechnol. Bioeng. 2016;113: 1814-1824. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Faculty of Biology and Earth Sciences, Department of Cell Biology and Imaging, Jagiellonian University, 30-387 Kraków, Malopolska, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Filipowska
authors.affiliation: Department of Materials Science and Engineering, INSIGNEO Institute for In Silico Medicine, University of Sheffield, Sheffield, United Kingdom.
authors.forename: Gwendolen C
authors.initials: GC
authors.lastname: Reilly
authors.affiliation: Faculty of Biology and Earth Sciences, Department of Cell Biology and Imaging, Jagiellonian University, 30-387 Kraków, Malopolska, Poland. a.m.osyczka@uj.edu.pl.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Osyczka
mesh_terms: D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology*; D010012:Osteogenesis / Q000502:physiology*; D010477:Perfusion; D011140:Polyurethanes; D016062:Porosity; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D011140:Polyurethanes
keywords: 3D culture; flow perfusion bioreactor; human bone marrow-derived mesenchymal stem cells; osteogenesis; polyurethane scaffolds; unidirectional flow perfusion