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  • Time-lapsed imaging of nanocomposite scaffolds reveals increased bone formation in dynamic compression bioreactors.

Time-lapsed imaging of nanocomposite scaffolds reveals increased bone formation in dynamic compression bioreactors.

Communications biology (2021-01-27)
Gian Nutal Schädli, Jolanda R Vetsch, Robert P Baumann, Anke M de Leeuw, Esther Wehrle, Marina Rubert, Ralph Müller
ABSTRACT

Progress in bone scaffold development relies on cost-intensive and hardly scalable animal studies. In contrast to in vivo, in vitro studies are often conducted in the absence of dynamic compression. Here, we present an in vitro dynamic compression bioreactor approach to monitor bone formation in scaffolds under cyclic loading. A biopolymer was processed into mechanically competent bone scaffolds that incorporate a high-volume content of ultrasonically treated hydroxyapatite or a mixture with barium titanate nanoparticles. After seeding with human bone marrow stromal cells, time-lapsed imaging of scaffolds in bioreactors revealed increased bone formation in hydroxyapatite scaffolds under cyclic loading. This stimulatory effect was even more pronounced in scaffolds containing a mixture of barium titanate and hydroxyapatite and corroborated by immunohistological staining. Therefore, by combining mechanical loading and time-lapsed imaging, this in vitro bioreactor strategy may potentially accelerate development of engineered bone scaffolds and reduce the use of animals for experimentation.

MATERIALS
Product Number
Brand
Product Description

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Arg-Gly-Asp, ≥97% (TLC)
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Phalloidin–Tetramethylrhodamine B isothiocyanate, sequence from Amanita phalloides(synthetic: peptide sequence)
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Dopamine hydrochloride
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Barium titanate(IV), nanopowder (cubic crystalline phase), <100 nm particle size (BET), ≥99% trace metals basis
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Direct Red 80, Dye content 25 %
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Chloroform, contains 100-200 ppm amylenes as stabilizer, ≥99.5%
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Sodium chloride, BioXtra, ≥99.5% (AT)
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L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate, ≥95%
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Hydroxyapatite, nanopowder, <200 nm particle size (BET), ≥97%, synthetic
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Trizma® base, BioXtra, pH 10.5-12.0 (1 M in H2O), ≥99.9% (titration)
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Magnesium chloride, powder, <200 μm
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bisBenzimide H 33342 trihydrochloride, ≥98% (HPLC)
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Dichloromethane, anhydrous, ≥99.8%, contains 40-150 ppm amylene as stabilizer
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Alizarin Red S, certified by the BSC
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Triton X-100, laboratory grade