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  • Induction of osteogenic differentiation of bone marrow stromal cells on 3D polyester-based scaffolds solely by subphysiological fluidic stimulation in a laminar flow bioreactor.

Induction of osteogenic differentiation of bone marrow stromal cells on 3D polyester-based scaffolds solely by subphysiological fluidic stimulation in a laminar flow bioreactor.

Journal of tissue engineering (2021-07-16)
Shuntaro Yamada, Mohammed Ahmed Yassin, Thomas Schwarz, Jan Hansmann, Kamal Mustafa
ABSTRACT

The fatal determination of bone marrow mesenchymal stem/stromal cells (BMSC) is closely associated with mechano-environmental factors in addition to biochemical clues. The aim of this study was to induce osteogenesis in the absence of chemical stimuli using a custom-designed laminar flow bioreactor. BMSC were seeded onto synthetic microporous scaffolds and subjected to the subphysiological level of fluid flow for up to 21 days. During the perfusion, cell proliferation was significantly inhibited. There were also morphological changes, with F-actin polymerisation and upregulation of ROCK1. Notably, in BMSC subjected to flow, mRNA expression of osteogenic markers was significantly upregulated and RUNX2 was localised in the nuclei. Further, under perfusion, there was greater deposition of collagen type 1 and calcium onto the scaffolds. The results confirm that an appropriate level of fluid stimuli preconditions BMSC towards the osteoblastic lineage on 3D scaffolds in the absence of chemical stimulation, which highlights the utility of flow bioreactors in bone tissue engineering.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Oil Red O, certified by the Biological Stain Commission
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Fibrinogen from bovine plasma, Type I-S, 65-85% protein (≥75% of protein is clottable)
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Aprotinin from bovine lung, lyophilized powder, 3-8 TIU/mg solid
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L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate, ≥95%
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Anti-SCA-1 Antibody, from rabbit, purified by affinity chromatography
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L-Ascorbic acid, 99%
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Fluoroshield with DAPI, histology mounting medium
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Sodium borohydride, powder, ≥98.0%
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Dexamethasone, powder, BioReagent, suitable for cell culture, ≥97%
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Human Adipose Mesenchymal Stem Cells, Human Adipose Mesenchymal Stem Cells (MSCs) are isolated from human adipose tissue and are cryopreserved at P2 to ensure optimal phenotype and the highest viability and plating efficiency.