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Calcified Tissue International
Published in: Calcified Tissue International 5/2011

01-05-2011 | Original Research

A Scaffold-Free Multicellular Three-Dimensional In Vitro Model of Osteogenesis

Authors: Umut A. Gurkan, Vipuil Kishore, Keith W. Condon, Teresita M. Bellido, Ozan Akkus

Published in: Calcified Tissue International | Issue 5/2011

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Abstract

In vitro models of osteogenesis are essential for investigating bone biology and the effects of pharmaceutical, chemical, and physical cues on bone formation. Osteogenesis takes place in a complex three-dimensional (3D) environment with cells from both mesenchymal and hematopoietic origins. Existing in vitro models of osteogenesis include two-dimensional (2D) single type cell monolayers and 3D cultures. However, an in vitro scaffold-free multicellular 3D model of osteogenesis is missing. We hypothesized that the self-inductive ossification capacity of bone marrow tissue can be harnessed in vitro and employed as a scaffold-free multicellular 3D model of osteogenesis. Therefore, rat bone marrow tissue was cultured for 28 days in three settings: 2D monolayer, 3D homogenized pellet, and 3D organotypic explant. The ossification potential of marrow in each condition was quantified by micro-computed tomography. The 3D organotypic marrow explant culture resulted in the greatest level of ossification with plate-like bone formations (up to 5 mm in diameter and 0.24 mm in thickness). To evaluate the mimicry of the organotypic marrow explants to newly forming native bone tissue, detailed compositional and morphological analyses were performed, including characterization of the ossified matrix by histochemistry, immunohistochemistry, Raman microspectroscopy, energy dispersive X-ray spectroscopy, backscattered electron microscopy, and micromechanical tests. The results indicated that the 3D organotypic marrow explant culture model mimics newly forming native bone tissue in terms of the characteristics studied. Therefore, this platform holds significant potential to be used as a model of osteogenesis, offering an alternative to in vitro monolayer cultures and in vivo animal models.
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Metadata
Title
A Scaffold-Free Multicellular Three-Dimensional In Vitro Model of Osteogenesis
Authors
Umut A. Gurkan
Vipuil Kishore
Keith W. Condon
Teresita M. Bellido
Ozan Akkus
Publication date
01-05-2011
Publisher
Springer-Verlag
Published in
Calcified Tissue International / Issue 5/2011
Print ISSN: 0171-967X
Electronic ISSN: 1432-0827
DOI
https://doi.org/10.1007/s00223-011-9467-3

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