Recapitulating cranial osteogenesis with neural crest cells in 3-D microenvironments
File(s)
Accepted version
Author(s)
Type
Journal Article
Abstract
The experimental systems that recapitulate the complexity of native tissues and enable precise control over the microenvironment are becoming essential for the pre-clinical tests of therapeutics and tissue engineering. Here, we described a strategy to develop an in vitro platform to study the developmental biology of craniofacial osteogenesis. In this study, we directly osteo-differentiated cranial neural crest cells (CNCCs) in a 3-D in vitro bioengineered microenvironment. Cells were encapsulated in the gelatin-based photo-crosslinkable hydrogel and cultured up to three weeks. We demonstrated that this platform allows efficient differentiation of p75 positive CNCCs to cells expressing osteogenic markers corresponding to the sequential developmental phases of intramembranous ossification. During the course of culture, we observed a decrease in the expression of early osteogenic marker Runx2, while the other mature osteoblast and osteocyte markers such as Osterix, Osteocalcin, Osteopontin and Bone sialoprotein increased. We analyzed the ossification of the secreted matrix with alkaline phosphatase and quantified the newly secreted hydroxyapatite. The Field Emission Scanning Electron Microscope (FESEM) images of the bioengineered hydrogel constructs revealed the native-like osteocytes, mature osteoblasts, and cranial bone tissue morphologies with canaliculus-like intercellular connections. This platform provides a broadly applicable model system to potentially study diseases involving primarily embryonic craniofacial bone disorders, where direct diagnosis and adequate animal disease models are limited.
Date Issued
2015-12-07
Date Acceptance
2015-12-02
Citation
Acta Biomaterialia, 2015, 31, pp.301-311
ISSN
1878-7568
Publisher
Elsevier
Start Page
301
End Page
311
Journal / Book Title
Acta Biomaterialia
Volume
31
Copyright Statement
© 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved. This manuscript is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Identifier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000370086100027&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
Subjects
Science & Technology
Technology
Engineering, Biomedical
Materials Science, Biomaterials
Engineering
Materials Science
3-D culture
Controlling 3-D tissue microenvironment
Ectodermal to mesodermal differentiation
Bone
STEM-CELLS
CRANIOFACIAL DEVELOPMENT
CARTILAGE FORMATION
BONE-FORMATION
TISSUE
SOX9
DIFFERENTIATION
CULTURE
TRANSCRIPTION
VERTEBRATES
Alkaline Phosphatase
Animals
Cell Culture Techniques
Cell Differentiation
Core Binding Factor Alpha 1 Subunit
Cryoelectron Microscopy
Culture Media
Freeze Fracturing
Gelatin
Hydrogels
Immunohistochemistry
Mice
Microscopy, Electron, Scanning
Neural Crest
Osteocalcin
Osteogenesis
Osteopontin
Skull
Tissue Engineering
Transcription Factors
Biomedical Engineering
MD Multidisciplinary
Publication Status
Published