Header

UZH-Logo

Maintenance Infos

Characterization and vascularization of a 3D-printed hydroxyapatite scaffold with different extracellular matrix coatings under perfusion culture


Burgio, Floriana; Rimmer, Natalie; Pieles, Uwe; Buschmann, Johanna; Beaufils-Hugot, Marina (2018). Characterization and vascularization of a 3D-printed hydroxyapatite scaffold with different extracellular matrix coatings under perfusion culture. Biology Open, 7(12):1-11.

Abstract

For the fabrication of appropriate bone tissue-engineered constructs several prerequisites should be fulfilled. They should offer long-term stability, allow proper cell attachment and proliferation and furthermore be osteoinductive and easy to be vascularized. Having these requirements as background, we fabricated a novel porous 3D-printed hydroxyapatite (HA) scaffold and treated it with oxygen plasma (OPT). MG-63 pre-osteoblast-seeded bone constructs allowed good cell attachment and proliferation, which was even better when cultivated in a perfusion flow bioreactor. Moreover, the deposition of extracellular matrix (ECM) on the otherwise inorganic surface changed the mechanical properties in a favourable manner: elasticity increased from 42.95±1.09 to 91.9±5.1 MPa (assessed by nanoindentation). Compared to static conditions, osteogenic differentiation was enhanced in the bioreactor, with upregulation of ALP, collagen I and osteocalcin gene expression. In parallel experiments, primary human bone marrow mesenchymal stromal cells (hBMSCs) were used and findings under dynamic conditions were similar; with a higher commitment towards osteoblasts compared to static conditions. In addition, angiogenic markers CD31, eNOS and VEGF were upregulated, especially when osteogenic medium was used rather than proliferative medium. To compare differently fabricated ECMs in terms of vascularization, decellularized constructs were tested in the chorioallantoic membrane (CAM) assay with subsequent assessment of the functional perfusion capacity by MRI in the living chick embryo. Here, vascularization induced by ECM from osteogenic medium led to a vessel distribution more homogenous throughout the construct, while ECM from proliferative medium enhanced vessel density at the interface and, to a lower extent, at the middle and top. We conclude that dynamic cultivation of a novel porous OPT HA scaffold with hBMSCs in osteogenic medium and subsequent decellularization provides a promising off-the-shelf bone tissue-engineered construct.

Abstract

For the fabrication of appropriate bone tissue-engineered constructs several prerequisites should be fulfilled. They should offer long-term stability, allow proper cell attachment and proliferation and furthermore be osteoinductive and easy to be vascularized. Having these requirements as background, we fabricated a novel porous 3D-printed hydroxyapatite (HA) scaffold and treated it with oxygen plasma (OPT). MG-63 pre-osteoblast-seeded bone constructs allowed good cell attachment and proliferation, which was even better when cultivated in a perfusion flow bioreactor. Moreover, the deposition of extracellular matrix (ECM) on the otherwise inorganic surface changed the mechanical properties in a favourable manner: elasticity increased from 42.95±1.09 to 91.9±5.1 MPa (assessed by nanoindentation). Compared to static conditions, osteogenic differentiation was enhanced in the bioreactor, with upregulation of ALP, collagen I and osteocalcin gene expression. In parallel experiments, primary human bone marrow mesenchymal stromal cells (hBMSCs) were used and findings under dynamic conditions were similar; with a higher commitment towards osteoblasts compared to static conditions. In addition, angiogenic markers CD31, eNOS and VEGF were upregulated, especially when osteogenic medium was used rather than proliferative medium. To compare differently fabricated ECMs in terms of vascularization, decellularized constructs were tested in the chorioallantoic membrane (CAM) assay with subsequent assessment of the functional perfusion capacity by MRI in the living chick embryo. Here, vascularization induced by ECM from osteogenic medium led to a vessel distribution more homogenous throughout the construct, while ECM from proliferative medium enhanced vessel density at the interface and, to a lower extent, at the middle and top. We conclude that dynamic cultivation of a novel porous OPT HA scaffold with hBMSCs in osteogenic medium and subsequent decellularization provides a promising off-the-shelf bone tissue-engineered construct.

Statistics

Citations

Dimensions.ai Metrics
8 citations in Web of Science®
7 citations in Scopus®
Google Scholar™

Altmetrics

Downloads

28 downloads since deposited on 13 Feb 2019
17 downloads since 12 months
Detailed statistics

Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > University Hospital Zurich > Clinic for Reconstructive Surgery
Dewey Decimal Classification:610 Medicine & health
Scopus Subject Areas:Life Sciences > General Biochemistry, Genetics and Molecular Biology
Life Sciences > General Agricultural and Biological Sciences
Language:English
Date:26 November 2018
Deposited On:13 Feb 2019 11:22
Last Modified:15 Apr 2020 22:49
Publisher:The Company of Biologists Ltd.
ISSN:2046-6390
OA Status:Gold
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1242/bio.034488
PubMed ID:30341104

Download

Gold Open Access

Download PDF  'Characterization and vascularization of a 3D-printed hydroxyapatite scaffold with different extracellular matrix coatings under perfusion culture'.
Preview
Content: Published Version
Filetype: PDF
Size: 1MB
View at publisher
Licence: Creative Commons: Attribution 4.0 International (CC BY 4.0)