Hypoxia decreases osteogenic differentiation potential but enhances vasculogenic differentiation potential of human adipose stem cells on biphasic calcium phosphate scaffolds in fibrin gels

Fransisca A.S. van Esterik, Jolanda M.A. Hogervorst, Pieter Koolwijk, Jenneke Klein-Nulend

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Background: In bone tissue engineering, biphasic calcium phosphate (BCP) with different hydroxyapatite (HA)/ß-tricalcium phosphate (ß-TCP) ratios can be combined with regeneration-competent stem cells, and bioscaffolds like fibrin that trigger neovascularization. We have shown recently that human adipose stem cells (hASCs) on BCP with a HA/ß-TCP ratio of 20/80 (BCP20/80) incorporated in fibrin gel show enhanced osteogenic and/or vasculogenic differentiation potential compared to BCP with a HA/ß-TCP ratio of 60/40 (BCP60/40) in vitro. A hypoxic microenvironment for the cells might occur in these hASC-seeded BCP-based composites after implantation in vivo. This limits oxygen supply which is essential for successful bone tissue engineering. Objectives: Here we aimed to test the effect of hypoxia on osteogenic and/or vasculogenic differentiation of hASC-seeded BCP60/40 or BCP20/80 scaffolds in fibrin gels as well as fibrin gel degradation. Methods: BCP60/40-based composites and BCP20/80-based composites were cultured under normoxia (20% O2) or hypoxia (1% O2) for 12 days, and total DNA content, alkaline phosphatase activity, nitric oxide production, vascular endothelial growth factor production, and fibrin gel degradation were assessed during culture. Results: Hypoxia similarly decreased total DNA content at day 7 (1.3-fold) and 12 (1.2-fold) in both composites. It also decreased alkaline phosphatase activity at day 5 (5.0 and 4.7-fold), day 7 (4.5 and 6-fold), and 12 (7.1 and 6.8-fold) in BCP60/40-based composites and BCP20/80-based composites. Hypoxia similarly increased vascular endothelial growth factor production at day 5 (3.9-fold) and day 7 (2.6-fold) by both BCP-based composites. Hypoxia did not affect nitric oxide production and fibrin gel degradation in both BCP-based composites at all-time points. Conclusion: Hypoxia decreased osteogenic differentiation but enhanced vasculogenic differentiation of cultured BCP-based composites. These in vitro results implicate that cell-seeded BCP-based composites implanted in vivo, might enhance vascular endothelial growth factor production resulting in sprouting and tube formation leading to increased survival of cells in these composites, and stimulation of bone formation.

Original languageEnglish
Pages (from-to)41-52
Number of pages12
JournalJournal of Stem Cells
Volume12
Issue number2
Publication statusPublished - 2017

Cite this

@article{e42664eb52634f4da4e7deddfe3836b7,
title = "Hypoxia decreases osteogenic differentiation potential but enhances vasculogenic differentiation potential of human adipose stem cells on biphasic calcium phosphate scaffolds in fibrin gels",
abstract = "Background: In bone tissue engineering, biphasic calcium phosphate (BCP) with different hydroxyapatite (HA)/{\ss}-tricalcium phosphate ({\ss}-TCP) ratios can be combined with regeneration-competent stem cells, and bioscaffolds like fibrin that trigger neovascularization. We have shown recently that human adipose stem cells (hASCs) on BCP with a HA/{\ss}-TCP ratio of 20/80 (BCP20/80) incorporated in fibrin gel show enhanced osteogenic and/or vasculogenic differentiation potential compared to BCP with a HA/{\ss}-TCP ratio of 60/40 (BCP60/40) in vitro. A hypoxic microenvironment for the cells might occur in these hASC-seeded BCP-based composites after implantation in vivo. This limits oxygen supply which is essential for successful bone tissue engineering. Objectives: Here we aimed to test the effect of hypoxia on osteogenic and/or vasculogenic differentiation of hASC-seeded BCP60/40 or BCP20/80 scaffolds in fibrin gels as well as fibrin gel degradation. Methods: BCP60/40-based composites and BCP20/80-based composites were cultured under normoxia (20{\%} O2) or hypoxia (1{\%} O2) for 12 days, and total DNA content, alkaline phosphatase activity, nitric oxide production, vascular endothelial growth factor production, and fibrin gel degradation were assessed during culture. Results: Hypoxia similarly decreased total DNA content at day 7 (1.3-fold) and 12 (1.2-fold) in both composites. It also decreased alkaline phosphatase activity at day 5 (5.0 and 4.7-fold), day 7 (4.5 and 6-fold), and 12 (7.1 and 6.8-fold) in BCP60/40-based composites and BCP20/80-based composites. Hypoxia similarly increased vascular endothelial growth factor production at day 5 (3.9-fold) and day 7 (2.6-fold) by both BCP-based composites. Hypoxia did not affect nitric oxide production and fibrin gel degradation in both BCP-based composites at all-time points. Conclusion: Hypoxia decreased osteogenic differentiation but enhanced vasculogenic differentiation of cultured BCP-based composites. These in vitro results implicate that cell-seeded BCP-based composites implanted in vivo, might enhance vascular endothelial growth factor production resulting in sprouting and tube formation leading to increased survival of cells in these composites, and stimulation of bone formation.",
keywords = "Adipose stem cells, Biphasic calcium phosphate, Hypoxia, Osteogenic differentiation, Vasculogenic differentiation",
author = "{van Esterik}, {Fransisca A.S.} and Hogervorst, {Jolanda M.A.} and Pieter Koolwijk and Jenneke Klein-Nulend",
year = "2017",
language = "English",
volume = "12",
pages = "41--52",
journal = "Journal of Stem Cells",
issn = "1556-8539",
publisher = "Nova Science Publishers, Inc.",
number = "2",

}

Hypoxia decreases osteogenic differentiation potential but enhances vasculogenic differentiation potential of human adipose stem cells on biphasic calcium phosphate scaffolds in fibrin gels. / van Esterik, Fransisca A.S.; Hogervorst, Jolanda M.A.; Koolwijk, Pieter; Klein-Nulend, Jenneke.

In: Journal of Stem Cells, Vol. 12, No. 2, 2017, p. 41-52.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Hypoxia decreases osteogenic differentiation potential but enhances vasculogenic differentiation potential of human adipose stem cells on biphasic calcium phosphate scaffolds in fibrin gels

AU - van Esterik, Fransisca A.S.

AU - Hogervorst, Jolanda M.A.

AU - Koolwijk, Pieter

AU - Klein-Nulend, Jenneke

PY - 2017

Y1 - 2017

N2 - Background: In bone tissue engineering, biphasic calcium phosphate (BCP) with different hydroxyapatite (HA)/ß-tricalcium phosphate (ß-TCP) ratios can be combined with regeneration-competent stem cells, and bioscaffolds like fibrin that trigger neovascularization. We have shown recently that human adipose stem cells (hASCs) on BCP with a HA/ß-TCP ratio of 20/80 (BCP20/80) incorporated in fibrin gel show enhanced osteogenic and/or vasculogenic differentiation potential compared to BCP with a HA/ß-TCP ratio of 60/40 (BCP60/40) in vitro. A hypoxic microenvironment for the cells might occur in these hASC-seeded BCP-based composites after implantation in vivo. This limits oxygen supply which is essential for successful bone tissue engineering. Objectives: Here we aimed to test the effect of hypoxia on osteogenic and/or vasculogenic differentiation of hASC-seeded BCP60/40 or BCP20/80 scaffolds in fibrin gels as well as fibrin gel degradation. Methods: BCP60/40-based composites and BCP20/80-based composites were cultured under normoxia (20% O2) or hypoxia (1% O2) for 12 days, and total DNA content, alkaline phosphatase activity, nitric oxide production, vascular endothelial growth factor production, and fibrin gel degradation were assessed during culture. Results: Hypoxia similarly decreased total DNA content at day 7 (1.3-fold) and 12 (1.2-fold) in both composites. It also decreased alkaline phosphatase activity at day 5 (5.0 and 4.7-fold), day 7 (4.5 and 6-fold), and 12 (7.1 and 6.8-fold) in BCP60/40-based composites and BCP20/80-based composites. Hypoxia similarly increased vascular endothelial growth factor production at day 5 (3.9-fold) and day 7 (2.6-fold) by both BCP-based composites. Hypoxia did not affect nitric oxide production and fibrin gel degradation in both BCP-based composites at all-time points. Conclusion: Hypoxia decreased osteogenic differentiation but enhanced vasculogenic differentiation of cultured BCP-based composites. These in vitro results implicate that cell-seeded BCP-based composites implanted in vivo, might enhance vascular endothelial growth factor production resulting in sprouting and tube formation leading to increased survival of cells in these composites, and stimulation of bone formation.

AB - Background: In bone tissue engineering, biphasic calcium phosphate (BCP) with different hydroxyapatite (HA)/ß-tricalcium phosphate (ß-TCP) ratios can be combined with regeneration-competent stem cells, and bioscaffolds like fibrin that trigger neovascularization. We have shown recently that human adipose stem cells (hASCs) on BCP with a HA/ß-TCP ratio of 20/80 (BCP20/80) incorporated in fibrin gel show enhanced osteogenic and/or vasculogenic differentiation potential compared to BCP with a HA/ß-TCP ratio of 60/40 (BCP60/40) in vitro. A hypoxic microenvironment for the cells might occur in these hASC-seeded BCP-based composites after implantation in vivo. This limits oxygen supply which is essential for successful bone tissue engineering. Objectives: Here we aimed to test the effect of hypoxia on osteogenic and/or vasculogenic differentiation of hASC-seeded BCP60/40 or BCP20/80 scaffolds in fibrin gels as well as fibrin gel degradation. Methods: BCP60/40-based composites and BCP20/80-based composites were cultured under normoxia (20% O2) or hypoxia (1% O2) for 12 days, and total DNA content, alkaline phosphatase activity, nitric oxide production, vascular endothelial growth factor production, and fibrin gel degradation were assessed during culture. Results: Hypoxia similarly decreased total DNA content at day 7 (1.3-fold) and 12 (1.2-fold) in both composites. It also decreased alkaline phosphatase activity at day 5 (5.0 and 4.7-fold), day 7 (4.5 and 6-fold), and 12 (7.1 and 6.8-fold) in BCP60/40-based composites and BCP20/80-based composites. Hypoxia similarly increased vascular endothelial growth factor production at day 5 (3.9-fold) and day 7 (2.6-fold) by both BCP-based composites. Hypoxia did not affect nitric oxide production and fibrin gel degradation in both BCP-based composites at all-time points. Conclusion: Hypoxia decreased osteogenic differentiation but enhanced vasculogenic differentiation of cultured BCP-based composites. These in vitro results implicate that cell-seeded BCP-based composites implanted in vivo, might enhance vascular endothelial growth factor production resulting in sprouting and tube formation leading to increased survival of cells in these composites, and stimulation of bone formation.

KW - Adipose stem cells

KW - Biphasic calcium phosphate

KW - Hypoxia

KW - Osteogenic differentiation

KW - Vasculogenic differentiation

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M3 - Article

VL - 12

SP - 41

EP - 52

JO - Journal of Stem Cells

JF - Journal of Stem Cells

SN - 1556-8539

IS - 2

ER -