<sup>18</sup>F- based Quantification of the Osteogenic Potential of hMSCs

In bone tissue engineering, there is a constant need to design new methods for promoting in vitro osteogenic differentiation. Consequently, there is a strong demand for fast, effective and reliable methods to track and quantify osteogenesis in vitro. In this study, we used the radiopharmacon fluorin...

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Main Authors: Tobias Grossner, Uwe Haberkorn, Tobias Gotterbarm
Format: Article
Language:English
Published: MDPI AG 2020-10-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/21/20/7692
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spelling doaj-8c076ae033e0473abb091235b4a8df182020-11-25T02:45:35ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-10-01217692769210.3390/ijms21207692<sup>18</sup>F- based Quantification of the Osteogenic Potential of hMSCsTobias Grossner0Uwe Haberkorn1Tobias Gotterbarm2Trauma Surgery and Paraplegiology, Clinic for Orthopedics and Trauma Surgery, Center for Orthopedics, University Hospital Heidelberg, 69120 Heidelberg, GermanyDepartment of Nuclear Medicine, University Hospital Heidelberg, 69120 Heidelberg, GermanyDepartment of Orthopedics and Traumatology, Kepler University Hospital, Linz 4020, AustriaIn bone tissue engineering, there is a constant need to design new methods for promoting in vitro osteogenic differentiation. Consequently, there is a strong demand for fast, effective and reliable methods to track and quantify osteogenesis in vitro. In this study, we used the radiopharmacon fluorine-18 (<sup>18</sup>F) to evaluate the amount of hydroxylapatite produced by mesenchymal stem cells (MSCs) in a monolayer cell culture in vitro. The hydroxylapatite bound tracer was evaluated using µ-positron emission tomography (µ-PET) scanning and activimeter analysis. It was therefore possible to determine the amount of synthesized mineral and thus to conclude the osteogenic potential of the cells. A Student’s <i>t</i>-test revealed a highly significant difference regarding tracer uptake between the osteogenic group and the corresponding control group (µ-PET <i>p</i> = 0.043; activimeter analysis <i>p</i> = 0.012). This tracer uptake showed a highly significant correlation with the gold standard of quantitative Alizarin Red staining (ARS) (r<sup>2</sup> = 0.86) as well as with the absolute calcium content detected by inductively coupled plasma mass spectrometry (r<sup>2</sup> = 0.81). The results showed that <sup>18</sup>F labeling is a novel method to prove and quantify hydroxyapatite content in MSC monolayer cultures. The mineral layer remains intact for further analysis. This non-destructive in vitro method can be used to rapidly investigate bone tissue engineering strategies in terms of hydroxylapatite production, and could therefore accelerate the process of implementing new strategies in clinical practice.https://www.mdpi.com/1422-0067/21/20/7692mesenchymal stem cellsosteogenic quantificationosteogenic differentiation<sup>18</sup>Fµ-PETactivimeter analysis
collection DOAJ
language English
format Article
sources DOAJ
author Tobias Grossner
Uwe Haberkorn
Tobias Gotterbarm
spellingShingle Tobias Grossner
Uwe Haberkorn
Tobias Gotterbarm
<sup>18</sup>F- based Quantification of the Osteogenic Potential of hMSCs
International Journal of Molecular Sciences
mesenchymal stem cells
osteogenic quantification
osteogenic differentiation
<sup>18</sup>F
µ-PET
activimeter analysis
author_facet Tobias Grossner
Uwe Haberkorn
Tobias Gotterbarm
author_sort Tobias Grossner
title <sup>18</sup>F- based Quantification of the Osteogenic Potential of hMSCs
title_short <sup>18</sup>F- based Quantification of the Osteogenic Potential of hMSCs
title_full <sup>18</sup>F- based Quantification of the Osteogenic Potential of hMSCs
title_fullStr <sup>18</sup>F- based Quantification of the Osteogenic Potential of hMSCs
title_full_unstemmed <sup>18</sup>F- based Quantification of the Osteogenic Potential of hMSCs
title_sort <sup>18</sup>f- based quantification of the osteogenic potential of hmscs
publisher MDPI AG
series International Journal of Molecular Sciences
issn 1661-6596
1422-0067
publishDate 2020-10-01
description In bone tissue engineering, there is a constant need to design new methods for promoting in vitro osteogenic differentiation. Consequently, there is a strong demand for fast, effective and reliable methods to track and quantify osteogenesis in vitro. In this study, we used the radiopharmacon fluorine-18 (<sup>18</sup>F) to evaluate the amount of hydroxylapatite produced by mesenchymal stem cells (MSCs) in a monolayer cell culture in vitro. The hydroxylapatite bound tracer was evaluated using µ-positron emission tomography (µ-PET) scanning and activimeter analysis. It was therefore possible to determine the amount of synthesized mineral and thus to conclude the osteogenic potential of the cells. A Student’s <i>t</i>-test revealed a highly significant difference regarding tracer uptake between the osteogenic group and the corresponding control group (µ-PET <i>p</i> = 0.043; activimeter analysis <i>p</i> = 0.012). This tracer uptake showed a highly significant correlation with the gold standard of quantitative Alizarin Red staining (ARS) (r<sup>2</sup> = 0.86) as well as with the absolute calcium content detected by inductively coupled plasma mass spectrometry (r<sup>2</sup> = 0.81). The results showed that <sup>18</sup>F labeling is a novel method to prove and quantify hydroxyapatite content in MSC monolayer cultures. The mineral layer remains intact for further analysis. This non-destructive in vitro method can be used to rapidly investigate bone tissue engineering strategies in terms of hydroxylapatite production, and could therefore accelerate the process of implementing new strategies in clinical practice.
topic mesenchymal stem cells
osteogenic quantification
osteogenic differentiation
<sup>18</sup>F
µ-PET
activimeter analysis
url https://www.mdpi.com/1422-0067/21/20/7692
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