Quantifying migration and polarization of murine mesenchymal stem cells on different bone substitutes by confocal laser scanning microscopy

Roldán, J.C.; Chang, E.; Kelantan, M.; Jazayeri, L.; Deisinger, U.; Detsch, R.; Reichert, T.E.; Gurtner, G.C.

doi:10.1016/j.jcms.2010.01.004

« Previous Next »Journal of Cranio-Maxillo-Facial Surgery
Volume 38, Issue 8 , Pages 580-588, December 2010

Quantifying migration and polarization of murine mesenchymal stem cells on different bone substitutes by confocal laser scanning microscopy

J.C. Roldán
- Department of Cranio-Maxillofacial Surgery, University of Regensburg, Germany
- J. Camilo ROLDÁN, MD, DMD, Department of Cranio-Maxillofacial Surgery, University of Regensburg, 93042 Regensburg, Germany.
E. Chang
- Laboratory for Microvascular Surgery and Vascular Tissue Engineering, Stanford University, USA
M. Kelantan
- Laboratory for Microvascular Surgery and Vascular Tissue Engineering, Stanford University, USA
L. Jazayeri
- Laboratory for Microvascular Surgery and Vascular Tissue Engineering, Stanford University, USA
U. Deisinger
- Friedrich-Baur-Research-Institute for Biomaterials, University of Bayreuth, Germany
R. Detsch
- BioCer Entwicklungs-GmbH, Bayreuth, Germany
T.E. Reichert
- Department of Cranio-Maxillofacial Surgery, University of Regensburg, Germany
G.C. Gurtner
- Laboratory for Microvascular Surgery and Vascular Tissue Engineering, Stanford University, USA

Received 19 September 2009; accepted 19 January 2010. published online 02 March 2010.

Summary

Introduction

Cell migration is preceded by cell polarization. The aim of the present study was to evaluate the impact of the geometry of different bone substitutes on cell morphology and chemical responses in vitro.

Materials and methods

Cell polarization and migration were monitored temporally by using confocal laser scanning microscopy (CLSM) to follow green fluorescent protein (GFP)±mesenchymal stem cells (MSCs) on anorganic cancellous bovine bone (Bio-Oss^®), β-tricalcium phosphate (β-TCP) (chronOS^®) and highly porous calcium phosphate ceramics (Friedrich-Baur-Research-Institute for Biomaterials, Germany). Differentiation GFP±MSCs was observed using pro-angiogenic and pro-osteogenic biomarkers.

Results

At the third day of culture polarized vs. non-polarized cellular sub-populations were clearly established. Biomaterials that showed more than 40% of polarized cells at the 3rd day of culture, subsequently showed an enhanced cell migration compared to biomaterials, where non-polarized cells predominated (p<0.003). This trend continued untill the 7th day of culture (p<0.003). The expression of vascular endothelial growth factor was enhanced in biomaterials where cell polarization predominated at the 7th day of culture (p=0.001).

Conclusions

This model opens an interesting approach to understand osteoconductivity at a cellular level. MSCs are promising in bone tissue engineering considering the strong angiogenic effect before differentiation occurs.

Keywords: bone substitutes, mesenchymal stem cells, green fluorescent protein, confocal scanning laser microscopy, cell migration, cell polarity