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The Influence of Adipose-Derived Stem Cells Induced into Endothelial Cells on Ectopic Vasculogenesis and Osteogenesis

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Abstract

Established vascular network has a crucial importance in bone regeneration. Adipose-derived stem cells (ADSCs) can be differentiated in vitro towards endothelial cells (ECs) that give a possibility for their application in bone tissue engineering (BTE). The aim of our study was to examine the influence of ADSCs in vitro induced into ECs on vascularization and osteogenic process in subcutaneous implants. Induced ADSCs were implanted subcutaneously into BALB/c mice, in combination with the bone mineral matrix carrier (BC) and platelet-rich plasma (PRP), parallel with the implants without the cells. The combination of BC, PRP and ADSCs induced into ECs increased vascularization in subcutaneous implants that was shown through endothelial-related gene expression, high percentage of vascularization and VEGFR-2 immunoexpression. Osteocalcin immunoexpression, relative expression of osteopontin gene, and histological analysis showed that osteogenic process was more pronounced when the carrier was loaded with ADSCs induced into ECs which was associated with strong vascularization in cellularized implants. In implants without the cells vasculogenesis was initially stimulated, but vascular network was unsustainable at later observation points. Therefore, the approach that includes ADSCs in vitro induced into ECs combined with BC and PRP can be a good strategy for improving vascularization in bone regeneration and BTE.

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Abbreviations

BTE:

Bone tissue engineering

ADSCs:

Adipose-derived stem cells

ECs:

Endothelial cells

BC:

Bone mineral matrix carrier

PRP:

Platelet-rich plasma

SVF:

Stromal vascular fraction

DMEM:

Dulbecco’s Modified Eagles Minimal Essential Medium

P03:

Third passage

ECP:

The implants consisted of ADSCs in vitro induced into ECs, BC and PRP

CP:

The implants consisted of BC and PRP

OC:

Osteocalcin

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Acknowledgments

This work was supported by Project Grant III41017, Ministry of Education, Science and Technological Development, Republic of Serbia. The authors would like to thank Tanja Prokić, a technician from Faculty of Medicine, University of Niš, for helping with animal procedures and tissue processing.

Conflict of Interest

Jelena G Najdanović, Vladimir J Cvetković, Sanja Stojanović, Marija Đ Vukelić-Nikolić, Milica N Stanisavljević, Jelena M Živković and Stevo J Najman declare that they have no conflicts of interest.

Ethical Standards

No human studies were carried out by the authors for this article. All animal procedures were approved by the Local Ethical Committee (approval number 01-2857-8) and conducted in accordance with the Animal Welfare Act (Republic of Serbia). The animals were treated conforming to the regulation of the “European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes (ETS no. 123 Appendix A)”.

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Authors and Affiliations

  1. Institute of Biology and Human Genetics; Department for Cell and Tissue Engineering, Faculty of Medicine, University of Niš, 18000, Niš, Serbia

    Jelena G. Najdanović, Sanja Stojanović, Marija Đ. Vukelić-Nikolić, Jelena M. Živković & Stevo J. Najman

  2. Department of Biology and Ecology, Faculty of Science and Mathematics, University of Niš, 18000, Niš, Serbia

    Vladimir J. Cvetković

  3. Scientific Research Center for Biomedicine, Faculty of Medicine, University of Niš, 18000, Niš, Serbia

    Milica N. Stanisavljević

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  1. Jelena G. Najdanović
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  2. Vladimir J. Cvetković
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Correspondence to Jelena G. Najdanović.

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Associate Editor Alyssa Panitch oversaw the review of this article.

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Najdanović, J.G., Cvetković, V.J., Stojanović, S. et al. The Influence of Adipose-Derived Stem Cells Induced into Endothelial Cells on Ectopic Vasculogenesis and Osteogenesis. Cel. Mol. Bioeng. 8, 577–590 (2015). https://doi.org/10.1007/s12195-015-0403-x

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