Endothelial microvascular networks affect gene-expression profiles and osteogenic potential of tissue-engineered constructs

Stem Cell Research & Therapy

Kamal Mustafa⁵  Inge Fristad⁵  Knut N Leknes⁵  James B Lorens²  Lars A Akslen¹  Anna Finne-Wistrand⁴  Yang Sun⁴  Ying Xue⁵  Zhe Xing⁵  Anna L Blois³  Torbjorn O Pedersen² 

[1] Department of Pathology, Haukeland University Hospital, Bergen, Norway;Department of Biomedicine, University of Bergen, Bergen, Norway;Centre for Cancer Biomarkers, The Gade Institute, University of Bergen, Bergen, Norway;Department of Fibre and Polymer Technology, KTH Royal Institute of Technology, Stockholm, Sweden;Department of Clinical Dentistry, Center for Clinical Dental Research, University of Bergen, Årstadveien 19, Bergen N-5009, Norway

关键词: Osteogenesis;    Copolymer;    Mesenchymal stem cells;    Endothelial cells;    Tissue engineering;   

Others  :  847530
DOI  :  10.1186/scrt202

 received in 2012-09-05, accepted in 2013-05-14,  发布年份 2013

【 摘 要 】

Introduction

A major determinant of the potential size of cell/scaffold constructs in tissue engineering is vascularization. The aims of this study were twofold: first to determine the in vitro angiogenic and osteogenic gene-expression profiles of endothelial cells (ECs) and mesenchymal stem cells (MSCs) cocultured in a dynamic 3D environment; and second, to assess differentiation and the potential for osteogenesis after in vivo implantation.

Methods

MSCs and ECs were grown in dynamic culture in poly(L-lactide-co-1,5-dioxepan-2-one) (poly(LLA-co-DXO)) copolymer scaffolds for 1 week, to generate three-dimensional endothelial microvascular networks. The constructs were then implanted in vivo, in a murine model for ectopic bone formation. Expression of selected genes for angiogenesis and osteogenesis was studied after a 1-week culture in vitro. Human cell proliferation was assessed as expression of ki67, whereas α-smooth muscle actin was used to determine the perivascular differentiation of MSCs. Osteogenesis was evaluated in vivo through detection of selected markers, by using real-time RT-PCR, alkaline phosphatase (ALP), Alizarin Red, hematoxylin/eosin (HE), and Masson trichrome staining.

Results

The results show that endothelial microvascular networks could be generated in a poly(LLA-co-DXO) scaffold in vitro and sustained after in vivo implantation. The addition of ECs to MSCs influenced both angiogenic and osteogenic gene-expression profiles. Furthermore, human ki67 was upregulated before and after implantation. MSCs could support functional blood vessels as perivascular cells independent of implanted ECs. In addition, the expression of ALP was upregulated in the presence of endothelial microvascular networks.

Conclusions

This study demonstrates that copolymer poly(LLA-co-DXO) scaffolds can be prevascularized with ECs and MSCs. Although a local osteoinductive environment is required to achieve ectopic bone formation, seeding of MSCs with or without ECs increases the osteogenic potential of tissue-engineered constructs.

【 授权许可】

   
2013 Pedersen et al.; licensee BioMed Central Ltd.

【 图 表 】

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