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Open Access Research article

Development of a new pre-vascularized tissue-engineered construct using pre-differentiated rADSCs, arteriovenous vascular bundle and porous nano-hydroxyapatide-polyamide 66 scaffold

Pei Yang1, Xin Huang2, Jacson Shen3, Chunsheng Wang1, Xiaoqian Dang1, Henry Mankin45, Zhenfeng Duan45 and Kunzheng Wang1*

Author Affiliations

1 Department of Orthopaedics, Second Affiliated Hospital of Medical College of Xi’an Jiaotong University, No. 157 Xiwu Road, 710004 Xi’an, Shaanxi, China

2 Department of Cardiology, First Affiliated Hospital of Medical College of Xi’an Jiaotong University, Ion Channel Disease Laboratory, Key Laboratory of Environment and Genes related to Diseases of Education Ministry, 710061 Xi’an, Shaanxi, P.R.China

3 Department of Biological Chemistry, Wellesley College, 02481 Wellesley, Massachusetts, USA

4 Department of Orthopaedic Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA

5 Sarcoma Biology Laboratory, Center for Sarcoma and Connective Tissue Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA

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BMC Musculoskeletal Disorders 2013, 14:318  doi:10.1186/1471-2474-14-318

Published: 8 November 2013

Abstract

Background

Development of a pre-vascularized tissue-engineered construct with intrinsic vascular system for cell growth and tissue formation still faces many difficulties due to the complexity of the vascular network of natural bone tissue. The present study was to design and form a new vascularized tissue-engineered construct using pre-differentiated rADSCs, arteriovenous vascular bundle and porous nHA-PA 66 scaffold.

Methods

rADSCs were pre-differentiated to endothelial cells (rADSCs-Endo) and then incorporated in nHA-PA 66 scaffolds in vitro. Subsequently, in vivo experiments were carried out according to the following groups: Group A (rADSCs-Endo/nHA-PA 66 scaffold with arteriovenous vascular bundle), Group B (rADSCs/nHA-PA 66 scaffold with arteriovenous vascular bundle); Group C (nHA-PA66 scaffold with arteriovenous vascular bundle), Group D (nHA-PA 66 scaffold only). The vessel density and vessel diameter were measured based on histological and immunohistochemical evaluation, furthermore, the VEGF-C, FGF-2 and BMP-2 protein expressions were also evaluated by western blot analysis.

Results

The results of in vivo experiments showed that the vessel density and vessel diameter in group A were significantly higher than the other three groups. Between Group B and C, no statistical difference was observed at each time point. In accordance with the results, there were dramatically higher expressions of VEGF-C and FGF-2 protein in Group A than that of Group B, C and D at 2 or 4 weeks. Statistical differences were not observed in VEGF-C and FGF-2 expression between Group B and C. BMP-2 was not expressed in any group at each time point.

Conclusions

Compared with muscular wrapping method, arteriovenous vascular bundle implantation could promote vascularization of the scaffold; and the angiogenesis of the scaffold was significantly accelerated when pre-differentiated rADSCs (endothelial differentiation) were added. These positive results implicate the combination of pre-differentiated rADSCs (endothelial differentiation) and arteriovenous vascular bundle may achieve rapidly angiogenesis of biomaterial scaffold.

Keywords:
Adipose-derived stem cells; Tissue engineering; Angiogenesis; Scaffolds; Prefabrication