IMPACT OF ADIPOSE-DERIVED STEM CELLS COMBINED WITH VASCULAR BUNDLE IMPLANTATION ONVASCULARIZED TISSUE ENGINEERING SCAFFOLDS IN VIVO_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

JI Wenchen , YANG Pei , ZHANG Yuelin , WANG Chunsheng , NI Jianlong , ZHANG Yongtao , WANG Kunzheng

1 No.1 Department of Orthopedics, the Second Affiliated Hospital of Medical School, Xi’an Jiaotong University, Xi’an Shaanxi, 710004, P.R.China;;
2Department of Neurosurgery, Shaanxi Provincial People’s Hospital, the Third Affiliated Hospital of Medical College, Xi’an Jiaotong University.;

Corresponding author：

Keywords：

Tissue engineering scaffold; Adipose-derived stem cells; Vascular bundle implantation; Vascularization; Rat

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【Abstract】 Objective To discuss the impact of adi pose-derived stem cells (ADSCs) combined with vascular bundle implantation on vascularized tissue engineering scaffolds in vivo so as to provide a theoretical basis for the repair of
avascular necrosis of the femoral head. Methods ADSCs were isolated from 4-month-old Sprague Dawley (SD) rats and
cultured, then were induced to osteogenesis and identified. ADSCs at the 3rd passage were seeded on the nano-hydroxyapatide/ polyamide-66 (nHA/PA66) to prepare the composite scaffolds. The compound condition of cells and scaffold materials were observed under scanning electronic microscope (SEM). Twenty-four 4-month-old SD rats (weighing 350-400 g) were randomly divided into 3 groups (n=8). In group A and group B, the inferior epigastric artery and vein of rats were implanted into composite scaffold cultured for 10 days or simple nHA/PA66 scaffold, respectively. In group C, two composite scaffolds cultured for 10 days were embedded into quadriceps femoris muscle of both thighs, respectively. After 2 and 4 weeks of operation, angiogenesis was observed by HE staining and CD34 immunohistochemical staining. Results Cells isolated from adi pose were identified as ADSCs. SEM showed that the number of cells increased after being cultured for 10 days, cell morphology stretched fully with a shape of long spindle. HE staining and immunohistochemical staining showed that a large number of angiogenesis was observed around the implanted artery and vein in group A, which was superior to groups B and C in the number of blood vessels and the maturity of blood vessel wall. After 2 and 4 weeks of operation, the blood vessel density and blood vessel diameter were significantly higher in group A than in group B and group C, and in group B than in group C (P lt; 0.05). Conclusion Combined application of ADSCs and vascular bundle implantation can promote the degree of vascularization, which could make the scaffold vascularization rel iable.

Citation： JI Wenchen,YANG Pei,ZHANG Yuelin,WANG Chunsheng,NI Jianlong,ZHANG Yongtao,WANG Kunzheng. IMPACT OF ADIPOSE-DERIVED STEM CELLS COMBINED WITH VASCULAR BUNDLE IMPLANTATION ONVASCULARIZED TISSUE ENGINEERING SCAFFOLDS IN VIVO. Chinese Journal of Reparative and Reconstructive Surgery, 2012, 26(2): 129-134. doi: Copy

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3.	Picinich SC, Mishra PJ, Mishra PJ, et al. The therapeutic potential of mesenchymal stem cell. Cell-tissue-based therapy. Expert Opin Biol Ther, 2007, 7(7): 965-973.
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10.	王簕, 裴国献, 高梁斌, 等. 血管化组织工程骨修复兔股骨干骨缺损. 中华医学杂志, 2010, 90(23): 1637-1641.
11.	王簕, 覃俊君, 裴国献, 等. 血管束植入组织工程骨修复兔股骨缺损对血管内皮生长因子表达的影响. 中华创伤骨科杂志, 2009, 11(6): 540-545.
12.	Li B, Zeng Q, Wang H, et al. Effect of cytokines secreted by human adipose stromal cells on endothelial cells. J Huazhong Univ Sci Technolog Med Sci, 2006, 26(4): 396-398.
13.	Sadat S, Gehmert S, Song YH, et al. The cardioprotective effect of stem cells is mediated by IGF-I and VEGF. Biochem Biophys Res Commun, 2007, 363(3): 674-679.
14.	Shigemura N, Ukumura M, Mizuno S, et al. Autologous transplantation of adipose tissue-derived stromal cells ameliorates pulmonary emphysema. Am J Transplant, 2006, 6(11): 2592-2600.
15.	Lu F, Mizuno H, Uysal CA, et al. Improved viability of random pattern skin flaps through the use of adipose-derived stem cells. Plast Reconstr Surg, 2008, 121(1): 50-58.
16.	Suga H, Shigeura T, Matsumoto D, et al. Rapid expansion of human adipose-derived stromal cells preserving multipotency. Cytotherapy, 2007, 9(8): 738-745.
17.	De Francesco F, Tirino V, Desiderio V, et al. Human CD34+/CD90+ ASCs are capable of growing as sphere clusters, producing high levels of VEGF and forming capillaries. PLoS One, 2009, 4(8): e6537.
18.	Rehman J, Traktuev D, Li J, et al. Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation, 2004, 109(10): 1292-1298.
19.	Sanz-Ruiz R, Santos ME, Muñoa MD, et al. Adipose tissue-derived stem cells: the friendly side of a classic cardiovascular foe. J Cardiovasc Transl Res, 2008, 1(1): 55-63.
20.	Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng, 2001, 7(2): 211-228.
21.	Jurgens WJ, Oedayrajsingh-Varma MJ, Helder MN, et al. Effect of tissue-harvesting site on yield of stem cells derived from adipose tissue: implications for cell-based therapies. Cell Tissue Res, 2008, 332(3): 415-426.

1. Mont MA, Jones LC, Hungerford DS, et al. Nontraumatic osteonecrosis of the femoral head: ten years later. J Bone Joint Surg (Am), 2006, 88(5): 1117-1132.
2. Wen Q, Ma L, Chen YP, et al. Treatment of avascular necrosis of the femoral head by hepatocyte growth factor-transgenic bone marrow stromal stem cells. Gene Ther, 2008, 15(23): 1523-1535.
3. Picinich SC, Mishra PJ, Mishra PJ, et al. The therapeutic potential of mesenchymal stem cell. Cell-tissue-based therapy. Expert Opin Biol Ther, 2007, 7(7): 965-973.
4. Kalita SJ, Bhardwaj A, Bhatt HA. Nanocrystalline calcium phosphate ceramics in biomeical engineering. Materials Science and Engineering, 2007, 27(3): 441-449.
5. 赵文, 江昭林, 魏朝阳, 等. 人工椎板(纳米羟基磷灰石/聚酰胺66)的临床应用. 临床骨科杂志, 2009, 12(4): 398-399.
6. Cai X, Tong H, Shen X, et al. Preparation and characterization of homogeneous chitosan-polylactic acid/hydroxyapatite nanocomposite for bone tissue engineering and evaluation of its mechanical properties. Acta Biomater, 2009, 5(7): 2693-2703.
7. 戚孟春, 邓久鹏, 董伟, 等. 骨髓间充质干细胞与支架材料nHA/PA66复合培养的生物活性研究. 口腔医学研究, 2010, 26(6): 805-808.
8. Doldercr JH, Abberton KM, Thompson EW, et al. Spontaneous large volume adipose tissue generation from a vascularized pedicled fat flap inside a chamber space. Tissue Eng, 2007, 13(4): 673-681.
9. Pelissier P, Villars F, Mathoulin-Pelissier S, et al. Influences of vascularization and osteogenic cells on heterotopic bone formation within a madreporic ceramic in rats. Plast Reconstr Surg, 2003, 111(6): 1932-1941.
10. 王簕, 裴国献, 高梁斌, 等. 血管化组织工程骨修复兔股骨干骨缺损. 中华医学杂志, 2010, 90(23): 1637-1641.
11. 王簕, 覃俊君, 裴国献, 等. 血管束植入组织工程骨修复兔股骨缺损对血管内皮生长因子表达的影响. 中华创伤骨科杂志, 2009, 11(6): 540-545.
12. Li B, Zeng Q, Wang H, et al. Effect of cytokines secreted by human adipose stromal cells on endothelial cells. J Huazhong Univ Sci Technolog Med Sci, 2006, 26(4): 396-398.
13. Sadat S, Gehmert S, Song YH, et al. The cardioprotective effect of stem cells is mediated by IGF-I and VEGF. Biochem Biophys Res Commun, 2007, 363(3): 674-679.
14. Shigemura N, Ukumura M, Mizuno S, et al. Autologous transplantation of adipose tissue-derived stromal cells ameliorates pulmonary emphysema. Am J Transplant, 2006, 6(11): 2592-2600.
15. Lu F, Mizuno H, Uysal CA, et al. Improved viability of random pattern skin flaps through the use of adipose-derived stem cells. Plast Reconstr Surg, 2008, 121(1): 50-58.
16. Suga H, Shigeura T, Matsumoto D, et al. Rapid expansion of human adipose-derived stromal cells preserving multipotency. Cytotherapy, 2007, 9(8): 738-745.
17. De Francesco F, Tirino V, Desiderio V, et al. Human CD34+/CD90+ ASCs are capable of growing as sphere clusters, producing high levels of VEGF and forming capillaries. PLoS One, 2009, 4(8): e6537.
18. Rehman J, Traktuev D, Li J, et al. Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation, 2004, 109(10): 1292-1298.
19. Sanz-Ruiz R, Santos ME, Muñoa MD, et al. Adipose tissue-derived stem cells: the friendly side of a classic cardiovascular foe. J Cardiovasc Transl Res, 2008, 1(1): 55-63.
20. Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng, 2001, 7(2): 211-228.
21. Jurgens WJ, Oedayrajsingh-Varma MJ, Helder MN, et al. Effect of tissue-harvesting site on yield of stem cells derived from adipose tissue: implications for cell-based therapies. Cell Tissue Res, 2008, 332(3): 415-426.

Chinese Journal of Reparative and Reconstructive Surgery

IMPACT OF ADIPOSE-DERIVED STEM CELLS COMBINED WITH VASCULAR BUNDLE IMPLANTATION ONVASCULARIZED TISSUE ENGINEERING SCAFFOLDS IN VIVO

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