Experimental Study of Compatibility of Endothelial Outgrowth Cells Cultured with Nanofibers PLLA Scaffold_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LU Huijun ¹ , FENG Zhangqi ² , GU Zhongze ² , LIU Changjian ¹

1.Department of Vascular Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, China;;
2.State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, ChinaCorresponding Author: LIU Chang-jian, E-mail: dr_cjliu@hotmail.com;

Corresponding author：

Keywords：

Scaffold; PLLA; Super-aligned nanofibrous; Endothelial outgrowth cell; Tissue repair

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective 　To study the cellular biocompatibility, adhesion and proliferation of endothelial outgrowth cells (EOCs) isolated and expanded from rabbit peripheral blood cultured with aligned poly-L-lactic acid (PLLA) nanofibrous scaffolds in vitro so as to provide a basis for the applications of scaffolds biomaterials in tissue repair.
Methods 　Nanofibrous scaffolds of PLLA by electrostatic spinning were modified by hypothermal plasmas body and type Ⅰ collagen was coated onto the materials physically. In vitro, EOCs were cultured on the modified PLLA scaffold. Adhesion and proliferation were surveyed and morphological changes and biocompatibility of seeding cells on PLLA scaffold were observed by growth curves of the cells, fluorescent microscope and scanning electron microscope respectively.
Results 　Fibers with diameters ranging from 300 nm to 400 nm were included in the nanofibrous scaffolds, whose porosities were more than 90%. Absorbance (A) of each scaffold increased gradually after EOCs grew in the absence or presence of random, aligned, or super-aligned PLLA nanofibrous scaffold. Although there was no detectable effect of the random PLLA scaffold on the growth EOCs (P gt;0.05), both aligned and super-aligned PLLA nanofibrous scaffold had significantly enhanced their growth since the 5th day (P＜0.05). The rates of adhesion in both aligned and super-aligned PLLA nanofibrous scaffold were significantly higher than those of random PLLA scaffold after 12 h and 24 h incubation (P＜0.01). The rates of proliferation after 1 d, 3 d and 7 d incubation in aligned and super-aligned PLLA nanofibrous scaffold were significantly higher than those of random PLLA nanofibrous scaffold (P＜0.05, P＜0.01). EOCs grew well with PLLA scaffold, yet confused and disorderly in random nanofibers. EOCs could attach, extend and proliferate following fibrous orientation in aligned and super-aligned PLLA nanofibrous scaffold, in majority of the fibers were oriented along the longitudinal axis so that a unique aligned topography was formed. Especially super-aligned PLLA nanofibrous had advantageous to keep well on cell morphology.
Conclusion 　EOCs are ideal seeding cells for tissue engineering. EOCs can be adhered well to aligned and super-aligned PLLA nanofibrous scaffold and proliferate, keep well on cell morphology. So this type of PLLA nanofibrous scallfold is proposed to be an optimal candidate material for EOCs transplantation in tissue repair.

Citation： LU Huijun,FENG Zhangqi,GU Zhongze,LIU Changjian. Experimental Study of Compatibility of Endothelial Outgrowth Cells Cultured with Nanofibers PLLA Scaffold. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2009, 16(4): 269-275. doi: Copy

1.	Barnes CP, Sell SA, Boland ED, et al. Nanofiber technology: designing the next generation of tissue engineering scaffolds [Ｊ]. Adv Drug Deliv Rev, 2007; 59(14): 1413-1433.
2.	Bashur CA, Dahlgren LA, Goldstein AS. Effect of fiber diameter and orientation on fibroblast morphology and proliferation on electrospun poly (D, L-lactic-co-glycolic acid) meshes [Ｊ]. Biomaterials, 2006; 27(33): 5681-5688.
3.	汪忠镐，李震. 我国血管外科的现状和发展 [Ｊ]. 中国普外基础与临床杂志， 2006； 13(6): 625-628.
4.	Asahara T, Murohara T, Sullivan A, et al. Isolation of putative progenitor endothelial cells for angiogenesis [Ｊ]. Science, 1997; 275(5302): 964-967.
5.	Kawamoto A, Losordo DW. Endothelial progenitor cells for cardiovascular regeneration [Ｊ]. Trends Cardiovasc Med, 2008; 18(1): 33-37.
6.	Ben-Shoshan J, George J. Endothelial progenitor cells as therapeutic vectors in cardiovascular disorders: from experimental models to human trials [Ｊ]. Pharmacol Ther, 2007; 115(1): 25-36.
7.	Li WJ, Mauck RL, Cooper JA, et al. Engineering controllable anisotropy in electrospun biodegradable nanofibrous scaffolds for musculoskeletal tissue engineering [Ｊ]. J Biomech, 2007; 40(8): 1686-1693.
8.	Yang F, Murugan R, Ramakrishna S, et al. Fabrication of nano-structured porous PLLA scaffold intended for nerve tissue engineering [Ｊ]. Biomaterials, 2004; 25(10): 1891-1900.
9.	Yao YF, Gu ZZ, Zhang JZ, et al. Fiber-oriented liquid crystal polarizers based on anisotropic electrospinning [Ｊ]. Adv Mater, 2007; 19(21): 3707-3711.
10.	Funcke F, Hoyer H, Brenig F, et al. Characterisation of the interaction between circulating and in vitro cultivated endothelial progenitor cells and the endothelial barrier [Ｊ]. Eur J Cell Biol, 2008; 87(2): 81-90.
11.	Fuchs S, Hermanns MI, Kirkpatrick CJ. Retention of a differentiated endothelial phenotype by outgrowth endothelial cells isolated from human peripheral blood and expanded in long-term cultures [Ｊ]. Cell Tissue Res, 2006; 326(1): 79-92.
12.	Xu CY, Inai R, Kotaki M, et al. Aligned biodegradable nanofibrous structure: a potential scaffold for blood vessel engineering [Ｊ]. Biomaterials, 2004; 25(5): 877-886.
13.	Liang D, Hsiao BS, Chu B. Functional electrospun nanofibrous scaffolds for biomedical applications [Ｊ]. Adv Drug Deliv Rev, 2007; 59(14): 1392-1412.
14.	Gulati R, Jevremovic D, Peterson TE, et al. Diverse origin and function of cells with endothelial phenotype obtained from adult human blood [Ｊ]. Circ Res, 2003; 93(11): 1023-1025.
15.	Mukai N, Akahori T, Komaki M, et al. A comparison of the tube forming potentials of early and late endothelial progenitor cells [Ｊ]. Exp Cell Res, 2008; 314(3): 430-440.
16.	Ingram DA, Mead LE, Tanaka H, et al. Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood [Ｊ]. Blood, 2004; 104(9): 2752-2760.
17.	Hur J, Yoon CH, Kim HS, et al. Characterization of two types of endothelial progenitor cells and their different contributions to neovasculogenesis [Ｊ]. Arterioscler Thromb Vasc Biol, 2004; 24(2): 288-293.
18.	Liang D, Hsiao BS, Chu B. Functional electrospun nanofibrous scaffolds for biomedical applications [Ｊ]. Adv Drug Deliv Rev, 2007; 59(14): 1392-1412.
19.	Choi JS, Lee SJ, Christ GJ, et al. The influence of electrospun aligned poly (epsilon-caprolactone)/collagen nanofiber meshes on the formation of self-aligned skeletal muscle myotubes [Ｊ]. Biomaterials, 2008; 29(19): 2899-2906.
20.	Ma Z, Kotaki M, Yong T, et al. Surface engineering of electrospun polyethylene terephthalate (PET) nanofibers towards development of a new material for blood vessel engineering [Ｊ]. Biomaterials, 2005; 26(15): 2527-2536.

1. Barnes CP, Sell SA, Boland ED, et al. Nanofiber technology: designing the next generation of tissue engineering scaffolds [Ｊ]. Adv Drug Deliv Rev, 2007; 59(14): 1413-1433.
2. Bashur CA, Dahlgren LA, Goldstein AS. Effect of fiber diameter and orientation on fibroblast morphology and proliferation on electrospun poly (D, L-lactic-co-glycolic acid) meshes [Ｊ]. Biomaterials, 2006; 27(33): 5681-5688.
3. 汪忠镐，李震. 我国血管外科的现状和发展 [Ｊ]. 中国普外基础与临床杂志， 2006； 13(6): 625-628.
4. Asahara T, Murohara T, Sullivan A, et al. Isolation of putative progenitor endothelial cells for angiogenesis [Ｊ]. Science, 1997; 275(5302): 964-967.
5. Kawamoto A, Losordo DW. Endothelial progenitor cells for cardiovascular regeneration [Ｊ]. Trends Cardiovasc Med, 2008; 18(1): 33-37.
6. Ben-Shoshan J, George J. Endothelial progenitor cells as therapeutic vectors in cardiovascular disorders: from experimental models to human trials [Ｊ]. Pharmacol Ther, 2007; 115(1): 25-36.
7. Li WJ, Mauck RL, Cooper JA, et al. Engineering controllable anisotropy in electrospun biodegradable nanofibrous scaffolds for musculoskeletal tissue engineering [Ｊ]. J Biomech, 2007; 40(8): 1686-1693.
8. Yang F, Murugan R, Ramakrishna S, et al. Fabrication of nano-structured porous PLLA scaffold intended for nerve tissue engineering [Ｊ]. Biomaterials, 2004; 25(10): 1891-1900.
9. Yao YF, Gu ZZ, Zhang JZ, et al. Fiber-oriented liquid crystal polarizers based on anisotropic electrospinning [Ｊ]. Adv Mater, 2007; 19(21): 3707-3711.
10. Funcke F, Hoyer H, Brenig F, et al. Characterisation of the interaction between circulating and in vitro cultivated endothelial progenitor cells and the endothelial barrier [Ｊ]. Eur J Cell Biol, 2008; 87(2): 81-90.
11. Fuchs S, Hermanns MI, Kirkpatrick CJ. Retention of a differentiated endothelial phenotype by outgrowth endothelial cells isolated from human peripheral blood and expanded in long-term cultures [Ｊ]. Cell Tissue Res, 2006; 326(1): 79-92.
12. Xu CY, Inai R, Kotaki M, et al. Aligned biodegradable nanofibrous structure: a potential scaffold for blood vessel engineering [Ｊ]. Biomaterials, 2004; 25(5): 877-886.
13. Liang D, Hsiao BS, Chu B. Functional electrospun nanofibrous scaffolds for biomedical applications [Ｊ]. Adv Drug Deliv Rev, 2007; 59(14): 1392-1412.
14. Gulati R, Jevremovic D, Peterson TE, et al. Diverse origin and function of cells with endothelial phenotype obtained from adult human blood [Ｊ]. Circ Res, 2003; 93(11): 1023-1025.
15. Mukai N, Akahori T, Komaki M, et al. A comparison of the tube forming potentials of early and late endothelial progenitor cells [Ｊ]. Exp Cell Res, 2008; 314(3): 430-440.
16. Ingram DA, Mead LE, Tanaka H, et al. Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood [Ｊ]. Blood, 2004; 104(9): 2752-2760.
17. Hur J, Yoon CH, Kim HS, et al. Characterization of two types of endothelial progenitor cells and their different contributions to neovasculogenesis [Ｊ]. Arterioscler Thromb Vasc Biol, 2004; 24(2): 288-293.
18. Liang D, Hsiao BS, Chu B. Functional electrospun nanofibrous scaffolds for biomedical applications [Ｊ]. Adv Drug Deliv Rev, 2007; 59(14): 1392-1412.
19. Choi JS, Lee SJ, Christ GJ, et al. The influence of electrospun aligned poly (epsilon-caprolactone)/collagen nanofiber meshes on the formation of self-aligned skeletal muscle myotubes [Ｊ]. Biomaterials, 2008; 29(19): 2899-2906.
20. Ma Z, Kotaki M, Yong T, et al. Surface engineering of electrospun polyethylene terephthalate (PET) nanofibers towards development of a new material for blood vessel engineering [Ｊ]. Biomaterials, 2005; 26(15): 2527-2536.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Experimental Study of Compatibility of Endothelial Outgrowth Cells Cultured with Nanofibers PLLA Scaffold

Abstract Full text Figures/Tables Video References Cited by

Format

Content