«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:

2015年第5期

页码:

547-552

栏目:

基础研究

出版日期:

2015-05-05

文章信息/Info

Title:

Biocompatibility of layer-by-layer structured silk fibroin/chitosan nanofibers serving as cardiac patches

作者:

陈江炜¹; 张建伟²; 王亚斌¹; 马 赛¹; 张 然³; 邓红兵²; 曹 丰¹; 3

（1.第四军医大学西京医院心内科，陕西 西安 710032；
2.武汉大学资源与环境科学学院，湖北 武汉 430079；
3.解放军总医院心内科，北京 100039）

Author(s):

CHEN Jiang-wei¹;  ZHANG Jian-wei²;  WANG Ya-bin¹;  MA Sai¹;  ZHANG Ran³;  DENG Hong-bing²;  CAO Feng¹; 3

(1.Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi, China；
2.School of Resource and Environmental Science, Wuhan University, Wuhan 430079, Hubei, China；
3.Department of Cardiology, PLA General Hospital, Beijing 100039, China)

关键词:

丝素蛋白/壳聚糖纳米纤维膜; 脂肪间充质干细胞; 心脏补片

Keywords:

silk fibroin/chitosan nanofibrous membrane;  adipose tissue-derived mesenchymal stem cells;  cardiac patchs

分类号:

R318.08

DOI:

-

文献标识码:

A

摘要:

目的 探讨丝素蛋白(SF)/壳聚糖(CS)复合纳米纤维膜对小鼠脂肪间充质干细胞（AD-MSCs）黏附和生长的影响，评估该材料用作心脏补片的可行性。 方法 通过静电纺丝技术制备纤维素纳米纤维底板，用层层自组装技术(LBL)，将SF和CS组装到纤维素纳米纤维表面，得到改性的SF/CS复合纳米纤维膜，用ζ- 电位和X射线光电子能谱(XPS)测定材料的表征。分离培养携带绿色荧光蛋白（GFP）和荧光素酶（Fluc）双重报告基因的小鼠AD-MSCs细胞，并接种于SF/CS复合纳米纤维膜上共培养，用激光共聚焦显微镜(CFM)观察细胞黏附和生长形态，扫描电子显微镜(SEM)观察超微结构，生物发光成像(BLI)及MTT比色法评估AD-MSCs细胞的数量和活性。结果 XPS测定的结果提示SF/CS复合纳米纤维膜构建成功。SEM观察可见纳米纤维直径均一，具有良好的三维多孔结构，细胞在SF/CS复合纳米纤维膜上增殖活跃，形态和生长状态良好。CFM、BLI及MTT比色法的结果提示细胞与SF/CS复合纳米纤维膜共培养后呈三维分布，增殖状态好。 结论 对纤维素纳米纤维用LBL进行改性后可明星促进AD-MSCs的三维生长，SF/CS复合纳米纤维膜是可用于组织工程心肌补片的理想材料。

Abstract:

AIM To investigate the biocompatibility of the synthesized silk fibroin(SF)/chitosan (CS) nanofibers with adipose tissue-derived mesenchymal stem cells (AD-MSCs) and the feasibility of nanofibers serving as cardiac patches. METHODS Cellulose nanofibers were fabricated by electrospinning and layer-by-layer (LBL) coated with SF and CS. Nanofibers characteristics were analyzed by ζ-potentials and X-ray photoelectron spectroscopy (XPS). AD-MSCs were constitutively expressed. Both firefly luciferase (Fluc) and green fluorescent protein (GFP) were isolated from Fluc-GFP transgenic mice and co-cultured with SF/CS nanofibrous membrane for 24 h. Cellular morphology and microstructure were observed using confocal microscopy (CFM) and scanning electron microscope (SEM). The linear relationship of the bioluminescent intensity and AD-MSCs count was detected using bioluminescence imaging (BLI) to assess the number of seeded cells. Cell viability was measured by MTT assay. RESULTS XPS displayed the alteration of surface element composition of the mats during LBL coating process. SEM showed that cellulose nanofibers and LBL structured nanofibers displayed porous and uniform nanofibrous 3D microstructures. The survival ratio, spread level and pseudopods of cells on SF/CS nanofibrous membrane were superior to those on cellulose nanofibrous mat. Results of CFM, BLI and MTT revealed cellular proliferation activity and 3D distribution of AD-MSC on SF/CS nanofibers. CONCLUSION Cellulose nanofibers LBL coated by SF and CS have improved biocompatibility.The application of nanofibers could promote cellular proliferation and 3D distribution of seeded AD-MSCs. LBL structured SF/CS nanofibers are ideal biomaterial materials for tissue-engineered cardiac patches.

参考文献/References

[1]van Amerongen MJ,Engel FB.Features of cardiomyocyte proliferation and its potential for cardiac regeneration[J]. J Cell Mol Med,2008,12 (6A):2233-2244.
[2]Velagaleti RS,Pencina MJ,Murabito JM,et al.Long-term trends in the incidence of heart failure after myocardial infarction[J].Circulation,2008,118(20):2057-2062.
[3]Orlic D,Kajstura J,Chimenti S,et al.Bone marrow cells regenerate infarcted myocardium[J].Nature,2001,410(6829):701-705.
[4]Hofmann M,Wollert KC,Meyer GP,et al.Monitoring of bone marrow cell homing into the infarcted human myocardium[J].Circulation,2005,111(17):2198-2202.
[5]Matsuura K,Haraguchi Y,Shimizu T,et al.Cell sheet transplantation for heart tissue repair[J].J Control Release,2013,169(3):336-340.
[6]Leor J,Aboulafia-Etzion S,Dar A,et al.Bioengineered cardiac grafts:A new approach to repair the infarcted myocardium?[J].Circulation, 2000,102(19 Suppl 3):III56-III61.
[7]Zimmermann WH,Melnychenko I,Wasmeier G,et al.Engineered heart tissue grafts improve systolic and diastolic function in infarcted rat hearts[J].Nat Med,2006,12(4):452-458.
[8] Dvir T,Timko BP,Kohane DS,et al.Nanotechnological strategies for engineering complex tissues[J].Nat Nanotechnol,2011,6(1):13-22.
[9]Zimmermann WH,Schneiderbanger K,Schubert P,et al.Tissue engineering of a differentiated cardiac muscle construct[J].Circ Res,2002,90 (2): 223-230.
[10]Kelley ST,Malekan R,Gorman JH,et al.Restraining infarct expansion preserves left ventricular geometry and function after acute anteroapical infarction[J].Circulation,1999,99(1):135-142.
[11]Aznar-Cervantes S,Roca MI,Martinez JG,et al.Fabrication of conductive electrospun silk fibroin scaffolds by coating with polypyrrole for biomedical applications[J].Bioelectrochemistry,2012,85:36-43.
[12]Acharya C,Ghosh SK,Kundu SC.Silk fibroin protein from mulberry and non-mulberry silkworms: cytotoxicity, biocompatibility and kinetics of L929 murine fibroblast adhesion[J].J Mater Sci Mater Med,2008,19(8):2827-2836.
[13]Ghadi A,Mahjoub S,Tabandeh F.Synthesis and optimization of chitosan nanoparticles: Potential applications in nanomedicine and biomedical engineering[J].Caspian J Intern Med,2014,5(3):156-161.
[14]Francis MP,Sachs PC,Elmore LW.Isolating adipose-derived mesenchymal stem cells from lipoaspirate blood and saline fraction[J].Organogenesis,2010,6(1):11-14.
[15]Jaffer FA,Weissleder R.Molecular imaging in the clinical arena[J].JAMA,2005,293(7):855-862.
[16]Wu JC,Chen IY,Sundaresan G,et al.Molecular imaging of cardiac cell transplantation in living animals using optical bioluminescence and positron emission tomography[J].Circulation,2003,108(11):1302-1305.

备注/Memo

备注/Memo:

收稿日期：2014-11-24.
基金资助：国家自然科学基金杰青项目资助(81325009);军队十二五重点项目资助（BWS12J037）
通讯作者：曹丰，主任医师，主要从事分子影像和干细胞治疗研究 Email:wind8828@gmail.com
共同通讯作者：邓红兵，副教授，主要从事纳米技术应用研究 Email：hbdeng@whu.edu.cn
作者简介：陈江炜，硕士生Email：cjwfmmu@gmail.com

常用功能

更新日期/Last Update: 2015-04-28