| 丝素蛋白/壳聚糖复合纳米纤维膜心脏补片的生物相容性研究 |
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| 引用本文： | 陈江炜,张建伟,王亚斌,马 赛,张 然,邓红兵,曹 丰.丝素蛋白/壳聚糖复合纳米纤维膜心脏补片的生物相容性研究[J].心脏杂志,2015,27(5):547-552. |
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| 作者姓名： | 陈江炜 张建伟 王亚斌 马 赛 张 然 邓红兵 曹 丰 |
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| 作者单位： | （1.第四军医大学西京医院心内科，陕西 西安 710032； |
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| 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。
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| 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 |
| 收稿时间： | 2014-11-24 |
Biocompatibility of layer-by-layer structured silk fibroin/chitosan nanofibers serving as cardiac patches |
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| 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. |
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