自聚肽纳米纤维支架对骨髓源性心肌干细胞的作用研究

中国临床解剖学杂志 ›› 2011, Vol. 29 ›› Issue (4): 428-432.

自聚肽纳米纤维支架对骨髓源性心肌干细胞的作用研究

崔晓军¹，　谭玉珍²，　李少华¹，　国海东²，　王　存²，　张剑凯¹

1.广东医学院人体解剖学教研室, 东莞 524808； 2.复旦大学上海医学院解剖学与组织胚胎学系，上海 200032

收稿日期:2011-01-18 发布日期:2011-07-26
作者简介:崔晓军（1975-），男，博士，主要从事心血管疾病防治方面的研究，Tel：（0769）22896359
基金资助:
国家自然科学基金（30971674）; 湛江市科技攻关项目（2010C3102004）

The effect of the self-assemble peptide nanofiber scaffold on the proliferation and differentiation of MCSCs in vitro

CUI Xiao-jun¹, TAN Yu-zhen², LI Shao-hua¹, GUO Hai-dong², WANG Cun², ZHANG Jian-kai¹

1.Department of Anatomy, Guangdong Medical College, Dongguan 524808, China; 2. Department of Anatomy and Histology and Embryology, Shanghai Medical School, Fudan University, Shanghai 200032, China

Received:2011-01-18 Published:2011-07-26

摘要/Abstract

摘要：

目的　研究自聚肽纳米纤维支架对骨髓源性心肌干细胞（MCSCs）生长和存活的作用。方法　设计和固相合成自聚肽，扫描电镜和原子力显微镜下观察其自组装后的形态结构和与MCSCs的相互关系；通过CCK-8法和免疫荧光染色检测纳米纤维支架对MCSCs增殖和分化情况。结果　1%多肽溶液自组装成直径约10 nm，长度为100 ~ 300 nm纳米纤维，并相互交织成孔径为50 ~ 200 nm的网状结构；MCSCs在纤维支架中培养1 d后，可见细胞呈长梭形，位于三维纳米纤维支架的网孔中，生长状态良好；生长曲线显示细胞在纤维支架中的增殖能力明显高于对照组；MCSCs在纤维支架中诱导培养4 周时，形态和排列方式均发生明显改变。结论　纤维支架与MCSCs有良好生物相容性，纤维支架有利于MCSCs的生长和定向分化。

关键词: 自聚肽纳米纤维支架, 骨髓源性心肌干细胞, 生物相容性, 分化

Abstract:

Objective　 To explore the effect of the self-assemble peptide nanofiber scaffold (NFS) on the proliferation and differentiation of marrow-derived cardiac stem cells (MCSCs).　Methods　The self-assemble peptides were designed and synthesized. The self-assemble state was viewed by AFM and SEM. The compatibility of MCSCs with self-assembling peptide nanofiber scarffolds was viewed by SEM. The effect of NFS on the growth of MCSCs was detected by CCK-8. Through cardiac-specific troponin cTnT staining, the effect of NFS was evaluated on myocardial cell differentiation of MCSCs.　Results　SEM showed the self-assembly of the peptides into the retiform interwoven nanofiber scaffold structure, with the stent diameter of 50 ~ 200 nm. MCSCs were seeded into the self-assemble peptide nanofibers 1 day later. Cells were detected in the network-like nanofiber, which closely linked with the nano-fibers under SEM. The viability of cells in NFS was significantly higher than that of control group. The cells in NFS contacted closely with each other, and demonstrated the same direction and arrangement. After 4 weeks' induction, all of cells expressed cTnT. Conclusions　NFS and MCSCs have good biocompatibility. The three-dimensional micro-environment of NFS is valuable for the growth and differentiation of MCSCs in vitro.

Key words: Self-assemble peptide nanofibers, Bone marrow-derived cardiac stem cells, Biocompatibility, Differentiation

中图分类号:

Q813.1

崔晓军, 谭玉珍, 李少华, 国海东, 王　存, 张剑凯. 自聚肽纳米纤维支架对骨髓源性心肌干细胞的作用研究[J]. 中国临床解剖学杂志, 2011, 29(4): 428-432.

CUI Xiao-Jun, TAN Yu-Zhen, LI Shao-Hua, GUO Hai-Dong, WANG 　Cun, ZHANG Jian-Kai. The effect of the self-assemble peptide nanofiber scaffold on the proliferation and differentiation of MCSCs in vitro[J]. Chinese Journal Of Clinical Anatomy, 2011, 29(4): 428-432.

参考文献

[1] Zhang S. Designer self-assembling peptide nanofiber scaffolds for study of 3D cell biology and beyond
[J]. Adv Cancer Res, 2008, 99: 335-362.

[2] Guo J, Su H, Zeng Y, et al. Reknitting the injured spinal cord by self-assembling peptide nanofiber scaffold
[J]. Nanomedicine, 2007, 3(4): 311-321.

[3] Tysseling-Mattiace VM, Sahni V, Niece KL, et al. Self-assembling nanofibers inhibit glial scar formation and promote axon elongation after spinal cord injury
[J]. J Neurosci, 2008, 28(14): 3814-3823.

[4] 贺其志，谭玉珍，王海杰，等.骨形态发生蛋白-2在骨髓源性心肌干细胞向心肌分化中的作用
[J].解剖学报，2005，36(5)：498-502.

[5] 张贵焘，谭玉珍，王海杰，等.骨髓源性心肌干细胞移植治疗心肌梗死的实验研究
[J].中华心血管病杂志，2007，35(10)：940-944.

[6] Cui XJ, Wang HJ, Guo HD, et al. Transplantation of mesenchymal stem cells preconditioned with diazoxide, a mitochondrial ATP-sensitive potassium channel opener, promotes repair of myocardial infarction in rats
[J]. Tohoku J Exp Med, 2010, (220):139-147.

[7] Lin X, Takahashi K, Liu Y, et al. Enhancement of cell attachment and tissue integration by a IKVAV containing multi-domain peptide
[J]. Biochim Biophys Acta, 2006, 1760(9): 1403-1410.

[8] Santiago LY, Nowak RW, Rubin JP, et al. Peptide-surface modification of poly(caprolactone) with laminin-derived sequences for adipose-derived stem cell applications
[J]. Biomaterials, 2006, 27(15): 2962-2969.

[9] Niece KL , Hartgerink JD , Donners J, et al. Self-assembly combining two bioactive peptide-amphiphile molecules into nanofibers by electrostatic attraction
[J]. J Am Chem Soc, 2003, 125(24): 7146-7147.

[10]Ryadnov MG, Woolfson DN. Fiber recruiting peptides: noncovalent decoration of an engineered protein scaffold
[J]. J Am Chem Soc, 2004, 126(24): 7454-7455.

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