Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (12): 1907-1913.doi: 10.3969/j.issn.2095-4344.2014.12.017
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Liu Xi1, 2, Wang Xiu-mei2
Revised:
2014-01-18
Online:
2014-03-19
Published:
2014-03-19
Contact:
Wang Xiu-mei, Professor, Institute for Regenerative Medicine and Biomimetic Materials, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084, China
About author:
Liu Xi, Ph.D., Assistant professor, National Engineering Laboratory for Modern Silk, College of Textile and Clothing Engineering, Soochow University, Suzhou 215123, Jiangsu Province, China; Institute for Regenerative Medicine and Biomimetic Materials, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084, China
Supported by:
Natural Science Foundation of the Higher Education Institutions of Jiangsu Province, No. 13KJB430019; China Postdoctoral Science Foundation, No. 2013M541724; Tsinghua University Initiative Scientific Research Program, No. 20121087982
CLC Number:
Liu Xi, Wang Xiu-mei. Application and characteristics of self-assembling peptide nanofiber hydrogel in three-dimensional cell culture systems[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(12): 1907-1913.
2.1 自组装多肽的发现 第1个自组装多肽EAK16-II (AEAEAKAKAEAEAKAK)的发现源于对酵母遗传学和结构蛋白学的研究,它由丙氨酸(alanine,A)、谷氨酸(glutamate,E)和赖氨酸(lysine,K)交替排列而成,并在水中能够形成稳定的β折叠结构[10-11]。由于该β折叠结构一侧是非极性疏水端,另一侧是由带有正负电氨基酸组成的亲水端,这类多肽分子又被称为“离子互补型自组装多肽分子”。在水溶液中,多肽分子发生自组装,通过分子间的疏水作用力,肽链的疏水端聚集在内侧,而外侧亲水端的氨基酸通过互补的离子相互作用横向交替排列,使自组装的多肽分子延长并形成稳定的直径约10 nm的纳米纤维。在改变多肽溶液pH值或加入碱性阳离子的情况下,自组装多肽分子开始凝胶化,形成含水量99%(5-10 g/L)以上的水凝胶。 基于这一发现,多种具有类似特征的多肽序列越来越受到人们的关注与研究,尽管设计方案和组装形态不尽相同,但在组织工程领域都取得了令人欣喜的结果。其中,Zhang等[12-17]设计的RADA16-I (AcN- RADARADARADARADA-CNH2)与RADA16-II (AcN-RARADADARARADADA-CNH2)研究和应用最为广泛,RADA16-I已被商品化(PuraMatrixTM,BD Biosciences)。这里由精氨酸(arginine,R)和天门冬氨酸(aspartate,D)分别代替赖氨酸K和谷氨酸E。除RADA外,自组装多肽分子家族仍在不断增加。在自组装多肽分子的设计中,疏水性和多肽序列长度是影响多肽自组装特性的重要因素。疏水性越高、多肽序列越长,自组装过程越容易发生。 2.2 功能化RADA自组装多肽水凝胶 以短肽为基本组成单元,不仅使得这类多肽材料具有更好的生物相容性、可降解性,更为突出的优点是其具有可设计性,可根据需求不同复合不同的生物活性分子,赋予该类材料“生物智能”的特性。在RADA自组装多肽的基础上,可以复合具有细胞特异性的功能短肽片段,显著提高材料的生物功能和特异性[3-5]。复合功能片段主要是在多肽固相合成过程中,通过延长C末端将自组装多肽和功能多肽通过几个甘氨酸连接起来(图1)。 其中,甘氨酸的作用是为了增加功能片段的灵活性,避免功能片段影响自组装多肽的自组装过程。一般将接枝有功能短肽的多肽与RADA多肽1∶1混合使用,实验表明功能片段并未影响自组装多肽的自组装过程,并且能够显著提高自组装多肽的生物功能。下面简要介绍一些常见的功能片段:"
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