West China Hospital, Laboratory for Nanobiomedical Technology, Sichuan University, Chengdu, Sichuan 610065, China.
Biomaterials. 2011 Mar;32(8):2013-20. doi: 10.1016/j.biomaterials.2010.11.049. Epub 2010 Dec 16.
We previously reported a class of designer self-assembling peptides that form 3-dimensional nanofiber scaffolds using only l-amino acids. Here we report that using d-amino acids, the chiral self-assembling peptide d-EAK16 also forms 3-dimensional nanofiber scaffold that is indistinguishable from its counterpart l-EAK16. These chiral peptides containing all d-amino acids, d-EAK16, self-assemble into well-ordered nanofibers. However with alternating d- and l-amino acids, EAK16 and EAK16, showed poor self-assembling properties. To fully understand individual molecular building blocks and their structures, assembly properties and dynamic behaviors for rapid hemostasis, we used circular dichroism, atomic force microscopy and scanning electron microscopy to study in detail the peptides. We also used rheological measurement to study the hydrogel gelation property. Furthermore, we used an erythrocyte-agglutination test and a rabbit liver wound healing model, particularly in the transverse rabbit liver experiments, to examine rapid hemostasis. We showed that 1% d-EAK16 for the liver wound hemostasis took ∼20 s, but using 1% of EAK16 and EAK16 that have alternating chiral d- and l-amino acids took ∼70 and ∼80 s, respectively. We here propose a plausible model not only to provide insights in understanding the chiral assembly properties for rapid hemostasis, but also to aid in further design of self-assembling d-form peptide scaffolds for clinical applications.
我们之前报道过一类设计自组装肽,它们仅使用 l-氨基酸形成三维纳米纤维支架。在这里,我们报告说使用 d-氨基酸时,手性自组装肽 d-EAK16 也能形成与 l-EAK16 无法区分的三维纳米纤维支架。这些含有所有 d-氨基酸的手性肽,d-EAK16,自组装成有序的纳米纤维。然而,具有交替的 d-和 l-氨基酸的 EAK16 和 EAK16 显示出较差的自组装性能。为了充分了解单个分子构建块及其结构、组装特性和快速止血的动态行为,我们使用圆二色性、原子力显微镜和扫描电子显微镜详细研究了这些肽。我们还使用流变学测量研究了水凝胶的凝胶化特性。此外,我们使用红细胞凝集试验和兔肝伤口愈合模型,特别是在兔肝横向实验中,检查了快速止血。我们表明,1%的 d-EAK16 用于肝脏伤口止血只需约 20 秒,但使用具有交替手性 d-和 l-氨基酸的 1%的 EAK16 和 EAK16 则分别需要约 70 秒和 80 秒。我们在这里提出了一个合理的模型,不仅有助于理解快速止血的手性组装特性,还有助于进一步设计用于临床应用的自组装 d 型肽支架。