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精氨酸-甘氨酸-谷氨酰胺和丝氨酸-异亮氨酸-赖氨酸-缬氨酸-丙氨酸-缬氨酸修饰的聚(L-丙交酯)薄膜:用于表面接枝以引导细胞收缩表型的生物活性分子。

Arginine-glycine-glutamine and serine-isoleucine-lysine-valine-alanine-valine modified poly(l-lactide) films: bioactive molecules used for surface grafting to guide cellular contractile phenotype.

作者信息

Boccafoschi Francesca, Fusaro Luca, Botta Margherita, Ramella Martina, Chevallier Pascale, Mantovani Diego, Cannas Mario

机构信息

Human Anatomy Laboratory, Department of Health Sciences, University of Piemonte Orientale "A. Avogadro", 28100 Novara, Italy.

Laboratory for Biomaterials and Bioengineering, Department of Min-Met-Materials Engineering, Laval University and University Hospital Research Center, Quebec City, G1V 0A6, Canada.

出版信息

Biointerphases. 2014 Jun;9(2):029002. doi: 10.1116/1.4864432.

Abstract

Tissue engineering is defined as "an interdisciplinary field that applies the principles of engineering and life sciences toward the development of biological substitutes that restore, maintain, or improve tissue function." The biological substitutes can be developed with the help of natural or synthetic materials. Polymeric materials are primarily used, because of the high variability in mechanical, physical, and chemical properties. Biodegradable polymers are object of the majority of studies, because of the ability to be degraded by the host organism, avoiding late stent thrombosis unlike permanent grafts. Poly-l-lactide acid (PLLA) is one of the most used polymers in research. In order to improve the material's bioactivity, in this work, PLLA surface was modified by grafted arginine-glycine-glutamine (RGD), a fibronectin derived adhesion motif, and serine-isoleucine-lysine-valine-alanine-valine (SIKVAV), a laminin derived motif, and rat cardiac (H9C2) and mouse (C2C12) myoblasts proliferation and differentiation on modified PLLA were evaluated. In order to verify the surface modification, x-ray photoelectron spectroscopy analysis was performed. After seeding, cells' viability was confirmed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay while proliferating cell nuclear antigen expression was used to investigate cell proliferation. Myf5, Myogenin and Myosin heavy chain were used to analyze cell differentiation. Moreover, RGD peptide slightly inhibited rat myoblast (H9C2) proliferation, whereas less strong effect was observed on C2C12. However, both cell lines showed to enhance the contractile phenotype in the presence of SIKVAV peptides. These results suggest that bioactive molecules grafting could be useful on polymeric scaffolds for guiding cell phenotype expression, and, to ultimately maintain adequate biological characteristics suitable for the tissue functional regeneration.

摘要

组织工程被定义为“一个跨学科领域,它将工程学和生命科学原理应用于生物替代品的开发,以恢复、维持或改善组织功能”。生物替代品可以借助天然或合成材料来开发。由于聚合物材料在机械、物理和化学性质方面具有高度可变性,因此主要使用聚合物材料。可生物降解的聚合物是大多数研究的对象,因为它们能够被宿主生物体降解,与永久性移植物不同,可避免晚期支架血栓形成。聚左旋乳酸(PLLA)是研究中最常用的聚合物之一。为了提高材料的生物活性,在这项工作中,PLLA表面通过接枝精氨酸 - 甘氨酸 - 谷氨酰胺(RGD)(一种纤连蛋白衍生的黏附基序)和丝氨酸 - 异亮氨酸 - 赖氨酸 - 缬氨酸 - 丙氨酸 - 缬氨酸(SIKVAV)(一种层粘连蛋白衍生的基序)进行了修饰,并评估了大鼠心肌(H9C2)和小鼠(C2C12)成肌细胞在修饰后的PLLA上的增殖和分化情况。为了验证表面修饰,进行了X射线光电子能谱分析。接种后,通过3 -(4,5 - 二甲基噻唑 - 2 - 基)- 2,5 - 二苯基四氮唑溴盐试验确认细胞活力,同时使用增殖细胞核抗原表达来研究细胞增殖。Myf5、肌细胞生成素和肌球蛋白重链用于分析细胞分化。此外,RGD肽略微抑制大鼠成肌细胞(H9C2)的增殖,而对C2C12的抑制作用较弱。然而,在存在SIKVAV肽的情况下,两种细胞系均显示出增强收缩表型。这些结果表明,生物活性分子接枝可能对聚合物支架引导细胞表型表达有用,并最终维持适合组织功能再生的适当生物学特性。

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