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基于微图案化的结冷胶水凝胶,通过整合层粘连蛋白衍生肽来用于骨骼肌组织工程。

Micropatterned gellan gum-based hydrogels tailored with laminin-derived peptides for skeletal muscle tissue engineering.

机构信息

3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal; ICVS/3B's-PT Government Associate Laboratory, 4710-057, Braga/Guimarães, Portugal.

3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal; ICVS/3B's-PT Government Associate Laboratory, 4710-057, Braga/Guimarães, Portugal.

出版信息

Biomaterials. 2021 Dec;279:121217. doi: 10.1016/j.biomaterials.2021.121217. Epub 2021 Oct 29.

Abstract

The efficacy of current therapies for skeletal muscle disorders/injuries are limited urging the need for new treatments. Skeletal muscle tissue engineered platforms represent a promising tool to shed light on the pathophysiology of skeletal muscle disorders/injuries and to investigate the efficacy of new therapies. Herein, we developed a skeletal muscle platform composed of aligned and differentiated myoblasts on micropatterned gellan gum (GG)-based hydrogels tailored with a laminin-derived peptide. To this aim, the binding of murine skeletal muscle cells (C2C12) to different laminin-derived peptides (CIKVAVS (V), KNRLTIELEVRTC (T), and RKRLQVQLSIRTC (Q)) and the binding of laminin-derived peptides to chemically functionalized GG was studied. C2C12-binding to peptide V, T and Q was 10%, 48% and 25%, whereas the peptide tethering to GG was 60%, 40% and 31%, respectively. Peptide-biofunctionalized hydrogels prepared with different polymer content showed different mechanics and peptide exposure at hydrogel surface. Cellular adhesion was detected in all hydrogel formulations, but spreading and differentiation was only promoted in peptide Q-biofunctionalized hydrogels and preferably in stiffer hydrogels. Myoblast alignment was promoted in micropatterned hydrogel surfaces. Overall, the engineered skeletal muscle herein proposed can be further explored as a platform to better understand skeletal muscle disorders/injuries and to screen new therapies.

摘要

当前治疗骨骼肌疾病/损伤的疗效有限,因此需要新的治疗方法。骨骼肌组织工程平台代表了一种有前途的工具,可以深入了解骨骼肌疾病/损伤的病理生理学,并研究新疗法的疗效。在此,我们开发了一种由微图案化的卡拉胶(GG)基水凝胶上排列和分化的成肌细胞组成的骨骼肌平台,该水凝胶经过精心设计,含有层粘连蛋白衍生肽。为此,研究了鼠骨骼肌细胞(C2C12)与不同层粘连蛋白衍生肽(CIKVAVS(V)、KNRLTIELEVRTC(T)和 RKRLQVQLSIRTC(Q))的结合以及层粘连蛋白衍生肽与化学功能化 GG 的结合。C2C12 与肽 V、T 和 Q 的结合率分别为 10%、48%和 25%,而肽与 GG 的结合率分别为 60%、40%和 31%。用不同聚合物含量制备的肽生物功能化水凝胶表现出不同的力学性能和水凝胶表面的肽暴露。在所有水凝胶配方中都检测到细胞黏附,但仅在肽 Q 生物功能化水凝胶中促进了细胞的铺展和分化,并且在更硬的水凝胶中更有利于促进细胞的铺展和分化。成肌细胞在微图案化水凝胶表面排列。总之,本文提出的工程化骨骼肌可进一步作为平台来深入了解骨骼肌疾病/损伤,并筛选新的治疗方法。

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