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生物打印明胶水凝胶平台促进平滑肌细胞收缩表型维持。

Bioprinted gelatin hydrogel platform promotes smooth muscle cell contractile phenotype maintenance.

作者信息

Tijore Ajay, Behr Jean-Marc, Irvine Scott Alexander, Baisane Vrushali, Venkatraman Subbu

机构信息

School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798, Singapore.

出版信息

Biomed Microdevices. 2018 Mar 28;20(2):32. doi: 10.1007/s10544-018-0274-8.

Abstract

Three dimensional (3D) bioprinting has been proposed as a method for fabricating tissue engineered small diameter vascular prostheses. This technique not only involves constructing the structural features to obtain a desired pattern but the morphology of the pattern may also be used to influence the behavior of seeded cells. Herein, we 3D bioprinted a gelatin hydrogel microchannel construct to promote and preserve the contractile phenotype of vascular smooth muscle cells (vSMCs), which is crucial for vasoresponsiveness. The microchanneled surface of a gelatin hydrogel facilitated vSMC attachment and an elongated alignment along the microchannel direction. The cells displayed distinct F-actin anisotropy in the direction of the channel. The vSMC contractile phenotype was confirmed by the positive detection of contractile marker gene proteins (α-smooth muscle actin (α-SMA) and smooth muscle-myosin heavy chain (SM-MHC)). Having demonstrated the effectiveness of the hydrogel channels bioprinted on a film, the bioprinting was applied radially to the surface of a 3D tubular construct by integrating a rotating mandrel into the 3D bioprinter. The hydrogel microchannels printed on the 3D tubular vascular construct also orientated the vSMCs and strongly promoted the contractile phenotype. Together, our study demonstrated that microchannels bioprinted using a transglutaminase crosslinked gelatin hydrogel, could successfully promote and preserve vSMC contractile phenotype. Furthermore, the hydrogel bioink could be retained on the surface of a rotating polymer tube to print radial cell guiding channels onto a vascular graft construct.

摘要

三维(3D)生物打印已被提议作为一种制造组织工程小直径血管移植物的方法。该技术不仅涉及构建结构特征以获得所需图案,而且图案的形态也可用于影响接种细胞的行为。在此,我们通过3D生物打印制备了一种明胶水凝胶微通道构建体,以促进和维持血管平滑肌细胞(vSMC)的收缩表型,这对血管反应性至关重要。明胶水凝胶的微通道表面促进了vSMC的附着以及沿微通道方向的伸长排列。细胞在通道方向上显示出明显的F-肌动蛋白各向异性。通过收缩标记基因蛋白(α-平滑肌肌动蛋白(α-SMA)和平滑肌肌球蛋白重链(SM-MHC))的阳性检测证实了vSMC的收缩表型。在证明了在薄膜上生物打印的水凝胶通道的有效性后,通过将旋转心轴集成到3D生物打印机中,将生物打印径向应用于3D管状构建体的表面。打印在3D管状血管构建体上的水凝胶微通道也使vSMC定向,并强烈促进收缩表型。总之,我们的研究表明,使用转谷氨酰胺酶交联的明胶水凝胶生物打印的微通道可以成功地促进和维持vSMC的收缩表型。此外,水凝胶生物墨水可以保留在旋转聚合物管的表面,以在血管移植物构建体上打印径向细胞引导通道。

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