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基于立体光刻技术的壳聚糖:聚乙二醇二丙烯酸酯混合凝胶支架的力学性能、细胞相容性及可制造性

Mechanical Properties, Cytocompatibility and Manufacturability of Chitosan:PEGDA Hybrid-Gel Scaffolds by Stereolithography.

作者信息

Morris Viola B, Nimbalkar Siddharth, Younesi Mousa, McClellan Phillip, Akkus Ozan

机构信息

Department of Mechanical and Aerospace Engineering, Case Western Reserve University, 10900 Euclid Ave, Cleveland, OH, 44106-7222, USA.

Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH, 44106, USA.

出版信息

Ann Biomed Eng. 2017 Jan;45(1):286-296. doi: 10.1007/s10439-016-1643-1. Epub 2016 May 10.

Abstract

Extracellular matrix mimetic hydrogels which hybridize synthetic and natural polymers offer molecularly-tailored, bioactive properties and tunable mechanical strength. In addition, 3D bioprinting by stereolithography allows fabrication of internal pores and defined macroscopic shapes. In this study, we formulated a hybrid biocompatible resin using natural and synthetic polymers (chitosan and polyethylene glycol diacrylate (PEGDA), respectively) by controlling molecular weight of chitosan, feed-ratios, and photo-initiator concentration. Ear-shaped, hybrid scaffolds were fabricated by a stereolithographic method using a 405 nm laser. Hybrid hydrogel scaffolds of chitosan (50-190 kDa) and PEGDA (575 Da) were mixed at varying feed-ratios. Some of the cationic, amino groups of chitosan were neutralized by dialysis in acidic solution containing chitosan in excess of sodium acetate solution to inhibit quenching of newly formed photoradicals. A feed-ratio of 1:7.5 was found to be the most appropriate of the formulations considered in this study in terms of mechanical properties, cell adhesion, and printability. The biofabricated hybrid scaffold showed interconnected, homogeneous pores with a nominal pore size of 50 µm and an elastic modulus of ~400 kPa. Moreover, long-term cell viability and cell spreading was observed via actin filament staining. Printability of the biocompatible resin was confirmed by printing thresholded MR images of an ear and the feed ratio of 1:7.5 provided the most faithful reproduction of the shape. To the best of our knowledge, this is the first report of stereolithographic printing hybridizing cell-adhesive properties of chitosan with mechanical robustness of PEG in scaffolds suitable for repair of complex tissue geometries, such as those of the human ear.

摘要

将合成聚合物与天然聚合物相结合的细胞外基质模拟水凝胶具有分子定制的生物活性特性和可调的机械强度。此外,通过立体光刻进行的3D生物打印能够制造内部孔隙和确定的宏观形状。在本研究中,我们通过控制壳聚糖的分子量、进料比和光引发剂浓度,使用天然和合成聚合物(分别为壳聚糖和聚乙二醇二丙烯酸酯(PEGDA))配制了一种混合生物相容性树脂。使用405nm激光通过立体光刻方法制造了耳形混合支架。壳聚糖(50 - 190 kDa)和PEGDA(575 Da)的混合水凝胶支架以不同的进料比混合。壳聚糖的一些阳离子氨基在含有过量醋酸钠溶液的壳聚糖酸性溶液中通过透析被中和,以抑制新形成的光自由基的猝灭。就机械性能、细胞粘附和可打印性而言,发现1:7.5的进料比是本研究中考虑的配方中最合适的。生物制造的混合支架显示出相互连接的均匀孔隙,标称孔径为50μm,弹性模量约为400 kPa。此外,通过肌动蛋白丝染色观察到长期细胞活力和细胞铺展。通过打印耳朵的阈值化磁共振图像证实了生物相容性树脂的可打印性,并且1:7.5的进料比提供了最逼真的形状再现。据我们所知,这是关于立体光刻打印的首次报道,该打印将壳聚糖的细胞粘附特性与PEG的机械稳健性在适合修复复杂组织几何形状(如人耳)的支架中相结合。

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