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两种不同海藻酸盐基生物墨水的特性及其对黑色素瘤生长的影响。

Characterization of two different alginate-based bioinks and the influence of melanoma growth within.

机构信息

Department of Plastic and Hand Surgery, Laboratory for Tissue Engineering and Regenerative Medicine, University Hospital Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg, 91054, Erlangen, Germany.

Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander-University Erlangen-Nürnberg, 91052, Erlangen, Germany.

出版信息

Sci Rep. 2024 Jun 5;14(1):12945. doi: 10.1038/s41598-024-63642-3.

Abstract

Extrusion-based bioprinting is an established method in biofabrication. Suitable bioinks have fundamentally different compositions and characteristics, which should be examined, in order to find a perfect model system. Here, we investigate the effect of two alginate-based, yet unalike 3D-printed bioinks, pre-crosslinked alginate-dialdehyde gelatin (ADA-GEL) and a mixture of alginate, hyaluronic acid, and gelatin (Alg/HA/Gel), on the melanoma cell line Mel Im and vice versa in terms of stiffness, shrinkage, cellular behavior and colony formation over 15 days. Rheological stiffness measurements revealed two soft gels with similar storage moduli. The cells did not have a significant impact on the overall stiffness, whereas ADA-GEL (2.5/2.5%) was significantly stiffer than Alg/HA/Gel (0.5/0.1/3%). Regarding the shrinkage of printed constructs, cells had a significant influence, especially in ADA-GEL, which has covalent bonds between the oxidized alginate and gelatin. Multi-photon microscopy exhibited proliferation, cell spreading and migration in ADA-GEL with cell-cell and cell-matrix interaction, dissimilarly to Alg/HA/Gel, in which cells formed spherical, encapsulated colonies. Scanning electron microscopy and histology showed degradation and multi-layered growth on ADA-GEL and fewer examples of escaped cells on Alg/HA/Gel. Both gels serve as proliferation bioink for melanoma with more necrosis in deeper Alg/HA/Gel colonies and differences in spreading and matrix interaction. These findings show the importance of proper characterization of the bioinks for different applications.

摘要

基于挤出的生物打印是生物制造中的一种既定方法。合适的生物墨水具有根本不同的组成和特性,应该进行检查,以找到一个完美的模型系统。在这里,我们研究了两种基于藻酸盐的、不同的 3D 打印生物墨水对黑色素瘤细胞系 Mel Im 的影响,以及反之亦然,即在刚度、收缩、细胞行为和 15 天内的菌落形成方面。流变学刚度测量显示两种柔软的凝胶具有相似的储能模量。细胞对整体刚度没有显著影响,而 ADA-GEL(2.5/2.5%)明显比 Alg/HA/Gel(0.5/0.1/3%)更硬。关于打印结构的收缩,细胞有显著的影响,特别是在 ADA-GEL 中,氧化藻酸盐和明胶之间有共价键。多光子显微镜显示,在 ADA-GEL 中,细胞增殖、细胞扩散和迁移伴随着细胞-细胞和细胞-基质相互作用,而在 Alg/HA/Gel 中则不同,细胞形成了球形、包裹的菌落。扫描电子显微镜和组织学显示,ADA-GEL 上有降解和多层生长,而在 Alg/HA/Gel 上则较少出现逃逸细胞的例子。这两种凝胶都可以作为黑色素瘤的增殖生物墨水,Alg/HA/Gel 深层的细胞坏死更多,而在细胞扩散和基质相互作用方面存在差异。这些发现表明,对于不同的应用,对生物墨水进行适当的特性描述非常重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb0f/11153560/caac9d147b76/41598_2024_63642_Fig1_HTML.jpg

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