Development of a Human Recombinant Collagen for Vat Polymerization-Based Bioprinting

IF 3.2 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Domenic Schlauch, Jan Peter Ebbecke, Johanna Meyer, Tabea Marie Fleischhammer, Hamidreza Pirmahboub, Lutz Kloke, Selin Kara, Antonina Lavrentieva, Iliyana Pepelanova
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Abstract

In light-based 3D-bioprinting, gelatin methacrylate (GelMA) is one of the most widely used materials, as it supports cell attachment, and shows good biocompatibility and degradability in vivo. However, as an animal-derived material, it also causes safety concerns when used in medical applications. Gelatin is a partial hydrolysate of collagen, containing high amounts of hydroxyproline. This causes the material to form a thermally induced gel at ambient temperatures, a behavior also observed in GelMA. This temperature-dependent gelation requires precise temperature control during the bioprinting process to prevent the gelation of the material. To avoid safety concerns associated with animal-derived materials and reduce potential issues caused by thermal gelation, a recombinant human alpha-1 collagen I fragment was expressed in Komagataella phaffii without hydroxylation. The resulting protein was successfully modified with methacryloyl groups and underwent rapid photopolymerization upon ultraviolet light exposure. The developed material exhibited slightly slower polymerization and lower storage modulus compared to GelMA, while it showed higher stretchability. However, unlike the latter, the material did not undergo physical gelation at ambient temperatures, but only when cooled down to below 10°C, a characteristic that has not been described for comparable materials so far. This gelation was not caused by the formation of triple-helical structures, as shown by the absence of the characteristic peak at 220 nm in CD spectra. Moreover, the developed recombinant material facilitated cell adherence with high cell viability after crosslinking via light to a 3D structure. Furthermore, desired geometries could be easily printed on a stereolithographic bioprinter.

Abstract Image

开发用于基于大桶聚合的生物打印的人类重组胶原蛋白。
在光基三维生物打印中,甲基丙烯酸明胶(GelMA)是使用最广泛的材料之一,因为它支持细胞附着,并在体内显示出良好的生物相容性和降解性。然而,作为一种动物源性材料,它在医疗应用中也会引起安全问题。明胶是胶原蛋白的部分水解物,含有大量羟脯氨酸。这导致该材料在环境温度下形成热诱导凝胶,在 GelMA 中也观察到这种行为。这种随温度变化的凝胶现象要求在生物打印过程中精确控制温度,以防止材料凝胶化。为了避免与动物源性材料相关的安全问题,并减少热凝胶化引起的潜在问题,在 Komagataella phaffii 中表达了重组人α-1 胶原 I 片段,但未进行羟基化。由此产生的蛋白质被成功地用甲基丙烯酰基修饰,并在紫外线照射下发生快速光聚合。与 GelMA 相比,所开发的材料聚合速度稍慢,储存模量较低,但拉伸性较高。然而,与后者不同的是,这种材料在环境温度下不会发生物理凝胶化,只有在冷却到 10°C 以下时才会发生凝胶化,这是迄今为止同类材料所没有描述过的特性。这种凝胶化不是由三螺旋结构的形成引起的,CD 光谱中 220 纳米处没有特征峰就说明了这一点。此外,所开发的重组材料在通过光交联成三维结构后,可促进细胞粘附,并具有很高的细胞存活率。此外,所需的几何形状可以很容易地在立体光刻生物打印机上打印出来。
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来源期刊
Biotechnology Journal
Biotechnology Journal Biochemistry, Genetics and Molecular Biology-Molecular Medicine
CiteScore
8.90
自引率
2.10%
发文量
123
审稿时长
1.5 months
期刊介绍: Biotechnology Journal (2019 Journal Citation Reports: 3.543) is fully comprehensive in its scope and publishes strictly peer-reviewed papers covering novel aspects and methods in all areas of biotechnology. Some issues are devoted to a special topic, providing the latest information on the most crucial areas of research and technological advances. In addition to these special issues, the journal welcomes unsolicited submissions for primary research articles, such as Research Articles, Rapid Communications and Biotech Methods. BTJ also welcomes proposals of Review Articles - please send in a brief outline of the article and the senior author''s CV to the editorial office. BTJ promotes a special emphasis on: Systems Biotechnology Synthetic Biology and Metabolic Engineering Nanobiotechnology and Biomaterials Tissue engineering, Regenerative Medicine and Stem cells Gene Editing, Gene therapy and Immunotherapy Omics technologies Industrial Biotechnology, Biopharmaceuticals and Biocatalysis Bioprocess engineering and Downstream processing Plant Biotechnology Biosafety, Biotech Ethics, Science Communication Methods and Advances.
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