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通过辣根过氧化物酶催化交联的甜菜果胶 3D 生物打印。

3D Bioprinting of Sugar Beet Pectin through Horseradish Peroxidase-Catalyzed Cross-Linking.

机构信息

Division of Chemical Engineering, Department of Materials Engineering Science, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka 560-8531, Japan.

出版信息

ACS Appl Bio Mater. 2024 May 20;7(5):3506-3514. doi: 10.1021/acsabm.4c00418. Epub 2024 May 2.

Abstract

Horseradish peroxidase (HRP)-mediated hydrogelation, caused by the cross-linking of phenolic groups in polymers in the presence of hydrogen peroxide (HO), is an effective route for bioink solidification in 3D bioprinting. Sugar beet pectin (SBP) naturally has cross-linkable phenols through the enzymatic reaction. Therefore, chemical modifications are not required, unlike the various polymers that have been used in the enzymatic cross-linking system. In this study, we report the application of SBP in extrusion-based bioprinting including HRP-mediated bioink solidification. In this system, HO necessary for the solidification of inks is supplied in the gas phase. Cell-laden liver lobule-like constructs could be fabricated using bioinks consisting of 10 U/mL HRP, 4.0 and 6.0 w/v% SBP, and 6.0 × 10 cells/mL human hepatoblastoma (HepG2) cells exposed to air containing 16 ppm of HO concurrently during printing and 10 min postprinting. The HepG2 cells enclosed in the printed constructs maintained their viability, metabolic activity, and hepatic functions from day 1 to day 7 of the culture, which indicates the cytocompatibility of this system. Taken together, this result demonstrates the potential of SBP and HRP cross-linking systems for 3D bioprinting, which can be applied in tissue engineering applications.

摘要

辣根过氧化物酶(HRP)介导的水凝胶化是一种有效的生物墨水固化方法,它是在存在过氧化氢(HO)的情况下,通过聚合物中酚类基团的交联实现的,这种方法在 3D 生物打印中被广泛应用。糖甜菜果胶(SBP)天然具有可通过酶反应交联的酚类基团,因此不需要进行化学修饰,这与各种已被用于酶交联系统的聚合物不同。在本研究中,我们报告了 SBP 在挤出式生物打印中的应用,包括 HRP 介导的生物墨水固化。在这个系统中,用于墨水固化的 HO 以气相的形式提供。使用包含 10 U/mL HRP、4.0 和 6.0 w/v% SBP 和 6.0×10 个细胞/mL 人肝癌细胞(HepG2)的生物墨水,可以制造出含有肝小叶样结构的细胞负载结构体。在打印过程中和打印后 10 分钟内,同时向空气中提供含有 16 ppm HO 的气体,可以使打印结构体中的 HepG2 细胞保持活力、代谢活性和肝功能,从培养的第 1 天到第 7 天。这表明了该系统的细胞相容性。综上所述,该结果表明 SBP 和 HRP 交联系统在 3D 生物打印中的应用潜力,该系统可应用于组织工程应用。

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