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在生理温度下对多材料脱细胞化肝脏基质水凝胶进行 3D 生物打印。

3D Bioprinting of Multi-Material Decellularized Liver Matrix Hydrogel at Physiological Temperatures.

机构信息

Science for Life Laboratory, Division of Nanobiotechnology, Department of Protein Science, KTH Royal Institute of Technology, 17165 Solna, Sweden.

Biopromic AB, 17165 Solna, Sweden.

出版信息

Biosensors (Basel). 2022 Jul 13;12(7):521. doi: 10.3390/bios12070521.

Abstract

Bioprinting is an acclaimed technique that allows the scaling of 3D architectures in an organized pattern but suffers from a scarcity of appropriate bioinks. Decellularized extracellular matrix (dECM) from xenogeneic species has garnered support as a biomaterial to promote tissue-specific regeneration and repair. The prospect of developing dECM-based 3D artificial tissue is impeded by its inherent low mechanical properties. In recent years, 3D bioprinting of dECM-based bioinks modified with additional scaffolds has advanced the development of load-bearing constructs. However, previous attempts using dECM were limited to low-temperature bioprinting, which is not favorable for a longer print duration with cells. Here, we report the development of a multi-material decellularized liver matrix (dLM) bioink reinforced with gelatin and polyethylene glycol to improve rheology, extrudability, and mechanical stability. This shear-thinning bioink facilitated extrusion-based bioprinting at 37 °C with HepG2 cells into a 3D grid structure with a further enhancement for long-term applications by enzymatic crosslinking with mushroom tyrosinase. The heavily crosslinked structure showed a 16-fold increase in viscosity (2.73 Pa s) and a 32-fold increase in storage modulus from the non-crosslinked dLM while retaining high cell viability (85-93%) and liver-specific functions. Our results show that the cytocompatible crosslinking of dLM bioink at physiological temperatures has promising applications for extended 3D-printing procedures.

摘要

生物打印是一种备受推崇的技术,可按组织模式对 3D 架构进行缩放,但存在生物墨水不足的问题。异种去细胞细胞外基质(dECM)作为一种生物材料,已被广泛用于促进组织特异性再生和修复。基于 dECM 的 3D 人工组织的开发前景受到其固有低机械性能的阻碍。近年来,通过添加额外支架对基于 dECM 的生物墨水进行 3D 生物打印,推动了承重结构的发展。然而,以前使用 dECM 的尝试仅限于低温生物打印,这不利于细胞的长时间打印。在这里,我们报告了一种多材料去细胞化肝脏基质(dLM)生物墨水的开发,该生物墨水通过添加明胶和聚乙二醇来改善流变学、可挤出性和机械稳定性。这种剪切稀化生物墨水可在 37°C 下进行基于挤出的生物打印,将 HepG2 细胞打印成 3D 网格结构,进一步通过蘑菇酪氨酸酶的酶交联进行长期应用增强。交联后的结构的粘度增加了 16 倍(2.73 Pa·s),储能模量增加了 32 倍,而非交联的 dLM 则保留了较高的细胞活力(85-93%)和肝脏特异性功能。我们的结果表明,在生理温度下对 dLM 生物墨水进行细胞相容的交联具有在扩展 3D 打印过程中的应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0c93/9313433/36c056b26a0a/biosensors-12-00521-g001.jpg

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