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一种可光交联的软骨衍生细胞外基质生物墨水,用于耳廓软骨组织工程。

A photo-crosslinkable cartilage-derived extracellular matrix bioink for auricular cartilage tissue engineering.

机构信息

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA; Department of Plastic, Reconstructive, and Hand Surgery, Amsterdam UMC, Amsterdam 1081HV, the Netherlands.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.

出版信息

Acta Biomater. 2021 Feb;121:193-203. doi: 10.1016/j.actbio.2020.11.029. Epub 2020 Nov 21.

Abstract

Three-dimensional (3D) bioprinting of patient-specific auricular cartilage constructs could aid in the reconstruction process of traumatically injured or congenitally deformed ear cartilage. To achieve this, a hydrogel-based bioink is required that recapitulates the complex cartilage microenvironment. Tissue-derived decellularized extracellular matrix (dECM)-based hydrogels have been used as bioinks for cell-based 3D bioprinting because they contain tissue-specific ECM components that play a vital role in cell adhesion, growth, and differentiation. In this study, porcine auricular cartilage tissues were isolated and decellularized, and the decellularized cartilage tissues were characterized by histology, biochemical assay, and proteomics. This cartilage-derived dECM (cdECM) was subsequently processed into a photo-crosslinkable hydrogel using methacrylation (cdECMMA) and mixed with chondrocytes to create a printable bioink. The rheological properties, printability, and in vitro biological properties of the cdECMMA bioink were examined. The results showed cdECM was obtained with complete removal of cellular components while preserving major ECM proteins. After methacrylation, the cdECMMA bioinks were printed in anatomical ear shape and exhibited adequate mechanical properties and structural integrity. Specifically, auricular chondrocytes in the printed cdECMMA hydrogel constructs maintained their viability and proliferation capacity and eventually produced cartilage ECM components, including collagen and glycosaminoglycans (GAGs). The potential of cell-based bioprinting using this cartilage-specific dECMMA bioink is demonstrated as an alternative option for auricular cartilage reconstruction.

摘要

三维(3D)生物打印患者特异性耳廓软骨构建体有助于创伤性损伤或先天性畸形耳软骨的重建过程。为此,需要一种基于水凝胶的生物墨水,该生物墨水可再现复杂的软骨微环境。组织来源的去细胞化细胞外基质(dECM)基水凝胶已被用作基于细胞的 3D 生物打印的生物墨水,因为它们包含组织特异性 ECM 成分,这些成分在细胞粘附、生长和分化中起着至关重要的作用。在这项研究中,从猪耳软骨组织中分离并去细胞化,然后通过组织学、生化分析和蛋白质组学对去细胞化的软骨组织进行表征。随后,使用甲基丙烯酰化(cdECMMA)将这种软骨衍生的 dECM(cdECM)加工成可光交联的水凝胶,并与软骨细胞混合以创建可打印的生物墨水。研究了 cdECMMA 生物墨水的流变性能、可打印性和体外生物学性能。结果表明,在保留主要 ECM 蛋白的同时,完全去除了细胞成分,获得了 cdECM。甲基丙烯酰化后,cdECMMA 生物墨水可按解剖学耳形状打印,具有足够的机械性能和结构完整性。具体而言,打印 cdECMMA 水凝胶构建体中的耳廓软骨细胞保持其活力和增殖能力,并最终产生软骨 ECM 成分,包括胶原蛋白和糖胺聚糖(GAG)。使用这种软骨特异性 dECMMA 生物墨水进行基于细胞的生物打印的潜力证明了它是耳廓软骨重建的另一种选择。

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