合成肽基生物链接(PeptiInk Alpha 1)用于软骨组织模型体外3D生物打印的评价

IF 6.8 3区 医学 Q1 ENGINEERING, BIOMEDICAL International Journal of Bioprinting Pub Date : 2023-09-06 DOI:10.36922/ijb.0899
Patricia Santos-Beato, Andrew A. Pitsillides, Alberto Saiani, Aline Miller, Ryo Torii, Deepak M. Kalaskar
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引用次数: 0

摘要

软骨病理在人类疾病的了解很少,需要进一步的研究。利用体外人软骨模型作为临床前研究的替代方法,已经进行了各种各样的尝试来研究软骨病理。三维(3D)生物打印是一种使用动物源性材料(如明胶或透明质酸)体外3D生物打印软骨组织模型的技术,但在可扩展性、可重复性和伦理问题方面存在挑战。我们提出了一种评估合成自组装肽作为生物打印人类体外软骨模型的生物墨水。将人原代软骨细胞与PeptiInk Alpha 1混合,3D打印培养14天,并与3D软骨细胞颗粒对照进行比较。通过LIVE/DEAD测定和DNA定量评估细胞活力。在PeptiInk培养中观察到高细胞活力,而在3D颗粒对照中观察到DNA水平快速下降。使用苏木精和伊红染色以及免疫荧光标记对SOX-9、II型胶原和聚集蛋白进行组织学评估显示,3d生物打印的PeptiInk中细胞分布均匀,并且在对照和PeptiInk培养物中软骨标志物的高表达。通过定量实时聚合酶链反应(qRT-PCR)评估mRNA表达水平证实了软骨细胞的行为。这些数据显示了PeptiInk Alpha 1作为人体软骨体外模型的生物可打印制造材料的潜力。
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Evaluation of a synthetic peptide-based bioink (PeptiInk Alpha 1) for in vitro 3D bioprinting of cartilage tissue models
Cartilage pathology in human disease is poorly understood and requires further research. Various attempts have been made to study cartilage pathologies using in vitro human cartilage models as an alternative for preclinical research. Three-dimensional (3D) bioprinting is a technique that has been used to 3D-bioprint cartilage tissue models in vitro using animal-derived materials such as gelatine or hyaluronan, which present challenges in terms of scalability, reproducibility, and ethical concerns. We present an assessment of synthetic self-assembling peptides as bioinks for bioprinted human in vitro cartilage models. Primary human chondrocytes were mixed with PeptiInk Alpha 1, 3D-bioprinted and cultured for 14 days, and compared with 3D chondrocyte pellet controls. Cell viability was assessed through LIVE/DEAD assays and DNA quantification. High cell viability was observed in the PeptiInk culture, while a fast decrease in DNA levels was observed in the 3D pellet control. Histological evaluation using hematoxylin and eosin staining and immunofluorescence labeling for SOX-9, collagen type II, and aggrecan showed a homogeneous cell distribution in the 3D-bioprinted PeptiInks as well as high expression of chondrogenic markers in both control and PeptiInk cultures. mRNA expression levels assessed by - qRT-PCR (quantitative real time-polymerase chain reaction) confirmed chondrogenic cell behavior. These data showed promise in the potential use of PeptiInk Alpha 1 as a bioprintable manufacturing material for human cartilage in vitro models.
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来源期刊
CiteScore
6.90
自引率
4.80%
发文量
81
期刊介绍: The International Journal of Bioprinting is a globally recognized publication that focuses on the advancements, scientific discoveries, and practical implementations of Bioprinting. Bioprinting, in simple terms, involves the utilization of 3D printing technology and materials that contain living cells or biological components to fabricate tissues or other biotechnological products. Our journal encompasses interdisciplinary research that spans across technology, science, and clinical applications within the expansive realm of Bioprinting.
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