基于 ColMA 的生物打印三维支架可用于研究人类肌腱干细胞的韧化事件

IF 6.1 2区 医学 Q1 ENGINEERING, BIOMEDICAL Bioengineering & Translational Medicine Pub Date : 2024-10-31 DOI:10.1002/btm2.10723
Giacomo Cortella, Erwin Pavel Lamparelli, Maria Camilla Ciardulli, Joseph Lovecchio, Emanuele Giordano, Nicola Maffulli, Giovanna Della Porta
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引用次数: 0

摘要

生物打印技术的出现使精确的三维(3D)细胞培养物成为可能,它适用于仿生体外模型。在这项研究中,我们开发了一种新的三维打印方法,将甲基丙烯酸胶原蛋白(ColMa 或 PhotoCol®)与从人体肌腱外植体中提取的肌腱干/祖细胞(hTSPCs)结合在一起。虽然纯 ColMa 以前从未被提议用作可打印的水凝胶,但本文概述了生物打印这种材料的稳健且高度可重现的管道。事实上,我们成功制作了三维生物工程支架,并在灌注条件下用补充了生长/分化因子-5(GDF-5)的培养基培养了 21 天。这种生物打印管道和培养条件创造了一个非常有利的三维环境,使细胞能够增殖,表现出韧性行为,并产生新的 I 型胶原基质,从而重塑周围环境。事实上,在灌注条件下进行的 21 天培养中,腱鞘蛋白的表达在第 7 天显著上调,与第 14 天和第 21 天相比增加了 2.3 倍。到第 14 天,I 型胶原蛋白基因表达上调了近 10 倍。这一趋势在 Western 印迹分析中得到了进一步证实,该分析表明,第 21 天与第 7 天和第 14 天之间的tenomodulin 表达有显著的统计学差异。就 I 型胶原蛋白而言,在第 0 天和第 21 天之间以及第 0 天和第 14 天之间观察到了显著差异,P 值为 0.01。这些结果表明 I 型胶原蛋白逐渐过度表达,反映了细胞向适当的腱鞘表型分化。细胞因子(如 IL-8 和 IL-6)的水平在第 7 天分别达到 8566 和 7636 pg/mL 的峰值,到第 21 天分别降至 54 和 46 pg/mL。总之,这些数据表明,新型 ColMa 生物打印方案有效地为 hTSPCs 的生长和适当分化提供了有利环境,展示了其在研究细胞行为和韧带分化方面的潜力。
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ColMA‐based bioprinted 3D scaffold allowed to study tenogenic events in human tendon stem cells
The advent of bioprinting has enabled the creation of precise three‐dimensional (3D) cell cultures suitable for biomimetic in vitro models. In this study, we developed a novel protocol for 3D printing methacrylated collagen (ColMa, or PhotoCol®) combined with tendon stem/progenitor cells (hTSPCs) derived from human tendon explants. Although pure ColMa has not previously been proposed as a printable hydrogel, this paper outlines a robust and highly reproducible pipeline for bioprinting this material. Indeed, we successfully fabricated a 3D bioengineered scaffold and cultured it for 21 days under perfusion conditions with medium supplemented with growth/differentiation factor‐5 (GDF‐5). This bioprinting pipeline and the culture conditions created an exceptionally favorable 3D environment, enabling the cells to proliferate, exhibit tenogenic behaviors, and produce a new collagen type I matrix, thereby remodeling the surrounding environment. Indeed, over the 21‐day culture period under perfusion condition, tenomodulin expression showed a significant upregulation on day 7, with a 2.3‐fold increase, compared to days 14 and 21. Collagen type I gene expression was upregulated nearly 10‐fold by day 14. This trend was further confirmed by western blot analysis, which revealed a statistically significant difference in tenomodulin expression between day 21 and both day 7 and day 14. For type I collagen, significant differences were observed between day 0 and day 21, as well as between day 0 and day 14, with a p‐value of 0.01. These results indicate a progressive over‐expression of type I collagen, reflecting cell differentiation towards a proper tenogenic phenotype. Cytokines, such as IL‐8 and IL‐6, levels peaked at 8566 and 7636 pg/mL, respectively, on day 7, before decreasing to 54 and 46 pg/mL by day 21. Overall, the data suggest that the novel ColMa bioprinting protocol effectively provided a conducive environment for the growth and proper differentiation of hTSPCs, showcasing its potential for studying cell behavior and tenogenic differentiation.
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来源期刊
Bioengineering & Translational Medicine
Bioengineering & Translational Medicine Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science
CiteScore
8.40
自引率
4.10%
发文量
150
审稿时长
12 weeks
期刊介绍: Bioengineering & Translational Medicine, an official, peer-reviewed online open-access journal of the American Institute of Chemical Engineers (AIChE) and the Society for Biological Engineering (SBE), focuses on how chemical and biological engineering approaches drive innovative technologies and solutions that impact clinical practice and commercial healthcare products.
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