3D bioprinting of in vitro porous hepatoma models: establishment, evaluation, and anticancer drug testing

IF 8.1 1区 医学 Q1 ENGINEERING, BIOMEDICAL
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Abstract

Traditional tumor models do not tend to accurately simulate tumor growth in vitro or enable personalized treatment and are particularly unable to discover more beneficial targeted drugs. To address this, this study describes the use of three-dimensional (3D) bioprinting technology to construct a 3D model with human hepatocarcinoma SMMC-7721 cells (3DP-7721) by combining gelatin methacrylate (GelMA) and poly(ethylene oxide) (PEO) as two immiscible aqueous phases to form a bioink and innovatively applying fluorescent carbon quantum dots for long-term tracking of cells. The GelMA (10%, mass fraction) and PEO (1.6%, mass fraction) hydrogel with 3:1 volume ratio offered distinct pore-forming characteristics, satisfactory mechanical properties, and biocompatibility for the creation of the 3DP-7721 model. Immunofluorescence analysis and quantitative real-time fluorescence polymerase chain reaction (PCR) were used to evaluate the biological properties of the model. Compared with the two-dimensional culture cell model (2D-7721) and the 3D mixed culture cell model (3DM-7721), 3DP-7721 significantly improved the proliferation of cells and expression of tumor-related proteins and genes. Moreover, we evaluated the differences between the three culture models and the effectiveness of antitumor drugs in the three models and discovered that the efficacy of antitumor drugs varied because of significant differences in resistance proteins and genes between the three models. In addition, the comparison of tumor formation in the three models found that the cells cultured by the 3DP-7721 model had strong tumorigenicity in nude mice. Immunohistochemical evaluation of the levels of biochemical indicators related to the formation of solid tumors showed that the 3DP-7721 model group exhibited pathological characteristics of malignant tumors, the generated solid tumors were similar to actual tumors, and the deterioration was higher. This research therefore acts as a foundation for the application of 3DP-7721 models in drug development research.

体外多孔肝癌模型的三维生物打印:建立、评估和抗癌药物测试
摘要 传统的肿瘤模型往往不能准确模拟肿瘤在体外的生长情况,也无法实现个性化治疗,特别是无法发现更多有益的靶向药物。针对这一问题,本研究介绍了利用三维(3D)生物打印技术,通过将甲基丙烯酸明胶(Gelatin Methacrylate,GelMA)和聚环氧乙烷(Poly(Ethylene oxide,PEO)作为两种不相溶的水相形成生物墨水,并创新性地应用荧光碳量子点对细胞进行长期追踪,从而构建出带有人肝癌SMMC-7721细胞的三维模型(3DP-7721)。体积比为 3:1 的 GelMA(10%,质量分数)和 PEO(1.6%,质量分数)水凝胶具有独特的成孔特性、令人满意的机械性能和生物相容性,可用于创建 3DP-7721 模型。免疫荧光分析和定量实时荧光聚合酶链反应(PCR)用于评估模型的生物特性。与二维培养细胞模型(2D-7721)和三维混合培养细胞模型(3DM-7721)相比,3DP-7721能显著改善细胞的增殖以及肿瘤相关蛋白和基因的表达。此外,我们还评估了三种培养模型之间的差异以及抗肿瘤药物在三种模型中的疗效,发现抗肿瘤药物的疗效因三种模型之间抗性蛋白和基因的显著差异而不同。此外,通过比较三种模型的肿瘤形成情况发现,3DP-7721 模型培养的细胞在裸鼠体内具有很强的致瘤性。对实体瘤形成相关生化指标水平的免疫组化评估显示,3DP-7721 模型组表现出恶性肿瘤的病理特征,生成的实体瘤与实际肿瘤相似,且恶化程度较高。因此,这项研究为 3DP-7721 模型在药物开发研究中的应用奠定了基础。
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来源期刊
Bio-Design and Manufacturing
Bio-Design and Manufacturing Materials Science-Materials Science (miscellaneous)
CiteScore
13.30
自引率
7.60%
发文量
148
期刊介绍: Bio-Design and Manufacturing reports new research, new technology and new applications in the field of biomanufacturing, especially 3D bioprinting. Topics of Bio-Design and Manufacturing cover tissue engineering, regenerative medicine, mechanical devices from the perspectives of materials, biology, medicine and mechanical engineering, with a focus on manufacturing science and technology to fulfil the requirement of bio-design.
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