Development of an In Vitro 3D Brain Tissue Model Mimicking In Vivo-Like Pro-inflammatory and Pro-oxidative Responses
Development of an In Vitro 3D Brain Tissue Model Mimicking In Vivo-Like Pro-inflammatory and Pro-oxidative Responses
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DOI：
10.1007/s10439-018-2004-z
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发表时间：
2018-06-01
影响因子：
3.8
通讯作者：
Lee, Yong W.
中科院分区：
文献类型：
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作者：
Cho, Hyung Joon;Verbridge, Scott S.;Lee, Yong W.
为了分析体外系统中的复杂炎症反应，我们构建了一个新的3D体外脑组织模型，该模型显示了对炎症刺激的体内样组织反应（例如免疫细胞表型和分子反应）。氧气扩散和细胞耗氧量的有限元建模预测了三维结构内的氧气分布，由嵌入小鼠小胶质细胞的I型胶原水凝胶组成。活力和细胞毒性分析支持数学分析，确定3D构建开发的最佳细胞生长条件。Real-time RT-PCR和ELISA显示，在脂多糖（LPS）刺激的体外细胞培养（2D和3D）和体内小鼠模型系统中，促炎介质如tnf - α、MCP-1、IL-6和IL-1 β显著上调。有趣的是，体外3D模型系统的炎症反应水平比体外2D模型更接近体内。此外，原位双氢乙啶（DHE）测定和免疫荧光染色显示，体外3D模型系统中lps刺激的活性氧（ROS）生成和小胶质细胞激活水平比体外2D更接近体内。这些结果表明，与体外2D模型相比，体外3D模型在大脑中提供了更多与生理相关的促氧化和促炎症环境。
To analyze complex inflammatory responses in an in vitro system, we constructed a new 3D in vitro brain tissue model that exhibits in vivo-like tissue responses (e.g. immune cell phenotypes, and molecular response) to inflammatory stimuli. Finite element modeling of oxygen diffusion and cellular oxygen consumption predicted the oxygen profile within 3D structures, consisting of Type I collagen hydrogel embedded with murine microglia. Viability and cytotoxicity analyses supported the mathematical analysis, determining optimal cell growth conditions for 3D construct development. Real-time RT-PCR and ELISA demonstrated significant up-regulation of pro-inflammatory mediators, such as TNF-alpha, MCP-1, IL-6 and IL-1 beta, in lipopolysaccharide (LPS)-stimulated in vitro cell culture (2D and 3D) and in vivo mouse model systems. Interestingly, levels of inflammatory responses from the in vitro 3D model system were more similar to in vivo than in vitro 2D. Additionally, in situ dihydroethidium (DHE) assay and immunofluorescence staining revealed that levels of LPS-stimulated reactive oxygen species (ROS) generation and microglial activation from in vitro 3D model system were closer to in vivo than in vitro 2D. These results demonstrated that an in vitro 3D model provides more physiologically relevant pro-oxidative and pro-inflammatory environments in brain than an in vitro 2D model.