在许多损伤和病理情况下都可以观察到大脑持续压缩性损伤 (SCI)，包括肿瘤、缺血性中风和创伤性脑损伤相关的组织肿胀。持续性压缩损伤的特点是随着时间的推移组织负荷，目前，很少有体外模型适合研究神经细胞对应变依赖性持续压缩性损伤的反应。在这里，我们提出了一种通过离心造成持续压迫性神经损伤的体外模型。球体由新生大鼠皮层细胞制成，以 4000 个细胞/球体接种并在体外培养 14 天。将一部分球体以 104、209、313 或 419 rad/s 的速度离心 2 分钟。通过有限元分析对球体的物理变形进行建模，我们发现以上述角速度离心的球体分别经历 10、38、84 和 149 kPa 的压力，以及 10% (5%) 的压缩（分别是拉伸）应变。 ）、18%（9%）、27%（14%）和35%（18%）。 LIVE-DEAD 测定和 Hoechst 33342 核染色的定量显示，在损伤后 2、8 和 24 小时，承受 10 kPa 以上压力的离心球体比对照球体表现出显着更高的 DNA 损伤。离心损伤后 2、8 和 24 小时的 β3-微管蛋白网络的免疫组织化学显示，随着时间的推移，随着应变的增加，微管的降解也在增加。我们的研究结果表明，细胞损伤是由于持续组织应变的特定水平和时间而发生的。该实验性 SCI 模型提供了一个高通量体外平台来检查细胞损伤，以深入了解可针对治疗策略的脑损伤。版权所有：© 2024 González-Cruz 等人。这是一篇根据知识共享署名许可条款分发的开放获取文章，允许在任何媒体上不受限制地使用、分发和复制，前提是注明原始作者和来源。

Sustained compressive injury (SCI) in the brain is observed in numerous injury and pathological scenarios, including tumors, ischemic stroke, and traumatic brain injury-related tissue swelling. Sustained compressive injury is characterized by tissue loading over time, and currently, there are few in vitro models suitable to study neural cell responses to strain-dependent sustained compressive injury. Here, we present an in vitro model of sustained compressive neural injury via centrifugation. Spheroids were made from neonatal rat cortical cells seeded at 4000 cells/spheroid and cultured for 14 days in vitro. A subset of spheroids was centrifuged at 104, 209, 313 or 419 rads/s for 2 minutes. Modeling the physical deformation of the spheroids via finite element analyses, we found that spheroids centrifuged at the aforementioned angular velocities experienced pressures of 10, 38, 84 and 149 kPa, respectively, and compressive (resp. tensile) strains of 10% (5%), 18% (9%), 27% (14%) and 35% (18%), respectively. Quantification of LIVE-DEAD assay and Hoechst 33342 nuclear staining showed that centrifuged spheroids subjected to pressures above 10 kPa exhibited significantly higher DNA damage than control spheroids at 2, 8, and 24 hours post-injury. Immunohistochemistry of β3-tubulin networks at 2, 8, and 24 hours post-centrifugation injury showed increasing degradation of microtubules over time with increasing strain. Our findings show that cellular injuries occur as a result of specific levels and timings of sustained tissue strains. This experimental SCI model provides a high throughput in vitro platform to examine cellular injury, to gain insights into brain injury that could be targeted with therapeutic strategies.Copyright: © 2024 González-Cruz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.