Department of Pharmacology and Molecular Therapeutics, Uniformed Services University, Bethesda, Maryland, USA.
Department of Biomedical Engineering, Tufts University, Medford, Massachusetts, USA.
J Neurosci Res. 2023 Jan;101(1):3-19. doi: 10.1002/jnr.25123. Epub 2022 Oct 6.
Primary blast injury is caused by the direct impact of an overpressurization wave on the body. Due to limitations of current models, we have developed a novel approach to study primary blast-induced traumatic brain injury. Specifically, we employ a bioengineered 3D brain-like human tissue culture system composed of collagen-infused silk protein donut-like hydrogels embedded with human IPSC-derived neurons, human astrocytes, and a human microglial cell line. We have utilized this system within an advanced blast simulator (ABS) to expose the 3D brain cultures to a blast wave that can be precisely controlled. These 3D cultures are enclosed in a 3D-printed surrogate skull-like material containing media which are then placed in a holder apparatus inside the ABS. This allows for exposure to the blast wave alone without any secondary injury occurring. We show that blast induces an increase in lactate dehydrogenase activity and glutamate release from the cultures, indicating cellular injury. Additionally, we observe a significant increase in axonal varicosities after blast. These varicosities can be stained with antibodies recognizing amyloid precursor protein. The presence of amyloid precursor protein deposits may indicate a blast-induced axonal transport deficit. After blast injury, we find a transient release of the known TBI biomarkers, UCHL1 and NF-H at 6 h and a delayed increase in S100B at 24 and 48 h. This in vitro model will enable us to gain a better understanding of clinically relevant pathological changes that occur following primary blast and can also be utilized for discovery and characterization of biomarkers.
原发性爆震伤是由超压波对身体的直接冲击引起的。由于当前模型的局限性，我们开发了一种新的方法来研究原发性爆震性颅脑损伤。具体来说，我们采用了一种由胶原蛋白注入丝蛋白甜甜圈状水凝胶组成的生物工程 3D 类脑组织培养系统，其中嵌入了人类诱导多能干细胞衍生的神经元、人类星形胶质细胞和人类小胶质细胞系。我们已经在先进的爆震模拟器（ABS）中使用该系统将 3D 脑培养物暴露于可精确控制的爆震波下。这些 3D 培养物被封闭在含有介质的 3D 打印替代颅骨状材料中，然后放置在 ABS 内的Holder 装置中。这使得可以单独暴露于爆震波下，而不会发生任何二次损伤。我们发现，爆震会引起培养物中乳酸脱氢酶活性和谷氨酸释放的增加，表明细胞损伤。此外，我们还观察到爆震后轴突的分支明显增加。这些分支可以用识别淀粉样前体蛋白的抗体染色。淀粉样前体蛋白沉积物的存在可能表明爆震诱导的轴突运输缺陷。在爆震损伤后，我们发现已知的 TBI 生物标志物 UCHL1 和 NF-H 在 6 小时时短暂释放，而 S100B 在 24 小时和 48 小时时延迟增加。这种体外模型将使我们能够更好地理解原发性爆震后发生的与临床相关的病理变化，也可以用于发现和表征生物标志物。
