MICORALIS (E.A. 7354), Faculty of Dental Surgery and Odontology, University Côte d'Azur, 06300 Nice, France.
IDUN Centre of Excellence, Department of Health Technology, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
Biosensors (Basel). 2024 Aug 20;14(8):401. doi: 10.3390/bios14080401.
Static well plates remain the gold standard to study viral infections in vitro, but they cannot accurately mimic dynamic viral infections as they occur in the human body. Therefore, we established a dynamic cell culture platform, based on centrifugal microfluidics, to study viral infections in perfusion. To do so, we used human primary periodontal dental ligament (PDL) cells and herpes simplex virus-1 (HSV-1) as a case study. By microscopy, we confirmed that the PDL cells efficiently attached and grew in the chip. Successful dynamic viral infection of perfused PDL cells was monitored using fluorescent imaging and RT-qPCR-based experiments. Remarkably, viral infection in flow resulted in a gradient of HSV-1-infected cells gradually decreasing from the cell culture chamber entrance towards its end. The perfusion of acyclovir in the chip prevented HSV-1 spreading, demonstrating the usefulness of such a platform for monitoring the effects of antiviral drugs. In addition, the innate antiviral response of PDL cells, measured by interferon gene expression, increased significantly over time in conventional static conditions compared to the perfusion model. These results provide evidence suggesting that dynamic viral infections differ from conventional static infections, which highlights the need for more physiologically relevant in vitro models to study viral infections.
静态微孔板仍然是研究病毒体外感染的金标准，但它们无法准确模拟人体中发生的动态病毒感染。因此，我们基于离心微流控技术建立了一个动态细胞培养平台，以在灌注条件下研究病毒感染。为此，我们使用人原发性牙周膜（PDL）细胞和单纯疱疹病毒-1（HSV-1）作为案例研究。通过显微镜观察，我们证实 PDL 细胞在芯片中有效附着和生长。通过荧光成像和基于 RT-qPCR 的实验成功监测了灌注 PDL 细胞的动态病毒感染。值得注意的是，在流动中发生的病毒感染导致 HSV-1 感染的细胞从细胞培养腔室入口逐渐向其末端呈梯度减少。芯片中阿昔洛韦的灌注阻止了 HSV-1 的传播，证明了这种平台在监测抗病毒药物效果方面的有用性。此外，与灌注模型相比，PDL 细胞的先天抗病毒反应（通过干扰素基因表达测量）在常规静态条件下随时间显著增加。这些结果表明，动态病毒感染与传统的静态感染不同，这突出表明需要更具生理相关性的体外模型来研究病毒感染。
