@article{SHAO2025, 
author = {Shuxian SHAO and Yanmei WANG and Yihan XU and Jiaxin ZHENG and Yufan ZHANG and Danning LIU and Yuan LI},
title = {Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein},
year = {2025},
journal = {Journal of Army Medical University},
volume = {47},
number = {11},
pages = {1199-1207},
keywords = {microfluidics, thrombosis, capillaries, retinal disease},
url = {https://www.sciopen.com/article/10.16016/j.2097-0927.202503005},
doi = {10.16016/j.2097-0927.202503005},
abstract = {ObjectiveTo develop an endothelialized microfluidic chip model that simulates the spatial architecture and bioactivity of retinal vasculature, enabling thrombosis modeling and thrombolytic efficacy validation.MethodsA tri-level microvascular network chip (300/200/100 μm diameters) with bifurcated architecture was fabricated using soft lithography. Human retinal microvascular endothelial cells (HRMECs) were perfused into channels, with endothelial coverage monitored via phase-contrast microscopy and F-actin staining. Cellular bioactivity was assessed using mitochondrial membrane potential probes (5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide, JC-1) and nitric oxide (NO) quantification. Fresh blood samples from 10 healthy donors (Yongchuan Hospital Affiliated to Chongqing Medical University, March to June 2024) were perfused with digital injection pump to mimic blood flow in human body into 3 experimental groups: normal whole blood, and TNF-α-activated endothelium+normal blood, TNF-α-activated endothelium+TNF-α-treated blood. Three inlet blood flow rates of 37.8、11.1 and 3.5 μL/min were set in each group. Two experimental groups, normal saline and recombinant human tissue-type plasminogen activator (rtPA), were established using the endothelialized microfluidic thrombosis model to validate thrombolytic efficacy. Endothelial functional impacts were assessed through integrated DAPI/NO staining and thrombosis model analysis across 3 intervention phases: pre-thrombosis, post-thrombosis, and post-thrombolysis.ResultsA tri-level microfluidic vascular model (300/200/100 μm diameters) was successfully constructed. In 72 h after endothelial cell perfusion, complete channel coverage was achieved, with phase-contrast microscopy and F-actin staining confirming confluent cellular alignment. JC-1/NO assays validated preserved endothelial bioactivity. Compared with the whole blood group, both TNF-α-activated endothelium+normal blood and TNF-α-activated endothelium+TNF-α-treated blood groups exhibited significantly increased thrombus occupancy rates at identical flow rates (all P<0.001). Notably, TNF-α-activated endothelium+TNF-α-treated blood group demonstrated the highest thrombus ratio at 3.5 μL/min (P<0.001). The rtPA group showed superior thrombolytic efficacy versus saline (P<0.001). Endothelial monolayer integrity was maintained across intervention phases, with thrombosis triggering significant NO elevation (P<0.001).ConclusionOur retinal vasculature-mimetic microfluidic model enables precise thrombosis modeling and drug evaluation, providing new methodology for studying retinal vascular occlusive diseases.}
}