Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein
Objective To develop an endothelialized microfluidic chip model that simulates the spatial architecture and bioactivity of retinal vasculature, enabling thrombosis modeling and thrombolytic efficacy validation. Methods A tri-level microvascular network chip (300/200/100 μm diameters) with bifurc...
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Editorial Office of Journal of Army Medical University
2025-06-01
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| Series: | 陆军军医大学学报 |
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| author | SHAO Shuxian WANG Yanmei XU Yihan |
| author_facet | SHAO Shuxian WANG Yanmei XU Yihan |
| author_sort | SHAO Shuxian |
| collection | DOAJ |
| description | Objective To develop an endothelialized microfluidic chip model that simulates the spatial architecture and bioactivity of retinal vasculature, enabling thrombosis modeling and thrombolytic efficacy validation. Methods A 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. Results A 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). Conclusion Our retinal vasculature-mimetic microfluidic model enables precise thrombosis modeling and drug evaluation, providing new methodology for studying retinal vascular occlusive diseases.
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| format | Article |
| id | doaj-art-b1f8126c0114474fb06daf7cf6a13d4a |
| institution | DOAJ |
| issn | 2097-0927 |
| language | zho |
| publishDate | 2025-06-01 |
| publisher | Editorial Office of Journal of Army Medical University |
| record_format | Article |
| series | 陆军军医大学学报 |
| spelling | doaj-art-b1f8126c0114474fb06daf7cf6a13d4a2025-08-20T03:10:07ZzhoEditorial Office of Journal of Army Medical University陆军军医大学学报2097-09272025-06-0147111199120710.16016/j.2097-0927.202503005Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis thereinSHAO Shuxian0WANG Yanmei 1XU Yihan2 Department of Ophthalmology， the Second Affiliated Hospital of Chongqing Medical University， Chongqing Department of Ophthalmology， the Second Affiliated Hospital of Chongqing Medical University， ChongqingDepartment of Ophthalmology， the Second Affiliated Hospital of Chongqing Medical University， ChongqingObjective To develop an endothelialized microfluidic chip model that simulates the spatial architecture and bioactivity of retinal vasculature, enabling thrombosis modeling and thrombolytic efficacy validation. Methods A 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. Results A 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). Conclusion Our retinal vasculature-mimetic microfluidic model enables precise thrombosis modeling and drug evaluation, providing new methodology for studying retinal vascular occlusive diseases. https://aammt.tmmu.edu.cn/html/202503005.htmlmicrofluidicsthrombosiscapillariesretinal disease |
| spellingShingle | SHAO Shuxian WANG Yanmei XU Yihan Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein 陆军军医大学学报 microfluidics thrombosis capillaries retinal disease |
| title | Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein |
| title_full | Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein |
| title_fullStr | Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein |
| title_full_unstemmed | Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein |
| title_short | Development of a microfluidic chip-based in vitro model of retinal microvasculature and thrombosis therein |
| title_sort | development of a microfluidic chip based in vitro model of retinal microvasculature and thrombosis therein |
| topic | microfluidics thrombosis capillaries retinal disease |
| url | https://aammt.tmmu.edu.cn/html/202503005.html |
| work_keys_str_mv | AT shaoshuxian developmentofamicrofluidicchipbasedinvitromodelofretinalmicrovasculatureandthrombosistherein AT wangyanmei developmentofamicrofluidicchipbasedinvitromodelofretinalmicrovasculatureandthrombosistherein AT xuyihan developmentofamicrofluidicchipbasedinvitromodelofretinalmicrovasculatureandthrombosistherein |