Publications

@article{nokey,

title = {Dimethyl Sulfoxide Acutely Induces Shear-Dependent Thrombosis in Flow Assays via Red Blood Cell-Dependent Mechanisms},

author = {Savindi De Zoysa Ramasundara; Allan Sun; Marc L. Ellis; Tiana Pelaia; Yunduo Charles Zhao; Mike Chia Lun Wu; Lining Arnold Ju},

doi = {https://doi.org/10.1016/j.bvth.2026.100149},

year  = {2026},

date = {2026-02-16},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{nokey,

title = {TMEM63 proteins act as mechanically activated cholesterol modulated lipid scramblases contributing to membrane mechano-resilience},

author = {Yiechang Lin; Zijing Zhou; Yaoyao Han; Delfine Cheng; Haoqing Wang; Lining Arnold Ju; Yixiao Zhang; Charles D. Cox; Ben Corry},

doi = {https://doi.org/10.1038/s41467-026-68919-x},

year  = {2026},

date = {2026-01-21},

urldate = {2026-01-21},

journal = {Nature Communications},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{nokey,

title = {An intelligent, compact wearable pressure-strain combo sensor system for continuous fetal movement monitoring},

author = {Lim Wei Yap; Arie Levin; Yiwen Jiang; Duong Nhu; Shu Gong; Ritesh Warty; David Vera Anaya; Qinhao Li; Yan Lu; Rui Gao; Xin Zhang; Talha Ilyas; Vinayak Smith; Allison Thomas; Aswandi Wibrianto; Yuxin Zhang; Jane Limas; Sharon A. McCracken; Jonathan M Morris; Ben W Mol; Euan M Wallace; Arnold Lining Ju; Zongyuan Ge; Faezeh Marzbanrad; Wenlong Cheng},

doi = {10.1126/sciadv.ady2661},

year  = {2025},

date = {2025-11-26},

urldate = {2025-11-26},

journal = {Science Advances},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{nokey,

title = {Rapid Glass‐Substrate Digital Light 3D Printing Enables Anatomically Accurate Stroke Patient‐Specific Carotid Artery‐on‐Chips for Personalized Thrombosis Investigation},

author = {Yunduo Charles Zhao; Zihao Wang; Arian Nasser; Allan Sun; Zhao Wang; Yingqi Zhang; Jianfang Ren; Haimei Zhao; Nicole Alexis Yap; Yinyan Wang; Zhiyong Li; Ken S Butcher; Freda Passam; Timothy Ang; Lining Arnold Ju},

doi = {https://doi.org/10.1002/adma.202508890},

year  = {2025},

date = {2025-11-11},

urldate = {2025-11-11},

journal = {Advanced Materials},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{nokey,

title = {Neutrophil Membrane Tether Coalescence: A Novel Mechanoadaptive Response to Abrupt Flow Acceleration},

author = {Laura Moldovan; Allan Sun; Tao Huang; Yao Wang; Haoqing Jerry Wang; Haoran Caroline Song; Qian Peter Su; Lining Arnold Ju},

doi = {https://doi.org/10.1002/anbr.202500113},

year  = {2025},

date = {2025-11-04},

urldate = {2025-11-04},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{nokey,

title = {Endo-chip laser-induced thrombus formation: a vessel-on-chip model for in vitro testing of antithrombotic agents},

author = {Alexander Dupuy; Miao Qi; Jemma CL Fenwick; Daisie M Yates; Paul R Coleman; Jennifer R Gamble; Lining Arnold Ju; Freda H Passam},

doi = {https://doi.org/10.1016/j.bvth.2025.100096},

year  = {2025},

date = {2025-11-01},

urldate = {2025-11-01},

journal = {Blood Vessels, Thrombosis & Hemostasis},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{nokey,

title = {The evolution of nanomedicine: The rise of next-generation nanomaterials in cancer nanomedicine},

author = {Helen Forgham; Yixin Chang; Yao Wang; Jiayuan Zhu; Liwei Liu; Heather Biggs; Aleksandr Kakinen; Yuhao Jiang; Xinru You; Kristofer J Thurecht; Shaohua Ma; Lining Arnold Ju; Wei Tao; Thomas P Davis; Joyce Y Wong; Ruirui Qiao},

doi = {10.1126/sciadv.adx1576},

year  = {2025},

date = {2025-10-22},

urldate = {2025-10-22},

journal = {Science Advances},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Chen2025,

title = {Shear-dependent platelet aggregation by ChAdOx1 nCoV-19 vaccine: a novel biophysical mechanism for arterial thrombosis},

author = {Yiyao Catherine Chen and Naveen Eugene Louis Richard Louis and Angela Huang and Allan Sun and Alexander Dupuy and Laura Moldovan and Tiana Pelaia and Jianfang Ren and Taylor S. Cohen and Sarah C. Gilbert and Huyen Tran and Karlheinz Peter and James D. McFadyen and Lining Arnold Ju},

doi = {10.1182/blood.2024027675},

issn = {1528-0020},

year  = {2025},

date = {2025-07-24},

volume = {146},

number = {4},

pages = {496--503},

publisher = {American Society of Hematology},

abstract = {

                  Abstract

                  Rare thrombotic events associated with ChAdOx1 nCoV-19 (ChAdOx1) vaccination have raised concerns; however, the underlying mechanisms remain elusive. Here, we report a novel biophysical mechanism by which ChAdOx1 directly interacts with platelets under arterial shear conditions, potentially contributing to postvaccination arterial thrombosis. Using microfluidic post assays, we demonstrate that ChAdOx1 induces shear-dependent platelet aggregation, distinct from conventional von Willebrand factor–mediated adhesion. This interaction is mediated by platelet integrin αIIbβ3 and requires biomechanical activation, explaining the absence of significant binding under static conditions. Molecular dynamics simulations and docking studies reveal preferential binding of ChAdOx1's penton arginine-glycine-aspartic acid (RGD) motif to the activated conformation of αIIbβ3. Inhibiting integrin αIIbβ3 completely abolishes ChAdOx1-induced platelet aggregation, whereas blocking glycoprotein (GP) Ib has minimal effect, confirming a mechanism that bypasses the conventional GPIb-dependent platelet adhesion pathway. Mutagenesis of the RGD motif to AAA eliminates platelet binding, verifying the specificity of this interaction. These findings provide a potential explanation for the association between ChAdOx1 vaccination and arterial thrombotic events, distinct from vaccine-induced immune thrombotic thrombocytopenia. Our results highlight the importance of considering biomechanical factors in vaccine-related thrombotic complications and suggest that shear-dependent integrin activation may be another determinant in the pathogenesis of these rare adverse events.

               },

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Wang2025,

title = {Wet chemically produced nanomaterials for soft wearable biosensors},

author = {Ren Wang and Guangzhao Mao and Dewei Chu and Noushin Nasiri and Yuling Wang and Marcela Bilek and Ken-Tye Yong and Wallace Wong and Stan Skafidas and Jefferson Zhe Liu and Yuri Kivshar and Madhu Bhaskaran and Yuerui Lu and Benjamin Eggleton and Arnold Ju and Qianqian Shi and Nam-Trung Nguyen and Chwee Teck Lim and Wenlong Cheng},

doi = {10.1039/d5nh00048c},

issn = {2055-6764},

year  = {2025},

date = {2025-07-21},

journal = {Nanoscale Horiz.},

volume = {10},

number = {8},

pages = {1517--1541},

publisher = {Royal Society of Chemistry (RSC)},

abstract = {This review covers wet chemically synthesized nanomaterials for soft wearable biosensors, covering materials, inks, fabrication and sensing applications, and concludes with challenges and opportunities in personalized healthcare.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Jiang2025,

title = {Intravasation‐On‐µDevice (INVADE): Engineering Dynamic Vascular Interfaces to Study Cancer Cell Intravasation},

author = {Fengtao Jiang and Yingqi Zhang and Guocheng Fang and Yao Wang and Alexander Dupuy and Jasmine Jin and Yi Shen and Khoon S Lim and Yinyan Wang and Yu Shrike Zhang and Ann‐Na Cho and Hongxu Lu and Lining Arnold Ju},

doi = {10.1002/adma.202501466},

issn = {1521-4095},

year  = {2025},

date = {2025-07-00},

journal = {Advanced Materials},

volume = {37},

number = {26},

publisher = {Wiley},

abstract = {AbstractCancer metastasis begins with intravasation, where cancer cells enter blood vessels through complex interactions with the endothelial barrier. Understanding this process remains challenging due to the lack of physiologically relevant models. Here, INVADE (Intravasation‐on‐µDevice), a biomimetic microfluidic platform, is presented, enabling high‐throughput analysis of cancer cell intravasation under controlled conditions. This engineered platform integrates 23 parallel niche chambers with an endothelialized channel, providing both precise microenvironmental control and optical accessibility for real‐time visualization. Using this platform, distinct intravasation mechanisms are uncovered: MCF‐7 cells exhibit collective invasion, while MDA‐MB‐231 cells demonstrate an interactive mode with three functionally distinct subpopulations. A previously unknown epithelial‐mesenchymal transition (EMT) and mesenchymal‐epithelial transition (MET) switch is We discovered during intravasation, where MDA‐MB‐231 cells initially increase Vimentin expression before undergoing a 2.3 fold decrease over 96 h alongside a 1.5 fold increase in epithelial cell adhesion molecule (EpCAM). Remarkably, endothelial cells directly suppress cancer cell mesenchymal properties, as evidenced by a 4.6 fold reduction in Vimentin expression compared to mono‐cultures. Additionally, bilateral cancer‐endothelial interactions are revealed, aggressive cancer cells induce significant intercellular adhesion molecule‐1 (ICAM‐1) upregulation in endothelium. The INVADE platform represents an engineering advancement for studying complex cell–cell interactions with implications for understanding metastatic mechanisms.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Li2025,

title = {Innovative breakthroughs in novel biomaterials for traumatic brain injury and cranial repair},

author = {Yuan-Min Li and Yang Zhang and Yi-Xin Fu and Yan-Rong Lu and Kai Jiang and Lan Li and Quan Liu and Alexander Dupuy and Lining Arnold Ju and Yin-Yan Wang and Jin-Wei Li},

doi = {10.1007/s12598-025-03333-8},

issn = {1867-7185},

year  = {2025},

date = {2025-07-00},

journal = {Rare Met.},

volume = {44},

number = {7},

pages = {4315--4345},

publisher = {Springer Science and Business Media LLC},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Dupuy2025b,

title = {Endo-chip laser-induced thrombus formation; a vessel-on-chip model for in vitro testing of antithrombotic agents},

author = {Alexander Dupuy and Miao Qi and Jemma Fenwick and Daisie Yates and Paul Coleman and Jennifer Gamble and Lining Arnold Ju and Freda Passam},

doi = {10.1016/j.bvth.2025.100096},

issn = {2950-3272},

year  = {2025},

date = {2025-07-00},

journal = {Blood Vessels, Thrombosis & Hemostasis},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Lun(Mike)Wu2025,

title = {Complement C9-mediated RBC hemolysis drives microvascular obstruction via endothelial necroptosis and hemolyzed RBC aggregation in COVID-19},

author = {Chia Lun (Mike) Wu and Lining Arnold Ju and Ethan Italiano and Rocko Jarvis-Child and Imala Alwis and Rhyll Smythe and Eduardo A. Albornoz and Jonathan Noonan and Marie Portelli and Marrisa Baptista and Jessica Maclean and Pashtana Norri and Jinglu Yang and John Lee and James D. McFadyen and Alexandra Sharland and Trent M. Woodruff and Andre Samson and Amy Rapkiewicz and Tessa Barrett and Alan Pham and Simone Schoenwaelder and Yuping Yuan and Shaun P. Jackson},

doi = {10.1016/j.imbio.2025.153045},

issn = {0171-2985},

year  = {2025},

date = {2025-07-00},

journal = {Immunobiology},

volume = {230},

number = {4},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Dupuy2025,

title = {Endothelial cell activation enhances thromboinflammation in vaccine-induced immune thrombotic thrombocytopenia},

author = {Alexander Dupuy and Xiaoming Liu and Yvonne Kong and Miao Qi and Jose Perdomo and Jemma Fenwick and Jessica Tieng and Bede Johnston and Qiyu Sara Shi and Mark Larance and Yingqi Zhang and Lining Arnold Ju and Paul Coleman and Jennifer Gamble and Elizabeth E. Gardiner and Mortimer Poncz and Huyen Tran and Vivien Chen and Freda H. Passam},

doi = {10.1182/bloodadvances.2024014165},

issn = {2473-9537},

year  = {2025},

date = {2025-06-24},

volume = {9},

number = {12},

pages = {2891--2906},

publisher = {American Society of Hematology},

abstract = {

                  Abstract

                  Vaccine-induced immune thrombotic thrombocytopenia (VITT) is a rare but serious complication of the ChAdOx1 nCOV-19 vaccine. In Australia, the diagnosis of VITT required the detection of antibodies against platelet factor 4 (PF4) in plasma using a PF4/polyanion enzyme-linked immunosorbent assay (ELISA). Half of the patients who fulfilled the clinical criteria for VITT tested positive when using this ELISA and another third tested positive when using platelet activation assays, highlighting limitations in the assays used for VITT. Using a microfluidic device coated with endothelial cells, the Endo-chip, we measured the effects of serum and immunoglobulin G (IgG) from patients with clinical VITT on endothelial thromboinflammation. Our cohort comprised 40 patients (21 ELISA-positive and 19 ELISA-negative patients as measured by PF4/polyanion ELISA), 12 vaccinated patients with venous thromboembolism without VITT, and 17 individuals who received the ChAdOx1 vaccine without adverse events (vax controls). Treatment with VITT serum, plasma, or IgG increased endothelial tissue factor (TF) expression and activity. Perfusion of blood from healthy donors labelled with fluorescent antibodies against platelets, neutrophils, and fibrin through Endo-chips treated with VITT serum or IgG induced a twofold to threefold increase in platelet, neutrophil, and fibrin deposition. Thromboinflammation was enhanced with addition of PF4 and reduced with an inhibitory antibody against TF. We conclude that endothelial activation contributes to thromboinflammation in patients with clinical features of VITT. The Endo-chip offers a platform for the study of endothelial responses in immune thrombosis.

               },

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Huang2025,

title = {Synthesizing vascular trees at speed},

author = {Yan Yan Shery Huang and Lining Arnold Ju},

doi = {10.1126/science.ady6122},

issn = {1095-9203},

year  = {2025},

date = {2025-06-12},

journal = {Science},

volume = {388},

number = {6752},

pages = {1133--1134},

publisher = {American Association for the Advancement of Science (AAAS)},

abstract = {A computational algorithm can render a complex artificial vascular structure in minutes},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Ren2025,

title = {Vorticity‐Facilitated Platelet Aggregation: a High Expansion‐Ratio Stenotic Microfluidic Platform Unravels the Role of Complex Flow Dynamics in Arterial Thrombosis},

author = {Jianfang Ren and Nurul Aisha Zainal Abidin and Allan Sun and Rui Gao and Yuxin Chen and Arian Nasser and Zihao Wang and Yunduo Charles Zhao and Alexander Dupuy and Anna Waterhouse and Qian Peter Su and Daniele Vigolo and Mike Chia Lun Wu and Lining Arnold Ju},

doi = {10.1002/adhm.202500436},

issn = {2192-2659},

year  = {2025},

date = {2025-06-04},

journal = {Adv Healthcare Materials},

publisher = {Wiley},

abstract = {AbstractComplex flow patterns play a critical role in arterial thrombosis, yet the specific contribution of vorticity—the rotational component of fluid flow—remains poorly understood. An innovative microfluidic platform with systematically varied expansion angles (β = 30°‐150°) in a double stenosis design is developed to isolate vorticity's effects under controlled conditions. The high expansion‐ratio device with sharp‐angled geometries successfully generates distinct vortical flow patterns, confirmed through computational and experimental flow visualizations. Real‐time confocal microscopy revealed a strong positive correlation (r = 0.6698) between vorticity magnitude and thrombus size, with high‐vorticity conditions producing thrombi up to four times larger than low‐vorticity settings. Mechanistic investigations demonstrated enhanced von Willebrand Factor (vWF) accumulation and platelet integrin activation in vortical environments. Platelets in high‐vorticity regions exhibited integrin αIIbβ3 intermediate activation states with significantly enhanced calcium signaling, suggesting vorticity amplifies platelet mechanosensing pathways. Inhibition of the interaction between vWF and platelet glycoprotein Ibα (GPIbα) receptor abolished biomechanical platelet aggregation in vortical regions. These findings provide valuable insights into platelet thrombosis in complex flow environments with significant implications for optimizing medical devices to minimize thrombotic complications associated with vortex formation.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Goh2025,

title = {Thrombotic response to mechanical circulatory support devices},

author = {Tiffany Goh and Lining Arnold Ju and Anna Waterhouse},

doi = {10.1016/j.jtha.2025.02.037},

issn = {1538-7836},

year  = {2025},

date = {2025-06-00},

journal = {Journal of Thrombosis and Haemostasis},

volume = {23},

number = {6},

pages = {1743--1757},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Zhao2025,

title = {Sensing the Future of Thrombosis Management: Integrating Vessel-on-a-Chip Models, Advanced Biosensors, and AI-Driven Digital Twins},

author = {Yunduo Charles Zhao and Zihao Wang and Haimei Zhao and Nicole Alexis Yap and Ren Wang and Wenlong Cheng and Xin Xu and Lining Arnold Ju},

doi = {10.1021/acssensors.4c02764},

issn = {2379-3694},

year  = {2025},

date = {2025-03-28},

journal = {ACS Sens.},

volume = {10},

number = {3},

pages = {1507--1520},

publisher = {American Chemical Society (ACS)},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Sun2025,

title = {3M engineering approaches to combat high-shear thrombosis: Integrating modeling, microfluidics, and mechanobiology},

author = {Allan Sun and Arian Nasser and Nicole Alexis Yap and Rui Gao and Lining Arnold Ju},

doi = {10.1016/j.cobme.2025.100576},

issn = {2468-4511},

year  = {2025},

date = {2025-03-00},

journal = {Current Opinion in Biomedical Engineering},

volume = {33},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Aye2025,

title = {Integrating microfluidics, hydrogels, and 3D bioprinting for personalized vessel-on-a-chip platforms},

author = {San Seint Seint Aye and Zhongqi Fang and Mike C. L. Wu and Khoon S. Lim and Lining Arnold Ju},

doi = {10.1039/d4bm01354a},

issn = {2047-4849},

year  = {2025},

date = {2025-02-25},

journal = {Biomater. Sci.},

volume = {13},

number = {5},

pages = {1131--1160},

publisher = {Royal Society of Chemistry (RSC)},

abstract = {Advancement of vascular models from simple 2D culture to complex vessel-on-a-chip platforms through integration of microfluidics, biomimetic hydrogels, and 3D bioprinting, enabling controlled investigation of thrombosis mechanisms.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Major2025,

title = {Photochemistry as a tool for dynamic modulation of hydrogel mechanics},

author = {Gretel S. Major and Habib Joukhdar and Yu Suk Choi and Jelena Rnjak-Kovacina and Steven G. Wise and Lining Arnold Ju and Thomas R. Cox and Chun Xu and Giselle C. Yeo and Jennifer L. Young and Khoon S. Lim},

doi = {10.1016/j.xcrp.2024.102366},

issn = {2666-3864},

year  = {2025},

date = {2025-01-00},

journal = {Cell Reports Physical Science},

volume = {6},

number = {1},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Lv2024,

title = {Protein disulfide isomerase cleaves allosteric disulfides in histidine-rich glycoprotein to regulate thrombosis},

author = {Keyu Lv and Shuai Chen and Xulin Xu and Joyce Chiu and Haoqing J. Wang and Yunyun Han and Xiaodan Yang and Sheryl R. Bowley and Hao Wang and Zhaoming Tang and Ning Tang and Aizhen Yang and Shuofei Yang and Jinyu Wang and Si Jin and Yi Wu and Alvin H. Schmaier and Lining A. Ju and Philip J. Hogg and Chao Fang},

doi = {10.1038/s41467-024-47493-0},

issn = {2041-1723},

year  = {2024},

date = {2024-12-00},

journal = {Nat Commun},

volume = {15},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {AbstractThe essence of difference between hemostasis and thrombosis is that the clotting reaction is a highly fine-tuned process. Vascular protein disulfide isomerase (PDI) represents a critical mechanism regulating the functions of hemostatic proteins. Herein we show that histidine-rich glycoprotein (HRG) is a substrate of PDI. Reduction of HRG by PDI enhances the procoagulant and anticoagulant activities of HRG by neutralization of endothelial heparan sulfate (HS) and inhibition of factor XII (FXIIa) activity, respectively. Murine HRG deficiency (Hrg−/−) leads to delayed onset but enhanced formation of thrombus compared to WT. However, in the combined FXII deficiency (F12−/−) and HRG deficiency (by siRNA or Hrg−/−), there is further thrombosis reduction compared to F12−/− alone, confirming HRG’s procoagulant activity independent of FXIIa. Mutation of target disulfides of PDI leads to a gain-of-function mutant of HRG that promotes its activities during coagulation. Thus, PDI-HRG pathway fine-tunes thrombosis by promoting its rapid initiation via neutralization of HS and preventing excessive propagation via inhibition of FXIIa.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Liu2024,

title = {Nanoscale insights into hematology: super-resolved imaging on blood cell structure, function, and pathology},

author = {Jinghan Liu and Yuping Yolanda Tan and Wen Zheng and Yao Wang and Lining Arnold Ju and Qian Peter Su},

doi = {10.1186/s12951-024-02605-2},

issn = {1477-3155},

year  = {2024},

date = {2024-12-00},

journal = {J Nanobiotechnol},

volume = {22},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {AbstractFluorescence nanoscopy, also known as super-resolution microscopy, has transcended the conventional resolution barriers and enabled visualization of biological samples at nanometric resolutions. A series of super-resolution techniques have been developed and applied to investigate the molecular distribution, organization, and interactions in blood cells, as well as the underlying mechanisms of blood-cell-associated diseases. In this review, we provide an overview of various fluorescence nanoscopy technologies, outlining their current development stage and the challenges they are facing in terms of functionality and practicality. We specifically explore how these innovations have propelled forward the analysis of thrombocytes (platelets), erythrocytes (red blood cells) and leukocytes (white blood cells), shedding light on the nanoscale arrangement of subcellular components and molecular interactions. We spotlight novel biomarkers uncovered by fluorescence nanoscopy for disease diagnosis, such as thrombocytopathies, malignancies, and infectious diseases. Furthermore, we discuss the technological hurdles and chart out prospective avenues for future research directions. This review aims to underscore the significant contributions of fluorescence nanoscopy to the field of blood cell analysis and disease diagnosis, poised to revolutionize our approach to exploring, understanding, and managing disease at the molecular level.

                Graphical Abstract},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Wang2024,

title = {Microscale geometrical modulation of PIEZO1 mediated mechanosensing through cytoskeletal redistribution},

author = {Haoqing Jerry Wang and Yao Wang and Seyed Sajad Mirjavadi and Tomas Andersen and Laura Moldovan and Parham Vatankhah and Blake Russell and Jasmine Jin and Zijing Zhou and Qing Li and Charles D. Cox and Qian Peter Su and Lining Arnold Ju},

doi = {10.1038/s41467-024-49833-6},

issn = {2041-1723},

year  = {2024},

date = {2024-12-00},

journal = {Nat Commun},

volume = {15},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {AbstractThe microgeometry of the cellular microenvironment profoundly impacts cellular behaviors, yet the link between it and the ubiquitously expressed mechanosensitive ion channel PIEZO1 remains unclear. Herein, we describe a fluorescent micropipette aspiration assay that allows for simultaneous visualization of intracellular calcium dynamics and cytoskeletal architecture in real-time, under varied micropipette geometries. By integrating elastic shell finite element analysis with fluorescent lifetime imaging microscopy and employing PIEZO1-specific transgenic red blood cells and HEK cell lines, we demonstrate a direct correlation between the microscale geometry of aspiration and PIEZO1-mediated calcium signaling. We reveal that increased micropipette tip angles and physical constrictions lead to a significant reorganization of F-actin, accumulation at the aspirated cell neck, and subsequently amplify the tension stress at the dome of the cell to induce more PIEZO1’s activity. Disruption of the F-actin network or inhibition of its mobility leads to a notable decline in PIEZO1 mediated calcium influx, underscoring its critical role in cellular mechanosensing amidst geometrical constraints.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Sun2024,

title = {Strategic reuse of rapid antigen tests for coagulation status assessment: an integrated machine learning approach},

author = {Allan Sun and Arian Nasser and Chaohao Chen and Yunduo Charles Zhao and Haimei Zhao and Zihao Wang and Wenlong Cheng and Pierre Qian and Lining Arnold Ju},

doi = {10.1007/s44258-024-00025-3},

issn = {2731-8710},

year  = {2024},

date = {2024-12-00},

journal = {Med-X},

volume = {2},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {AbstractAddressing the pressing demand for rapid and inexpensive coagulation testing in cardiovascular care, this study introduces a novel application of repurposed COVID-19 rapid antigen tests (RATs) as paper-based lateral flow assays (LFAs) combined with machine learning for coagulation status evaluation. By further developing a mobile app prototype, we present a platform that enables clinicians to perform immediate and accurate anticoagulant dosing adjustments using existing post-pandemic resources. Our proof-of-concept employs a random forest machine learning classifier to interpret image feature variations on RAT NC membrane, correlating red blood cell (RBC) wicked diffusion distance in recalcified citrated whole blood with changes in coagulative viscosity, easily interpreted. Enhanced by confocal imaging studies of paper microfluidics, our approach provides insights into the mechanisms dissecting coagulation components, achieving high classification precision, recall, and F1-scores. The inverse relationship between RBC wicked diffusion distance and enoxaparin concentration paves the way for machine learning to inform real-time dose prescription adjustments, aligning with individual patient profiles to optimize therapeutic outcomes. This study not only demonstrates the potential of leveraging surplus RATs for coagulation management but also exemplifies a cost-effective, rapid, and smart strategy to enhance clinical decision-making in the post-pandemic era.

                Graphical Abstract},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Din2024b,

title = {Multi-parametric thrombus profiling microfluidics detects intensified biomechanical thrombogenesis associated with hypertension and aging},

author = {Misbahud Din and Souvik Paul and Sana Ullah and Haoyi Yang and Rong-Guang Xu and Nurul Aisha Zainal Abidin and Allan Sun and Yiyao Catherine Chen and Rui Gao and Bari Chowdhury and Fangyuan Zhou and Stephenie Rogers and Mariel Miller and Atreyee Biswas and Liang Hu and Zhichao Fan and Christopher Zahner and Jing Fan and Zi Chen and Megan Berman and Lingzhou Xue and Lining Arnold Ju and Yunfeng Chen},

doi = {10.1038/s41467-024-53069-9},

issn = {2041-1723},

year  = {2024},

date = {2024-12-00},

journal = {Nat Commun},

volume = {15},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {AbstractArterial thrombosis is a leading cause of death and disability worldwide with no effective bioassay for clinical prediction. As a symbolic feature of arterial thrombosis, severe stenosis in the blood vessel creates a high-shear, high-gradient flow environment that facilitates platelet aggregation towards vessel occlusion. Here, we present a thrombus profiling assay that monitors the multi-dimensional attributes of thrombi forming in such biomechanical conditions. Using this assay, we demonstrate that different receptor–ligand interactions contribute distinctively to the composition and activation status of the thrombus. Our investigation into hypertensive and older individuals reveals intensified biomechanical thrombogenesis and multi-dimensional thrombus profile abnormalities, endorsing the diagnostic potential of the assay. Furthermore, we identify the hyperactivity of GPIbα-integrin αIIbβ3 mechanosensing axis as a molecular mechanism that contributes to hypertension-associated arterial thrombosis. By studying drug-disease interactions and inter-individual variability, our work reveals a need for personalized anti-thrombotic drug selection that accommodates each patient’s pathological profile.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Liu2024b,

title = {Harnessing the power of bioprinting for the development of next-generation models of thrombosis},

author = {Yanyan Liu and Tao Huang and Nicole Alexis Yap and Khoon Lim and Lining Arnold Ju},

doi = {10.1016/j.bioactmat.2024.08.040},

issn = {2452-199X},

year  = {2024},

date = {2024-12-00},

journal = {Bioactive Materials},

volume = {42},

pages = {328--344},

publisher = {Elsevier BV},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Dupuy2024,

title = {Platelet Endoplasmic Reticulum Stress Promotes Thrombosis in an \textit{in Vitro} Laser-Induced Endothelial Injury Model},

author = {Alexander Dupuy and Paul Coleman and Jennifer Gamble and Lining Ju and Freda H Passam},

doi = {10.1182/blood-2024-212327},

issn = {1528-0020},

year  = {2024},

date = {2024-11-05},

volume = {144},

number = {Supplement 1},

pages = {1233--1233},

publisher = {American Society of Hematology},

abstract = {

                  

                  Aim: Platelet endoplasmic reticulum (ER) stress has recently been associated with platelet activation (Jain et al. Circ Res 2022). ER stress is triggered by calcium depletion from the ER which rapidly triggers downstream activation pathways. On the other hand, platelet ER stress activates the ER stress sensor, inositol-requiring enzyme 1 α (IRE1), which increases platelet secretion and reactivity (Lay et al. Blood Adv 2023). However, the role of IRE1 activation in platelet calcium mobilization is underexplored in the context of vascular thrombosis. Here, we utilized an endothelialized microfluidic platform that we have developed to study the relation of ER stress and platelet calcium mobilization in real time, under conditions closely mimicking the development of a vascular thrombus. This was achieved by perfusing whole blood across an endothelialized microfluidic device and inducing a site-specific endothelial injury using a high-powered laser and comparing with a standardized in vivo laser-induced venous thrombosis model. We further investigated the role of IRE1 using a chemical inhibitor of IRE1.

                  Method: Initially, we standardized the kinetics of platelet, fibrin and neutrophil accumulation in a laser-induced mesenteric vein thrombosis in 3-month-old C57/BL6 mice. This was used as a comparator for the laser-induced thrombus formation on our endothelialized device at shear rates of 100 s-1 and 800 s-1. The laser injury was performed using a straight channel microfluidic device fabricated from polydimethylsiloxane and endothelialised with human umbilical vein endothelial cells (HUVECs). Citrated (0.32% w/v) whole blood was collected from healthy volunteers and stained with anti-CD41-FITC (0.5 μg/mL), anti-fibrin-Alexa Fluor 594 (0.5 μg/mL) antibodies, and Annexin V-Alexa Fluor 647 (1:200) to visualize platelets, fibrin, and phosphatidylserine exposure respectively. For calcium mobilization, whole blood was stained with an anti-CD42a-PE (0.5 μg/mL) antibody and Cal520 (2 μM). To examine the effects of anticoagulants in our endothelialized device, whole blood was treated with heparin (0.5 U/mL) or thrombin inhibitor argatroban (0.5 μg/mL). We then compared with the effects of IRE1 activator IXA4 (10- 300 μM). IXA4 was incubated with whole blood for 2 hrs prior to perfusion following whole blood recalcification with CaCl2 (10 mM). A localized injury was then induced at the junction of endothelial cells by pulsing a 300 mW, 355nm laser at 10% power, 5 times over 2 sec, in a 1 um square area parfocal to the endothelial monolayer. Subsequent platelet adhesion, fibrin formation, and phosphatidylserine exposure was imaged over 15 min and quantified based on the surface area of fluorescence. Statistical analysis was by paired student t-test after confirming normality.

                  Results: Laser injury of the endothelial monolayer resulted in the Annexin V binding to endothelial cells directly adjacent to the injury site with no loss of endothelial cell junctions. Platelet adhesion and fibrin formation was restricted to Annexin V positive endothelial cells, forming a tear drop-shaped thrombus within 6 mins of injury. This closely followed the kinetics formation in vivo. Compared to vehicle, both argatroban and heparin inhibited platelet adhesion (argatroban: 2856 vs 226.0 um2; heparin: 2856 vs 56.70 um2, P<0.05) and fibrin formation (argatroban: 8421 vs 1176 um^2; heparin: 8421 vs 9.651 um2, P<0.05), without affecting Annexin V binding. IXA4 enhanced the accumulation of calcium in platelets after laser injury (0.07080 vs 0.1470 AU, P<0.05). Furthermore, it enhanced platelet adhesion (1709 vs 1957 um2; P<0.05) and fibrin formation (3045 vs 7716 um2; P< 0.05).

                  Conclusion: The kinetics of our in vitro, laser-induced vascular thrombosis model closely mirrors laser-induced thrombus formation in vivo, as measured by intravital microscopy. Thrombus formation in the endothelialized device was Factor Xa and/or thrombin dependent. We propose our model offers a robust assay for the study of calcium kinetics in platelets under conditions of vascular thrombosis. This model offers a feasible alternative to mouse models of thrombosis using laser injury for the study of calcium and novel therapeutic agents. Activation of IRE1 increased calcium mobilization in the platelets and thrombus formation. The above introduce the novel concept of ER stress-dependent calcium mobilization in vascular thrombosis.

               },

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Goh2024,

title = {Platelet Adhesion and Activation in an ECMO Thrombosis‐on‐a‐Chip Model},

author = {Tiffany Goh and Lingzi Gao and Jasneil Singh and Richard Totaro and Ruaidhri Carey and Kevin Yang and Bruce Cartwright and Mark Dennis and Lining Arnold Ju and Anna Waterhouse},

doi = {10.1002/advs.202401524},

issn = {2198-3844},

year  = {2024},

date = {2024-08-00},

journal = {Advanced Science},

volume = {11},

number = {30},

publisher = {Wiley},

abstract = {AbstractUse of extracorporeal membrane oxygenation (ECMO) for cardiorespiratory failure remains complicated by blood clot formation (thrombosis), triggered by biomaterial surfaces and flow conditions. Thrombosis may result in ECMO circuit changes, cause red blood cell hemolysis, and thromboembolic events. Medical device thrombosis is potentiated by the interplay between biomaterial properties, hemodynamic flow conditions and patient pathology, however, the contribution and importance of these factors are poorly understood because many in vitro models lack the capability to customize material and flow conditions to investigate thrombosis under clinically relevant medical device conditions. Therefore, an ECMO thrombosis‐on‐a‐chip model is developed that enables highly customizable biomaterial and flow combinations to evaluate ECMO thrombosis in real‐time with low blood volume. It is observed that low flow rates, decelerating conditions, and flow stasis significantly increased platelet adhesion, correlating with clinical thrombus formation. For the first time, it is found that tubing material, polyvinyl chloride, caused increased platelet P‐selectin activation compared to connector material, polycarbonate. This ECMO thrombosis‐on‐a‐chip model can be used to guide ECMO operation, inform medical device design, investigate embolism, occlusion and platelet activation mechanisms, and develop anti‐thrombotic biomaterials to ultimately reduce medical device thrombosis, anti‐thrombotic drug use and therefore bleeding complications, leading to safer blood‐contacting medical devices.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Ren2024,

title = {Charting the course of blood flow: vessel-on-a-chip technologies in thrombosis studies},

author = {Jianfang Ren and Zhao Wang and Nixon Du and Wenlong Cheng and Lining Arnold Ju},

doi = {10.20517/microstructures.2023.106},

issn = {2770-2995},

year  = {2024},

date = {2024-06-12},

journal = {Microstructures},

volume = {4},

number = {3},

publisher = {OAE Publishing Inc.},

abstract = {Cardiovascular diseases, primarily driven by thrombosis, remain the leading cause of global mortality. Although traditional cell culture and animal models have provided foundational insights, they often fail to capture the complex pathophysiology of thrombosis, which hinders the development of targeted therapies for cardiovascular diseases. The advent of microfluidics and vascular tissue engineering has propelled the advancement of vessel-on-a-chip technologies, which enable the simulation of the key aspects of Virchow’s Triad: hypercoagulability, alteration in blood flow, and endothelial wall injury. With the ability to replicate patient-specific vascular architectures and hemodynamic conditions, vessel-on-a-chip models offer unprecedented insights into the mechanisms underlying thrombosis formation and progression. This review explores the evolution of microfluidic technologies in thrombosis research, highlighting breakthroughs in endothelialized devices and their roles in emulating conditions such as vessel stenosis, flow reversal, and endothelial damage. The limitations and challenges of the current vessel-on-a-chip systems are addressed, and future perspectives on the potential for personalized medicine and targeted therapies are presented. Vessel-on-a-chip technology holds immense potential for revolutionizing thrombosis research, enabling the development of targeted, patient-specific diagnostic tools and therapeutic strategies. Realizing this potential will require interdisciplinary collaboration and continued innovation in the fields of microfluidics and vascular tissue engineering.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Din2024,

title = {Abstract 153: Multi-parametric Thrombus Profiling Microfluidics Uncovers Mechanobiology Of Hypertension And Aging-related Thrombosis},

author = {Misbahud Din and Souvik Paul and Sana Ullah and Haoyi Yang and Rongguang Xu and Nurul A Abidin and Bari Chowdhury and Fangyuan Zhou and Stephenie Rogers and Mariel Miller and Atreyee Biswas and Liang Hu and Zhichao Fan and Christopher Zahner and Megan Berman and Lingzhou Xue and Lining Ju and Yunfeng Chen},

doi = {10.1161/atvb.44.suppl_1.153},

issn = {1524-4636},

year  = {2024},

date = {2024-05-00},

journal = {ATVB},

volume = {44},

number = {Suppl_1},

publisher = {Ovid Technologies (Wolters Kluwer Health)},

abstract = {Arterial thrombosis, which represents a critical complication of cardiovascular diseases, is a leading cause of death and disability worldwide. A symbolic feature of arterial thrombosis is large-scale platelet aggregation due to high shear stress generated by stenosis. However, current hematological tests cannot evaluate arterial thrombosis in such physiological settings.

          

            To bridge this gap, we developed a microfluidics-based thrombus profiling assay (Fig. 1A) that combines multi-fluorescence imaging with a stenotic channel design to comprehensively characterize thrombi formed in high shear blood flow (Fig. 1B, C). Thrombi are characterized by a 7-dimensional profile indicating the thrombus size (Plt) and levels of fibrinogen (Fg), VWF, P-selectin, phosphatidylserine (PS) and extended (E

            +

            ) and activated α

            IIb

            β

            3

            in the thrombus (Fig. 1D).

          

          

            Using this assay, we discovered intensified thrombus formation in subjects with hypertension and/or aging (Fig. 1D). We also found extended integrin α

            IIb

            β

            3

            as a superior biomarker for platelet hyperreactivity than P-selectin and activated integrin α

            IIb

            β

            3

            and a potential predictor for arterial thrombosis (Fig. 1E). By studying the effects of anti-thrombotic agents NMC4 and 7E3 on hypertension patients’ blood, our work reveals a ‘treatment mismatch’ phenomenon that can seriously affect the efficacy and safety of anti-thrombotic drugs (Fig. 1F).

          

          Our findings provide mechanistic insights into the high risk of arterial thrombosis and antiplatelet resistance in populations with hypertension and aging. The discovered ‘treatment mismatch’ heralds a new era in thrombosis management where patients’ thrombus profiles dictate personalized therapeutic decisions to optimize treatment efficacy and reduce adverse outcomes. The outstanding performance of our assay underscores its translational potential for anti-thrombotic drug screening, thrombosis diagnosis, and personalized anti-thrombotic regimen selection.

          

            

          },

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Li2024,

title = {Endoplasmic Reticulum Protein 72 Regulates Integrin Mac-1 Activity to Influence Neutrophil Recruitment},

author = {Yaofeng Li and Xulin Xu and Haoqing Jerry Wang and Yiyao Catherine Chen and Yaobing Chen and Joyce Chiu and Li Li and Lei Wang and Jinyu Wang and Zhaoming Tang and Lehao Ren and Hongliang Li and Xuanbin Wang and Si Jin and Yi Wu and Mingdong Huang and Lining Arnold Ju and Chao Fang},

doi = {10.1161/atvbaha.123.319771},

issn = {1524-4636},

year  = {2024},

date = {2024-03-00},

journal = {ATVB},

volume = {44},

number = {3},

publisher = {Ovid Technologies (Wolters Kluwer Health)},

abstract = {

            BACKGROUND:

            Integrins mediate the adhesion, crawling, and migration of neutrophils during vascular inflammation. Thiol exchange is important in the regulation of integrin functions. ERp72 (endoplasmic reticulum–resident protein 72) is a member of the thiol isomerase family responsible for the catalysis of disulfide rearrangement. However, the role of ERp72 in the regulation of Mac-1 (integrin αMβ2) on neutrophils remains elusive.

          

          

            METHODS:

            Intravital microscopy of the cremaster microcirculation was performed to determine in vivo neutrophil movement. Static adhesion, flow chamber, and flow cytometry were used to evaluate in vitro integrin functions. Confocal fluorescent microscopy and coimmunoprecipitation were utilized to characterize the interactions between ERp72 and Mac-1 on neutrophil surface. Cell-impermeable probes and mass spectrometry were used to label reactive thiols and identify target disulfide bonds during redox exchange. Biomembrane force probe was performed to quantitatively measure the binding affinity of Mac-1. A murine model of acute lung injury induced by lipopolysaccharide was utilized to evaluate neutrophil-associated vasculopathy.

          

          

            RESULTS:

            ERp72-deficient neutrophils exhibited increased rolling but decreased adhesion/crawling on inflamed venules in vivo and defective static adhesion in vitro. The defect was due to defective activation of integrin Mac-1 but not LFA-1 (lymphocyte function-associated antigen-1) using blocking or epitope-specific antibodies. ERp72 interacted with Mac-1 in lipid rafts on neutrophil surface leading to the reduction of the C654-C711 disulfide bond in the αM subunit that is critical for Mac-1 activation. Recombinant ERp72, via its catalytic motifs, increased the binding affinity of Mac-1 with ICAM-1 (intercellular adhesion molecule-1) and rescued the defective adhesion of ERp72-deficient neutrophils both in vitro and in vivo. Deletion of ERp72 in the bone marrow inhibited neutrophil infiltration, ameliorated tissue damage, and increased survival during murine acute lung injury.

          

          

            CONCLUSIONS:

            Extracellular ERp72 regulates integrin Mac-1 activity by catalyzing disulfide rearrangement on the αM subunit and may be a novel target for the treatment of neutrophil-associated vasculopathy.

          },

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Chen2023,

title = {Force-Regulated Spontaneous Conformational Changes of Integrins α_{5}β_{1} and α_{V}β_{3}},

author = {Yunfeng Chen and Zhenhai Li and Fang Kong and Lining Arnold Ju and Cheng Zhu},

doi = {10.1021/acsnano.3c06253},

issn = {1936-086X},

year  = {2024},

date = {2024-01-09},

journal = {ACS Nano},

volume = {18},

number = {1},

pages = {299--313},

publisher = {American Chemical Society (ACS)},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Jin2024,

title = {Fluorescence-coupled Micropipette Aspiration Assay to Investigate Red Blood Cell Mechanosensing},

author = {Jasmine Jin and Haoqing Jerry Wang and Yiyao Catherine Chen and Blake Russell and Allan Sun and Yao Wang and Lining Arnold Ju},

doi = {10.3791/66265},

issn = {1940-087X},

year  = {2024},

date = {2024-00-00},

journal = {JoVE},

number = {203},

publisher = {MyJove Corporation},

keywords = {},

pubstate = {published},

tppubtype = {article}

}