Surface Functionalization and Plasma-Based Approaches in Microfluidic Models of Leukocyte Adhesion in Atherosclerosis

Inam, Shehla; Waqar, Muhammad; Ikram, Sehrish

doi:10.48309/ijabbr.2026.2064200.1623

Surface Functionalization and Plasma-Based Approaches in Microfluidic Models of Leukocyte Adhesion in Atherosclerosis

Document Type : Review Article

Authors

Shehla Inam ¹

Muhammad Waqar ²

Sehrish Ikram ¹

¹ Department of Biomedical Engineering, Faculty of Engineering and Applied Sciences, Riphah International University, Islamabad, Pakistan

² Department of Biomedical Engineering, Faculty of Engineering, Balochistan University of Engineering and Technology, Khuzdar, Balochistan, Pakistan

https://doi.org/10.48309/ijabbr.2026.2064200.1623

Abstract

Atherosclerosis is a chronic inflammatory disease characterized by endothelial dysfunction, with leukocyte adhesion primarily driven by interactions between P-selectin and PSGL-1, which play a crucial role. Capturing these rolling interactions under physiological shear stress is essential for understanding disease progression. However, traditional in vivo models and static assays often fail to replicate the dynamic blood flow conditions. Microfluidic platforms have become valuable tools, enabling real-time studies of leukocyte behaviour under precisely controlled flow environments. A major challenge remains achieving stable and functional protein immobilization on microchannel surfaces. This review thoroughly examines both covalent and non-covalent surface functionalization techniques designed to incorporate reactive chemistries that boost ligand retention, reduce detachment, and maintain biological activity. When combined with protein micropatterning, these methods allow spatial control over adhesion molecules, better mimicking the complex heterogeneity of endothelial surfaces. Despite their promise, current approaches still face issues with reproducibility, long-term stability, and application in complex biological systems, such as live-cell rolling assays or synthetic leukocyte models using nanoparticles. Nonetheless, plasma-assisted microfluidic platforms present an exciting, largely unexplored opportunity for high-fidelity modelling of cardiovascular inflammation and leukocyte–endothelial interactions.

Graphical Abstract

Keywords

Leukocyte Adhesion

Microfluidics

P-Selectin

PSGL-1

Endothelial Dysfunction

Plasma Surface Modification

Atherosclerosis

Subjects

Bioengineering

OPEN ACCESS

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[1]. Maringanti, R., Van Dijk, C.G., Meijer, E.M., Brandt, M.M., Li, M., Tiggeloven, V.P., Krebber, M.M., Chrifi, I., Duncker, D.J., Verhaar, M.C., Atherosclerosis on a chip: A 3-dimensional microfluidic model of early arterial events in human plaques. Arteriosclerosis, Thrombosis, and Vascular Biology, 2024, 44(12), 2453-2472.

[2]. Henein, M.Y., Vancheri, S., Longo, G., Vancheri, F., The role of inflammation in cardiovascular disease. International Journal of Molecular Sciences, 2022, 23(21), 12906.

[3]. Drożdż, D., Drożdż, M., Wójcik, M., Endothelial dysfunction as a factor leading to arterial hypertension. Pediatric Nephrology, 2023, 38(9), 2973-2985.

[4]. Lundberg, J.O., Weitzberg, E., Nitric oxide signaling in health and disease. Cell, 2022, 185(16), 2853-2878.

[5]. Jebari-Benslaiman, S., Galicia-García, U., Larrea-Sebal, A., Olaetxea, J.R., Alloza, I., Vandenbroeck, K., Benito-Vicente, A., Martín, C., Pathophysiology of atherosclerosis. International Journal of Molecular Sciences, 2022, 23(6), 3346.

[6]. Leick, M., Azcutia, V., Newton, G., Luscinskas, F.W., Leukocyte recruitment in inflammation: Basic concepts and new mechanistic insights based on new models and microscopic imaging technologies. Cell and Tissue Research, 2014, 355(3), 647-656.

[7]. Tvaroška, I., Selvaraj, C., Koča, J., Selectins—the two dr. Jekyll and mr. Hyde faces of adhesion molecules—a review. Molecules, 2020, 25(12), 2835.

[8]. Sperandio, M., Pickard, J., Unnikrishnan, S., Acton, S.T., Ley, K., Analysis of leukocyte rolling in vivo and in vitro. Methods in Enzymology, 2006, 416, 346-371.

[9]. Zuidema, M.Y., Korthuis, R.J. Intravital microscopic methods to evaluate anti-inflammatory effects and signaling mechanisms evoked by hydrogen sulfide. Methods in Enzymology, 2015, 555, 93-125.

[10]. Boyden, S., The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. The Journal of Experimental Medicine, 1962, 115(3), 453-466.

[11]. Flores-Espinosa, P., Mancilla-Herrera, I., Olmos-Ortiz, A., Díaz, L., Zaga-Clavellina, V. Evaluation of leukocyte chemotaxis induced by human fetal membranes in an in vitro model. Maternal Placental Interface: Methods and Protocols, 2024, 27-37.

[12]. Zen, K., Guo, Y.L., Li, L.M., Bian, Z., Zhang, C.Y., Liu, Y., Cleavage of the cd11b extracellular domain by the leukocyte serprocidins is critical for neutrophil detachment during chemotaxis. Blood, The Journal of the American Society of Hematology, 2011, 117(18), 4885-4894.

[13]. Chaubey, S., Ridley, A.J., Wells, C.M. Using the dunn chemotaxis chamber to analyze primary cell migration in real time. Cell migration: Developmental Methods and Protocols, 2011, 41-51.

[14]. Sundd, P., Zou, X., Goetz, D.J., Tees, D.F., Leukocyte adhesion in capillary-sized, p-selectin-coated micropipettes. Microcirculation, 2008, 15(2), 109-122.

[15]. Wiese, G., Barthel, S.R., Dimitroff, C.J., Analysis of physiologic e-selectin-mediated leukocyte rolling on microvascular endothelium. Journal of Visualized Experiments: JoVE, 2009, (24), 1009.

[16]. Levy, O., Anandakumaran, P., Ngai, J., Karnik, R., Karp, J.M., Systematic analysis of in vitro cell rolling using a multi-well plate microfluidic system. Journal of visualized experiments: JoVE, 2013, (80), 50866.

[17]. Canetti, E.F., Keane, J., McLellan, C., Gray, A., Comparison of capillary and venous blood in the analysis of concentration and function of leucocyte sub-populations. European Journal of Applied Physiology, 2016, 116(8), 1583-1593.

[18]. Jones, C.N., Dalli, J., Dimisko, L., Wong, E., Serhan, C.N., Irimia, D., Microfluidic chambers for monitoring leukocyte trafficking and humanized nano-proresolving medicines interactions. Proceedings of the National Academy of Sciences, 2012, 109(50), 20560-20565.

[19]. Regmi, S., Poudel, C., Adhikari, R., Luo, K.Q., Applications of microfluidics and organ-on-a-chip in cancer research. Biosensors, 2022, 12(7), 459.

[20]. Nielsen, J.B., Hanson, R.L., Almughamsi, H.M., Pang, C., Fish, T.R., Woolley, A.T., Microfluidics: Innovations in materials and their fabrication and functionalization. Analytical Chemistry, 2019, 92(1), 150-168.

[21]. Hameed, S.F., Turkie, D., A novel approach for study of surface morphology & roughness analysis for characterization of precipitation product at a nanoscale level via the reaction of fluconazole with phosphomolybidic acid. Chemical Methodologies, 2022.

[22]. Dion, I., Roques, X., More, N., Labrousse, L., Caix, J., Lefebvre, F., Rouais, F., Gautreau, J., Baquey, C., Ex vivo leucocyte adhesion and protein adsorption on tin. Biomaterials, 1993, 14(9), 712-719.

[23]. van Engeland, N.C., Pollet, A.M., den Toonder, J.M., Bouten, C.V., Stassen, O.M., Sahlgren, C.M., A biomimetic microfluidic model to study signalling between endothelial and vascular smooth muscle cells under hemodynamic conditions. Lab on a Chip, 2018, 18(11), 1607-1620.

[24]. Siddique, A., Meckel, T., Stark, R.W., Narayan, S., Improved cell adhesion under shear stress in pdms microfluidic devices. Colloids and Surfaces B: Biointerfaces, 2017, 150, 456-464.

[25]. Coghill, P.A., Kesselhuth, E.K., Shimp, E.A., Khismatullin, D.B., Schmidtke, D.W., Effects of microfluidic channel geometry on leukocyte rolling assays. Biomedical Microdevices, 2013, 15(1), 183-193.

[26]. Yu, Z., Chen, Y., Li, J., Chen, C., Lu, H., Chen, S., Zhang, T., Guo, T., Zhu, Y., Jin, J., A tempo-spatial controllable microfluidic shear-stress generator for in-vitro mimicking of the thrombus. Journal of Nanobiotechnology, 2024, 22(1), 187.

[27]. Siddique, A., Pause, I., Narayan, S., Kruse, L., Stark, R.W., Endothelialization of pdms-based microfluidic devices under high shear stress conditions. Colloids and surfaces B: Biointerfaces, 2021, 197, 111394.

[28]. Singh, J., Gambhir, T., Goh, T., van Vuuren, I., Gao, L., Wise, S.G., Waterhouse, A., Spatiotemporally mapped endothelial dysfunction at bifurcations in a coronary artery‐on‐a‐chip. Advanced Materials Technologies, 2024, 9(8), 2301596.

[29]. Ashok, D., Singh, J., Jiang, S., Waterhouse, A., Bilek, M., Reagent‐free covalent immobilization of biomolecules in a microfluidic organ‐on‐a‐chip. Advanced Functional Materials, 2024, 34(30), 2313664.

[30]. Pierres, A., Touchard, D., Benoliel, A.-M., Bongrand, P., Dissecting streptavidin-biotin interaction with a laminar flow chamber. Biophysical Journal, 2002, 82(6), 3214-3223.

[31]. Plouffe, B.D., Kniazeva, T., Mayer Jr, J.E., Murthy, S.K., Sales, V.L., Development of microfluidics as endothelial progenitor cell capture technology for cardiovascular tissue engineering and diagnostic medicine. The FASEB Journal, 2009, 23(10), 3309.

[32]. Shimp, E.A., Alsmadi, N.Z., Cheng, T., Lam, K.H., Lewis, C.S., Schmidtke, D.W., Effects of shear on p-selectin deposition in microfluidic channels. Biomicrofluidics, 2016, 10(2).

[33]. Nalayanda, D.D., Kalukanimuttam, M., Schmidtke, D.W., Micropatterned surfaces for controlling cell adhesion and rolling under flow. Biomedical Microdevices, 2007, 9(2), 207-214.

[34]. Andersen, A.S., Zheng, W., Sutherland, D., Jiang, X., Versatile multiple protein nanopatterning within a microfluidic channel for cell recruitment studies. Lab on a Chip, 2015, 15(24), 4524-4532.

[35]. Ashok, D., Singh, J., Howard, H.R., Cottam, S., Waterhouse, A., Bilek, M.M., Interfacial engineering for biomolecule immobilisation in microfluidic devices. Biomaterials, 2024, 123014.

[36]. Ferrari, E., Ugolini, G.S., Piutti, C., Marzorati, S., Rasponi, M., Plasma-enhanced protein patterning in a microfluidic compartmentalized platform for multi-organs-on-chip: A liver-tumor model. Biomedical Materials, 2021, 16(4), 045032.

[37]. Kim, D., Herr, A.E., Protein immobilization techniques for microfluidic assays. Biomicrofluidics, 2013, 7(4).

[38]. Yap, F.L., Zhang, Y., Protein micropatterning using surfaces modified by self-assembled polystyrene microspheres. Langmuir, 2005, 21(12), 5233-5236.

[39]. Haddadi, M., Mousavi, M.J., Regenerative medicine: Highlight on the significance of therapeutics with novel strategies. Internationa Journal of Advanced Biological and Biomedical Research, 2020, 8(4), 370-387.