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Research Article: Flow-based imaging reveals dissociated monocyte adhesion and transmigration patterns between TNF-? and diabetes-induced vascular inflammation

Date Published: 2026-07-10

Abstract:
Monocyte–endothelial interactions drive both acute inflammation and chronic vascular disease in type 2 diabetes mellitus (T2DM), yet whether TNF-?–induced and diabetes-associated monocyte trafficking engage similar or distinct molecular programs remains unclear. Using a physiological flow-based imaging system, we quantified CD14 + monocyte adhesion, transendothelial migration (TEM), and abluminal residence on HUVEC monolayers activated with TNF-? or T2DM serum under controlled shear stress. Both stimuli induced comparable monocyte adhesion (~40 cells by 3 min), but TEM efficiency diverged markedly: TNF-? promoted robust TEM (~18% by 5 min) whereas T2DM conditions showed severely impaired TEM (~5%). Under T2DM conditions, monocytes exhibited prolonged abluminal retention (median 85 min vs. 25 min; p<0.0001), a phenotype recapitulated by endothelial hyperglycemic exposure alone. Comparative RNA-seq analysis of T2DM (GSE92724) and TNF-?–stimulated (GSE134489) endothelial cells revealed near-zero transcriptional correlation (r = 0.018) between conditions. TNF-? drove coordinate NF-?B–dependent upregulation of VCAM1, ICAM1, E-selectin, and junctional molecules with organized junctional remodeling, whereas T2DM produced VCAM1-biased adhesion with ICAM1 downregulation (0.61-fold), claudin suppression, and junctional disorganization. KEGG pathway mapping confirmed organized endothelial–leukocyte integrin co-activation under TNF-? versus discoordination junctional loss under T2DM. qPCR validation identified selective RAGE and JAM3 upregulation under T2DM—consistent with AGE-RAGE signaling and JAM-3–MAC-1 monocyte trapping—without classical NF-?B activation. These findings define two fundamentally distinct paradigms: a TNF-? “recruitment model” enabling efficient trafficking and resolution, versus a T2DM “retention model” characterized by VCAM1-biased adhesion, junctional disorganization, and impaired reverse transmigration driving chronic monocyte accumulation, identifying AGE-RAGE signaling and JAM-3–MAC-1 interactions as potential therapeutic targets for diabetic vascular inflammation.

Introduction:
Type 2 diabetes mellitus (T2DM) represents one of the most significant global health challenges of the 21st century. According to the International Diabetes Federation (IDF), approximately 537 million adults were living with diabetes worldwide in 2021, a figure projected to rise to 783 million by 2045, with T2DM accounting for over 90% of cases ( 1 ). Diabetes is the leading cause of preventable blindness, end-stage renal disease, and non-traumatic lower limb amputation, and it confers a two- to four-fold…

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