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Updated: Aug 23, 2026
Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions
Published on: August 23, 2013
Sided Stimulation of Endothelial Cells Modulates Neutrophil Trafficking in an In Vitro Sepsis Model
Danial Ahmad1, Isabelle Linares1, Anthony Pietropaoli2
1Department of Biomedical Engineering, University of Rochester, Rochester, NY, 14627, USA.
Abstract:
While the role of dysregulated polymorphonuclear leukocyte (PMN) transmigration in septic mediated tissue damage is well documented, strategies to mitigate aberrant transmigration across endothelium have yet to yield viable therapeutics. Recently, microphysiological systems (MPS) have emerged as novel in vitro mimetics that facilitate the development of human models of disease. With this advancement, aspects of endothelial physiology that are difficult to assess with other models can be directly probed. In this study, the role of endothelial cell (EC) apicobasal polarity on leukocyte trafficking response is evaluated with the µSiM-MVM (microphysiological system enabled by a silicon membrane - microvascular mimetic). Here, ECs are stimulated either apically or basally with a cytokine cocktail to model a septic-like challenge before introducing healthy donor PMNs into the device. Basally oriented stimulation generated a stronger PMN transmigratory response versus apical stimulation. Importantly, healthy PMNs are unable to migrate towards a bacterial peptide chemoattractant when ECs are apically stimulated, which mimics the attenuated PMN chemotaxis seen in sepsis. Escalating the apical inflammatory stimulus by a factor of five is necessary to elicit high PMN transmigration levels across endothelium. These results demonstrate that EC apicobasal polarity modulates PMN transmigratory behavior and provides insight into the mechanisms underlying sepsis.
Insights
Dysregulated polymorphonuclear leukocyte (PMN) transmigration contributes to sepsis-induced tissue damage. Endothelial cell apicobasal polarity significantly impacts PMN trafficking, offering new therapeutic targets for sepsis.
Area of Science:
- Cell Biology
- Immunology
- Biomedical Engineering
Background:
- Dysregulated polymorphonuclear leukocyte (PMN) transmigration is a key factor in sepsis-mediated tissue damage.
- Current therapeutic strategies to control aberrant transmigration remain limited.
- Microphysiological systems (MPS) offer advanced in vitro models for studying human diseases, including endothelial physiology.
Purpose of the Study:
- To evaluate the role of endothelial cell (EC) apicobasal polarity in leukocyte trafficking.
- To investigate PMN transmigration using a novel microphysiological system (µSiM-MVM).
- To model sepsis-like conditions and assess their impact on PMN behavior.
Main Methods:
- Utilized the µSiM-MVM, a microphysiological system with a silicon membrane, to mimic microvasculature.
- Stimulated endothelial cells (ECs) apically or basally with a cytokine cocktail to simulate a septic-like challenge.
- Introduced healthy donor PMNs to assess transmigration in response to different EC stimulation conditions and bacterial peptide chemoattractants.
Main Results:
- Basal EC stimulation resulted in a stronger PMN transmigratory response compared to apical stimulation.
- Apical EC stimulation with a septic-like challenge attenuated PMN chemotaxis, preventing migration towards bacterial chemoattractants.
- A five-fold increase in apical inflammatory stimulus was required to achieve high PMN transmigration levels.
Conclusions:
- Endothelial cell apicobasal polarity is a critical regulator of PMN transmigratory behavior.
- The findings provide novel insights into the mechanisms underlying sepsis-associated leukocyte trafficking.
- This study highlights the potential of MPS in dissecting complex cellular interactions in disease models.
