Biology subjects

Hind, L. E.

Publications and source records attributed to Hind, L. E..

4 recordsLinked to original sources

bioRxiv2026

Endothelial Heterogeneity Across Vascular Beds Impacts Inflammatory Signaling and Neutrophil Adhesion

Endothelial cells (ECs) are key players in maintaining homeostasis and coordinating immune responses, activating during acute inflammation to recruit immune cells. Endothelial heterogeneity has been found to impact transcription level differences across EC sources, but how these differences drive downstream effects in inflammatory signaling and immune interactions remains unclear. Here, we employed multiplexed ELISA to quantify secretion for 19 inflammatory factors following tumor necrosis factor (TNF) or Pseudomonas aeruginosa activation of four primary human EC sources: umbilical artery (HUAEC), umbilical vein (HUVEC), dermal microvascular (HDMEC), and pulmonary microvascular (HPMEC) endothelial cells. We also quantified changes in neutrophil adhesion to each EC source and used partial least squares regression (PLSR) to identify key inflammatory proteins associated with changes in neutrophil adhesion. We found distinct inflammatory secretion profiles across all cell types, with veinous ECs showing the highest basal secretion of most inflammatory proteins and pulmonary ECs exhibiting the lowest. Arterial ECs exhibited the lowest sensitivity to inflammatory stimulus, while pulmonary ECs exhibited dynamic responses following activation. Furthermore, inflammatory stimulus caused large differences in expression across cell sources for six factors: GM-CSF, IL-1{beta}, IL-6, IP-10, E-selectin, and ICAM-1. We found endothelial heterogeneity also contributed to differences in neutrophil adhesion to unstimulated ECs. Our PLSR analysis revealed five secreted factors most indicative of changes in neutrophil adhesion: E-selectin, ICAM-1, PECAM1, IL-6, and IL-8. Collectively, our findings strengthen the emerging view that vascular-bed specific differences in EC phenotype can impact downstream immune responses.

bioengineering↗

bioRxiv2026

Fibronectin and laminin differentially affect the inflammatory environment in microphysiological systems

Tissue engineered constructs are increasingly used for both modeling organs and disease in vitro as well as for therapeutic intervention. In addition to collagen, these constructs commonly include native extracellular matrix proteins (ECM), such as fibronectin and laminin. Given the critical role of inflammatory pathways in disease and in response to implanted materials, it is important to understand the role these proteins play in regulating the inflammatory environment. Fibronectin and laminin influence neutrophil function and endothelial activation in 2D, but their regulation of the inflammatory environment in 3D engineered constructs is not clear. For this study, we used an inflammation-on-a-chip device that includes a model blood vessel surrounded by a collagen I hydrogel with fibronectin and/or laminin. We investigated the additive effects of both proteins and a range of concentrations for each protein to determine concentration dependence. Both fibronectin and laminin have concertation dependent effects on neutrophils and the endothelium. High concentrations (50 {micro}g/mL) of fibronectin reduced neutrophil migration, while 20 {micro}g/mL laminin reduced neutrophil extravasation and migration, potentially due to lower ICAM-1 expression by the endothelium. Interestingly, 50 {micro}g/mL of laminin significantly disrupted endothelial vessel formation and reduced ICAM-1 and VE-cadherin expression, likely due to significant changes in the collagen architecture. The inclusion of fibronectin and laminin, even at physiological levels, results in significant effects on neutrophil behavior, endothelial vessel formation, and collagen architecture. These proteins impact the inflammatory environment and thus need to be considered when modeling diseases and designing therapeutics, especially when neutrophils or an endothelium are involved. Translational Impact StatementThis work uses an inflammation-on-a-chip device to study how fibronectin and laminin impact neutrophil behavior and vascular inflammation as these proteins are commonly used in engineered constructs. We found that fibronectin impairs neutrophil migration, while laminin decreases neutrophil extravasation and migration and at higher concentrations also prevents endothelial vessel formation. Therefore, researchers should be aware that these proteins will alter the inflammatory environment when including them in engineered constructs.

bioengineering↗

bioRxiv2026

Interleukin-6 Concentration Regulates the Neutrophil Response to Diverse Bacteria

Neutrophils are critical mediators of the innate immune response, and their antimicrobial functions are tightly regulated by a myriad of cytokines. Interleukin-6 (IL-6) is known to be essential for an effective immune response; however, how varying IL-6 concentrations affect the neutrophil response remains poorly understood. Because IL-6 concentrations can vary greatly across different disease states, we investigated the concentration dependent effects of IL-6 on the neutrophil response to diverse bacterial pathogens using an infection-on-a-chip microfluidic device. We found that a high exogenous IL-6 concentration (100 ng/mL) reduced neutrophil extravasation, migration speed, and displacement compared to conditions without exogenous IL-6. In contrast, a lower exogenous IL-6 concentration (10 ng/mL) produced pathogen-specific effects on neutrophil extravasation: exogenous IL-6 increased neutrophil extravasation in response to Pseudomonas aeruginosa, did not change in response to Listeria monocytogenes, and decreased in response to Staphylococcus aureus relative to controls. We then determined the potential endothelial cell contributions to these responses. We found that increasing IL-6 concentration resulted in decreased VE-cadherin expression and that 100 ng/mL exogenous IL-6 resulted in lower ICAM-1 expression than 10 ng/mL exogenous IL-6 in an endothelium exposed to P. aeruginosa. Together, these results demonstrate that IL-6 exerts concentration- and pathogen-dependent effects on neutrophil recruitment and migration, supporting a dual role for IL-6 as both pro-inflammatory and anti-inflammatory, with higher IL-6 concentrations resulting in a more anti-inflammatory neutrophil response.

immunology↗

bioRxiv2025

Synovial fibroblasts support vascular function in an acute injury-on-a-chip model

Most patients who sustain an acute joint injury develop degenerative joint disease, or osteoarthritis (OA). Animal models have informed the design of OA therapeutics; however, no disease-modifying therapy has successfully translated to human patients. Thus, there is a strong motivation to develop humanized in vitro platforms to fill a critical gap in knowledge of disease progression post-injury. Here, we develop an acute injury-on-a-chip model of the synovium, a vascularized, joint-lining tissue that has been implicated in OA progression and as a key driver of joint disease. We apply this chip-based system to investigate crosstalk between endothelial cells, lining an engineered vessel, and synovial fibroblasts, embedded within an extracellular matrix hydrogel. Our data indicate that synovial fibroblasts, rather than initiating disease, attempt to support and maintain vascular function in the presence of acute inflammation (i.e., interleukin-1{beta}). Such knowledge may provide new targets for OA therapeutics, preventing the progression from joint injury to disease in patients. TeaserIn the presence of inflammation, a hallmark of acute injury, synovial fibroblasts work to maintain vascular health.

bioengineering↗