Microfluidic devices for neutrophil chemotaxis studies

Zhao, Wenjie; Zhao, Haiping; Li, Mingxiao; Huang, Chengjun

doi:10.1186/s12967-020-02335-7

Cited by 21 publications

(11 citation statements)

References 95 publications

(207 reference statements)

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“…We also examined the chemotactic properties of PMNs using a microfluidic PMN migration chip. This system potentially minimizes the gravitational interference that may contribute to chemokinetic transport of PMNs or cellular debris down the transwell, as may occur in conventional chemotaxis experiments ( 34 , 35 ). Although no significant differences were observed, in the sepsis-mimicking medium, the fraction of chemotactic PMNs and their migrating velocity appeared to be reduced compared to those in the control medium.…”

Section: Discussionmentioning

confidence: 99%

Circulating mitochondrialN-formyl peptides contribute to secondary nosocomial infection in patients with septic shock

Kwon

Suh

Jung

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

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Secondary infections typically worsen outcomes of patients recovering from septic shock. Neutrophil [polymorphonuclear leukocytes (PMNs)] migration to secondarily inoculated sites may play a key role in inhibiting progression from local bacterial inoculation to secondary infection. Mitochondrial N-formyl peptide (mtFP) occupancy of formyl peptide receptor-1 (FPR1) has been shown to suppress PMN chemotaxis. Therefore, we studied the association between circulating mtFPs and the development of secondary infection in patients with septic shock. We collected clinical data and plasma samples from patients with septic shock admitted to the intensive care unit for longer than 72 h. Impacts of circulating nicotinamide adenine dinucleotide dehydrogenase subunit-6 (ND6) upon clinical outcomes were analyzed. Next, the role of ND6 in PMN chemotaxis was investigated using isolated human PMNs. Studying plasma samples from 97 patients with septic shock, we found that circulating ND6 levels at admission were independently and highly associated with the development of secondary infection (odds ratio = 30.317, 95% CI: 2.904 to 316.407, P = 0.004) and increased 90-d mortality (odds ratio = 1.572, 95% CI: 1.002 to 2.465, P = 0.049). In ex vivo experiments, ND6 pretreatment suppressed FPR1-mediated PMN chemotactic responses to bacterial peptides in the presence of multiple cytokines and chemokines, despite increased nondirectional PMN movements. Circulating mtFPs appear to contribute to the development of secondary infection and increased mortality in patients with septic shock who survive their early hyperinflammatory phase. The increased susceptibility to secondary infection is probably partly mediated by the suppression of FPR1-mediated PMN chemotaxis to secondary infected sites.

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Section: Discussionmentioning

confidence: 99%

Circulating mitochondrialN-formyl peptides contribute to secondary nosocomial infection in patients with septic shock

Kwon

Suh

Jung

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

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“…fMLP has been demonstrated to induce neutrophil chemotaxis [ 23 ]. Thus, we first validated the function of F4-Chip by testing the neutrophil transmigration in the fMLP gradients (10, 100, and 1000 nM).…”

Section: Resultsmentioning

confidence: 99%

“…Microfluidic devices have been widely used to study cell migration and chemotaxis during the past 20 years due to the superiority of miniaturization and micro-environmental control [ 20 , 21 , 22 , 23 ]. In addition, microfluidic devices have been developed and improved to be diagnostic tools for neutrophil migration-related diseases [ 24 , 25 , 26 ].…”

Section: Introductionmentioning

confidence: 99%

A Novel Microfluidic Device for the Neutrophil Functional Phenotype Analysis: Effects of Glucose and Its Derivatives AGEs

et al. 2021

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Neutrophil dysfunction is closely related to the pathophysiology of patients with diabetes mellitus, but existing immunoassays are difficult to implement in clinical applications, and neutrophil’s chemotaxis as a functional biomarker for diabetes mellitus prognostic remains largely unexplored. Herein, a novel microfluidic device consisted of four independent test units with four cell docking structures was developed to study the neutrophil chemotaxis, which allowed multiple cell migration observations under a single field of view (FOV) and guaranteed more reliable results. In vitro studies, the chemotaxis of healthy neutrophils to N-Formyl-Met-Leu-Phe (fMLP) gradient (0, 10, 100, and 1000 nM) was concentration-dependent. The distinct promotion or suppression in the chemotaxis of metformin or pravastatin pretreated cells were observed after exposure to 100 nM fMLP gradient, indicating the feasibility and efficiency of this novel microfluidic device for clinically relevant evaluation of neutrophil functional phenotype. Further, the chemotaxis of neutrophils pretreated with 25, 50, or 70 mM of glucose was quantitatively lower than that of the control groups (i.e., 5 mM normal serum level). Neutrophils exposed to highly concentrated advanced glycation end products (AGEs) (0.2, 0.5, or 1.0 μM; 0.13 μM normal serum AGEs level), a product of prolonged hyperglycemia, showed that the higher the AGEs concentration was, the weaker the migration speed became. Specifically, neutrophils exposed to high concentrations of glucose or AGEs also showed a stronger drifting along with the flow, further demonstrating the change of neutrophil chemotaxis. Interestingly, adding the N-benzyl-4-chloro-N-cyclohexylbenzamide (FPS-ZM1) (i.e., high-affinity RAGE inhibitor) into the migration medium with AGEs could hinder the binding between AGEs and AGE receptor (RAGE) located on the neutrophil, thereby keeping the normal chemotaxis of neutrophils than the ones incubated with AGEs alone. These results revealed the negative effects of high concentrations of glucose and AGEs on the neutrophil chemotaxis, suggesting that patients with diabetes should manage serum AGEs and also pay attention to blood glucose indexes. Overall, this novel microfluidic device could significantly characterize the chemotaxis of neutrophils and have the potential to be further improved into a tool for risk stratification of diabetes mellitus.

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“…[ 6 ] Alternatives to the transwell assay were introduced to address some of its limitations, including tracking and monitoring single cells (as in Dunn chambers), [ 7 ] and detecting cell reversibility or fugetaxis (as in under‐agarose migration assays). [ 8 ] More recently, microfluidic systems have been developed [ 9 ] that enable control of stable gradients, [ 10 ] distinction between different types of movement (e.g., chemotaxis, chemokinesis—non‐directional cell migration, and fugetaxis [ 11 ] ), tracking individual cells in real‐time, [ 12 ] and increased throughput [ 13 ] —sometimes achieved with less reliance on specialized equipment. [ 14 ] While microfluidic approaches show great promise, their uptake in biomedical research has been impeded by the technical complexity required to operate devices, long fabrication and prototyping times, the problematic biocompatibility of the plastic often used (i.e., polydimethylsiloxane, PDMS), and failure rates associated with air bubbles disturbing flow patterns.…”

Section: Introductionmentioning

confidence: 99%