Dataset Information

Model Particulate Drug Carriers Modulate Leukocyte Adhesion in Human Blood Flows.

ABSTRACT: Drug carriers have been widely explored as a method of improving the efficacy of therapeutic drugs for a variety of diseases, including those involving inflammation. However, few of these formulations have advanced past clinical trials. There are still major gaps in our understanding of how drug carriers impact leukocytes, particularly in inflammatory conditions. In this work, we investigated how targeted and nontargeted drug carriers affect the function of leukocytes in blood flow. We explored three primary mechanisms: (1) collisions in blood flow disrupt leukocyte adhesion, (2) specific binding to the endothelium competes with leukocytes for binding sites, and (3) particle phagocytosis alters leukocyte phenotype, resulting in reduced adhesion. We find that each of these mechanisms contributes to significantly reduced leukocyte adhesion to an inflamed endothelium, and that particle phagocytosis may be the most significant driver of this effect. These results are crucial for understanding the totality of the impact of drug carriers on leukocyte behavior and response to inflammation and should inform the future design of any such drug carriers.

SUBMITTER: Kelley WJ

PROVIDER: S-EPMC9588176 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Model Particulate Drug Carriers Modulate Leukocyte Adhesion in Human Blood Flows.

Kelley William J WJ Onyskiw Peter J PJ Fromen Catherine A CA Eniola-Adefeso Omolola O

ACS biomaterials science & engineering 20191106 12

Drug carriers have been widely explored as a method of improving the efficacy of therapeutic drugs for a variety of diseases, including those involving inflammation. However, few of these formulations have advanced past clinical trials. There are still major gaps in our understanding of how drug carriers impact leukocytes, particularly in inflammatory conditions. In this work, we investigated how targeted and nontargeted drug carriers affect the function of leukocytes in blood flow. We explored ...[more]

PMID: 33417805

Similar Datasets

Project description:Targeted drug carriers are attractive for the delivery of therapeutics directly to the site of a disease, reducing systemic side effects and enhancing the efficacy of therapeutic molecules. However, the use of particulate carriers for drug delivery comes with its own set of challenges and barriers. Among these, a great deal of research effort has focused on protecting carriers from clearance by phagocytes via altering carrier surface chemistry, mostly with the use of polyethylene glycol (PEG) chain coatings. However, few papers have explored the effects of PEGylation on uptake by freshly-obtained primary human phagocytes in physiological conditions. In this work, we investigate the effect of PEGylation on particle uptake by primary human neutrophils in vitro and compare these effects to several cell lines and other model phagocytic cells systems. We find that human neutrophils in whole blood preferentially phagocytose PEGylated particles (e.g., ∼40% particle positive neutrophils for PEGylated versus ∼20% for carboxylated polystyrene microspheres) and that this effect is linked to factors present in human plasma. Model phagocytes internalized PEGylated particles less efficiently or equivalently to carboxylated particles in culture medium but preferentially phagocytosed PEGylated particles in the human plasma (e.g., ∼86% versus ∼63% PEGylated versus carboxylated particle positive cells, respectively). These findings have significant implications for the efficacy of PEGylation in designing long-circulating drug carriers, as well as the need for thorough characterization of drug carrier platforms in a wide array of in vitro and in vivo assays.Statement of significanceThe work in this manuscript is highly significant to the field of drug delivery, as it explores in-depth the effects of polyethylene glycol (PEG) coatings, which are frequently used to prevent phagocytic clearance of particulate drug carriers, on the phagocytosis of such carriers by neutrophils, the most abundant leukocyte in blood circulation. Surprisingly, we find that PEGylation enhances uptake by primary human neutrophils, specifically in the presence of human plasma. This result suggests that PEGylation may not confer the benefits in humans once thought, and may help to explain why PEG has not become the "magic bullet" it was once thought to be in the field of particulate drug delivery.

Project description:Multiple sclerosis is a chronic neuroinflammatory disorder characterized by demyelination, oligodendrocyte damage/loss and neuroaxonal injury in the context of immune cell infiltration in the central nervous system. No neuroprotective therapy is available to promote the survival of oligodendrocytes and protect their myelin processes in immune-mediated demyelinating diseases. Pro-inflammatory CD4 T helper (Th)17 cells can directly interact with oligodendrocytes in multiple sclerosis and its animal model, causing injury to myelinating processes and cell death through direct contact. However, the molecular mechanisms underlying the close contact and subsequent detrimental interaction of Th17 cells with oligodendrocytes remain unclear. In this study we conducted single cell RNA sequencing of human primary oligodendrocytes cultured alone, in contact or separated by an insert with primary Th17 polarized T cells and identified several cell adhesion molecules that may modulate T cell contact mediated damage of oligodendrocytes. Furthermore, we used flow cytometry and immunofluorescence studies on central nervous system tissue from multiple sclerosis subjects, its animal model and controls to characterize the expression of cell adhesion molecules by mature oligodendrocytes. We found that a significant proportion of human and murine mature oligodendrocytes express melanoma cell adhesion molecule and activated leukocyte cell adhesion molecule in multiple sclerosis, EAE and controls although their regulation differ between human and mouse. We observed that exposure to pro-inflammatory cytokines or to human activated T cells are associated with a marked downregulation of the expression of melanoma cell adhesion molecule but not activated leukocyte cell adhesion molecule at the surface of human primary oligodendrocytes. Furthermore, we used in vitro live-imaging, immunofluorescence, and flow cytometry to determine the contribution of these molecules to Th17-polarized cell adhesion and cytotoxicity towards human oligodendrocytes. Silencing and blocking activated leukocyte cell adhesion molecule but not melanoma cell adhesion molecule limited prolonged interactions between oligodendrocytes and Th17-polarized cells, resulting in decreased adhesion of Th17-polarized cells to oligodendrocytes and conferred significant protection of oligodendrocytic processes.

Project description:Dendritic cells (DCs) are promising targets for drug delivery, as they can induce immunity or tolerance. The current study aims to examine the potential of using nanostructured lipid carriers (NLC) as delivery systems for human DC by evaluating nanoparticle internalization, cell labeling, and drug activity. NLC were formulated incorporating the fluorochrome fluorescein isothiocyanate (FITC-NLC) or the natural anti-inflammatory molecule resveratrol (rsv-NLC). Primary human DCs were differentiated from peripheral blood monocytes, and the innovative imaging flow cytometry technique was used to examine FITC-NLC internalization. The capacity of rsv-NLC to inhibit DC activation in response to proinflammatory cytokine tumor necrosis factor-α (TNF- α) was investigated by conventional flow cytometry. A combination of imaging and conventional flow cytometry was used to assess NLC cytotoxicity. The results obtained indicate that both NLC formulations were stable over time, with mean diameter <200 nm and highly negative zeta potential (about -30 mV). When DCs were placed in contact with NLC, imaging flow cytometry clearly showed that DCs efficiently internalized FITC-NLC, with nearly 100% of cells internalizing nanoparticles upon 1 hour of incubation. Both immature and mature DCs internalized NLC to high and comparable levels, and without cytotoxicity. Stimulating DC with TNF-α in the presence of rsv-NLC revealed that, using these nanoparticles, very small concentrations of rsv were sufficient to significantly decrease surface expression of activation marker CD83 (5 µM) and major histocompatibility complex-class II molecule human leukocyte antigen - antigen D related (10 µM), both upregulated in response to TNF-α stimulation. Rsv-NLC were compared with free rsv; at 5 µM, rsv-NLC were able to inhibit nuclear factor κ beta phosphorylation and significantly decrease the level of interleukin-12/23, both upregulated in response to TNF-α, while 10 µM free rsv were needed to promote a similar effect. Taken together, the results presented show that NLC are suitable carriers of fluorescent labels or bioactive molecules for human DCs, leading to inflammation modulation.

Project description:Simple Summary Intercellular adhesion molecule 1 (ICAM-1) is a protein produced by cells in blood vessel walls, as well as many other cell types. It localizes to the surface of these cells to allow them to communicate with adjacent cells. The production of ICAM-1 is induced by inflammation, such as occurs during infection, when one of its main functions is to direct white blood cells to the site of the infection. However, in autoimmune diseases and certain types of infection, ICAM-1 can be involved in directing cells to damage healthy tissue. ICAM-1 can also promote or protect against cancer, depending on the type of cancer and the cell populations that are producing it. In this review article, the ICAM1 gene and ICAM-1 protein are introduced, and their regulation is summarized. The roles of ICAM-1 in the immune system and a spectrum of diseases, as well as therapeutics that target ICAM-1, are discussed. Abstract Intercellular adhesion molecule 1 (ICAM-1) is a transmembrane protein in the immunoglobulin superfamily expressed on the surface of multiple cell populations and upregulated by inflammatory stimuli. It mediates cellular adhesive interactions by binding to the β2 integrins macrophage antigen 1 and leukocyte function-associated antigen 1, as well as other ligands. It has important roles in the immune system, including in leukocyte adhesion to the endothelium and transendothelial migration, and at the immunological synapse formed between lymphocytes and antigen-presenting cells. ICAM-1 has also been implicated in the pathophysiology of diverse diseases from cardiovascular diseases to autoimmune disorders, certain infections, and cancer. In this review, we summarize the current understanding of the structure and regulation of the ICAM1 gene and the ICAM-1 protein. We discuss the roles of ICAM-1 in the normal immune system and a selection of diseases to highlight the breadth and often double-edged nature of its functions. Finally, we discuss current therapeutics and opportunities for advancements.

Dataset Information

Model Particulate Drug Carriers Modulate Leukocyte Adhesion in Human Blood Flows.

Publications

Model Particulate Drug Carriers Modulate Leukocyte Adhesion in Human Blood Flows.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets