Understanding how to modulate immune pathways is attracting great interest to develop new therapeutic strategies. In recent years, nanomedicine has demonstrated promising results in modulating the immune response. To boost the application of nanomedicine in immunotherapy, it is fundamental to understand the mechanisms of interaction between immune cells and nanomaterials (NMs). Since the extracellular matrix (ECM) modulates the behaviour of immune cells in healthy and diseased tissues, the study of interactions between immune cells and NMs should consider the extracellular microenvironment where these interactions occur. In this work, we aim at studying the role of the ECM in influencing macrophage response to NMs. To address this issue, we prepared ECM-like substrates based on denatured collagen, the most abundant protein present in the ECM, and then loaded with increasing concentrations of carbon nanotubes (CNTs), known to affect macrophage activity when dispersed in cell culture media. Macrophage-like cells, derived from the human monocytic THP-1 cell line, were grown on the substrates up to 5 days. Preliminary data demonstrated the ability of macrophages to adhere to all the substrates and no effect on cell viability was observed after 24 h from seeding, regardless the presence of CNTs. However, at higher CNT concentrations and culture times, cell morphological changes were observed with an increase in filopodia, a decrease in spreading area by assuming a round shape. Furthermore, scanning electron microscopy analysis indicated that macrophages remodelled the ECM-like substrates, making bioavailable the CNTs that are internalized by THP-1 cells. Finally, conditioned media by THP-1 cells grown on CNT-loaded substrates promote cell migration of endothelial cells in wound healing assay compared to conditioned media of control cells. Taken altogether these findings indicate that the presence of NMs embedded in the matrices can modulate the macrophage response.
Investigating on the biological mechanisms underlying the macrophages-ECM crosstalk.
Naym Blal;D. Guarnieri
2024
Abstract
Understanding how to modulate immune pathways is attracting great interest to develop new therapeutic strategies. In recent years, nanomedicine has demonstrated promising results in modulating the immune response. To boost the application of nanomedicine in immunotherapy, it is fundamental to understand the mechanisms of interaction between immune cells and nanomaterials (NMs). Since the extracellular matrix (ECM) modulates the behaviour of immune cells in healthy and diseased tissues, the study of interactions between immune cells and NMs should consider the extracellular microenvironment where these interactions occur. In this work, we aim at studying the role of the ECM in influencing macrophage response to NMs. To address this issue, we prepared ECM-like substrates based on denatured collagen, the most abundant protein present in the ECM, and then loaded with increasing concentrations of carbon nanotubes (CNTs), known to affect macrophage activity when dispersed in cell culture media. Macrophage-like cells, derived from the human monocytic THP-1 cell line, were grown on the substrates up to 5 days. Preliminary data demonstrated the ability of macrophages to adhere to all the substrates and no effect on cell viability was observed after 24 h from seeding, regardless the presence of CNTs. However, at higher CNT concentrations and culture times, cell morphological changes were observed with an increase in filopodia, a decrease in spreading area by assuming a round shape. Furthermore, scanning electron microscopy analysis indicated that macrophages remodelled the ECM-like substrates, making bioavailable the CNTs that are internalized by THP-1 cells. Finally, conditioned media by THP-1 cells grown on CNT-loaded substrates promote cell migration of endothelial cells in wound healing assay compared to conditioned media of control cells. Taken altogether these findings indicate that the presence of NMs embedded in the matrices can modulate the macrophage response.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
