Dynamic alteration of poroelastic attributes as determinant membrane nanorheology for endocytosis of organ specific targeted gold nanoparticles

Abstract

Background Efficacy of targeted drug delivery using nanoparticles relies on several factors including the uptake mechanisms such as phagocytosis, macropinocytosis, micropinocytosis and receptor mediated endocytosis. These mechanisms have been studied with respect to the alteration in signaling mechanisms, cellular morphology, and linear nanomechanical properties (NMPs). Commonly employed classical contact mechanics models to address cellular NMPs fail to address mesh like structure consisting of bilayer lipids and proteins of cell membrane. To overcome this technical challenge, we employed poroelastic model which accounts for the biphasic nature of cells including their porous behavior exhibiting both solid like (fluid storage) and liquid like (fluid dissipate) behavior. Results In this study, we employed atomic force microscopy to monitor the influence of surface engineering of gold nanoparticles (GNPs) to the alteration of nonlinear NMPs such as drained Poisson’s ratio, effective shear stress, diffusion constant and pore dimensions of cell membranes during their uptake. Herein, we used pancreatic cancer (PDAC) cell lines including Panc1, AsPC-1 and endothelial cell (HUVECs) to understand the receptor-dependent and -independent endocytosis of two different GNPs derived using plectin-1 targeting peptide (PTP-GNP) and corresponding scrambled peptide (sPEP-GNP). Compared to untreated cells, in case of receptor dependent endocytosis of PTP-GNPs diffusion coefficient altered ~ 1264-fold and ~ 1530-fold and pore size altered ~ 320-fold and ~ 260-fold in Panc1 and AsPC-1 cells, respectively. Whereas for receptor independent mechanisms, we observed modest alteration in diffusion coefficient and pore size, in these cells compared to untreated cells. Effective shear stress corresponding to 7.38 ± 0.15 kPa and 20.49 ± 0.39 kPa in PTP-GNP treatment in Panc1 and AsPC-1, respectively was significantly more than that for sPEP-GNP. These results demonstrate that with temporal recruitment of plectin-1 during receptor mediated endocytosis affects the poroelastic attributes of the membrane. Conclusion This study confirms that nonlinear NMPs of cell membrane are directly associated with the uptake mechanism of nanoparticles and can provide promising insights of the nature of endocytosis mechanism involved for organ specific drug delivery using nanoparticles. Hence, nanomechanical analysis of cell membrane using this noninvasive, label-free and live-cell analytical tool can therefore be instrumental to evaluate therapeutic benefit of nanoformulations. Graphical Abstract

Biophysical, Molecular and Proteomic Profiling of Human Retinal Organoid-Derived Exosomes

“…Atomic force microscope (AFM) has become an attractive characterization tool as it is a label free, non-invasive technique ideal for soft biological samples such as cells, tissues including the EVs [36][37][38]. The state-of-the-art technology allows for both morphological and nanomechanical characterizations of these samples in fluid environment, thus preserving their overall characteristics [39]. AFM has been used to evaluate the morphology of mesenchymal stem cell (MSC)-derived EVs ornated with LJM-3064 aptamer, a myelin-specific DNA aptamer on remyelination processes and immunomodulatory activity in myelin oligodendrocyte glycoprotein (MOG35-55)-induced mouse multiple sclerosis model [39].…”

Behavior of Citrate-Capped Ultrasmall Gold Nanoparticles on a Supported Lipid Bilayer Interface at Atomic Resolution

“…18,22,23,25 The lipids not only play a passive role in hosting functional biomolecules but also play an active role in locally modulating the membrane mechanical properties in response to external stimuli and by being involved in adapting the molecular composition of both lipids and proteins to best support the needs of the cell. 26,27 It is therefore crucial to derive a comprehensive understanding of the nanomaterial−lipid interactions (i.e., adsorption, 12 desorption, 8 translocation, 28 and internalization/uptake 9,29 ) at the molecular level to achieve the desired application goal. Several experimental studies have outlined interactions between gold NPs (AuNPs) and model membranes, including their adsorption dynamics.…”

“…In all of these systems, − the approach often relies on mimicking the hydrophilicity profile of the membrane or finding ways to interact with or avoid specific cellular components (proteins, lysosomes, organelles, etc. ). − The underlying challenge is to achieve control of the membrane–nanomaterials interactions, a task complicated by the large number of different biomolecules present in most biomembranes.…”

“…Phospholipids make up the bulk of the biomembrane, forming a continuous bilayer that is the universal component of all cell membranes. ,,, Other biomolecules such as proteins and sugars can be embedded or anchored in this bilayer structure, ensuring the functionality of the membrane for cellular-scale processes such as transmembrane trafficking, budding, endocytosis, and protein-mediated channel regulation. ,,, The lipids not only play a passive role in hosting functional biomolecules but also play an active role in locally modulating the membrane mechanical properties in response to external stimuli and by being involved in adapting the molecular composition of both lipids and proteins to best support the needs of the cell. , It is therefore crucial to derive a comprehensive understanding of the nanomaterial–lipid interactions (i.e., adsorption, desorption, translocation, and internalization/uptake , ) at the molecular level to achieve the desired application goal. Several experimental studies have outlined interactions between gold NPs (AuNPs) and model membranes, including their adsorption dynamics. − In tandem, computational simulations have analyzed interfacial nanomaterial–supported lipid bilayer (SLB) interactions. − Results highlight the importance of the particle composition, ligand-cap dynamics, and the underlying membrane composition.…”

Vascular Endothelial Growth Factor Receptor-1 Modulates Hypoxia-Mediated Endothelial Senescence and Cellular Membrane Stiffness via YAP-1 Pathways

“…These parameters cumulatively are used to quantify NPs of various biological entities (Kasas and Dietler, 2008;Redondo-Morata et al, 2014;Kenchappa et al, 2020). In our previous studies, we have shown the importance of optimizing the ramping parameters behind the acquisition of useful and informative F-S curves (Kulkarni et al, 2021;Kulkarni et al, 2022;Kulkarni et al, 2019a). In the present study, we exposed HUVECs to various durations of hypoxia (24, 48, 72 h) and performed nanoindentation using optimized ramping parameters.…”