Protein Corona Research Articles

Complement opsonized protein corona activated by precoated immunoglobulin enables neutrophil-hitchhiking for rapid and enhanced drug delivery for acute liver failure recovery

In acute inflammation, the heightened activation and recruitment of immune cells present an opportunity to leverage them as natural carriers for efficient transport of diagnostic probes and nanotherapeutics. Although complement opsonization facilitates the internalization of nanomedicines by activated neutrophils, the development of strategies to specifically augment complement deposition remains a challenge. Herein, we engineer silymarin-loaded liposomes (Lips) coated with immunoglobulin G (IgG) to enhance complement 3 (C3) deposition in the protein corona, thereby enabling neutrophil-mediated, precise targeting to the site of inflammation. Through the examination of various serum proteins, we discover that IgG adsorption, particularly with its Fc portion exposed, prominently promotes C3 enrichment in the protein corona, resulting in C3 cleavage into iC3b. This facilitates the uptake of C3-enriched Lips by activated neutrophils with elevated C3 receptor expression, thus improving the efficiency and specificity of nanomedicine delivery to inflammatory site. Following the formation of neutrophil extracellular traps, the released nanomedicine effectively mitigates hepatocyte damage by eliminating accumulated reactive oxygen species and inducing a shift in macrophage polarization towards the anti-inflammatory M2 phenotype. Our IgG-modified nanomedicine demonstrates significant therapeutic efficacy against acute liver failure by regulating the protein corona and hitchhiking neutrophils, offering a promising strategy for efficient and precise treatment of inflammation.

How to describe the time-dependent dissolution of engineered nanomaterials?

Numerous processes such as solubility, agglomeration/aggregation, or protein corona formation may change over time and significantly affect engineered nanomaterial (ENM) structure, property, and availability, resulting in their reduced or increased toxicological activity. Therefore, understanding the dynamics of these processes is essential for assessing and managing the risks of ENMs during their lifecycle, ensuring safety by design. Of these processes, the importance of solubility (i.e., the ability to release ions from the surface) is undeniable. Thus, we propose a practical approach, the Kalapus equation (KEq), to determine ENMs' dissolution over time. As a proof-of-concept, the KEq was applied to determine the solubility of six commercially used metal and metal oxide nanoparticles over time. The KEq exhibited a higher coefficient of determination (R2=0.995-0.999) than the logarithmic equation (R2=0.835-0.986), and the pseudo-first-order equation (R2=0.915-0.994) over a range of experimental data. The newly introduced Kalapus equation outperformed the logarithmic and pseudo-first-order equations when extrapolating beyond the time range in which solubility was experimentally determined. The mean absolute error in solubility prediction for the KEq was 3.29% and 4.28% for the first and second data points, respectively, significantly lower than the 13.46% and 18.05% observed for the pseudo-first-order/first-order equation. The proposed equation can be used as a part of New Generation Risk Assessment (NGRA) methodology, especially new Integrated Approaches to Testing and Assessments (IATAs).

The formation of protein coronas and the interaction of soy protein isolate and whey protein isolate with starch nanoparticles

The influence of food components on the physicochemical properties and biological responses of nanoparticles is pivotal. Since soy protein isolate (SPI) and whey protein isolate (WPI) are commonly used as plant and animal proteins in food, it is imperative to gain comprehensive knowledge of the interaction between SPI/WPI and nanoparticles. Accordingly, we examined the adsorption of SPI and WPI onto starch nanoparticles (SNPs) and their associated interactions. SNP aggregation was observed in the SPI/WPI, and coronas formed around the surfaces of the SNPs. The binding of the protein isolates reduced the zeta potential of the SNPs from −10.34 to −26.5 mV for the SPI and to −21.37 mV for the WPI. However, mixing the SNPs with the protein isolates did not alter the microenvironment of aromatic amino acids and had no substantial impact on the secondary structure of the proteins. Thermogravimetric analysis revealed that the formation of SPI/WPI coronas enhanced the thermal stability of the SNPs. Isothermal titration calorimetry indicated that the adsorption of the SPI and WPI onto the SNPs was driven primarily by weak van der Waals forces and hydrogen bonds. This study facilitates the predicting of organic nanoparticle behaviours in complex food matrix and human gastrointestinal environments.

The role of patient-specific variables in protein corona formation and therapeutic efficacy in nanomedicine.

Despite their potential, the adoption of nanotechnology in therapeutics remains limited, with only around eighty nanomedicines approved in the past 30years. This disparity is partly due to the "one-size-fits-all" approach in medical design, which often overlooks patient-specific variables such as biological sex, genetic ancestry, disease state, environment, and age that influence nanoparticle behavior. Nanoparticles (NPs) must be transported through systemic, microenvironmental, and cellular barriers that vary across heterogeneous patient populations. Key patient-dependent properties impacting NP delivery include blood flow rates, body fat distribution, reproductive organ vascularization, hormone and protein levels, immune responses, and chromosomal differences. Understanding these variables is crucial for developing effective, patient-specific nanotechnologies. The formation of a protein corona around NPs upon exposure to biological fluids significantly alters NP properties, affecting biodistribution, pharmacokinetics, cytotoxicity, and organ targeting. The dynamics of the protein corona, such as time-dependent composition and formation of soft and hard coronas, depend on NP characteristics and patient-specific serum components. This review highlights the importance of understanding protein corona formation across different patient backgrounds and its implications for NP design, including sex, ancestry, age, environment, and disease state. By exploring these variables, we aim to advance the development of personalized nanomedicine, improving therapeutic efficacy and patient outcomes.

Contribution of Cancer-Specific Protein Coronas to the Pro-Tumor Effects of Nanoplastics through Enhanced Cellular Interactions.

The potential impact of nanoplastics (NPs) on human carcinogenic processes is a matter of growing concern, particularly in light of the global plastic pollution crisis. Although the potential effects of NPs on human health have been well investigated, many uncertainties remain regarding their role in tumor behavior. Upon exposure, NPs can enter the bloodstream and are prone to interacting with plasma proteins to form a protein corona (PC), which can influence their interactions with cancer cells. However, how the PCs adsorbed on NPs affect the particle-to-tumor cell interaction and their effect on the tumor biological behavior remain unclear. To better understand the formation of PCs following NPs exposure in the bloodstream under various clinical conditions, we investigated the PC compositions of NPs derived from thyroid cancer (TC) patients and healthy volunteers. Our data revealed a significant enrichment of fibrinogen in the PCs formed on NPs derived from TC patient plasma, which in turn accelerated the endocytosis of NPs into TC cells. In addition, the uptake pathway of NPs into TC cells differed substantially between the two groups studied due to the different PC compositions in cancer patients and healthy individuals. Moreover, intriguingly alterations in the PCs induced by the clinical pathology status were also found to promote NPs engulfment by human macrophages, resulting in potent pro-inflammatory effects, in turn exerting pro-tumor effects. These findings emphasize the importance of considering the significance of a realistic biological identity on NPs and their interactions with cancer cells and also pinpoint the implications of the carcinogenesis outcomes of NPs exposure in humans.

Abstract PR013: Highly accurate detection of early-stage colorectal cancer using tumor and immune extracellular vesicles biomarkers

Abstract Introduction: Colorectal cancer (CRC) is one of the most common cancers worldwide, and early detection is critical for successful treatment and improved survival rates. However, precancerous and early-stage CRC present significant diagnostic challenges due to the small size of lesions and the very low expression of tumor-specific biomarkers in the bloodstream. Current genomics-based diagnostic methods struggle to detect these lesions, making it imperative to develop more sensitive and specific approaches. Circulating extracellular vesicles (EVs) are emerging as a promising solution for early-stage CRC detection, since EVs are produced by tumor cells as well as the tumor microenviroment and host immune cells. Thus, EVs may offer the means to detect and monitor small early-stage tumors through both direct detection of tumor- associated biomarkers as well through tumor specific host response and tumor microenvironment biomarkers. Methods: To identify novel early-stage CRC specific biomarkers, we performed proteomics analysis on EVs purified from patient plasma using size exclusion chromatography and a proprietary buffer system that enhances EV and corona protein recovery. TrueDiscovery™ Data-independent acquisition (DIA) mass spectrometry (MS) analysis was conducted on 24 pre- cancer/stage 0 and 25 stage 1 CRC patients and 75 normal patient plasma samples. An in-house developed machine learning pipeline was used to identify differentially expressed proteins and model candidate multiplexes to identify those with extremely high diagnostic accuracy (> 0.99). Results: An average of ∼2,500 proteins were identified per sample and included in bioinformatics analysis. Comparative analysis between control patient EVs and either pre- cancerous/Stage 0, or Stage 1 colon cancer EVs using an in-house Machine Learning pipeline identified 336 and 493 differentially expressed proteins for each group (FDR adjusted p-value < 0.001). Of these, the best 17 pre/stage 0 proteins and 53 stage 1 proteins were trained and tested using Support Vector Machine to assess all potential 3-plexes in order to identify those with mean diagnostic accuracy > 99%. This yielded 5 3-plexes in precancer/stage 0 and 34 3-plexes in stage 1 with near perfect diagnostic accuracy. These plexes are currently being assessed in single analyte and multiplex immunoassays with the goal of translating candidate 3-plexes to the clinical. Conclusions: Key biomarkers from CRC patient EVs indicate the potential to detect and diagnose early-stage and low tumor burden cancers using our methods. Interestingly, the most accurate 3-plexes consist of proteins from immune, inflammatory and metabolic processes, suggesting that for the detection of early stage cancer it may be critical to include both tumor and host specific biomarkers. Citation Format: Poorva Mudgal, Cheryl Bandoski, Zachary Opheim, Martin Mehnert, Valesca Anschau, Issa Isaac, Alan Ezrin, Todd Hembrough. Highly accurate detection of early-stage colorectal cancer using tumor and immune extracellular vesicles biomarkers [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr PR013.

Small molecule modulation of protein corona for deep plasma proteome profiling

The protein corona formed on nanoparticles (NPs) has potential as a valuable diagnostic tool for improving plasma proteome coverage. Here, we show that spiking small molecules, including metabolites, lipids, vitamins, and nutrients into plasma can induce diverse protein corona patterns on otherwise identical NPs, significantly enhancing the depth of plasma proteome profiling. The protein coronas on polystyrene NPs when exposed to plasma treated with an array of small molecules allows for the detection of 1793 proteins marking an 8.25-fold increase in the number of quantified proteins compared to plasma alone (218 proteins) and a 2.63-fold increase relative to the untreated protein corona (681 proteins). Furthermore, we discovered that adding 1000 µg/ml phosphatidylcholine could singularly enable the detection of 897 proteins. At this specific concentration, phosphatidylcholine selectively depletes the four most abundant plasma proteins, including albumin, thus reducing the dynamic range of plasma proteome and enabling the detection of proteins with lower abundance. Employing an optimized data-independent acquisition approach, the inclusion of phosphatidylcholine leads to the detection of 1436 proteins in a single plasma sample. Our molecular dynamics results reveal that phosphatidylcholine interacts with albumin via hydrophobic interactions, H-bonds, and water bridges. The addition of phosphatidylcholine also enables the detection of 337 additional proteoforms compared to untreated protein corona using a top-down proteomics approach. Given the critical role of plasma proteomics in biomarker discovery and disease monitoring, we anticipate the widespread adoption of this methodology for the identification and clinical translation of biomarkers.

Recent Advances in Simulation Studies on the Protein Corona

When flowing through the blood stream, drug carriers such as nanoparticles encounter hundreds of plasma proteins, forming a protein layer on the nanoparticle surface, known as the “protein corona”. Since the protein corona influences the size, shape, and surface properties of nanoparticles, it can modulate their circulating lifetime, cytotoxicity, and targeting efficiency. Therefore, understanding the mechanism of protein corona formation at the atomic scale is crucial, which has become possible due to advances in computer power and simulation methodologies. This review covers the following topics: (1) the structure, dynamics, and composition of protein corona on nanoparticles; (2) the effects of protein concentration and ionic strength on protein corona formation; (3) the effects of particle size, morphology, and surface properties on corona formation; (4) the interactions among lipids, membranes, and nanoparticles with the protein corona. For each topic, mesoscale, coarse-grained, and all-atom molecular dynamics simulations since 2020 are discussed. These simulations not only successfully reproduce experimental observations but also provide physical insights into the protein corona formation. In particular, these simulation findings can be applied to manipulate the formation of a protein corona that can target specific cells, aiding in the rational design of nanomedicines for drug delivery applications.

Synthetic Nanocapsules with Tailored Surface Chemistry for Lung-Specific Protein Delivery and Cancer Immunotherapy.

Efficient delivery of therapeutic proteins remains a major challenge in developing effective immunotherapies and treatments for genetic disorders due to the limited tissue targeting capability of native proteins. In this study, the design and synthesis of protein nanocapsules (NCs) that achieve lung-specific delivery of therapeutic proteins are reported. These NCs are synthesized through a surface modification process that involves coating protein with functional monomers and cross-linkers, followed by in situ polymerization to create a protective shell on the protein surface with tailored surface chemistry. This approach preserves protein integrity and significantly enhances delivery efficiency and tissue specificity. Notably, it is shown that protein@NC with guanidine-rich surfaces exhibit exceptional lung-targeting capabilities. This is likely attributed to the formation of a vitronectin-rich protein corona, which facilitates receptor-mediated endocytosis by lung cells. The platform effectively delivers various proteins, such as ovalbumin, to antigen-presenting cells (APCs) in the lung, thereby enhancing antigen presentation and offering a promising strategy for cancer immunotherapy. These findings provide a significant advancement in tissue-specific protein delivery and hold the potential for targeted cancer immunotherapy.

Study of Hard Protein Corona on Lipid Surface of Composite Nanoconstruction.

The composition of the protein corona covering any nanoparticle (NP) when it enters a biological fluid determines the parameters of the NP's interaction with the body. To "control" these parameters, it is important to know the composition of the protein corona, the determination of which is a complex task associated with the two-layer organization of the corona (hard and soft coronas). In a previous publication, we reported obtaining lipid-coated NPs with a full protein corona, isolating them, and proving the presence of the corona on the surface of the NPs. This work reports on the preparation, isolation, and purification of lipid-coated NPs bearing a hard corona. The protein corona composition was determined by using the LC-MS/MS method. Thirty-seven serum proteins were identified with a high degree of reliability. The hard corona contained various apolipoproteins, including apolipoprotein E, which can potentially affect the penetration of NPs into the cell.

Essential protocols for decoding the composition and the functional effects of the nanoparticle protein corona

Identifying the function and composition of the protein corona (i.e., the set of host proteins interacting with nanoparticles) is considered a crucial step in the development of nanoparticles for medical and pharmacological applications. Evidence suggests that host proteins can alter NP stability, biocompatibility, and pharmacokinetics features. Therefore, in this review, we provide an updated conceptual, methodological, and experimental guideline for the study of the NP protein corona. We surveyed recent literature (2009–2024) focusing on in vitro and in vivo studies. We show that several methods, including shot-gun proteomics, protein identification after in-gel digestion, and TMT proteomics, must be carefully applied and integrated to shed light on this complex phenomenon. Hence, we discuss in detail the relative protocols, highlighting the importance of the experimental conditions, ranging from the administration route to basic, but determinant, parameters like the kind of biological host fluids, the incubation times and the NP concentrations. Additionally, we propose a series of protocols that involve studying the protein corona using purified serum or plasma proteins, as well as sera depleted of specific complement proteins, to investigate the role of their deposition on the nanoparticle surface. We also explore how the role of the protein corona in inducing uptake by phagocytic cells can be examined; finally, we discuss several methodological approaches to study the effects of different coatings on the composition of the protein corona. Available data indicated that it is possible to characterize and punctually study the differential adsorption of specific proteins onto the nanoparticle surface. This allows designing NP chemical coatings features to actively guide the protein corona formation, thus improving nanotheranostic development.

Structuring lipid nanoparticles, DNA, and protein corona into stealth bionanoarchitectures for in vivo gene delivery

Lipid nanoparticles (LNPs) play a crucial role in addressing genetic disorders, and cancer, and combating pandemics such as COVID-19 and its variants. Yet, the ability of LNPs to effectively encapsulate large-size DNA molecules remains elusive. This is a significant limitation, as the successful delivery of large-size DNA holds immense potential for gene therapy. To address this gap, the present study focuses on the design of PEGylated LNPs, incorporating large-sized DNA, departing from traditional RNA and ionizable lipids. The resultant LNPs demonstrate a unique particle morphology. These particles were further engineered with a DNA coating and plasma proteins. This multicomponent bionanoconstruct exhibits enhanced transfection efficiency and safety in controlled laboratory settings and improved immune system evasion in in vivo tests. These findings provide valuable insights for the design and development of bionanoarchitectures for large-size DNA delivery, opening new avenues for transformative gene therapies.

Glycopolymeric Nanoparticles Enrich Less Immunogenic Protein Coronas, Reduce Mononuclear Phagocyte Clearance, and Improve Tumor Delivery Compared to PEGylated Nanoparticles.

Nanoparticles (NPs) offer significant promise as drug delivery vehicles; however, their in vivo efficacy is often hindered by the formation of a protein corona (PC), which influences key physiological responses such as blood circulation time, biodistribution, cellular uptake, and intracellular localization. Understanding NP-PC interactions is crucial for optimizing NP design for biomedical applications. Traditional approaches have utilized hydrophilic polymer coatings like polyethylene glycol (PEG) to resist protein adsorption, but glycopolymer-coated nanoparticles have emerged as potential alternatives due to their biocompatibility and ability to reduce the adsorption of highly immunogenic proteins. In this study, we synthesized and characterized glycopolymer-based poly[2-(diisopropylamino)ethyl methacrylate-b-poly(methacrylamidoglucopyranose) (PDPA-b-PMAG) NPs as an alternative to PEGylated NPs. We characterized the polymers using a range of techniques to establish their molecular weight and chemical composition. PMAG and PEG-based NPs showed equivalent physicochemical properties with sizes of ∼100 nm, spherical morphology, and neutral surface charges. We next assessed the magnitude of protein adsorption on both NPs and catalogued the identity of the adsorbed proteins using mass spectrometry-based techniques. The PMAG NPs were found to adsorb fewer proteins in vitro as well as fewer immunogenic proteins such as Immunoglobulins and Complement proteins. Flow cytometry and confocal microscopy were employed to examine cellular uptake in RAW 264.7 macrophages and MDA-MB-231 tumor cells, where PMAG NPs showed higher uptake into tumor cells over macrophages. In vivo studies in BALB/c mice with orthotopic 4T1 breast cancer xenografts showed that PMAG NPs exhibited prolonged circulation times and enhanced tumor accumulation compared to PEGylated NPs. The biodistribution analysis also revealed greater selectivity for tumor tissue over the liver for PMAG NPs. These findings highlight the potential of glycopolymeric NPs to improve tumor targeting and reduce macrophage uptake compared to PEGylated NPs, offering significant advancements in cancer nanomedicine and immunotherapy.

Microplastics interactions and transformations during in vitro digestion with milk

Despite increasing awareness of microplastic contamination in food, the specific interactions and transformations of microplastics during digestion remain poorly understood. The study investigates the behavior of microplastics during in vitro digestion processes and their interaction with components of milk. The in vitro digestion studies are conducted to study the changes in microplastics in simulated digestive fluids, with and without milk. The study revealed that microplastics undergo significant changes in size, surface morphology, and chemical properties when subjected to digestion with and without milk. Notably, microplastics digested with milk exhibited a 15–25% increase in aggregation due to protein corona formation, enhancing their potential for interactions within biological systems. FTIR analysis revealed the formation of OH and CO groups in digested microplastics, indicating hydrolysis and structural changes. The stronger peaks in the 1630–1650 cm−1 range suggest significant adsorption of milk proteins, highlighting the complex interactions during digestion. Additionally, the chemicals and additives leached from microplastics into digesta raising the concerns about their potential health effects. The study emphasizes the necessity for additional research and regulatory measures to address the risks associated with microplastic contamination in food.

Nanoetched Stainless Steel Architecture Enhances Cell Uptake of Biomacromolecules and Alters Protein Corona Abundancy.

Nanotexture on biocompatible surfaces promotes cell adhesion and proliferation. High aspect ratio nanoachitecture serves as an ideal interface between implant materials and host cells that is well-suited for localized therapeutic delivery. Despite this potential, nanotextured surfaces have not been widely applied for biomacromolecule delivery. Here, we employed a low-cost, industrially relevant nanoetching process to modify the surface of biocompatible stainless steel 316 (SS316L), creating nanotextured SS316L (NT-SS316L) as a material for intracellular biomacromolecule delivery. As biomacromolecule cargoes are adsorbed to the steel and ultimately would be used in protein-rich environments, we performed serum protein corona analysis on unmodified SS316L and NT-SS316L using tandem mass spectrometry. We observed an increase in proteins associated with cell adhesion on the surface of NT-SS316L compared to that of SS316L, supporting literature reports of enhanced adhesion on nanotextured materials. For delivery to adherent cells, a "hard corona" of model biomacromolecule cargoes including superfolder green fluorescent protein (sfGFP) charge variants, cytochrome c, and siRNA was adsorbed on NT-SS316L to assess delivery. Nanotextured surfaces enhanced cellular biomacromolecule uptake and delivered cytosolic-functional proteins and nucleic acids through energy-dependent endocytosis. Collectively, these findings indicate that NT-SS316L holds potential as a surface modification for implants to achieve localized drug delivery for a variety of biomedical applications.

PEGylation Can Effectively Strike a Balance in siRNA Delivery Performances of Guanidinylated Linear Synthetic Polypeptides with Potential Use for Transcriptional Gene Silencing.

The prevailing design philosophy for polymeric vectors delivering siRNA is rooted in the post-transcriptional gene silencing (PTGS) mechanism. Yet, the transcriptional gene silencing (TGS) mechanism offers a potentially more durable silencing effect, which necessitates efficient siRNA delivery into the nucleus. However, it remains a challenge for the polymeric vectors to efficiently deliver siRNA into the nucleus. We have explored guanidinylated cyclic synthetic polypeptides (GCSPs) to enhance the nuclear delivery of siRNA, but an increased cytotoxicity and difficulty in producing the GCSPs on a large scale limit their utility. Herein, we simply prepare PEGylated guanidinylated linear synthetic polypeptides (PGLSPs) exhibiting improved membrane penetration, direct siRNA transport to the nucleus, reduced toxicity, high cellular uptake, and mitigation of protein corona formation. The PEGylation can effectively balance the vector's nuclear delivery capacity with other critical aspects of performances for siRNA delivery. Therefore, the PGLSPs hold promise as TGS-based delivery vectors, offering potential for future therapeutic applications.

Impact of Protein Coronas on Lipid Nanoparticle Uptake and Endocytic Pathways in Cells.

Lipid nanoparticles (LNPs), widely used in disease diagnosis and drug delivery, face the challenge of being surrounded by biological macromolecules such as proteins upon entering the human body. These molecules compete for binding sites on the nanoparticle surfaces, forming a protein corona. The impact of different types of protein coronas on LNP delivery remains unclear. In this study, we employed a newly developed, highly sensitive LNP labeling platform and analyzed the endocytosis of HeLa cells under different nutritional conditions using proteomics to address this critical issue. Our research found that under conditions of complete medium and amino acid starvation, most DNA-FITC vesicles in HeLa cells were located in the perinuclear region 4 h after transfection. In contrast, under serum starvation conditions, only a small portion of DNA-FITC vesicles were in the perinuclear region. On the other hand, through proteomics, we discovered that cells that were enriched in amino acids and complete medium contained more proteins, whereas those under serum starvation had relatively fewer enriched proteins. Through KEGG pathway enrichment analysis, we identified the phagosome and endocytosis pathways as particularly important. Lastly, differential analysis of proteins in these pathways revealed that proteins such as F-actin, Coronin, vATPase, TUBA, TUBB, MHCII, and TSP may have significant impacts on cellular endocytosis. Our research findings indicate that it is necessary to regulate cellular endocytosis based on different protein coronas to achieve optimal cytoplasmic release.

Machine Learning-Aided 3D Dynamic SERS Strategy for Physiological Mapping: Biotoxicity of Environmentally Dimensional Aged Nanoplastics and Corresponding Protein Corona Complexes.

Nanoplastics (NPs) are emerging pollutants that undergo inevitable aging in the environment, raising concerns about human exposure and health hazards. Research on the cytotoxicity of various polymer types of NPs, aged nanoplastics (aNPs), and their interactions with proteins (aNPs-protein corona) is still nascent. Traditional cytotoxicity detection methods often rely on end point assays with restricted temporal resolution and analysis of single or multiple biomarkers. Here, we propose a novel approach integrating the 3D dynamic SERS strategy (DSS) with machine learning to rapidly analyze the cell fate and death modes induced by NPs, aNPs, and aNPs-protein corona complexes at the molecular level. PS, PVC, PMMA, and PC products from the water environment were used to prepare the corresponding NPs, and the impact of UV irradiation on their physicochemical properties was examined. DSS systematically maps the molecular changes in the cellular secretome caused by these NPs. Machine learning effectively extracts information from complex spectra, differentiating between biological samples. Results show prolonged UV exposure increases cell sensitivity to ferroptosis and cytotoxicity in various aNPs, while the protein corona on aNPs significantly mitigates toxicity associated with surface oxygen-containing functional groups, resulting in a reduced similarity to ferroptosis signatures. 3D DSS with machine learning technique analyzes the overall metabolite profile at the molecular level rather than individual biomarkers. This study is the first attempt to compare the biotoxicity of diverse polymer NPs, aNPs, and aNPs-protein coronas at cellular and molecular levels in human hepatocytes, enhancing our understanding of the complex biological impacts of NPs.

Dual-functionalized architecture enables stable and tumor cell-specific SiO2NPs in complex biological fluids.

Most commercial anticancer nanomedicines are administered intravenously. This route is fast and precise as the drug enters directly into the systemic circulation, without undergoing absorption processes. When nanoparticles come into direct contact with the blood, however, they interact with physiological components that can induce colloidal destabilization and/or changes in their original biochemical identity, compromising their ability to selectively accumulate at target sites. In this way, these systems usually lack active targeting, offering limited therapeutic effectiveness. In the literature, there is a paucity of in-depth studies in complex environments to evaluate nanoparticle stability, protein corona formation, hemolytic activity, and targeting capabilities. To address this issue, fluorescent silica nanoparticles (SiO2NPs) are here functionalized with zwitterionic (kinetic stabilizer) and folate groups (targeting agent) to provide selective interaction with tumor cell lines in biological media. The stability of these dually functionalized SiO2NPs is preserved in unprocessed human plasma while yielding a decrease in the number of adsorbed proteins. Experiments in murine blood further proved that these nanoparticles are not hemolytic. Remarkably, the functionalized SiO2NPs are more internalized by tumor cells than their healthy counterparts. Investigations of this nature play a crucial role in garnering results with greater reliability, allowing the development of nanoparticle-based pharmaceutical drugs that exhibit heightened efficacy and reduced toxicity for medical purposes.

Preferred Lung Accumulation of Polystyrene Nanoplastics with Negative Charges.

With the increasing presence of nanoplastics (NPs) in the human bloodstream, it is urgent to investigate their tissue accumulation and potential health risks. This study examines the effects of the size and surface charges of polystyrene (PS) NPs on lung accumulation. Using magnetic separation, we identified the protein corona composition on iron-core PS NPs, revealing the enrichment of vitronectin and fibrinogen. The corona promotes integrin αIIbβ3 receptor-mediated uptake by lung endothelial cells, explaining that both the corona composition and protein structure determine preferred localization of negatively charged PS NPs in the lung. This study uncovers the role of protein corona in NP uptake and the way NPs enter the lung, emphasizing the need to consider interactions between nanoplastics with varying surface characteristics and biological molecules in the nanotoxicological field.