Corona Formation Research Articles

Bio- and eco-corona related to plants: Understanding the formation and biological effects of plant protein coatings on nanoparticles

The thriving nano-enabled agriculture facilitates the interaction of nanomaterials with plants. Recently, these interactions and their biological effects are receiving increasing attention. Upon entering plants via leaves, roots, stems, and other organs, nanoparticles adsorb numerous biomolecules inside plants and form bio-corona. In addition, nanoparticles that enter plants through roots may have formed eco-corona with root exudates in the rhizosphere environment before contacting with plant exogenous proteins. The most significant biological effects of plant protein corona include changes in protein structure and function, as well as changes in nanoparticle toxicity and targeting ability. However, the mechanisms, particularly how protein corona affects plant protein function, plant development and growth, and rhizosphere environment properties, require further investigation. Our review summarizes the current understanding of the formation and biological effects of nanoparticle-plant protein corona and provides an outlook on future research.

Hemolytic Activity of Nanoparticles as a Marker of Their Hemocompatibility.

The potential use of nanomaterials in medicine offers opportunities for novel therapeutic approaches to treating complex disorders. For that reason, a new branch of science, named nanotoxicology, which aims to study the dangerous effects of nanomaterials on human health and on the environment, has recently emerged. However, the toxicity and risk associated with nanomaterials are unclear or not completely understood. The development of an adequate experimental strategy for assessing the toxicity of nanomaterials may include a rapid/express method that will reliably, quickly, and cheaply make an initial assessment. One possibility is the characterization of the hemocompatibility of nanomaterials, which includes their hemolytic activity as a marker. In this review, we consider various factors affecting the hemolytic activity of nanomaterials and draw the reader's attention to the fact that the formation of a protein corona around a nanoparticle can significantly change its interaction with the red cell. This leads us to suggest that the nanomaterial hemolytic activity in the buffer does not reflect the situation in the blood plasma. As a recommendation, we propose studying the hemocompatibility of nanomaterials under more physiologically relevant conditions, in the presence of plasma proteins in the medium and under mechanical stress.

Biological Features of Nanoparticles: Protein Corona Formation and Interaction with the Immune System.

Nanoparticles (NPs) are versatile candidates for nanomedical applications due to their unique physicochemical properties. However, their clinical applicability is hindered by their undesirable recognition by the immune system and the consequent immunotoxicity, as well as their rapid clearance in vivo. After injection, NPs are usually covered with layers of proteins, called protein coronas (PCs), which alter their identity, biodistribution, half-life, and efficacy. Therefore, the characterization of the PC is for in predicting the fate of NPs in vivo. The aim of this review was to summarize the state of the art regarding the intrinsic factors closely related to the NP structure, and extrinsic factors that govern PC formation in vitro. In addition, well-known opsonins, including complement, immunoglobulins, fibrinogen, and dysopsonins, such as histidine-rich glycoprotein, apolipoproteins, and albumin, are described in relation to their role in NP detection by immune cells. Particular emphasis is placed on their role in mediating the interaction of NPs with innate and adaptive immune cells. Finally, strategies to reduce PC formation are discussed in detail.

Thermo-sensitive amylase-starch double-layer polymer nanoparticles with self-polishing and protein corona-free property for drug delivery applications

Protein corona formation can lead to obstructive screening of targeting groups of nanoparticles (NPs). Also, the targeting groups are subjected to physiochemical interactions when exposed to solvents. Here, these two factors can influence NP targeting efficiency. Therefore, it is necessary to prepare a general method of preparing an anti-fouling NPs with protected targeting groups. Here, we designed α-amylase-starch double-layer coated poly (methyl methacrylate-co-acrylic acid) NPs (α-ams-SCMMA NPs), functionalized with aptamer targeting groups and doped with Tetrakis(para-hydraoxylphenyl) porphyrin (TPPOH) as a payload drug. Natural polysaccharide starch and enzyme α-amylase were applied here for thermo-sensitive activation of starch hydrolyzation in order to render the NPs' self-polishing from protein corona effects. During incubation with serum media, the protein corona was formed at the exterior shell of NPs, while the self-polishing process was activated to remove the “protein fouling” when the incubation temperature increased to 37 °C (body temperature). Mechanistically, the starch layer of α-ams-SCMMA NPs was readily hydrolysed by α-amylase, whereby the adsorbed protein corona could be efficiently eliminated and the targeting groups were then presented. With this unique self-polishing NP design, we believe our method can be applied for potential NP applications in cancer therepy due to excellent antifouling property and protected targeting groups.

Insight into the formation and biological effects of natural organic matter corona on silver nanoparticles in water environment using biased cyclical electrical field-flow fractionation

Natural organic matter (NOM) readily interacts with nanoparticles, leading to the formation of NOM corona structures on their surface. NOM corona formation is closely related to the surface coatings and bioavailability of nanoparticles. However, the mechanism underlying NOM corona formation on silver nanoparticles (AgNPs) remains largely unknown due to the lack of effective analytical methods for identifying the changes in the AgNP surface. Herein, the separation ability of biased cyclical electrical field-flow fractionation (BCyElFFF) for same-sized polyvinyl pyrrolidone-coated and poly(ethylene glycol)-coated silver nanoparticles (AgNPs) with different electrophoretic mobilities was evaluated under various electrical conditions. Then, the mechanism behind the NOM corona formation on these AgNP surfaces was elucidated based on the changes in the elution time and off-line characterization of the collected fractions during their elution time in a BCyElFFF run. Finally, the survival rates of E. coli exposed to polyvinyl pyrrolidone-coated and poly(ethylene glycol)-coated AgNPs with or without NOM collected during repeated BCyElFFF runs were observed to increase with increasing NOM concentration, clearly demonstrating the negative effect of NOM corona structures on the bioavailability of AgNPs. These findings highlight the powerful separation and isolation ability of BCyElFFF in studying the transformation and fate of nanoparticles in aqueous environments.

Mimicking Pseudo-Virion Interactions with Abiotic Surfaces: Deposition of Polymer Nanoparticles with Albumin Corona.

Adsorption of human serum albumin (HSA) molecules on negatively charged polystyrene microparticles was studied using the dynamic light scattering, the electrophoretic and the solution depletion methods involving atomic force microscopy. Initially, the physicochemical characteristics of the albumin comprising the hydrodynamic diameter, the zeta potential and the isoelectric point were determined as a function of pH. Analogous characteristics of the polymer particles were acquired, including their size and zeta potential. The formation of albumin corona on the particles was investigated in situ by electrophoretic mobility measurements. The size, stability and electrokinetic properties of the particles with the corona were also determined. The particle diameter was equal to 125 nm, which coincides with the size of the SARS-CoV-2 virion. The isoelectric point of the particles appeared at a pH of 5. The deposition kinetics of the particles was determined by atomic force microscopy (AFM) under diffusion and by quartz microbalance (QCM) under flow conditions. It was shown that the deposition rate at a gold sensor abruptly vanished with pH following the decrease in the zeta potential of the particles. It is postulated that the acquired results can be used as useful reference systems mimicking virus adsorption on abiotic surfaces.

Sex-Specific Silica Nanoparticle Protein Corona Compositions Exposed to Male and Female BALB/c Mice Plasmas.

As various nanoparticles (NPs) are increasingly being used in nanomedicine products for more effective and less toxic therapy and diagnosis of diseases, there is a growing need to understand their biological fate in different sexes. Herein, we report a proof-of-concept result of sex-specific protein corona compositions on the surface of silica NPs as a function of their size and porosity upon incubation with plasma proteins of female and male BALB/c mice. Our results demonstrate substantial differences between male and female protein corona profiles on the surface of silica nanoparticles. By comparing protein abundances between male and female protein coronas of mesoporous silica nanoparticles and Stöber silica nanoparticles of ∼100, 50, and 100 nm in diameter, respectively, we detected 17, 4, and 4 distinct proteins, respectively, that were found at significantly different concentrations for these constructs. These initial findings demonstrate that animal sex can influence protein corona formation on silica NPs as a function of the physicochemical properties. A more thorough consideration of the role of plasma sex would enable nanomedicine community to design and develop safer and more efficient diagnostic and therapeutic nanomedicine products for both sexes.

Assessment of the Cytotoxicity of Nano Gallium Liquid Metal Droplets for Biomedical Applications

Gallium-based liquid metals (LMs) have emerged as novel materials for biomedical applications. Gallium (Ga) is considered significantly less toxic than other LMs, such as mercury. However, Ga droplets have not been thoroughly assessed for their cytotoxicity. Here, we report the concentration-dependent cytotoxic effects of Ga nano- and microdroplets on HaCaT cells, a human keratinocyte cell line. We found that Ga droplets demonstrated substantial cytotoxic effects at 1000 μg/mL after 24 h exposure, while lower concentrations such as 500 μg/mL indicated only a slight reduction in cell viability at longer exposure times. At higher concentrations, it was observed that the cells were unable to form confluent layers and the cytoplasmic membrane became damaged. The formation of a protein corona on the Ga droplets in cell media is also described, which will be an important consideration for future research on Ga droplets for bio-applications. Importantly, this work provides an indication of the concentration at which Ga droplets become damaging to human cells, which will be useful information for a wide range of researchers and regulatory bodies.

Protein corona mediated liposomal drug delivery for bacterial infection management

Liposomes have been widely investigated as a class of promising antibiotic delivery systems for the treatment of life-threatening bacterial infections. However, the inevitable formation of protein corona on the liposomal surface can heavily impact in vivo performance. A better understanding of the effects of protein corona on liposomal behavior can significantly improve antibacterial liposomal drug development. Here, the critical role of protein corona in mediating liposome-bacteria interactions was elucidated. Adsorption of negatively charged protein on cationic liposome weakened electrostatic attraction-enhanced liposomal binding to the bacteria. Cumulative complement deposition on anionic liposome composed of phosphatidylglycerol (DSPG sLip) contributed to a superior binding affinity of DSPG sLip to planktonic bacteria and biofilms, which was exploited to enhance bacteria-targeted drug delivery. In both S. aureus-related osteomyelitis and pneumonia mice models, DSPG sLip was demonstrated as a promising antibiotic nanocarrier for managing MRSA infection, indicating the benefits of lipid composition-based protein corona modulation in liposomal antibiotic delivery for bacterial infection treatment.

Immunotoxicity of nanomaterials in health and disease: Current challenges and emerging approaches for identifying immune modifiers in susceptible populations.

Nanosafety assessment has experienced an intense era of research during the past decades driven by a vivid interest of regulators, industry, and society. Toxicological assays based on in vitro cellular models have undergone an evolution from experimentation using nanoparticulate systems on singular epithelial cell models to employing advanced complex models more realistically mimicking the respective body barriers for analyzing their capacity to alter the immune state of exposed individuals. During this phase, a number of lessons were learned. We have thus arrived at a state where the next chapters have to be opened, pursuing the following objectives: (1) to elucidate underlying mechanisms, (2) to address effects on vulnerable groups, (3) to test material mixtures, and (4) to use realistic doses on (5) sophisticated models. Moreover, data reproducibility has become a significant demand. In this context, we studied the emerging concept of adverse outcome pathways (AOPs) from the perspective of immune activation and modulation resulting in pro-inflammatory versus tolerogenic responses. When considering the interaction of nanomaterials with biological systems, protein corona formation represents the relevant molecular initiating event (e.g., by potential alterations of nanomaterial-adsorbed proteins). Using this as an example, we illustrate how integrated experimental-computational workflows combining in vitro assays with in silico models aid in data enrichment and upon comprehensive ontology-annotated (meta)data upload to online repositories assure FAIRness (Findability, Accessibility, Interoperability, Reusability). Such digital twinning may, in future, assist in early-stage decision-making during therapeutic development, and hence, promote safe-by-design innovation in nanomedicine. Moreover, it may, in combination with in silico-based exposure-relevant dose-finding, serve for risk monitoring in particularly loaded areas, for example, workplaces, taking into account pre-existing health conditions. This article is categorized under: Toxicology and Regulatory Issues in Nanomedicine > Toxicology of Nanomaterials.

A Systematic Review on the Impact of Micro-Nanoplastics Exposure on Human Health and Diseases

Plastic production is continuously increasing worldwide for daily use. Micro-plastics and nano-plastics remain major emerging pollutants and threaten the environment, ecosystem, human health, and well-being. Micro-nanoplastics (MNPs) are also exposed to humans through cosmetics, inhalation, ingestion, drinking water, dietary sources, and drug formulations. Oral uptake is the major among the different exposure routes of MNPs to humans. After entry, it gets absorbed due to its nano size (˂100 nm) and easily distributed to all parts of the body through blood, affecting multiple organs, especially vital organs of the human body leading to severe diseases. It causes cancer, heart, liver, and kidney diseases, crosses the blood-brain barrier, and affects the brain. Its adsorption with protein leads to multi-layered corona formation in human blood plasma. MNPs interact with immune cells and induce pro-inflammatory mediators, inflammatory reactions, reactive oxygen species (ROS) production, and associated cytotoxicity. MNPs suppress T lymphocyte activity which results in a lack of immune regulation leading to autoimmune diseases. Hence, it is necessary to understand the impact of MNPs exposure on humans. Strict control measures for the production and use of plastics and developing appropriate strategies for safe disposal would prevent MNPs-mediated toxicity in humans.

Peptide ligands on the PEGylated nanoparticle surface and human serum composition are key factors for the interaction between immune cells and nanoparticles

The architecture of a nanoparticles’ surface formed due to a modification with a ligand and protein corona formation in biofluids is critical for interactions with cells in vivo. Here we studied interactions of immune cells with magnetic nanoparticles (MNPs) covalently modified with polyethylene glycol (PEG) and their counterparts conjugated with peptides: a pH (low) insertion peptide (pHLIP) and cycloRGD as a targeting ligand in human serum. The conjugation of MNPs-PEG with pHLIP, but not with cycloRGD, enhanced the association of these particles with mononuclear phagocytic cells in vitro and in vivo. We did not find a clear difference in protein corona composition between the pHLIP-modified and parental PEGylated nanoparticles. Analysis of the effect of autologous human serum on MNP uptake by monocytes showed that the efficiency of endocytosis varies among healthy donors and depends on intrinsic properties of serum. Nevertheless, using classic blood, coagulation, biochemical tests, and anti-PEG IgG serum level, we failed to identify the cause of the observed interdonor variation. These individual differences should be taken into consideration during testing of nanotherapeutics.

Impact of Vroman’s Effect on Pharmacodynamics and Pharmacokinetics on Nanoparticulate Drug Delivery Systems

Published on: October 2022Journal of Young Pharmacists, 2022; 14(4):355-358Review Article | doi:10.5530/jyp.2022.14.72 Authors:Chinthaginjala Haranath1,*, Krishnam Sai Vamsi1, Yousef AM Qarmout1, Hindustan Abdul Ahad2, Samuluru Chandana31Department of Pharmaceutics, Raghavendra Institute of Pharmaceutical Education and Research (RIPER)- Autonomous, Ananthapuramu, Andhra Pradesh, INDIA.2Department of Industrial Pharmacy, Raghavendra Institute of Pharmaceutical Education and Research (RIPER)- Autonomous, Ananthapuramu, Andhra Pradesh, INDIA.3Department of Quality Assurance, Raghavendra Institute of Pharmaceutical Education and Research (RIPER)- Autonomous, Ananthapuramu, Andhra Pradesh, INDIA. Abstract:Nanoparticle drug delivery can produce a unique pharmacokinetic and pharmacodynamic effect and unveiled better safety in vivo and in vitro toxicity profiles when compared with conventional drugs. In 1960, Vroman revealed that competitive protein exchange is a general phenomenon that occurs when a protein mixture adsorbs to a surface is called ‘Vroman’s effect’ which is responsible for the fate of the nanoparticles. The development and formation of protein corona on nanoparticles is due to the adsorption of proteins on the surface. It is vital in specific targeting and alter the size and composition of the nanoparticle. The entropy-driven binding manages this adsorption of protein on the surface. Binding of polyethylene glycol to the nanoparticles decreases the non-specific binding and increases the targeting of tissue via enhanced permeability and retention effect. An individual disease state and components in biofluids can modify the structure and shape of protein corona on nanoparticles. By label-free transducers, we can detect targets using Vroman’s effect. The biological identification of nanoparticles can lead to deleterious consequences, such as immunotoxicity, due to the formation of the protein corona.Keywords: Vromans effect, Nanoparticles, Protein corona, Disease state, Biofluids.

Diffusion 19F-NMR of Nanofluorides: In Situ Quantification of Colloidal Diameters and Protein Corona Formation in Solution.

The NMR-detectability of elements of organic ligands that stabilize colloidal inorganic nanocrystals (NCs) allow the study of their diffusion characteristics in solutions. Nevertheless, these measurements are sensitive to dynamic ligand exchange and often lead to overestimation of diffusion coefficients of dispersed colloids. Here, we present an approach for the quantitative assessment of the diffusion properties of colloidal NCs based on the NMR signals of the elements of their inorganic cores. Benefiting from the robust 19F-NMR signals of the fluorides in the core of colloidal CaF2 and SrF2, we show the immunity of 19F-diffusion NMR to dynamic ligand exchange and, thus, the ability to quantify, with high accuracy, the colloidal diameters of different types of nanofluorides in situ. With the demonstrated ability to characterize the formation of protein corona at the surface of nanofluorides, we envision that this study can be extended to additional formulations and applications.

SARS-CoV-2 Spike Protein (RBD) Subunit Adsorption at Abiotic Surfaces and Corona Formation at Polymer Particles.

The adsorption kinetics of the SARS-CoV-2 spike protein subunit with the receptor binding domain at abiotic surfaces was investigated. A combination of sensitive methods was used such as atomic force microscopy yielding a molecular resolution, a quartz microbalance, and optical waveguide lightmode spectroscopy. The two latter methods yielded in situ information about the protein adsorption kinetics under flow conditions. It was established that at pH 3.5–4 the protein adsorbed on mica and silica surfaces in the form of compact quasi-spherical aggregates with an average size of 14 nm. The maximum coverage of the layers was equal to 3 and 1 mg m−2 at pH 4 and 7.4, respectively. The experimental data were successfully interpreted in terms of theoretical results derived from modeling. The experiments performed for flat substrates were complemented by investigations of the protein corona formation at polymer particles carried out using in situ laser Doppler velocimetry technique. In this way, the zeta potential of the protein layers was acquired as a function of the coverage. Applying the electrokinetic model, these primary data were converted to the dependence of the subunit zeta potential on pH. It was shown that a complete acid-base characteristic of the layer can be acquired only using nanomolar quantities of the protein.

Proteomic Analysis Reveals Distinct Protein Corona Compositions of Citrate- and Riboflavin-Coated SPIONs.

Superparamagnetic iron oxide nanoparticles (SPIONs) are recognized as one of the most beneficial tools for biomedicine, especially in theranostic applications. Even though SPIONs have excellent properties regarding their biocompatibility and unique magnetic properties, they lack stability in biological fluids. To stabilize and increase the specificity of the SPIONs to target desirable cells or tissues, several surface coatings have been introduced. These surface coatings can lead to different preferences of serum protein bindings, which ultimately determine their behaviors in vitro and in vivo. Thus, understanding the interaction of SPIONs with biological systems is important for their biocompatible design and clinical applications. In this study, using proteomic analyses, we analyzed the protein corona fingerprints on SPIONs with two different coatings, including citrate and riboflavin, that have been widely used as surface coatings and ligands for enhancing cellular uptake in breast cancer cells. Though both citrate-coated SPIONs (C-SPIONs) and riboflavin-coated SPIONs (Rf-SPIONs) showed similar sizes and zeta potentials, we found that Rf-SPIONs adsorbed more serum proteins than bare SPIONs (B-SPIONs) or C-SPIONs, which was likely due to the higher hydrophobicity of the riboflavin. The enriched proteins consisted mainly of immune-responsive and blood coagulation proteins with different fingerprint profiles. Cellular uptake studies in MCF-7 breast cancer cells comparing the activities of preformed and in situ coronas showed different uptake behaviors, suggesting the role of protein corona formation in promoting the interaction between the SPIONs and the cells. The results obtained here provide the essential information for further development of the potential strategy to reduce or stimulate immune response in vivo to increase therapeutic applications of both C-SPIONs and Rf-SPIONs.

High Drug Loading Nanoparticles Stabilized with Autologous Serum Proteins Passively Inhibits Tumor Growth.

We report drug nanocrystals stabilized with host-specific serum proteins with high loading (∼63% w/w). The human serum derived curcumin nanoparticles (Cur-NanoSera) showed superior in vitro anticancer efficiency compared to a free drug with substantial hemocompatibility. The preadsorbed protein coating impeded further protein corona formation, even with repeated serum exposures. Acute and subacute toxicity evaluations post single and dual injections of C57BL/6 mice indicated that Cur-NanoSera showed no prominent inflammatory response or organ damage in the in-bred mice. Passive accumulation of Cur-NanoSera in tumor tissue significantly suppressed its growth in a syngeneic breast tumor model in addition to controlling tumor burden associated splenomegaly.

Studying the Interaction Behavior of Protein Coronated Gold Nanorods with Polystyrene Nanoplastics

AbstractThe formation of protein corona and their modulatory effects on the physicochemical and biological activity of the nanomaterial is well established. The active protein corona on the nanoparticles can further mediate interactions with trace amounts of exogenous entities present in the systemic circulation. In the present study, we utilize an in vitro simulation system wherein, protein corona is strategically engineered by varying the serum concentration on a model nanoparticle i. e. cationic‐gold nanorods (GNRs). Quantitative analysis indicated that the 4 nm cationic GNRs can accumulate as high as ∼1 mg/mL protein corona upon incubation with serum. The size distribution pattern of the bound corona and their subsequent impact on the physicochemical properties of GNRs were determined. To understand how these coronated nanorods would interact with a potent pollutant, the second layer of interaction was further introduced by interacting the coronated‐GNRs with 1 ppm nanoplastic. The impact of differently functionalized polystyrene beads (plain, aminated, and carboxylated) on the core size, hydrodynamic diameter, surface charge, and optical properties of the coronated GNRs is highlighted. A complex interplay of several interactive forces and amino acid‐mediated binding of the GNRs to the styrene and benzene rings of the nanoplastic leads to the formation of a coronated‐GNR‐PS complex (>400 nm). Further, upon subjecting the hybrid‐conjugate system to acidic pH conditions (pH‐5.6 and 4.3), a considerable increase in protein leaching was observed. The novel approach of the present study aims to look into the layered interactive patterns for nanomaterials that are utilized for in vivo applications.

Genetically Engineered Protein Corona‐based Cascade Nanozymes for Enhanced Tumor Therapy

AbstractThe emergence of nanomedicine has provided a promising strategy to greatly enhance the therapeutic efficacy of O2‐dependent photodynamic therapy (PDT). However, plasma‐derived protein corona and/or discontinuous O2 supply substantially limit their tumor delivery efficiency and therapeutic outcomes. Herein, protein corona cloaking‐based cascade nanozymes are developed using genetically engineered human ferritin heavy chain nanocages (FTn) as unique pre‐coated protein corona and cascade nanozymes as steady O2 suppliers. Specifically, FTn is coated onto mesoporous silica nanoparticles (MSNs) to form FTn‐based protein corona, providing active targeting of tumor cells by binding with its receptor. In situ synthesis of ultra‐small Au nanoparticles in MSNs, and biomimetic incorporation of Ru nanoclusters into FTn inner cavity showed glucose oxidase‐like activity and catalase‐like activity, respectively. The two nanozymes are incorporated into MSNs nanoplatform to induce cascade and circular catalytic reactions by consuming glucose and H2O2 within the tumor microenvironment. Compared to MSNs alone, the FTn‐based protein corona is capable of efficiently diminishing plasma‐derived protein corona formation to prolong blood circulation time and improving in vitro tumor cell uptake and in vivo tumor accumulation, thereby providing significantly enhanced therapeutic benefits of PDT by combining with the continuously produced O2 of cascade nanozymes.

Tailor-Made Protein Corona Formation on Polystyrene Microparticles and its Effect on Epithelial Cell Uptake.

Microplastic particles are pollutants in the environment with a potential impact on ecology and human health. As soon as microplastic particles get in contact with complex (biological) environments, they will be covered by an eco- and/or protein corona. In this contribution, protein corona formation was conducted under defined laboratory conditions on polystyrene (PS) microparticles to investigate the influence on surface properties, protein corona evolution, particle-cell interactions, and uptake in two murine epithelial cells. To direct protein corona formation, PS particles were preincubated with five model proteins, namely, bovine serum albumin (BSA), myoglobin, β-lactoglobulin, lysozyme, and fibrinogen. Subsequently, the single-protein-coated particles were incubated in a cell culture medium containing a cocktail of serum proteins to analyze changes in the protein corona profile as well as in the binding kinetics of the model proteins. Therein, we could show that the precoating step has a critical impact on the final composition of the protein corona. Yet, since proteins building the primary corona were still detectable after additional incubations in a protein-containing medium, backtracking of the particle's history is possible. Interestingly, whereas the precoating history significantly disturbs particle-cell interactions (PCIs), the cellular response (i.e., metabolic activity, MTT assay) stays unaffected. Of note, lysozyme precoating revealed one of the highest rates in PCI for both epithelial cell lines. Taken together, we could show that particle history has a significant impact on protein corona formation and subsequently on the interaction of particles with murine intestinal epithelial-like cells. However, as this study was limited to one cell type, further work is needed to assess if these observations can be generalized to other cell types.