The influence of protein corona on Graphene Oxide: implications for biomedical theranostics

: Graphene-based nanomaterials have attracted significant attention in the field of nanomedicine due to their unique atomic arrangement which allows for manifold applications. However, their inherent high hydrophobicity poses challenges in biological systems, thereby limiting their usage in biomedical areas. To address this limitation, one approach involves introducing oxygen functional groups on graphene surfaces, resulting in the formation of graphene oxide (GO). This modification enables improved dispersion, enhanced stability, reduced toxicity, and tunable surface properties. In this review, we aim to explore the interactions between GO and the biological fluids in the context of theranostics, shedding light on the formation of the "protein corona" (PC) i.e., the protein-enriched layer that formed around nanosystems when exposed to blood. The presence of the PC alters the surface properties and biological identity of GO, thus influencing its behavior and performance in various applications. By investigating this phenomenon, we gain insights into the bio-nano interactions that occur and their biological implications for different intents such as nucleic acid and drug delivery, active cell targeting, and modulation of cell signalling pathways. Additionally, we discuss diagnostic applications utilizing biocoronated GO and personalized PC analysis, with a particular focus on the detection of cancer biomarkers. By exploring these cutting-edge advancements, this comprehensive review provides valuable insights into the rapidly evolving field of GO-based nanomedicine for theranostic applications

Protein corona-enabled serological tests for early stage cancer detection

Abstract Early stage cancer detection is a major issue in current medicine. In recent years, nanotechnology is providing new alternatives for early diagnosis. Upon exposure to human plasma, several nanoparticle types (e.g. gold nanoparticles) are surrounded by a protein layer referred to as protein corona (PC). The PC changes the original identity of the nanoparticle conferring a new biological character. It is now accepted that slight variations in the composition of a protein source significantly varies the PC composition. Thus, nanomaterials incubated with plasma proteins of individuals with different physiological conditions generate PCs with different compositions. This gives rise to the new concept of personalised PC. Therefore, since protein patterns of subjects affected by certain pathologies differ from those of healthy ones, diagnostic technologies based on the evaluation of personalised PC could represent a fascinating opportunity for early disease detection. Herein, we review the concept of personalised PC along with recent advances on the topic, giving an overview of some innovative analytical approaches for early stage cancer detection

Mechanistic insights into the release of doxorubicin from graphene oxide in cancer cells

Liposomal doxorubicin (L-DOX) is a popular drug formulation for the treatment of several cancer types (e.g., recurrent ovarian cancer, metastatic breast cancer, multiple myeloma, etc.), but poor nuclear internalization has hampered its clinical applicability so far. Therefore, novel drug-delivery nanosystems are actively researched in cancer chemotherapy. Here we demonstrate that DOX-loaded graphene oxide (GO), GO-DOX, exhibits much higher anticancer efficacy as compared to its L-DOX counterpart if administered to cellular models of breast cancer. Then, by a combination of live-cell confocal imaging and fluorescence lifetime imaging microscopy (FLIM), we suggest that GO-DOX may realize its superior performances by inducing massive intracellular DOX release (and its subsequent nuclear accumulation) upon binding to the cell plasma membrane. Reported results lay the foundation for future exploitation of these new adducts as high-performance nanochemotherapeutic agents

Inhibiting the growth of 3D brain cancer models with bio-coronated liposomal temozolomide

Nanoparticles (NPs) have emerged as an effective means to deliver anticancer drugs into the brain. Among various forms of NPs, liposomal temozolomide (TMZ) is the drug-of-choice for the treatment and management of brain tumours, but its therapeutic benefit is suboptimal. Although many possible reasons may account for the compromised therapeutic efficacy, the inefficient tumour penetration of liposomal TMZ can be a vital obstacle. Recently, the protein corona, i.e., the layer of plasma proteins that surround NPs after exposure to human plasma, has emerged as an endogenous trigger that mostly controls their anticancer efficacy. Exposition of particular biomolecules from the corona referred to as protein corona fingerprints (PCFs) may facilitate interactions with specific receptors of target cells, thus, promoting efficient internalization. In this work, we have synthesized a set of four TMZ-encapsulating nanomedicines made of four cationic liposome (CL) formulations with systematic changes in lipid composition and physical−chemical properties. We have demonstrated that precoating liposomal TMZ with a protein corona made of human plasma proteins can increase drug penetration in a 3D brain cancer model derived from U87 human glioblastoma multiforme cell line leading to marked inhibition of tumour growth. On the other side, by fine-tuning corona composition we have also provided experimental evidence of a non-unique effect of the corona on the tumour growth for all the complexes investigated, thus, clarifying that certain PCFs (i.e., APO-B and APO-E) enable favoured interactions with specific receptors of brain cancer cells. Reported results open new perspectives into the development of corona-coated liposomal drugs with enhanced tumour penetration and antitumour efficacy

Investigating the mechanism of action of DNA-loaded PEGylated lipid nanoparticles

PEGylated lipid nanoparticles (LNPs) are commonly used to deliver bioactive molecules, but the role of PEGylation in DNA-loaded LNP interactions at the cellular and subcellular levels remains poorly understood. In this study, we investigated the mechanism of action of DNA-loaded PEGylated LNPs using gene reporter technologies, dynamic light scattering (DLS), synchrotron small angle X-ray scattering (SAXS), and fluorescence confocal microscopy (FCS). We found that PEG has no significant impact on the size or nanostructure of DNA LNPs but reduces their zeta potential and interaction with anionic cell membranes. PEGylation increases the structural stability of LNPs and results in lower DNA unloading. FCS experiments revealed that PEGylated LNPs are internalized intact inside cells and largely shuttled to lysosomes, while unPEGylated LNPs undergo massive destabilization on the plasma membrane. These findings can inform the design, optimization, and validation of DNA-loaded LNPs for gene delivery and vaccine development

Nanotechnology Meets Oncology: A Perspective on the Role of the Personalized Nanoparticle-Protein Corona in the Development of Technologies for Pancreatic Cancer Detection

In recent years nanotechnology has opened exciting opportunities in the struggle against cancer. In 2007 Dawson and coworkers demonstrated that nanomaterials exposed to biological fluids are coated with plasma proteins that form the so-called “protein corona”. A few years later our joint research team made of physicists, chemists, biotechnologists, surgeons, oncologists, and bioinformaticians introduced the concept of “personalized protein corona” and demonstrated that it is unique for each human condition. This concept paved the way for the development of nano-enabled blood (NEB) tests for the diagnosis of pancreatic ductal adenocarcinoma (PDAC). These studies gave an impetus to serious work in the field that came to maturity in the late 2010s. In this special issue, we provide the reader with a comprehensive overview of the most significant discoveries of our research team in the field of PDAC detection. We focus on the main achievements with an emphasis on the fundamental aspects of this arena and how they shaped the integration of different scientific backgrounds towards the development of advanced diagnostic technologies. We conclude the review by outlining future perspectives and opportunities to transform the NEB tests into a reliable clinical diagnostic technology for early diagnosis, follow-up, and management of PDAC patients

Magnetic Levitation of Personalized Nanoparticle–Protein Corona as an Effective Tool for Cancer Detection

Unprecedented opportunities for early stage cancer detection have recently emerged from the characterization of the personalized protein corona (PC), i.e., the protein cloud that surrounds nanoparticles (NPs) upon exposure to a patients’ bodily fluids. Most of these methods require “direct characterization” of the PC., i.e., they necessitate protein isolation, identification, and quantification. Each of these steps can introduce bias and affect reproducibility and inter-laboratory consistency of experimental data. To fulfill this gap, here we develop a nanoparticle-enabled blood (NEB) test based on the indirect characterization of the personalized PC by magnetic levitation (MagLev). The MagLev NEB test works by analyzing the levitation profiles of PC-coated graphene oxide (GO) NPs that migrate along a magnetic field gradient in a paramagnetic medium. For the test validation, we employed human plasma samples from 15 healthy individuals and 30 oncological patients affected by four cancer types, namely breast cancer, prostate cancer, colorectal cancer, and pancreatic ductal adenocarcinoma (PDAC). Over the last 15 years prostate cancer, colorectal cancer, and PDAC have continuously been the second, third, and fourth leading sites of cancer-related deaths in men, while breast cancer, colorectal cancer, and PDAC are the second, third and fourth leading sites for women. This proof-of-concept investigation shows that the sensitivity and specificity of the MagLev NEB test depend on the cancer type, with the global classification accuracy ranging from 70% for prostate cancer to an impressive 93.3% for PDAC. We also discuss how this tool could benefit from several tunable parameters (e.g., the intensity of magnetic field gradient, NP type, exposure conditions, etc.) that can be modulated to optimize the detection of different cancer types with high sensitivity and specificity

Test sierologico per coadiuvare la diagnosi e il monitoraggio del glioblastoma multiforme

L’invenzione riguarda un metodo per la diagnosi del glioblastoma multiforme che comprende l’utilizzo di nanoparticelle di ossido di grafene e una successiva analisi di immagini, un metodo di screening o monitoraggio di pazienti patologici e non per individuare pazienti patologici a rischio di glioblastoma multiforme che comprende l’utilizzo di nanoparticelle di ossido di grafene e una successiva analisi di immagini, un kit comprendente reagenti per la diagnosi precoce del glioblastoma multiform

Functionalization of gold nanoparticles with peptidomimetics as a targeted therapy for multiple sclerosis

Póster presentado a la 2nd Spanish Conference on Biomedical Applications of Nanomaterials (SBAN), celebrada en Madrid del 6 al 7 de junio de 2019.Multiple Sclerosis (MS) is an inflammatory autoimmune disease of the central nervous system that can lead to severe disability. The wide variety of symptoms characteristic for MS such as numbness, weakness, paresis, visual problems, severe fatigue and depression and also intellectual and cognitive impairments are caused by by brain and spinal cord's inflammation that damages the myelin sheath. During the last years, significant progress has been made in understanding how pathogenic leukocytes migrate from the periphery to the CNS, and the critical roles played by the α4ß1and α4ß7 integrins have been studied extensively in animal models, and in patients in clinical trials. Thus, the counter ligands of these integrins (VCAM1 and MAdCAM1) are a perfect target to prevent the inflammatory process. Gold nanoparticles (AuNPs) show low toxicity and excellent optical properties, they represent a very powerful tool in imaging and therapy applications. In this work, we propose the functionalization of AuNPs with five different integrins ligand-like peptidomimetics to target the pathogenic leukocytes and create a challenging device for imaging and therapy of MS. Preliminary outcomes in in vitro experiments will be presented in order to lay the foundations for future study.This project has received funding from MCINN (Spain) (project "SAF2014-54763-C2-1-R"), from Gobierno de Aragón (predoctoral contract of Rafael Ramírez Jiménez) and from Fondo Social of Gobierno de Aragón (Spain)

Funcionalización de nanopartículas de oro con peptidomiméticos como posible terapia para la esclerosis múltiple

Resumen del póster presentado a la 8ª Jornada de Jóvenes Investigadores (Química y Física) de Aragón, celebrada en Zaragoza el dia 22 de noviembre de 2018.La esclerosis múltiple es una enfermedad crónica autoinmune que se caracteriza por la desmielinización del sistema nervioso central lo que daña severamente su funcionalidad. Se relaciona con procesos de inflamación causados por linfocitos T autorreactivos que sobreexpresan las integrinas α4β1 y α4β7, gracias a las cuales llegan al cerebro cruzando la barrera hematoencefálica. Así, los ligandos de estas integrinas (VCAM1 and MAdCAM1) pueden ser una diana perfecta para prevenir estos procesos de inflamación. Las nanopartículas de oro presentan baja toxicidad y propiedades que las hacen idóneas para aplicaciones en terapia y diagnóstico. En este trabajo proponemos la funcionalización con seis peptidomiméticos de los ligandos VCAM1 and MAdCAM1 para que interaccionen con las integrinas presentes en células T autorreactivas y crear un instrumento de imagen y terapia de la esclerosis múltiple. Los peptidomiméticos se acoplaron sobre nanoesferas de oro de 14 nm recubiertas con ácido ω-mercaptopolietilenglicol-α-carboxílico. La funcionalización se llevó a cabo a través de formación de un enlace amida para lo que se estudiaron diferentes condiciones de reacción. Se realizó la caracterización físico-química de las nanopartículas funcionalizadas y se estudió su internalización en células Jurkat (línea inmortalizada de linfocitos T humanos) encontrando una mayor internalización de las nanopartículas funcionalizadas.Peer Reviewe