Apoferritin nanocage as drug reservoir: is it a reliable drug delivery system?

Apoferritin is a complex protein with a number of possibilities for drug delivery and drug targeting technologies, as it could be considered as the future self-assembling, not-toxic protein drug delivery carrier. Few years ago, this concept was a reality; nowadays, after more than 10 years of research, a clear painting of Apoferritin, loaded with drugs, is lacking, in terms of protocols of formulation, characterization, drug release and application. Therefore, a critical evaluation and overall understanding of Apoferritin is due to speed up the possibilities for its translatability into clinical application

Potential Use of Nanomedicine for Drug Delivery Across the Blood-Brain Barrier in Healthy and Diseased Brain

The research of efficacious non-invasive therapies for the treatment of brain diseases represents a huge challenge, as people affected by disorders of the central nervous system (CNS) will significantly increase. Moreover, the blood-brain barrier is a key factor in hampering a number of effective drugs to reach the CNS. This review is therefore focusing on possible interventions of nanomedicine-based approaches in selected diseases affecting the CNS. A wide overview of the most outstanding results on preclinical evaluations of the potential of nanomedicine in brain diseases (i.e. brain tumor, Alzheimer, Parkinson, epilepsy and others) is given, with highlights on the data with relevant interest and real possibility in translation from bench-to-bedside. Moreover, a critical evaluation on the rationale in planning nanosystems to target specific brain pathologies is described, opening the path to a more structured and pathology-tailored design of nanocarriers

Protein cage nanostructure as drug delivery system: magnifying glass on apoferritin

New frontiers in nanomedicine are moving towards the research of new biomaterials. Apoferritin (APO), is a uniform regular self-assemblies nano-sized protein with excellent biocompatibility and a unique structure that affords it the ability to stabilize small active molecules in its inner core. Areas covered: APO can be loaded by applying a passive process (mainly used for ions and metals) or by a unique formulative approach based on disassemby/reassembly process. In this article, we aim to organize the experimental evidence provided by a number of studies on the loading, release and targeting. Attention is initially focused on the most investigated antineoplastic drug and contrast agents up to the most recent application in gene therapy. Expert opinion: Various preclinical studies have demonstrated that APO improved the potency and selectivity of some chemotherapeutics. However, in order to translate the use of APO into therapy, some issues must be solved, especially regarding the reproducibility of the loading protocol used, the optimization of nanocarrier characterization, detailed understanding of the final structure of loaded APO, and the real mechanism and timing of drug release

Nanoparticle transport across the blood brain barrier

ABSTRACT: While the role of the blood-brain barrier (BBB) is increasingly recognized in the (development of treatments targeting neurodegenerative disorders, to date, few strategies exist that enable drug delivery of non-BBB crossing molecules directly to their site of action, the brain. However, the recent advent of Nanomedicines may provide a potent tool to implement CNS targeted delivery of active compounds. Approaches for BBB crossing are deeply investigated in relation to the pathology: among the main important diseases of the CNS, this review focuses on the application of nanomedicines to neurodegenerative disorders (Alzheimer, Parkinson and Huntington's Disease) and to other brain pathologies as epilepsy, infectious diseases, multiple sclerosis, lysosomal storage disorders, strokes

Current Strategies for the Delivery of Therapeutic Proteins and Enzymes to Treat Brain Disorders

Brain diseases and injuries are growing to be one of the most deadly and costly medical conditions in the world. Unfortunately, current treatments are incapable of ameliorating the symptoms let alone curing the diseases. Many brain diseases have been linked to a loss of function in a protein or enzyme, increasing research for improving their delivery. This is no easy task due to the delicate nature of proteins and enzymes in biological conditions, as well as the many barriers that exist in the body ranging from those in circulation to the more specific barriers to enter the brain. Several main techniques are being used (physical delivery, protein/enzyme conjugates, and nanoparticle delivery) to overcome these barriers and create new therapeutics. This review will cover recently published data and highlights the benefits and deficits of possible new protein or enzyme therapeutics for brain diseases

Apoferritin nanocage as streptomycin drug reservoir: Technological optimization of a new drug delivery system

The aim of this study is to formulate and characterize streptomycin-loaded apoferritin nanoparticles (ApoStrep NPs) for their potential therapeutic use in bacterial resistant infections (i.e. tuberculosis). ApoStrep NPs were prepared by disassembly/reassembly process via pH method and changing apoferritin/drug molar ratio, purified by dialyses process also associated with gel filtration chromatography and characterized in their chemico-physical and technological parameters as yield, size distribution, polidispersivity, morphology, internal structure, zeta potential and loading efficacy. The results showed that spherical reproducible NPs could be obtained by using apoferritin/drug molar ratio lower than 1:25 and purification based on the combination of dialysis and gel filtration chromatography. Photon correlation spectroscopy, Uv–visible detection and electron microscopy showed the maintenance of the native apoferritin chemico-physical properties and structure. When formulated with apoferritin/drug 1:10 and 1:25 molar ratio, ApoStrep NPs showed remarkable encapsulation efficacy (35% and 28%, respectively) along with kinetic profile of drug delivery, approximately 15% at 37 °C in 72 h, as evidenced by “in vitro” release experiments

AFM/TEM complementary structural analysis of surface-functionalized nanoparticles

In the field of nanomedicine, the characterization of functionalized drug delivery systems, introduced on market as efficacious and selective therapeutics, represents a pivotal aspect of great importance. In particular, the morphology of polymeric nanoparticles, the most studied nanocarriers, is frequently assessed by transmission electron microscopy (TEM). Despite of TEM high resolution and versatility, this technology is frequently hampered by both the complicated procedure for sample preparation and the operative condition of analysis. Considering the scanning probe microscopies, atomic force microscopy (AFM) represents an extraordinary tool for the detailed characterization of submicron-size structure as the surface functionalization at the atomic scale. In this paper we discussed the advantage and limits of these microscopies applied to the characterization of PLGA nanoparticles functionalized with three different kinds of ligands (carbohydrate ligand, an antibody and quantum dots crystals) intentionally designed, created and tailored with specific physico-chemical properties to meet the needs of specific applications (targeting or imaging)

Quantitative comparison of the protein corona of nanoparticles with different matrices

: Nanoparticles (NPs) are paving the way for improved treatments for difficult to treat diseases diseases; however, much is unknown about their fate in the body. One important factor is the interaction between NPs and blood proteins leading to the formation known as the "protein corona" (PC). The PC, consisting of the Hard (HC) and Soft Corona (SC), varies greatly based on the NP composition, size, and surface properties. This highlights the need for specific studies to differentiate the PC formation for each individual NP system. This work focused on comparing the HC and SC of three NPs with different matrix compositions: a) polymeric NPs based on poly(lactic-co-glycolic) acid (PLGA), b) hybrid NPs consisting of PLGA and Cholesterol, and c) lipidic NPs made only of Cholesterol. NPs were formulated and characterized for their physico-chemical characteristics and composition, and then were incubated in human plasma. In-depth purification, identification, and statistical analysis were then performed to identify the HC and SC components. Finally, similar investigations demonstrated whether the presence of a targeting ligand on the NP surface would affect the PC makeup. These results highlighted the different PC fingerprints of these NPs, which will be critical to better understand the biological influences of the PC and improve future NP designs

PEGylated siRNA lipoplexes for silencing of BLIMP-1 in Primary Effusion Lymphoma: In vitro evidences of antitumoral activity

Silencing of the B lymphocyte-induced maturation protein 1 (Blimp-1), a pivotal transcriptional regulator during terminal differentiation of B cells into plasma cells with siRNAs is under investigation as novel therapeutic approach in Primary Effusion Lymphoma (PEL), a HHV-8 related and aggressive B cell Lymphoma currently lacking of an efficacious therapeutic approach. The clinical application of small interfering RNA (siRNA) in cancer therapy is limited by the lack of an efficient systemic siRNA delivery system. In this study we aim to develop pegylated siRNA lipoplexes formed using the cationic lipid DOTAP and DSPE-PEG2000, capable to effectively stabilize anti-Blimp-1 siRNA and suitable for systemic administration. Two types of pegylated lipoplexes using a classic (C-PEG Lipoplexes) or a post-pegylation method (P-PEG-Lipoplexes) were formulated and compared in their physicochemical properties (size, zeta potential, morphology and structure) and efficiency on PEL cell lines. A stable siRNAs protection was obtained with post pegylation approach (2% molar of DSPE-PEG2000 with respect to lipid) resulting in structures with diameters of 300 nm and a complexation efficiency higher that 80% (0.08 nmol/10 nmol of lipid). In vitro studies on PEL cell lines suggested that empty liposomes were characterized by a low cell toxicity also after PEG modification (cell viability and cell density over 85% after treatment with 10 μM of lipid). We demonstrated that P-PEG-Lipoplexes were able to significantly reduce the levels of BLIMP-1 protein leading to reduction of viability (less that 15% after transfection with 100 nM of complexed siRNAs) and activation of apoptosis. In vitro efficiency encourages us to further test the in vivo potential of P-PEG-Lipoplexes in PEL therapy

Clinical severity of ischemic stroke and neural damage biomarkers in the acute setting: the STROke MArkers (STROMA) study

BACKGROUND: Stroke is a leading cause of long-term morbidity and mortality affecting several hundred-thousand people annually in the Western Countries. Various panels of biomarkers of neural damage have been developed and validated. The primary objective of this investigation was to measure the correlation between the clinical severity of stroke and the serum/plasma concentrations of neural damage biomarkers. METHODS: A prospective investigation was conducted on a panel of biomarkers composed of S100β, matrix metalloproteinase-9 (MMP-9), N-terminal pro-B-type natriuretic peptide (NT pro-BNP) and D-dimer at admission and after 24 hours, in a cohort patients with a confirmed diagnosis of stroke in an emergency setting (STROke-MArkers STROMA). RESULTS: A total of 58 consecutive patients were enrolled, no participant was excluded; according to clinical severity measured by National Institute of Health Stroke Scale (NIHSS) there were 29 minor strokes, 24 moderate, 3 moderate-severe, 2 severe. The Spearman's rank correlation test was used to assess the relationship between the baseline NIHSS value and the concentrations of the four biomarkers: all the studied biomarkers showed a statistically significant correlation with baseline NIHSS at 24 hours. A multivariate ordinal regression model was used to analyze the correlation of markers with stroke severity, stratified, according to NIHSS score: MMP-9 and S100β showed a statistically significant correlation after 24 hours. CONCLUSION: MMP-9, S100β, NT pro-BNP and D-dimer showed a good correlation with the clinical severity of stroke which may become an additional resource in the acute patient evaluation and potentially follow-up