Solvent-free nanoparticles synthesis for encapsulation of water-soluble compounds

The increasing concerns on green manufacturing practices with low environmental impact have put pressure on the pharmaceutical industry. Of particular concern is the large number of organic solvents used in a wide range of pharmaceutical products, posing a significant risk on human and environmental health. Recently, nanoparticles(NPs) emerged as advantageous drug-delivery systems with the potential to maximize drug efficacy and minimize side effects. Unfortunately, NPs synthesis processes are still environmentally unsustainable, due to the large amount of organic solvent involved. This contribution describes the synthesis and characterization of NPs encapsulating proteins or nucleic acids for metastatic melanoma treatment using alternative synthesis methods that replace organic solvents with water-based solutions. Two different green synthesis techniques have been investigated; (i)Ionic gelation was used to prepare chitosan(CS) NPs, exploiting the electrostatic interaction between the CS amino groups and tripolyphosphate(TPP), (ii)self-assembly technique to prepare siRNA/phosphate-poly(allylamine-hydrochloride)(PAH) NPs, exploiting the interactions between primary amines in the polymer and siRNAs to form stable complexes. CS and PAH NPs with the appropriate hydrodynamic diameters (~200 nm), polydispersity index, and Z potential for high cell internalization and tissue extravasation were obtained. Preliminary in vitro tests demonstrated that particles are well tolerated by human fibroblast which has shown high viability even when treated with the highest NPs concentration (viability ~85% at 48h from the treatment). Additional tests are currently ongoing to demonstrate the efficacy of the drug- loaded system on human fibroblasts. Carlotta Mattioda acknowledges PON "Ricerca e Innovazione" 2014-2020 Azione IV.R "dottorati su tematiche green" for co-financing her Ph.D scholarship

Nanomedicine for Imaging and Therapy of Pancreatic Adenocarcinoma

Pancreatic adenocarcinoma has the worst outcome among all cancer types, with a 5-year survival rate as low as 10%. The lethal nature of this cancer is a result of its silent onset, resistance to therapies, and rapid spreading. As a result, most patients remain asymptomatic and present at diagnosis with an already infiltrating and incurable disease. The tumor microenvironment, composed of a dense stroma and of disorganized blood vessels, coupled with the dysfunctional signal pathways in tumor cells, creates a set of physical and biological barriers that make this tumor extremely hard-to-treat with traditional chemotherapy. Nanomedicine has great potential in pancreatic adenocarcinoma, because of the ability of nano-formulated drugs to overcome biological barriers and to enhance drug accumulation at the target site. Moreover, monitoring of disease progression can be achieved by combining drug delivery with imaging probes, resulting in early detection of metastatic patterns. This review describes the latest development of theranostic formulations designed to concomitantly treat and image pancreatic cancer, with a specific focus on their interaction with physical and biological barriers

Designing multifunctional devices for regenerative pharmacology based on 3D scaffolds, drug-loaded nanoparticles, and thermosensitive hydrogels: a proof-of-concept study

Regenerative pharmacology combines tissue engineering/regenerative medicine (TERM) with drug delivery with the aim to improve the outcomes of traditional TERM approaches. In this work, we aimed to design a multicomponent TERM platform comprising a three-dimensional scaffold, a thermosensitive hydrogel, and drug-loaded nanoparticles. We used a thermally induced phase separation method to obtain scaffolds with anisotropic mechanical properties, suitable for soft tissue engineering. A thermosensitive hydrogel was developed using a Poloxamer® 407-based poly(urethane) to embed curcumin-loaded nanoparticles, obtained by the single emulsion nanoprecipitation method. We found that encapsulated curcumin could retain its antioxidant activity and that embedding nanoparticles within the hydrogel did not affect the hydrogel gelation kinetics nor the possibility to progressively release the drug. The porous scaffold was easily loaded with the hydrogel, resulting in significantly enhanced (4-fold higher) absorption of a model molecule of nutrients (fluorescein isothiocyanate dextran 4kDa) from the surrounding environment compared to pristine scaffold. The developed platform could thus represent a valuable alternative in the treatment of many pathologies affecting soft tissues, by concurrently exploiting the therapeutic effects of drugs, with the 3D framework acting as a physical support for tissue regeneration and the cell-friendly environment represented by the hydrogel

Small molecules targeting endocytic uptake and recycling pathways

Over the past years a growing number of studies highlighted the pivotal role of intracellular trafficking in cell physiology. Among the distinct transport itineraries connecting the endocytic system, both internalization (endocytosis) and recycling (endocytic recycling) pathways were found fundamental to ensure cellular sensing, cell-to-cell communication, cellular division, and collective cell migration in tissue specific-contexts. Consistently, the dysregulation of endocytic trafficking pathways is correlated with several human diseases including both cancers and neurodegeneration. Aimed at suppress specific intracellular trafficking routes involved in disease onset and progression, huge efforts have been made to identify small molecule inhibitors with suitable pharmacological properties for in vivo administration. Here, we review most used drugs and recently discovered small molecules able to block endocytosis and endocytic recycling pathways. We characterize such pharmacological inhibitors by emphasizing their target specificity, molecular affinity, biological activity and efficacy in both in vitro and in vivo experimental models

In vitro models of molecular and nano-particle transport across the blood-brain barrier

The blood-brain barrier (BBB) is the tightest endothelial barrier in humans. Characterized by the presence of tight endothelial junctions and adherens junctions, the primary function of the BBB is to maintain brain homeostasis through the control of solute transit across the barrier. The specific features of this barrier make for unique modes of transport of solutes, nanoparticles, and cells across the BBB. Understanding the different routes of traffic adopted by each of these is therefore critical in the development of targeted therapies. In an attempt to move towards controlled experimental assays, multiple groups are now opting for the use of microfluidic systems. A comprehensive understanding of bio-transport processes across the BBB in microfluidic devices is therefore necessary to develop targeted and efficient therapies for a host of diseases ranging from neurological disorders to the spread of metastases in the brain

Dual-function nanoparticles enzymatically conjugated with a custom-made polyurethane hydrogel for chronic wound treatment

Hydrogels are attractive drug delivery systems with the potential to protect their cargo and control its release. In particular, hydrogels based on synthetic polymers are gaining increasing interest by virtue of their controllable chemistry, ease of modification, and reproducibility. Moreover, the presence of specific side chains and pending functional groups in the polymer structure allows for the conjugation of drugs and other compounds resulting in improved control over drug release. Enzymes that catalyse reactions in a very specific way could also be used to control the conjugation of compounds to the polymeric chains to improve reproducibility and biocompatibility of the conjugation process. This contribution describes an innovative system for drug delivery comprising a bioartificial supramolecular hydrogel based on a customised polyurethane and α- cyclodextrins, and nanoparticles, for application in the treatment of chronic wounds. The system has the potential to reduce inflammation and eradicate infection by virtue of dual-function nanoparticles which incorporate cobalt as antimicrobial agent, and phenolated lignin as antioxidant. The nanoparticles are enzymatically conjugated to the hydrogel by means of the amine side groups exposed along the backbone of the ad-hoc synthesised polyurethane. The oxidase enzyme laccase is exploited to oxidize the phenol groups of lignin, to allow their interaction with the amines on the hydrogel. The effects of nanoparticles conjugation to the hydrogel are studied through gelification tests, stability tests, and rheology. Moreover, the release of nanoparticles from the hydrogel and their effects on patients’ wound fluids and against relevant bacterial strains are analysed in vitro