Schiff Base-Based Hydrogel Embedded with In Situ Generated Silver Nanoparticles Capped by a Hyaluronic Acid-Diethylenetriamine Derivative for Wound Healing Application.

In this study, hydrogels were produced using a Schiff base reaction between two hyaluronic acid derivatives: one containing aldehyde groups (HA-Ald) and the other holding a diethylenetriamine with terminal amino groups (HA-DETA). The DETA portion promotes the in situ growth, complexation, and stabilization of silver nanoparticles (AgNPs), eliminating the need for external reducing agents. The reaction between HA-DETA and HA-Ald leads to the formation of imine bonds, which results in dynamically pH-responsive cross-linking. While the DETA capping ability helped in embedding the AgNPs, the on/off pH environmental responsivity of the hydrogel allows for a controlled and on-demand release of the drug, mainly when bacterial infections cause pH variation of the wound bed. The injectable hydrogels resulted in being highly compatible in contact with blood red cells, fibroblasts, and keratinocytes and capable of having a proliferative effect on an in vitro wound scratch model. The pH-responsive hydrogels showed proper antibacterial activity againstPseudomonas aeruginosaandStaphylococcus aureus, common bacterial strains presented in wound infections. Finally, in vivo wound model studies demonstrated an overall speeding up in the wound healing rate and advanced wound conditions in the experimental group treated with the hydrogels compared to control samples.

PFAS-free superhydrophobic chitosan coating for fabrics.

In view of health and environmental concerns, together with the upcoming restrictive regulations on per- and polyfluoroalkyl substances (PFAS), less impactful materials must be explored for the hydrophobization of surfaces. Polysaccharides, and especially chitosan, are being explored for their desirable properties of film formation and ease of modification. We present a PFAS-free chitosan superhydrophobic coating for textiles deposited through a solvent-free method. By contact angle analysis and drop impact, we observe that the coating imparts hydrophobicity to the fabrics, reaching superhydrophobicty (θA = 151°, θR = 136°) with increased amount of coating (from 1.6 g/cm2). This effect is obtained by the combination of chemical water repellency of the modified chitosan and the nano- and micro-roughness, assessed by SEM analysis. We perform a comprehensive study on the durability of the coatings, showing good results especially for acidic soaking where the hydrophobicity is maintained until the 8th cycle of washing. We assess the degradation of the coating by a TGA-IR investigation to define the compounds released with thermal degradation, and we confirm the coating's biodegradability by biochemical oxygen consumption. Finally, we demonstrate its biocompatibility on keratinocytes (HaCaT cell line) and fibroblasts (HFF-1 cell line), confirming that the coating is safe for human skin cells.

Vision-based omnidirectional indoor robots for autonomous navigation and localization in manufacturing industry.

In this paper, we present a new generation of omnidirectional automated guided vehicles (omniagv) used for transporting materials within a manufacturing factory with the ability to navigate autonomously and intelligently by interacting with the environment, including people and other entities. This robot has to be integrated into the operating environment without significant changes to the current facilities or heavy redefinitions of the logistics processes already running. For this purpose, different vision-based systems and advanced methods in mobile and cognitive robotics are developed and integrated. In this context, vision and perception are key factors. Different developed modules are in charge of supporting the robot during its navigation in the environment. Specifically, the localization module provides information about the robot pose by using visual odometry and wheel odometry systems. The obstacle avoidance module can detect obstacles and recognize some object classes for adaptive navigation. Finally, the tag detection module aids the robot during the picking phase of carts and provides information for global localization. The smart integration of vision and perception is paramount for effectively using the robot in the industrial context. Extensive qualitative and quantitative results prove the capability and effectiveness of the proposed AGV to navigate in the considered industrial environment.

Electrosprayed zein nanoparticles as antibacterial and anti-thrombotic coatings for ureteral stents.

Ureteral stents are among the most frequently used human implants, with urothelium trauma, blood clots, and bacterial colonization being their main reasons for failure. In this study, berberine-loaded zein (ZB) nanoparticles with high drug encapsulation efficiency (>90 %) were fabricated via electrospray on flat and 3D stainless steel structures. Physico-chemical characterization revealed that the ZB nanoparticles created a highly hydrophilic, antioxidant, and scratch-resistant continuous coating over the metal structure. Results showed that the drug release rate was faster at neutral pH (i.e., PBS pH 7.4) than in an artificial urine medium (pH 5.3) due to the different swelling behavior of the zein polymeric matrix. In vitro evaluation of ZB particles onto human dermal fibroblasts and blood cells demonstrated good cell proliferation and enhanced anti-thrombotic properties compared to bare stainless steel. The ability of the electrosprayed zein particles to resist bacterial adherence and proliferation was evaluated with Gram-negative (Escherichia coli) bacteria, showing high inhibition rates (-29 % and -46 % for empty and berberine-loaded particles, respectively) compared to the medical-grade metal substrates. Overall, the proposed composite coating fulfilled the requirements for ureteral applications, and can advance the development of innovative biocompatible, biodegradable, and antibacterial coatings for drug-eluting stents.

Combining Alginate/PVPI-Based Film with Frequency Rhythmic Electrical Modulation System (FREMS) Technology as an Advanced Strategy for Diabetic Wounds.

Diabetes is rising as one of the most diffused diseases of the century with the related urgent necessity to face its systemic and local effects on the patients, such as cardiovascular problems, degeneration of limbs, and dysfunction of the wound healing process. The diffusion of leg ulcers has been estimated to be 1.51 for 1000 population, and these non-resolved wounds can produce several social, economic, and mental health issues in diabetic patients. At the same time, these people experience neuropathic pain that causes morbidity and a further decrease in their quality of life. Here, a new study is presented where asodium alginate/Polyvinylpyrrolidone-Iodine complex (PVPI)-based wound dressing is combined with the Frequency Rhythmic Electrical Modulation System (FREMS) technology, an established medical device for the treatment of neuropathic pain and diabetic ulcers. The produced Alginate/PVPI-based films are characterized in terms of morphology, chemistry, wettability, bio-/hemo-compatibility, and clotting capacity. Next, the Alginate/PVPI-based films are used together with FREMS technology in diabetic mice models, and synergism of their action in the wound closure rate and anti-inflammatory properties is found. Hence, how the combination of electrical neurostimulation devices and advanced wound dressings can be a new approach to improve chronic wound treatment is demonstrated.

Label-Free Intracellular Multi-Specificity in Yeast Cells by Phase-Contrast Tomographic Flow Cytometry.

In-flow phase-contrast tomography provides a 3D refractive index of label-free cells in cytometry systems. Its major limitation, as with any quantitative phase imaging approach, is the lack of specificity compared to fluorescence microscopy, thus restraining its huge potentialities in single-cell analysis and diagnostics. Remarkable results in introducing specificity are obtained through artificial intelligence (AI), but only for adherent cells. However, accessing the 3D fluorescence ground truth and obtaining accurate voxel-level co-registration of image pairs for AI training is not viable for high-throughput cytometry. The recent statistical inference approach is a significant step forward for label-free specificity but remains limited to cells' nuclei. Here, a generalized computational strategy based on a self-consistent statistical inference to achieve intracellular multi-specificity is shown. Various subcellular compartments (i.e., nuclei, cytoplasmic vacuoles, the peri-vacuolar membrane area, cytoplasm, vacuole-nucleus contact site) can be identified and characterized quantitatively at different phases of the cells life cycle by using yeast cells as a biological model. Moreover, for the first time, virtual reality is introduced for handling the information content of multi-specificity in single cells. Full fruition is proofed for exploring and interacting with 3D quantitative biophysical parameters of the identified compartments on demand, thus opening the route to a metaverse for 3D microscopy.

Galantamine-memantine hybrids for Alzheimer's disease: The influence of linker rigidity in biological activity and pharmacokinetic properties.

Neurodegenerative processes characterizing Alzheimer's disease (AD) are strictly related to the impairment of cholinergic and glutamatergic neurotransmitter systems which provoke synaptic loss. These experimental evidences still represent the foundation of the actual standard-of-care treatment for AD, albeit palliative, consisting on the coadministration of an acetylcholinesterase inhibitor and the NMDAR antagonist memantine. In looking for more effective treatments, we previously developed a series of galantamine-memantine hybrids where compound 1 (ARN14140) emerged with the best-balanced action toward the targets of interest paired to neuroprotective efficacy in a murine AD model. Unfortunately, it showed a suboptimal pharmacokinetic profile, which required intracerebroventricular administration for in vivo studies. In this work we designed and synthesized new hybrids with fewer rotatable bonds, which is related to higher brain exposure. Particularly, compound 2, bearing a double bond in the tether, ameliorated the biological profile of compound 1 in invitro studies, increasing cholinesterases inhibitory potencies and selective antagonism toward excitotoxic-related GluN1/2B NMDAR over beneficial GluN1/2A NMDAR. Furthermore, it showed increased plasma stability and comparable microsomal stability in vitro, paired with lower half-life and faster clearance in vivo. Remarkably, pharmacokinetic evaluations of compound 2 showed a promising increase in brain uptake in comparison to compound 1, representing the starting point for further chemical optimizations.

Plant-Based, Hydrogel-like Microfibers as an Antioxidant Platform for Skin Burn Healing.

Natural polymers from organic wastes have gained increasing attention in the biomedical field as resourceful second raw materials for the design of biomedical devices which can perform a specific bioactive function and eventually degrade without liberating toxic residues in the surroundings. In this context, patches and bandages, that need to support the skin wound healing process for a short amount of time to be then discarded, certainly constitute good candidates in our quest for a more environmentally friendly management. Here, we propose a plant-based microfibrous scaffold, loaded with vitamin C (VitC), a bioactive molecule which acts as a protecting agent against UV damages and as a wound healing promoter. Fibers were fabricated via electrospinning from various zein/pectin formulations, and subsequently cross-linked in the presence of Ca2+ to confer them a hydrogel-like behavior, which we exploited to tune both the drug release profile and the scaffold degradation. A comprehensive characterization of the physico-chemical properties of the zein/pectin/VitC scaffolds, either pristine or cross-linked, has been carried out, together with the bioactivity assessment with two representative skin cell populations (human dermal fibroblast cells and skin keratinocytes, HaCaT cells). Interestingly, col-1a gene expression of dermal fibroblasts increased after 3 days of growth in the presence of the microfiber extraction media, indicating that the released VitC was able to stimulate collagen mRNA production overtime. Antioxidant activity was analyzed on HaCaT cells via DCFH-DA assay, highlighting a fluorescence intensity decrease proportional to the amount of loaded VitC (down to 50 and 30%), confirming the protective effect of the matrices against oxidative stress. Finally, the most performing samples were selected for the in vivo test on a skin UVB-burn mouse model, where our constructs demonstrated to significantly reduce the inflammatory cytokines expression in the injured area (50% lower than the control), thus constituting a promising, environmentally sustainable alternative to skin patches.

Design, Synthesis, In Vitro and In Vivo Characterization of CDC42 GTPase Interaction Inhibitors for the Treatment of Cancer.

CDC42 GTPases (RHOJ, CDC42, and RHOQ) are overexpressed in multiple tumor types and activate pathways critical for tumor growth, angiogenesis, and metastasis. Recently, we reported the discovery of a novel lead compound, ARN22089, which blocks the interaction of CDC42 GTPases with specific downstream effectors. ARN22089 blocks tumor growth in BRAF mutant mouse melanoma models and patient-derived xenografts (PDXs) in vivo. ARN22089 also inhibits tumor angiogenesis in three-dimensional vascularized microtumor models in vitro. Notably, ARN22089 belongs to a novel class of trisubstituted pyrimidines. Based on these results, we describe an extensive structure-activity relationship of ∼30 compounds centered on ARN22089. We discovered and optimized two novel inhibitors (27, ARN25062, and 28, ARN24928), which are optimal back-up/follow-up leads with favorable drug-like properties and in vivo efficacy in PDX tumors. These findings further demonstrate the potential of this class of CDC42/RHOJ inhibitors for cancer treatment, with lead candidates ready for advanced preclinical studies.

Copper nano-architecture topical cream for the accelerated recovery of burnt skin.

Skin burns are debilitating injuries with significant morbidity and mortality associated with infections and sepsis, particularly in immunocompromised patients. In this context, nanotechnology can provide pioneering approaches for the topical treatment of burnt skin. Herein, the significant recovery of radiation-damaged skin by exploiting copper ultrasmall-in-nano architectures (CuNAs) dispersed in a home-made cosmetic cream is described and compared to other noble metals (such as gold). Owing to their peculiar design and components, CuNAs elicit a substantial recovery from burned skin in in vivo models after one topical application, and a significant anti-inflammatory effect is highlighted by reducing cytokine expression. The treatment exhibited neither significant toxicity nor the alteration of copper metabolism in the target organs because of the CuNA biocompatibility. This study may open new horizons in the treatment of radiation dermatitis and skin burns caused by other external events.

Dynamic investigation of zein-based degradable and hemocompatible coatings for drug-eluting stents: a microfluidic approach.

Biodegradable stent coatings have shown great potential in terms of delivering drugs to a damaged vessel wall, and their release profiles are key elements governing the overall performance of drug-eluting stents (DESs). However, release and degradation kinetics are usually not tested under simulated physiological conditions or in dynamic environments, both essential aspects in the design of novel DESs. To bridge this gap, fused silica-based microfluidic systems, with either round or square channel cross-sections, were designed to mimic the microenvironment of a stented vessel. In particular, we fabricated and characterized microfluidic chips based on customizable channels, which were spray-coated with a naturally-derived, rutin-loaded zein solution, to perform a comprehensive study under flow conditions. Dynamic assays after 6 hours showed how the degradation of the zein matrix was affected by the cross-sectional conformation (∼69% vs. ∼61%, square and round channel, respectively) and the simulated blood fluid components (∼55%, round channel with a more viscous solution). The released amount of rutin was ∼81% vs. ∼77% and ∼78% vs. ∼74% from the square and round channels, using the less and more viscous blood-simulated fluids, respectively. Fitting the drug release data to Korsmeyer-Peppas and first-order mathematical models provided further insight into the mechanism of rutin release and coating behavior under flowing conditions. More importantly, whole blood tests with our newly developed microfluidic platforms confirmed the hemocompatibility of our zein-based coating. In detail, in-flow and static studies on the blood cell behavior showed a significant reduction of platelet adhesion (∼73%) and activation (∼93%) compared to the stainless-steel substrate, confirming the benefits of using such naturally-derived coatings to avoid clogging. Overall, our microfluidic designs can provide a key practical tool for assessing polymer degradation and drug release from degradable matrices under flowing conditions, thus aiding future studies on the development of hemocompatible, controlled-release coatings for DESs.

The Fate of Intranasally Instilled Silver Nanoarchitectures.

The intranasal administration of drugs allows an effective and noninvasive therapeutic action on the respiratory tract. In an era of rapidly increasing antimicrobial resistance, new approaches to the treatment of communicable diseases, especially lung infections, are urgently needed. Metal nanoparticles are recognized as a potential last-line defense, but limited data on the biosafety and nano/biointeractions preclude their use. Here, we quantitatively and qualitatively assess the fate and the potential risks associated with the exposure to a silver nanomaterial model (i.e., silver ultrasmall-in-nano architectures, AgNAs) after a single dose instillation. Our results highlight that the biodistribution profile and the nano/biointeractions are critically influenced by both the design of the nanomaterial and the chemical nature of the metal. Overall, our data suggest that the instillation of rationally engineered nanomaterials might be exploited to develop future treatments for (non)communicable diseases of the respiratory tract.

Innovative Non-PrP-Targeted Drug Strategy Designed to Enhance Prion Clearance.

Prion diseases are a group of neurodegenerative disorders characterized by the accumulation of misfolded prion protein (called PrPSc). Although conversion of the cellular prion protein (PrPC) to PrPSc is still not completely understood, most of the therapies developed until now are based on blocking this process. Here, we propose a new drug strategy aimed at clearing prions without any direct interaction with neither PrPC nor PrPSc. Starting from the recent discovery of SERPINA3/SerpinA3n upregulation during prion diseases, we have identified a small molecule, named compound 5 (ARN1468), inhibiting the function of these serpins and effectively reducing prion load in chronically infected cells. Although the low bioavailability of this compound does not allow in vivo studies in prion-infected mice, our strategy emerges as a novel and effective approach to the treatment of prion disease.

Self-Adhesive and Antioxidant Poly(vinylpyrrolidone)/Alginate-Based Bilayer Films Loaded with Malva sylvestris Extracts as Potential Skin Dressings.

Malva sylvestris (MS) is a medicinal herb known worldwide for its beneficial effects due to the several active molecules present in its leaves and flowers. These compounds have shown antioxidant and anti-inflammatory properties and thus can be helpful in treatments of burns and chronic wounds, characterized mainly by high levels of free radicals and impairments of the inflammatory response. In this work, we propose bilayer films as wound dressings, based on poly(vinylpyrrolidone) (PVP) and sodium alginate loaded with M. sylvestris extracts from leaves and flowers and fabricated by combining solvent-casting and rod-coating methods. The top layer is produced in two different PVP/alginate ratios and loaded with the MS flowers' extract, while the bottom layer is composed of PVP and MS leaves' extract. The bilayers were characterized morphologically, chemically, and mechanically, while they showed superior self-adhesive properties on human skin compared to a commercial skin patch. The materials showed antioxidant activity, release of the bioactive compounds, and water uptake property. Moreover, the anthocyanin content of the flower extract provided the films with the ability to change color when immersed in buffers of different pH levels. In vitro tests using primary keratinocytes demonstrated the biocompatibility of the MS bilayer materials and their capacity to enhance the proliferation of the cells in a wound scratch model. Finally, the best performing MS bilayer sample with a PVP/alginate ratio of 70:30 was evaluated in mice models, showing suitable resorption properties and the capacity to reduce the level of inflammatory mediators in UVB-induced burns when applied to an open wound. These outcomes suggest that the fabricated bilayer films loaded with M. sylvestris extracts are promising formulations as active and multifunctional dressings for treating skin disorders.

Structure-based design of CDC42 effector interaction inhibitors for the treatment of cancer.

CDC42 family GTPases (RHOJ, RHOQ, CDC42) are upregulated but rarely mutated in cancer and control both the ability of tumor cells to invade surrounding tissues and the ability of endothelial cells to vascularize tumors. Here, we use computer-aided drug design to discover a chemical entity (ARN22089) that has broad activity against a panel of cancer cell lines, inhibits S6 phosphorylation and MAPK activation, activates pro-inflammatory and apoptotic signaling, and blocks tumor growth and angiogenesis in 3D vascularized microtumor models (VMT) in vitro. Additionally, ARN22089 has a favorable pharmacokinetic profile and can inhibit the growth of BRAF mutant mouse melanomas and patient-derived xenografts in vivo. ARN22089 selectively blocks CDC42 effector interactions without affecting the binding between closely related GTPases and their downstream effectors. Taken together, we identify a class of therapeutic agents that influence tumor growth by modulating CDC42 signaling in both the tumor cell and its microenvironment.

Evaluation of a Multifunctional Polyvinylpyrrolidone/Hyaluronic Acid-Based Bilayer Film Patch with Anti-Inflammatory Properties as an Enhancer of the Wound Healing Process.

The management of acute and chronic wounds is still a socioeconomic burden for society due to the lack of suitable tools capable of supporting all the healing phases. The exponential spread of diabetes worldwide and the consequent increase of complicated diabetic ulcers require further efforts to develop scalable, low-cost, and easy-to-use treatments for tackling this emergency. Recently, we explored the fabrication of a polyvinylpyrrolidone/hyaluronic acid-based bilayer wound dressing, characterizing its physicochemical features and detailing its excellent antimicrobial activity. Here, we further demonstrate its biocompatibility on fibroblasts, keratinocytes, and red blood cells. The bilayer shows anti-inflammatory properties, statistically reducing the level of IL-6, IL-1ß, and TNF-α, and a capacity to accelerate wound healing in vitro and in healthy and diabetic mice models compared to untreated mice. The outcomes suggest that this bilayer material can be an effective tool for managing different skin injuries.

Discovery and SAR Evolution of Pyrazole Azabicyclo[3.2.1]octane Sulfonamides as a Novel Class of Non-Covalent N-Acylethanolamine-Hydrolyzing Acid Amidase (NAAA) Inhibitors for Oral Administration.

Inhibition of intracellular N-acylethanolamine-hydrolyzing acid amidase (NAAA) activity is a promising approach to manage the inflammatory response under disabling conditions. In fact, NAAA inhibition preserves endogenous palmitoylethanolamide (PEA) from degradation, thus increasing and prolonging its anti-inflammatory and analgesic efficacy at the inflamed site. In the present work, we report the identification of a potent, systemically available, novel class of NAAA inhibitors, featuring a pyrazole azabicyclo[3.2.1]octane structural core. After an initial screening campaign, a careful structure-activity relationship study led to the discovery of endo-ethoxymethyl-pyrazinyloxy-8-azabicyclo[3.2.1]octane-pyrazole sulfonamide 50 (ARN19689), which was found to inhibit human NAAA in the low nanomolar range (IC50 = 0.042 µM) with a non-covalent mechanism of action. In light of its favorable biochemical, in vitro and in vivo drug-like profile, sulfonamide 50 could be regarded as a promising pharmacological tool to be further investigated in the field of inflammatory conditions.

Hydroxycinnamic Acids and Derivatives Formulations for Skin Damages and Disorders: A Review.

Alterations of skin homeostasis are widely diffused in our everyday life both due to accidental injuries, such as wounds and burns, and physiological conditions, such as late-stage diabetes, dermatitis, or psoriasis. These events are locally characterized by an intense inflammatory response, a high generation of harmful free radicals, or an impairment in the immune response regulation, which can profoundly change the skin tissue' repair process, vulnerability, and functionality. Moreover, diabetes diffusion, antibiotic resistance, and abuse of aggressive soaps and disinfectants following the COVID-19 emergency could be causes for the future spreading of skin disorders. In the last years, hydroxycinnamic acids and derivatives have been investigated and applied in several research fields for their anti-oxidant, anti-inflammatory, and anti-bacterial activities. First, in this study, we give an overview of these natural molecules' current source and applications. Afterwards, we review their potential role as valid alternatives to the current therapies, supporting the management and rebalancing of skin disorders and diseases at different levels. Also, we will introduce the recent advances in the design of biomaterials loaded with these phenolic compounds, specifically suitable for skin disorders treatments. Lastly, we will suggest future perspectives for introducing hydroxycinnamic acids and derivatives in treating skin disorders.

Design, Synthesis, and Biological Evaluation of a Series of Oxazolone Carboxamides as a Novel Class of Acid Ceramidase Inhibitors.

Acid ceramidase (AC) is a cysteine hydrolase that plays a crucial role in the metabolism of lysosomal ceramides, important members of the sphingolipid family, a diversified class of bioactive molecules that mediate many biological processes ranging from cell structural integrity, signaling, and cell proliferation to cell death. In the effort to expand the structural diversity of the existing collection of AC inhibitors, a novel class of substituted oxazol-2-one-3-carboxamides were designed and synthesized. Herein, we present the chemical optimization of our initial hits, 2-oxo-4-phenyl-N-(4-phenylbutyl)oxazole-3-carboxamide 8a and 2-oxo-5-phenyl-N-(4-phenylbutyl)oxazole-3-carboxamide 12a, which resulted in the identification of 5-[4-fluoro-2-(1-methyl-4-piperidyl)phenyl]-2-oxo-N-pentyl-oxazole-3-carboxamide 32b as a potent AC inhibitor with optimal physicochemical and metabolic properties, showing target engagement in human neuroblastoma SH-SY5Y cells and a desirable pharmacokinetic profile in mice, following intravenous and oral administration. 32b enriches the arsenal of promising lead compounds that may therefore act as useful pharmacological tools for investigating the potential therapeutic effects of AC inhibition in relevant sphingolipid-mediated disorders.