More than fifty years after the 3Rs definition and despite the continuous implementation of regulatory measures, animals continue to be widely used in basic research. Their use comprises not only in vivo experiments with animal models, but also the production of a variety of supplements and products of animal origin for cell and tissue culture, cell-based assays, and therapeutics. The animal-derived products most used in basic research are fetal bovine serum (FBS), extracellular matrix proteins such as Matrigel™, and antibodies. However, their production raises several ethical issues regarding animal welfare. Additionally, their biological origin is associated with a high risk of contamination, resulting, frequently, in poor scientific data for clinical translation. These issues support the search for new animal-free products able to replace FBS, Matrigel™, and antibodies in basic research. In addition, in silico methodologies play an important role in the reduction of animal use in research by refining the data previously to in vitro and in vivo experiments. In this review, we depicted the current available animal-free alternatives in in vitro research.

Helichrysum italicum essential oil (H. italicum EO) is recognized for its anti-inflammatory, antimicrobial and wound-healing properties. The main goal of the present work was the development and characterization of a gel formulation comprising H. italicum EO loaded in chitosan nanoparticles (NPs) for dermatological applications. H. italicum EO-loaded chitosan NPs presented hydrodynamic diameter and PdI of about 300 nm and 0.28, respectively, and a surface charge of +19 mV. The H. italicum EO-loaded chitosan NPs were prepared by means of ionic gelation and then incorporated into a thermal water gel formulation. The organoleptic and physicochemical properties of the developed gel were studied. The gel remained stable under accelerated test conditions, maintaining pH, viscosity and organoleptic properties. In addition, the formulation presented pH, viscosity and spreadability properties suitable for topical application. Finally, the performance of the gel in topical application was evaluated on the skin of volunteers using non-invasive methods, particularly, by means of biometric evaluation. These assays showed that the properties of the developed thermal water-based gel formulation with H. italicum EO-loaded chitosan NPs can improve skin hydration and maintain healthy skin conditions, demonstrating its putative role for distinct dermatological applications.

The interest in the pharmacological applications of cannabinoids is largely increasing in a wide range of medical areas. Recently, research on its potential role in eye conditions, many of which are chronic and/or disabling and in need of new alternative treatments, has intensified. However, due to cannabinoids' unfavorable physicochemical properties and adverse systemic effects, along with ocular biological barriers to local drug administration, drug delivery systems are needed. Hence, this review focused on the following: (i) identifying eye disease conditions potentially subject to treatment with cannabinoids and their pharmacological role, with emphasis on glaucoma, uveitis, diabetic retinopathy, keratitis and the prevention of Pseudomonas aeruginosa infections; (ii) reviewing the physicochemical properties of formulations that must be controlled and/or optimized for successful ocular administration; (iii) analyzing works evaluating cannabinoid-based formulations for ocular administration, with emphasis on results and limitations; and (iv) identifying alternative cannabinoid-based formulations that could potentially be useful for ocular administration strategies. Finally, an overview of the current advances and limitations in the field, the technological challenges to overcome and the prospective further developments, is provided.

Caffeine is a naturally occurring alkaloid found in various plants. It acts as a stimulant, antioxidant, anti-inflammatory, and even an aid in pain management, and is found in several over-the-counter medications. This naturally derived bioactive compound is the best-known ingredient in coffee and other beverages, such as tea, soft drinks, and energy drinks, and is widely consumed worldwide. Therefore, it is extremely important to research the effects of this substance on the human body. With this in mind, caffeine and its derivatives have been extensively studied to evaluate its ability to prevent diseases and exert anti-aging and neuroprotective effects. This review is intended to provide an overview of caffeine's effects on cancer and cardiovascular, immunological, inflammatory, and neurological diseases, among others. The heavily researched area of caffeine in sports will also be discussed. Finally, recent advances in the development of novel nanocarrier-based formulations, to enhance the bioavailability of caffeine and its beneficial effects will be discussed.

Senescent cells accumulation can contribute to the development of several age-related diseases, including cancer. Targeting and eliminating senescence cells, would allow the development of new therapeutic approaches for the treatment of different diseases. The 4N1Ks peptide, a 10 amino acid peptide derived from TSP1 protein, combines both features by targeting the CD47 receptor present in the surface of senescent cells and demonstrating senolytic activity, thereby representing a new strategy to take into account. Nonetheless, peptide drugs are known for their biopharmaceutical issues, such as low short half-life and tendency to aggregate, which reduces their bioavailability and limits their therapeutic potential. In order to overcome this problem, herein we propose the use of biodegradable and biocompatible sphingomyelin nanosystems (SNs), decorated with this peptide for the targeting of senescent cells. In order to efficiently associate the 4N1Ks peptide to the nanosystems while exposing it on their surface for an effective targeting of senescent cells, the 4N1Ks peptide was chemically conjugated to a PEGylated hydrophobic chain. The resulting SNs-4N1Ks (SNs-Ks), were extensively characterized for their physicochemical properties, by dynamic light scattering, multiple-angle dynamic light scattering, nanoparticle tracking analysis and atomic force microscopy. The SNs-Ks demonstrated suitable features in terms of size (similar to 100 nm), association efficiency (87.2 +/- 6.9%) and stability in different biorelevant media. Cell toxicity experiments in MCF7 cancer cells indicated an improved cytotoxic effect of SNs-Ks, decreasing cancer cells capacity to form colonies, with respect to free peptide, and an improved hemocompatibility. Lastly, senescence escape preliminary experiments demonstrated the improvement of SNs-Ks senolytic activity of in chemotherapyinduced senescence model of breast cancer cells. Therefore, these results demonstrate for the first time the potential of the combination of SNs with 4N1Ks peptide for the development of innovative senolytic therapies to battle cancer.

Long non-coding RNAs (lncRNAs) are an emerging group of RNAs with a crucial role in cancer pathogenesis. In gastrointestinal cancers, TP53 target 1 (TP53TG1) is an epigenetically regulated lncRNA that represents a promising therapeutic target due to its tumor suppressor properties regulating the p53-mediated DNA damage and the intracellular localization of the oncogenic YBX1 protein. However, to translate this finding into the clinic as a gene therapy, it is important to develop effective carriers able to deliver exogenous lncRNAs to the targeted cancer cells. Here, we propose the use of biocompatible sphingomyelin nanosystems comprising DOTAP (DSNs) to carry and deliver a plasmid vector encoding for TP53TG1 (pc(TP53TG1)-DSNs) to a colorectal cancer cell line (HCT-116). DSNs presented a high association capacity and convenient physicochemical properties. In addition, pc(TP53TG1)-DSNs showed anti-tumor activities in vitro, specifically a decrease in the proliferation rate, a diminished colony-forming capacity, and hampered migration and invasiveness of the treated cancer cells. Consequently, the proposed strategy displays a high potential as a therapeutic approach for colorectal cancer.

Triple negative breast cancer (TNBC) is known for being very aggressive, heterogeneous and highly metastatic. The standard of care treatment is still chemotherapy, with adjacent toxicity and low efficacy, highlighting the need for alternative and more effective therapeutic strategies. Edelfosine, an alkyl-lysophospholipid, has proved to be a promising therapy for several cancer types, upon delivery in lipid nanoparticles. Therefore, the objective of this work was to explore the potential of edelfosine for the treatment of TNBC. Edelfosine nanoemulsions (ET-NEs) composed by edelfosine, Miglyol 812 and phosphatidylcholine as excipients, due to their good safety profile, presented an average size of about 120 nm and a neutral zeta potential, and were stable in biorelevant media. The ability of ET-NEs to interrupt tumor growth in TNBC was demonstrated both in vitro, using a highly aggressive and invasive TNBC cell line, and in vivo, using zebrafish embryos. Importantly, ET-NEs were able to penetrate through the skin barrier of MDA-MB 231 xenografted zebrafish embryos, into the yolk sac, leading to an effective decrease of highly aggressive and invasive tumoral cells' proliferation. Altogether the results demonstrate the potential of ET-NEs for the development of new therapeutic approaches for TNBC.

In recent years, the synthesis of polymer electrolyte systems derived from biopolymers for the development of sustainable green electrochemical devices has attracted great attention. Here electrolytes based on the red seaweeds-derived polysaccharide κ-carrageenan (κ-Cg) doped with neodymium triflate (NdTrif3) and glycerol (Gly) were obtained by means of a simple, clean, fast, and low-cost procedure. The aim was to produce near-infrared (NIR)-emitting materials with improved thermal and mechanical properties, and enhanced ionic conductivity. Cg has a particular interest, due to the fact that it is a renewable, cost-effective natural polymer and has the ability of gelling in the presence of certain alkali- and alkaline-earth metal cations, being good candidates as host matrices for accommodating guest cations. The as-synthesised κ-Cg-based membranes are semi-crystalline, reveal essentially a homogeneous texture, and exhibit ionic conductivity values 1–2 orders of magnitude higher than those of the κ-Cg matrix. A maximum ionic conductivity was achieved for 50 wt.% Gly/κ-Cg and 20 wt.% NdTrif3/κ-Cg (1.03 × 10−4, 3.03 × 10−4, and 1.69 × 10−4 S cm−1 at 30, 60, and 97 °C, respectively). The NdTrif-based κ-Cg membranes are multi-wavelength emitters from the ultraviolet (UV)/visible to the NIR regions, due to the κ-Cg intrinsic emission and to Nd3+, 4F3/2→4I11/2-9/2.

Electrochromic devices (ECDs) combining visible/near-infrared (NIR) transparent amorphous indium zinc oxide (a-IZO) external layers with innovative NIR-emitting electrolytes composed of red seaweed-derived kappa-carrageenan (kappa-Cg) polysaccharide, glycerol (Gly), and erbium triflate (ErTrif(3)center dot xH(2)O) are proposed as a valuable technological solution for the development of smart windows providing less heating demand, less glare and more indoors human comfort for the new generation of energy-efficient buildings. The electrolyte preparation is cheap, clean, and fast. The optimized sample including 50 wt% Gly/kappa-Cg and 40 wt% ErTrif(3)center dot xH(2)O/kappa-Cg exhibits the highest ionic conductivity (1.5 X 10(-4) S cm(-1) at 20 degrees C) and displays ultraviolet (UV)/blue and NIR emissions associated with the kappa-Cg-based host and the Er3+ ions (I-4(15/2) -> I-4(13/2)), respectively. The 5-layer configuration ECD tested demonstrated fast switching time (50 s), high electrochromic contrast (transmittance variations of 46/51% at 550/1000 nm), high optical density change (0.89/0.75 at 550/1000 nm), outstanding coloration efficiency (450th cycle = -15902/-13400 cm(2) C-1 and +3072/+2589 cm(2) C-1 at 550/1000 nm for coloration and bleaching, respectively), excellent electrochemical stability, and self-healing after mechanical damage. The ECD encompasses two voltage-operated modes: semibright warm (+3.0 V, transmittances of 52/61% at 550/1000 nm) and dark cold (-3.0 V, transmittances of 7/11% at 550/1000 nm).

A set of new 1-vinylidene-naphthofurans substituted by styryl groups were synthetized. These compounds show photochromic properties when adsorbed in silica gel, or in acidified THF solutions, developing intense colours after exposure to the UV light and returning to the uncoloured state, in the dark, in several minutes. The introduction of the styryl chain extends the conjugation of the photo products and shifts their absorption bathochromically relative to the parent unsubstituted naphthofuran. For styryl-naphthofuran 5a, in an acidified THF solution, an intense grey colouration can be achieved after 1 min of UV exposure (or direct sunlight) and the decolouration, in the dark, occurs in less than 10 min at room temperature. (C) 2017 Elsevier Ltd. All rights reserved.

This review is a comprehensive analysis of the progress made so far on the delivery of polynucleotide-based therapeutics to the eye, using synthetic nanocarriers. Attention has been addressed to the capacity of different nanocarriers for the specific delivery of polynucleotides to both, the anterior and posterior segments of the eye, with emphasis on their ability to (i) improve the transport of polynucleotides across the different eye barriers; (ii) promote their intracellular penetration into the target cells; (iii) protect them against degradation and, (iv) deliver them in a long-term fashion way. Overall, the conclusion is that despite the advantages that nanotechnology may offer to the area of ocular polynucleotide-based therapies (especially AS-ODN and siRNA delivery), the knowledge disclosed so far is still limited. This fact underlines the necessity of more fundamental and product-oriented research for making the way of the said nanotherapies towards clinical translation.

In the area of regenerative medicine different approaches have been studied to restore the native structure of damaged tissues. Herein, the suitability of a photocrosslinkable hydrogel for tissue engineering applications was studied.

Fungal infections affecting human beings have increased during the last years and the currently available treatments, when administered for long periods, trigger microbial resistance. Such demands the development of new viable therapeutic alternatives. Silver is known since the antiquity by its antimicrobial properties and, herein, it was used to produce two types of nanoparticles (NPs), uncoated and coated with polyvinylpyrrolidone (PVP), which were aimed to be used in fungal infection treatment. NPs properties were characterized by Transmission electron microscopy, X-ray diffraction, UV–Vis, Dynamic light scattering, Fourier transform infrared, and Energy-dispersive X-ray spectroscopy. Furthermore, in vitro studies were also performed to evaluate NPs cytotoxic profile and antifungal activity. The results obtained revealed that the produced nanoparticles are biocompatible and have a good potential for being used in the treatment of common skin infections caused by Trichophyton rubrum and Trichophyton mentagrophytes, being PVP-coated silver NPs the most suitable ones.

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