Boronate-ester crosslinked hyaluronic acid hydrogels for dihydrocaffeic acid delivery and fibroblasts protection against UVB irradiation.

Herein, we exploit the dynamic nature and pH dependence of complexes between phenylboronic acid and diol-containing molecules to control the release of an anti-photoaging agent, dihydrocaffeic acid (DHCA), from a dynamic covalent hydrogel (HG). The HG is prepared by reversible formation of boronate ester crosslinks between hyaluronic acid (HA) modified with saccharide (GLU) residues and HA functionalized with 3-aminophenylboronic acid (APBA), part of which is involved in complexation with DHCA. The hydrogel exhibited increased dynamic moduli and a lower relaxation time at pH 7.4 in comparison to pH 6, and greater amount of DHCA was incorporated at pH 7.4. Moreover, this hydrogel prolonged DHCA release at pH 7.4 through drug reversible complexation/decomplexation, while the rate of release was fastest in acidic (skin) conditions. Very interestingly, the incorporation of DHCA into the network enhances its protection against UVB-induced L929 fibroblast death. Therefore, this smart hydrogel can contribute to photoaging prevention.

Development of chitosan nanocapsules containing essential oil of Matricaria chamomilla L. for the treatment of cutaneous leishmaniasis.

Matricaria chamomilla L. has been used for centuries in many applications, including antiparasitic activity. Leishmaniasis is a parasitic disease, with limited treatments, due to high cost and toxicity. Thus, there is a need to develop new treatments, and in this context, natural products are targets of these researches. We report the development of chitosan nanocapsules containing essential oil of M. chamomilla (CEO) from oil-in-water emulsions using chitosan modified with tetradecyl chains as biocompatible shell material. The nanocapsules of CEO (NCEO) were analyzed by optical microscopy and dynamic light scattering, which revealed spherical shape and an average size of 800 nm. Successful encapsulation of CEO was further confirmed by fluorescence microscopy observations taking advantage of the autofluorescence properties of CEO. The encapsulation efficiency was around 90%. The entrapment of CEO reduced its cytotoxicity towards normal cells. On the other hand, the CEO was active against promastigotes and intracellular amastigotes, exhibiting IC50 of 3.33 µg/mL and 14.56 µg/mL, respectively, while NCEO showed IC50 for promastigotes of 7.18 µg/mL and for intracellular amastigotes of 14.29 µg/mL. These results demonstrate that encapsulation of CEO in nanocapsules using an alkylated chitosan biosurfactant as a "green" stabilizer is a promising therapeutic strategy to treat leishmaniasis.

Liposome-based nanocarrier loaded with a new quinoxaline derivative for the treatment of cutaneous leishmaniasis.

The use of nanocarriers for drug delivery is a strategy aimed to improve therapeutic indices through changes in their pharmacokinetic and pharmacodynamic characteristics. Liposomes are well-investigated nanocarriers for drug delivery to macrophage-targeted therapy, the main hosts of intracellular pathogens of some infectious diseases, such as leishmaniasis. In this study, we developed hyaluronic acid (HA)-coated liposomes by different methods that can encapsulate a new quinoxaline derivative, the LSPN331, to increase its solubility and improve its bioavailability. The surface modification of liposomes and their physicochemical characteristics may depend on the coating method, which may be a critical parameter with regard to the route of administration of the antileishmanial drug. Liposomes with identical phospholipid composition containing the same drug were developed, and different biological responses were verified, and our hypothesis is that it is related to the type of modification of the surface. Different physicochemical characterization techniques (dynamic light scattering, transmission electron microscopy and UV-vis quantification of labeled-HA) were used to confirm the successful modification of liposomes as well as their stability upon storage. The encapsulation of LSPN331 was performed using HPLC method, and the entrapment efficiency (EE%) was satisfatory in all formulations, considering results of similar formulations in the literature. Furthermore, in vitro and in vivo studies were carried out to evaluate the efficacy against the parasite Leishmania amazonensis. The in vitro activity was maintained or even improved and HA-coated liposomes showed the ability to target to the site of action by the proposed routes of administration, topically and intravenously. Both formulations are promising for future tests of antileishmania activity in vivo.

Dihydrocaffeic Acid Prevents UVB-Induced Oxidative Stress Leading to the Inhibition of Apoptosis and MMP-1 Expression via p38 Signaling Pathway.

Chronic UVB exposure promotes oxidative stress, directly causes molecular damage, and induces aging-related signal transduction, leading to skin photoaging. Dihydrocaffeic acid (DHCA) is a phenolic compound with potential antioxidant capacity and is thus a promising compound for the prevention of UVB-induced skin photodamage. The aim of this study was to evaluate the antioxidant and protective effect of DHCA against oxidative stress, apoptosis, and matrix metalloproteinase (MMP) expression via the mitogen-activated protein kinase (MAPK) signaling pathway on L929 fibroblasts irradiated with UVB. DHCA exhibited high antioxidant capacity on 2,2-diphenyl-1-picrylhydrazyl (DPPH•), 2,2-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS•+), and xanthine/luminol/xanthine oxidase (XOD) assays and reduced UVB-induced cell death in the neutral red assay. DHCA also modulated oxidative stress by decreasing intracellular reactive oxygen species (ROS) and extracellular hydrogen peroxide (H2O2) production, enhancing catalase (CAT) and superoxide dismutase (SOD) activities and reduced glutathione (GSH) levels. Hence, cellular damage was attenuated by DHCA, including lipid peroxidation, apoptosis/necrosis and its markers (loss of mitochondria membrane potential, DNA condensation, and cleaved caspase 9 expression), and MMP-1 expression. Furthermore, DHCA reduced the phosphorylation of MAPK p38. These findings suggest that DHCA can be used in the development of skin care products to prevent UVB-induced skin damage.

Physical nanocomposite hydrogels filled with low concentrations of TiO2 nanoparticles: Swelling, networks parameters and cell retention studies.

Physical nanocomposite hydrogels composed of poly(2-hydroxyethylmethacrylate) and titanium oxide nanoparticles at low concentrations (<1.0 wt%) were synthesized. The effect of the nanoparticle content on the water swelling and mechanical properties of the hydrogels was investigated. Additionally, to study the influence of the polymer-nanoparticle interactions, a second type of nanocomposite was synthesized using surface functionalized nanoparticles with 3-methacryloxypropyltrimethoxysilane as the filler. The pristine nanoparticles increased the swelling capacity, especially at short time scales, and greater solvent diffusion coefficients and initial swelling rates were achieved. In contrast, the nanocomposite filled with functionalized nanoparticles exhibited a diminished swelling capacity, a constant diffusion coefficient and a significant decrease in the initial swelling rate. The mechanical properties were studied by dynamic mechanical analyses using stress-relaxation tests. Two Maxwell models in parallel agreed well with the curves of the relaxation modulus as a function of time and indicated that at short relaxation times, the nanoparticles did not cause an effect, but that at longer times, the nanoparticles decreased the relaxation time. Finally, hydrogel network parameters determined by swelling measurements and mechanical experiments indicated that the hydrogel with well distributed nanoparticles decreases the molar mass between crosslink point and the mesh size, while poorly distributed nanoparticles lead to larger mesh size. Our functional studies show that the addition of titanium oxide nanoparticles improves the ability of nanocomposite hydrogels to retain aggregates of skeletal muscle cells, revealing their potential use as suitable scaffolds for tissue repair strategies.

A3K2A3-induced apoptotic cell death of Leishmania amazonensis occurs through caspase- and ATP-dependent mitochondrial dysfunction.

Leishmaniasis is a neglected tropical disease that affects millions of people worldwide. Current therapies mainly rely on antimonial drugs that are inadequate because of their high toxicity and increased drug resistance. An urgent need exists to discover new, more effective, more affordable, and more target-specific drugs. Pathways that are associated with apoptosis-like cell death have been identified in unicellular eukaryotes, including protozoan parasites. In the present study, we studied the mechanism of cell death that is induced by A3K2A3 against L. amazonensis. A3K2A3 is a dibenzylideneacetone that has an acyclic dienone that is attached to aryl groups in both ß-positions, which is similar to curcuminoids and chalcone structures. This compound was previously shown to be safe with regard to cytotoxicity and active against the parasite. Biochemical and morphological approaches were used in the present study. The results suggested that A3K2A3 caused mitochondrial dysfunction in L. amazonensis promastigotes, leading to mechanisms of cell death that share some common phenotypic features with metazoan apoptosis, such as an increase in reactive oxygen species production, a decrease in the adenosine triphosphate ratio, phosphatidylserine exposure, a decrease in cell volume, caspase production, and DNA fragmentation. Altogether, these findings indicate that apoptosis can indeed be triggered by chemotherapeutic agents.

Thermoresponsive hyaluronic acid nanogels as hydrophobic drug carrier to macrophages.

Delivery systems for macrophages are particularly attractive since these phagocytic cells play a important role in immunological and inflammatory responses, also acting as host cells for microorganisms that are involved in deadly infectious diseases, such as leishmaniasis. Hyaluronic acid (HA) is specifically recognized by macrophages that are known to express HA receptors. Therefore, in this study, we focused on HA-based nanogels as drug carriers for these cells. The drug delivery was validated in an in vivo study on mice using intravital two-photon laser scanning microscopy. HA derivatives were modified with a biocompatible oligo(ethylene glycol)-based thermoresponsive polymer to form nanogels. These HA conjugates were readily prepared by varying the molar mass of initial HA and the degree of substitution via radical-mediated thiol-ene chemistry in aqueous solution. The derivatives were shown to self-assemble into spherical gel particles with diameters ranging from 150 to 214 nm above 37 °C. A poorly water-soluble two-photon dye was successfully loaded into the nanogels during this self-assembly process. In vitro cellular uptake tests using a RAW 264.7 murine macrophage cell line showed successful intracellular delivery of the hydrophobic dye. After intravenous injection in mice, the nanogels circulated freely in the blood but were rapidly phagocytized within 13 min by circulating macrophages and stored in the liver and spleen, as observed by two-photon microscopy. Benefit can be thus expected in using such a delivery system for the liver and spleen macrophage-associated diseases.