Enhanced performance of gelatin 5-fluorouracil-containing nanoparticles against squamous cell carcinoma in simulated chronic wounds conditions

Chronic wounds are considered a silent epidemic affecting a significant fraction of the World population. Their treatment supposes a large fraction of the public spending on the health of developed countries. In chronic wounds secondary to burns, trauma, UV light radiation, and diabetes, among others, the development of squamous cell carcinoma (SCC) has been reported. If detected early, 95% of SCC are most comfortable to be treated and cured; however, 5% of advanced SCC is more dangerous and challenging to treat. It has been reported that the pH value within the wound-milieu influences indirectly and directly all biochemical reactions taking place in this process of healing. Differences in pH values between normal skin and chronic cutaneous wounds could be considered in designing and developing stimuli-responsive nanomaterials. In this work, the anticancer drug 5-fluorouracil (5-FU) inclusion on gelatin-based NPs for SCC treatment has been projected. The present work goal is to prepare and characterize physicochemical and biological properties of new therapeutic-containing NPs for the sustainable delivery of 5-FU under simulated chronic wound conditions. In vitro experiments have been performed to assess the biocompatible character of these gelatin-based NPs in terms of their hemolytic and cytotoxicity properties. Due to hyperglycemia impact on both the chronicity of the wounds and chemotherapy efficacy, cellular responses have been determined under euglycemic and hyperglycemic conditions. In vitro cytotoxicity studies have reported good selective toxicity against the A431 cell line, demonstrating that gelatin-based NPs are promising dual-responsive delivery systems to SCC targeting under simulated chronic wound conditions

Regenerable Superhydrophobic Coatings for Biomedical Fabrics

Coatings with high water repellence represent a promising field for biomedical applications. Superhydrophobicity (SH) can be used for preventing adhesion, controlling cell deposition, and spreading by inhibition of adsorption processes at liquid-solid interfaces. The recyclability of medical aids like fabrics can open the way for lower cost and more environmentally-friendly solutions. In this case, two different coatings form recyclable and low global warming potential materials and green solvents have been prepared and characterized based on their wettability properties. The resulting substrates have been used for the adhesion and spreading of representative skin cell lines, both tumoral and non-tumoral, showing a strong decrease in cell viability with values <10%. The coated substrates showed a complete recovery on initial SH properties after rinsing with suitable solvents

Mammalian Cell Behavior on Hydrophobic Substrates: Influence of Surface Properties

The influence of different surface properties holding to a modification of the substrate towards hydrophobic or superhydrophobic behavior was reviewed in this paper. Cell adhesion, their communication, and proliferation can be strongly manipulated, acting on interfacial relationship involving stiffness, surface charge, surface chemistry, roughness, or wettability. All these features can play mutual roles in determining the final properties of biomedical applications ranging from fabrics to cell biology devices. The focus of this work is the mammalian cell viability in contact with moderate to highly water repellent coatings or materials and also in combination with hydrophilic areas for more specific application. Few case studies illustrate a range of examples in which these surface properties and design can be fruitfully matched to the specific aim

Cytotoxic Assessment of 3,3-Dichloro-β-Lactams Prepared through Microwave-Assisted Benzylic C-H Activation from Benzyl-Tethered Trichloroacetamides Catalyzed by RuCl2(PPh3)3

Natural and synthetic ß -lactam derivatives constitute an interesting class of compounds due to their diverse biological activity. Mostly used as antibiotics, they were also found to have antitubercular, anticancer and antidiabetic activities, among others. In this investigation, six new 3,3-dichloro-ß -lactams prepared in a previous work were evaluated for their hemolytic and cytotoxic properties. The results showed that the proposed compounds have non-hemolytic properties and exhibited an interesting cytotoxic activity toward squamous cell carcinoma (A431 cell line), which was highly dependent on the structure and concentration of these -lactams. Among the molecules tested, 2b was the most cytotoxic, with the lowest IC50 values (30-47 µg/mL) and a promising selectivity against the tumor cells compared with non-tumoral cells

Mammalian cell viability on hydrophobic and superhydrophobic fabrics

fecting its growth, survival and also for its communication with other cells on fabrics. The combination of low surface energy and a specific surface morphology (micro/nano-roughness) leads to significantly less wettable surfaces, known as superhydrophobic characterized by high contact angle above 150° and a very small hysteresis. Such high water repellent coatings feature small area available to be exploited in many applications where interactions with aqueous environment are strongly to be avoided. In this work, the authors have investigated the influence of coating polyester fabric at different degree of hydrophobicity by mixed organicinorganic coating with moderated to highly water repellence. Depending on the coating composition and structure, the hydrophobicity of the fabric can be finely modulated by an easy-to-prepare method applicable to commercial, low cost fabric substrates providing advanced performance. In vitro experiments have been performed in order to establish the influence of surface modification on adhesion of representative model mammalian cell lines such as 3T3 fibroblasts, HaCaT keratinocytes and HeLa epithelial carcinoma cells. The obtained results suggested that, in addition to the chemistry and morphology of the coating, the characteristics of the substrate are important parameters on the final cell viabilities

Dual responsive gelatin-based nanoparticles for enhanced 5-fluorouracil efficiency

The very slow progress in the therapeutic efficacy of the treatment of severe diseases has suggested the use of a growing need for a multidisciplinary approach to the delivery of therapeutics to targets tissues. There has been increasing effort in the design of stimuli-responsive nanomaterials that they will be developed into effective drug delivery vehicles. Most commonly, effective drug delivery is associated with nanomaterial-facilitated accumulation and/or cellular internalization. Recent studies in our lab have demonstrated that gelatin-based NPs can be considered suitable pH responsive devices for the effective intracellular delivery of drugs. Concerning cancer treatment, ligands recognizing tumour-associated antigens expressed on the surface of the tumour cells have been employed. Some of the target structures suitable for tumour targeting belong to integrins which mediate cell adhesion to extracellular matrix and other cells. Interestingly, gelatin chains contain motifs such as RGD sequences that can be recognised by integrins. In this work the inclusion of the anticancer drug 5-fluorouracil (5-FU) on these gelatin-based NPs has been projected. These NPs may provide an opportunity to increase the therapeutic effect using a dual approach by: i) targeting the therapeutic drug to the tumour cells by the action of the naturally occurring RGD-motif on gelatin and ii) minimizing the non-productive trafficking from endosomes to lysosomes by releasing the cargo using the charge reversal approach after cellular internalization. In vitro cytotoxicity experiments of NPs on tumoral and non-tumoral cell lines have reported selectivity indexes higher than 30 demonstrating a great selectivity on the mode of action as a function of the cell line and the imposed compositions. Keywords: 5-fluorouracil; Gelatin; In vitro cytotoxicity; Loading efficiency; Nanoparticles; Selectivity. Copyright © 2018 Elsevier B.V. All rights reserved

Aggregation Behavior, Antibacterial Activity and Biocompatibility of Catanionic Assemblies Based on Amino Acid-Derived Surfactants

Abstract: The surface activity, aggregates morphology, size and charge characteristics of binary catanionic mixtures containing a cationic amino acid-derived surfactant N(π), N(τ)-bis(methyl)- L-Histidine tetradecyl amide (DMHNHC14) and an anionic surfactant (the lysine-based surfactant Nα-lauroyl-Nεacetyl lysine (C12C3L) or sodium myristate) were investigated for the first time. The cationic surfactant has an acid proton which shows a strong pKa shift irrespective of aggregation. The resulting catanionic mixtures exhibited high surface activity and low critical aggregation concentration as compared with the pure constituents. Catanionic vesicles based on DMHNHC14/sodium myristate showed a monodisperse population of medium-size aggregates and good storage stability. According to Small-Angle X-Ray Scattering (SAXS), the characteristics of the bilayers did not depend strongly on the system composition for the positively charged vesicles. Negatively charged vesicles (cationic surfactant:myristate ratio below 1:2) had similar bilayer composition but tended to aggregate. The DMHNHC14-rich vesicles exhibited good antibacterial activity against Gram-positive bacteria and their bactericidal effectivity declined with the decrease of the cationic surfactant content in the mixtures. The hemolytic activity and cytotoxicity of these catanionic formulations against non-tumoral (3T3, HaCaT) and tumoral (HeLa, A431) cell lines also improved by increasing the ratio of cationic surfactant in the mixture. These results indicate that the biological activity of these systems is mainly governed by the cationic charge density, which can be modulated by changing the cationic/anionic surfactant ratio in the mixtures. Remarkably, the incorporation of cholesterol in those catanionic vesicles reduces their cytotoxicity and increases the safety of future biomedical applications of these systems. Keywords: amino acid-based surfactants; catanionic vesicles; SAXS; self-assembly; antibacterial activity; cytotoxicit

Effect of PEGylation on Ligand-Targeted Magnetoliposomes: A Missed Goal

We tested the targeting efficiency of magnetoliposomes (MLPs) labeled with tripeptide arginine-glycine-aspartic acid (RGD) on two types of cells: HeLa cells expressing RGD receptors and 3T3 cells lacking RGD receptors. The targeting ability of RGD-MLPs was compared to that of bare MLPs and MLPs stabilized with poly(ethylene glycol) (PEG). Cellular internalization of these liposomes was determined by flow cytometry and confocal microscopy, which showed that both types of cells took up more nontargeting MLPs than targeting RGD-MLPs or PEG-MLPs, with PEG-MLPs showing the lowest degree of internalization. The presence of specific receptors on HeLa cells did not facilitate the binding of RGD-MLPs, probably due to the presence of PEG chains on the liposomal surface. The polymer increases the circulation time of the liposomes in the organism but reduces their interactions with cells. Despite the localization of the RGD peptide on the tip of PEG in RGD-MLPs, the interaction between the liposome and cell was still limited. To avoid this drawback, targeting drug delivery systems can be prepared with two types of PEG: one of a short length to enable biocompatibility and the other of a longer chain to carry the ligand

Membrane-destabilizing activity of pH-responsive cationic lysine-based surfactants: role of charge position and alkyl chain length

Many strategies for treating diseases require the delivery of drugs into the cell cytoplasm following internalization within endosomal vesicles. Thus, compounds triggered by low pH to disrupt membranes and release endosomal contents into the cytosol are of particular interest. Here, we report novel cationic lysine-based surfactants (hydrochloride salts of N¿- and N¿-acyl lysine methyl ester) that differ in the position of the positive charge and the length of the alkyl chain. Amino acid-based surfactants could be promising novel biomaterials in drug delivery systems, given their biocompatible properties and low cytotoxic potential. We examined their ability to disrupt the cell membrane in a range of pH values, concentrations and incubation times, using a standard hemolysis assay as a model of endosomal membranes. Furthermore, we addressed the mechanism of surfactant-mediated membrane destabilization, including the effects of each surfactant on erythrocyte morphology as a function of pH. We found that only surfactants with the positive charge on the ¿-amino group of lysine showed pH-sensitive hemolytic activity and improved kinetics within the endosomal pH range, indicating that the positive charge position is critical for pH-responsive behavior. Moreover, our results showed that an increase in the alkyl chain length from 14 to 16 carbon atoms was associated with a lower ability to disrupt cell membranes. Knowledge on modulating surfactant-lipid bilayer interactions may help us to develop more efficient biocompatible amino acid-based drug delivery devices