Development of a thermosensitive hydrogel based on Polaxamer 407 and gellan gum with inclusion complexes (Sulfobutylated-ß-cyclodextrin-Farnesol) as a local drug delivery system.

This work proposes the development of a thermosensitive local drug release system based on Polaxamer 407, also known as Pluronic® F-127 (PF-127), Gellan Gum (GG) and the inclusion complex Sulfobutylated-ß-cyclodextrin (CD) with Farnesol (FOH). Rheological properties of the hydrogels and their degradation were studied. According to the rheological results, a solution of 20% w/v of PF-127 forms a strong gel with a gelling temperature of about 25 °C (storage modulus of 15,000 Pa). The addition of the GG increased the storage modulus (optimal concentration of 0.5 % w/v) twofold without modifying the gelling temperature. Moreover, including 0.5% w/v of GG also increased 6 times the degradation time of the hydrogel. Regarding the inclusion complex, the addition of free CD decreased the viscosity and the gel strength since polymer chains were included in CD cavity without affecting the gelling temperature. Contrarily, the inclusion complex CD-FOH did not significantly modify any property of the formulation because the FOH was hosted in the CD. Furthermore, a mathematical model was developed to adjust the degradation time. This model highlights that the addition of the GG decreases the number of released chains from the polymeric network (which coincides with an increase in the storage modulus) and that the free CD reduces the degradation rate, protecting the polymeric chains. Finally, FOH release was quantified with a specific device, that was designed and printed for this type of system, observing a sustainable drug release (similar to FOH aqueous solubility, 8 µM) dependent on polymer degradation.

Tryptophan association in water driven by charge-transfer interactions with electron-deficient aromatic haptens.

The ability of a series of electron-deficient aromatic compounds to form charge-transfer complexes with tryptophan in water has been evaluated by X-ray diffraction studies, UV-vis spectra and NMR. As dinitrophenyl (DNP) ligands are well-known to generate antibody-mediated responses and the π-π stacking interactions with tryptophan residues of the antibody Fab fragment have been reported, most of the aromatic receptors studied here are nitro derivatives. Charge-transfer interactions between the rich indole ring of tryptophan and the electron-deficient aromatic receptors have been observed in the solid state, as four crystal structures of the complexes were obtained. The aromatic donor-acceptor interactions in solution were also verified by UV-vis and NMR spectroscopy. The association of the tripeptide Trp-Gly-Trp, a motif found in antigen Ag43, with the electron-deficient aromatic diimide was also studied by UV-vis and NMR spectroscopy. Our results show that these simple electron-deficient molecules could potentially behave as novel haptens and be incorporated in more elaborated drugs targeting protein-protein interactions, due to the synergistic effect of multiple non-covalent interactions.

Biodegradable gellan gum hydrogels loaded with paclitaxel for HER2+ breast cancer local therapy.

Hydrogels loaded with chemotherapeutics are promising tools for local tumor treatment. In this work, redox-responsive implantable hydrogels based on gellan gum were prepared as paclitaxel carriers for HER2-positive breast cancer therapy. To achieve different degrees of chemical crosslinking, hydrogels were synthesized in both acetate buffer and phosphate buffer and crosslinked with different concentrations of l-cysteine. It was shown that both, the type of buffer and the l-cysteine concentration used, conditioned the dynamic modulus, equilibrium swelling rate, porosity, and thermal stability of the hydrogels. Then, the biocompatibility of the hydrogels with the most suitable porosity for drug delivery applications was assessed. Once confirmed, these hydrogels were loaded with paclitaxel:ß-cyclodextrin inclusion complexes, and they showed a glutathione-responsive controlled release of the taxane. Moreover, when tested in vitro, paclitaxel-loaded hydrogels exhibited great antitumor activity. Thus, they could act as excellent local tailored carriers of paclitaxel for future, post-surgical treatment of HER2-overexpressing breast tumors.

An Adjustable Cleft Based on an 8-Sulfonamide-2-Naphthoic Acid with Oxyanion Hole Geometry.

Cleft type receptors showing the oxyanion hole motif have been prepared in a straightforward synthesis starting from the commercial 3,7-dihidroxy-2-naphthoic acid. The double H-bond donor pattern is achieved by the introduction of a sulfonamide group in the C-8 position of naphthalene and a carboxamide at the C-2 position. This cleft, for which the geometry resembles that of an oxyanion hole, is able to adjust to different guests, as shown by the analysis of the X-ray crystal structures of associates with methanol or acetic acid. Combination of hydrogen bonds and charge-transfer interactions led to further stabilization of the complexes, in which the electron-rich aromatic ring of the receptor was close in space to the electron-deficient dinitroaromatic guests. Modelling studies and bidimensional NMR experiments have been carried out to provide additional information.

Tailored-Made Polydopamine Nanoparticles to Induce Ferroptosis in Breast Cancer Cells in Combination with Chemotherapy.

Ferroptosis is gaining followers as mechanism of selective killing cancer cells in a non-apoptotic manner, and novel nanosystems capable of inducing this iron-dependent death are being increasingly developed. Among them, polydopamine nanoparticles (PDA NPs) are arousing interest, since they have great capability of chelating iron. In this work, PDA NPs were loaded with Fe3+ at different pH values to assess the importance that the pH may have in determining their therapeutic activity and selectivity. In addition, doxorubicin was also loaded to the nanoparticles to achieve a synergist effect. The in vitro assays that were performed with the BT474 and HS5 cell lines showed that, when Fe3+ was adsorbed in PDA NPs at pH values close to which Fe(OH)3 begins to be formed, these nanoparticles had greater antitumor activity and selectivity despite having chelated a smaller amount of Fe3+. Otherwise, it was demonstrated that Fe3+ could be released in the late endo/lysosomes thanks to their acidic pH and their Ca2+ content, and that when Fe3+ was co-transported with doxorubicin, the therapeutic activity of PDA NPs was enhanced. Thus, reported PDA NPs loaded with both Fe3+ and doxorubicin may constitute a good approach to target breast tumors.

Steady and Oscillatory Shear Flow Behavior of Different Polysaccharides with Laponite®.

The rheological behavior, in terms of steady and oscillatory shear flow, of Laponite® with different polysaccharides (alginate, chitosan, xanthan gum and levan) in salt-free solutions was studied. Results showed that a higher polymer concentration increased the zero-rate viscosity and decreased the critical strain rate (Cross model fit) as well as increasing the elastic and viscous moduli. Those properties (zero-rate viscosity and critical strain rate) can be a suitable indicator of the effect of the Laponite® on the shear flow behavior for the different solutions. Specifically, the effect of the Laponite® predominates for solutions with large critical strain rate and low zero-rate viscosity, modifying significantly the previous parameters and even the yield stress (if existing). On the other hand, larger higher polymeric concentration hinders the formation of the platelet structure, and polymer entanglement becomes predominant. Furthermore, the addition of high concentrations of Laponite® increases the elastic nature, but without modifying the typical mechanical spectra for polymeric solutions. Finally, Laponite® was added to (previously crosslinked) gels of alginate and chitosan, obtaining different results depending on the material. These results highlight the possibility of predicting qualitatively the impact of the Laponite® on different polymeric solutions depending on the solutions properties.

Antineoplastic behavior of polydopamine nanoparticles prepared in different water/alcohol media.

Polydopamine nanoparticles (PD NPs) have been synthesized in the present work through the oxidative polymerization of dopamine in aqueous media containing five different types of alcohol in a constant solvent volume ratio. We have shown that the type of alcohol, along with the ammonium hydroxide concentration used in the synthesis process, conditions particle size. Additionally, it has been found that the type of alcohol employed influences the well-known capacity of polydopamine nanoparticles to adsorb iron. As a consequence, since a ferroptosis-like mechanism may account for the cytotoxicity of these nanoparticles, the type of alcohol could also have a determining role in their antineoplastic activity. Here, the existence of a correlation between the ability of polydopamine nanoparticles to load Fe3+ and their toxic effect on breast cancer cells has been proven. For instance, nanoparticles synthesized using 2-propanol adsorbed more Fe3+ and had the greatest capacity to reduce breast tumor cell viability. Moreover, none of the nanoparticle synthesized with the different alcohols significantly decreased normal cell survival. Cancer cells present greater iron-dependence than healthy cells and this fact may explain why polydopamine nanoparticles toxicity, in which Fenton chemistry could be implicated, seems tumor-specific.

Trastuzumab: More than a Guide in HER2-Positive Cancer Nanomedicine.

HER2 overexpression, which occurs in a fifth of diagnosed breast cancers as well as in other types of solid tumors, has been traditionally linked to greater aggressiveness. Nevertheless, the clinical introduction of trastuzumab has helped to improve HER2-positive patients' outcomes. As a consequence, nanotechnology has taken advantage of the beneficial effects of the administration of this antibody and has employed it to develop HER2-targeting nanomedicines with promising therapeutic activity and limited toxicity. In this review, the molecular pathways that could be responsible for trastuzumab antitumor activity will be briefly summarized. In addition, since the conjugation strategies that are followed to develop targeting nanomedicines are essential to maintaining their efficacy and tolerability, the ones most employed to decorate drug-loaded nanoparticles and liposomes with trastuzumab will be discussed here. Thus, the advantages and disadvantages of performing this trastuzumab conjugation through adsorption or covalent bindings (through carbodiimide, maleimide, and click-chemistry) will be described, and several examples of targeting nanovehicles developed following these strategies will be commented on. Moreover, conjugation methods employed to synthesized trastuzumab-based antibody drug conjugates (ADCs), among which T-DM1 is well known, will be also examined. Finally, although trastuzumab-decorated nanoparticles and liposomes and trastuzumab-based ADCs have proven to have better selectivity and efficacy than loaded drugs, trastuzumab administration is sometimes related to side toxicities and the apparition of resistances. For this reason also, this review focuses at last on the important role that newer antibodies and peptides are acquiring these days in the development of HER2-targeting nanomedicines.

Size Matters in the Cytotoxicity of Polydopamine Nanoparticles in Different Types of Tumors.

Polydopamine has acquired great relevance in the field of nanomedicine due to its physicochemical properties. Previously, it has been reported that nanoparticles synthetized from this polymer are able to decrease the viability of breast and colon tumor cells. In addition, it is well known that the size of therapeutic particles plays an essential role in their effect. As a consequence, the influence of this parameter on the cytotoxicity of polydopamine nanoparticles was studied in this work. For this purpose, polydopamine nanoparticles with three different diameters (115, 200 and 420 nm) were synthetized and characterized. Their effect on the viability of distinct sorts of human carcinomas (breast, colon, liver and lung) and stromal cells was investigated, as well as the possible mechanisms that could be responsible for such cytotoxicity. Moreover, polydopamine nanoparticles were also loaded with doxorubicin and the therapeutic action of the resulting nanosystem was analyzed. As a result, it was demonstrated that a smaller nanoparticle size is related to a more enhanced antiproliferative activity, which may be a consequence of polydopamine's affinity for iron ions. Smaller nanoparticles would be able to adsorb more lysosomal Fe3+ and, when they are loaded with doxorubicin, a synergistic effect can be achieved.

Paclitaxel-Trastuzumab Mixed Nanovehicle to Target HER2-Overexpressing Tumors.

Paclitaxel is one of the most widely used chemotherapeutic agents thanks to its effectiveness and broad spectrum of antitumor activity. However, it has a very poor aqueous solubility and a limited specificity. To solve these handicaps, a novel paclitaxel-trastuzumab targeted transport nanosystem has been developed and characterized in this work to specifically treat cancer cells that overexpress the human epidermal growth factor receptor-2 (HER2). METHODS: Alginate and piperazine nanoparticles were synthetized and conjugated with paclitaxel:ß-cyclodextrins complexes and trastuzumab. Conjugated nanoparticles (300 nm) were characterized and their internalization in HER2-overexpressing tumor cells was analyzed by immunofluorescence. Its specific antitumor activity was studied in vitro using human cell lines with different levels of HER2-expression. RESULTS: In comparison with free paclitaxel:ß-cyclodextrins complexes, the developed conjugated nanovehicle presented specificity for the treatment of HER2-overpressing cells, in which it was internalized by endocytosis. CONCLUSIONS: It seems that potentially avoiding the conventional adverse effects of paclitaxel treatment could be possible with the use of the proposed mixed nanovehicle, which improves its bioavailability and targets HER2-positive cancer cells.

Color Engineering of Silicon Nitride Surfaces to Characterize the Polydopamine Refractive Index.

A simple methodology to generate polydopamine (PDA) surfaces featured with color due to thin-film interference phenomena is presented. It is based on depositing ultra-thin films of polydopamine on a Si/Si3 N4 wafer that exhibits an interferential reflectance maximum right at the visible/UV boundary (∼400 nm). Therefore, a small deposit of PDA modifies the optical path, in such manner that the wavelength of the maximum of reflectance red shifts. Because the human eye is very sensitive to any change of the light spectral distribution at the visible region, very small film thickness changes (∼30 nm) are enough to notably modify the perceived color. Consequently, a controlled deposit of PDA, tune the color along the whole visible spectrum. Additionally, good quality of PDA deposits allowed us to determine the refractive index of polydopamine by ellipsometry spectroscopy. This data can be crucial in confocal skin microscopic techniques, presently used in diagnosis of skin tumors.

Cytotoxicity of paramagnetic cations-Loaded polydopamine nanoparticles.

Polydopamine (PD) is a synthetic melanin pigment of great importance in biomedicine, where its affinity for metallic cations, especially paramagnetic ions, has sparked interest in its use in the development of magnetic resonance imaging (MRI) contrast agents. In this work, we report the cytotoxicity of metal-enriched PD nanoparticles on NIH3T3, a healthy cell line and BT474, a breast cancer cell line. Remarkably, it was found that the metal- enriched PD particles (Mn+ = Fe3+, Fe2+ and Cu2+) were highly cytotoxic to the breast cancer cells, even after 24 h of treatment. Although, this effect was not selective systems, since an acute cytotoxic effect was also observed on the healthy cell line, this system can be considered as starting point for designing advanced antineoplastic agents.

Polydopamine nanoparticles kill cancer cells.

Polydopamine (PD) is a synthetic melanin analogue of growing importance in the field of biomedicine, especially with respect to cancer research, due, in part, to its biocompatibility. But little is known about the cytotoxic effects of PD on cancer cell lines. PD is a UV-vis absorbing material whose absorbance overlaps with that of formazan salts, which are used to assess cell viability in MTT assays. In this study, a protocol has been established to eliminate the contributing absorbance of PD at 550 nm, and has been applied to characterize the cytotoxicity of PD nanoparticles in both healthy and breast cancer cell lines. Once the protocol is applied, it was found that PD is per se an antineoplastic system, meaning it selectively kills cancer cells, especially those of breast cancer, but it has no toxic effect on healthy cells. The mechanism of action could be related to the production of ROS and the alteration of iron homeostasis in lysosomes. To the best of our knowledge there are only a few examples of nanoparticle systems devoid of drugs that selectively kill cancer cells.

Effect of bacteria type and sucrose concentration on levan yield and its molecular weight.

BACKGROUND: Levan has been traditionally produced from microorganism. However, there is a continuous effort in looking for new strains that improve levan production yield and uses alternative sugar sources for growth. Despite having a wide range of data about levan yield, there are not papers which allow controlling molecular weight, and that plays an essential role for further applications. RESULTS: The effect of the sucrose concentration on levan yield (and its molecular weight) from Bacillus atrophaeus and Acinetobacter nectaris (Gram positive and Gram negative respectively) was studied in this work. It was found that A. nectaris growth (from 3 to 1.5 g L-1 in 40 h) and its levan production (from 3 to 1.5 g L-1) decreases by increasing sucrose concentration (best results at a concentration of 120 g L-1) whereas B. atrophaeus growth (3.5 g L-1 in 30 h) and its levan production (also 3.5 g L-1) were not affected by modifying that parameter. Levan molecular weight from A. nectaris decreases by increasing sucrose concentration (from 8000 to 2000 kDa) whereas levan molecular weight from B. Atrophaeus remains always around 50 kDa. By performing a kinetic study, it was shown that A. nectaris growth follows a substrate-inhibition model, whereas Monod equation provided a good fit for B. atrophaeus growth. Finally, wastes from orange juice industry were used as a medium culture to cultivate those microorganism, obtaining good results with B. atrophaeus (growth 3 g L-1 in 30 h). CONCLUSIONS: Levan production kinetics was determined and compared between different bacteria types.

Molecular Approach to the Synergistic Effect on Astringency Elicited by Mixtures of Flavanols.

The interactions between salivary proteins and wine flavanols (catechin, epicatechin, and mixtures thereof) have been studied by HPLC-DAD, isothermal titration microcalorimetry, and molecular dynamics simulations. Chromatographic results suggest that the presence of these flavanol mixtures could facilitate the formation of precipitates to the detriment of soluble aggregates. Comparison between the thermodynamic parameters obtained showed remarkably higher negative values of ΔG in the system containing the mixture of both flavanols in comparison to the systems containing individual flavanols, indicating a more favorable scenario in the mixing system. Also, the apparent binding constants were higher in this system. Furthermore, molecular dynamics simulations suggested a faster and greater cooperative binding of catechin and epicatechin to IB714 peptides when both types of flavanols are present simultaneously in solution.

Development of a nanoparticle system based on a fructose polymer: Stability and drug release studies.

New drug delivery systems (DDSs) with levan or its carboxymethylated form, as carriers, and 5-fluorouracil as a drug, are produced in this work. Levan is obtained after cultivating A. nectaris and polymer nanoparticles are created in water by a self-assembled process. The effect of pH and the ionic strength on polymer nanoparticles aggregation is studied. Basic pHs produces a particle size between 300 and 400nm with a Z-potential around -20mV because a basic medium promotes repulsion forces. DDSs of 300-400nm and a Z-potential about -25mV are prepared by taking advantage of the amphiphilic properties of the levan. The drug is bound to either levan or carboxymethyllevan surfaces by electrostatic interactions, obtaining the best results at basic pHs. 45-70% of the drug is released from the levan in 23h depending on the pH preparation, whereas only a low percentage of the drug is released from the carboxymethyllevan.

Understanding and optimizing the addition of phytohormones in the culture of microalgae for lipid production.

Some studies have described the use of phytohormones in microalgal culture for the production of biodiesel or selected fatty acids. However, no study has determined the amount of phytohormones that maximizes lipid yield. We determined the optimal concentration of auxins and gibberellins (which is between 40 and 60 µM) in two strains (Scenedemus abundans and Chlorella ellipsoidea) suitable for biodiesel production. More than 3-fold increment was reached with S. abundans and near 7-fold increment with C. ellipsoidea. Furthermore, this work suggests that the improved growth of the microalgae in the presence of the phytohormones was due to a reduction in the level of reactive oxygen species (ROS) in the cells. An economic analysis showed that, due to its low cost, auxin offers a positive cost-benefit balance and therefore could be used at large scale. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1203-1211, 2016.

Effect of nitrogen source on growth and lipid accumulation in Scenedesmus abundans and Chlorella ellipsoidea.

Discovering microalgae strains containing a high lipid yield and adequate fatty acid composition is becoming a crucial fact in algae-oil factories. In this study, two unknown strains, named Scenedesmus abundans and Chlorella ellipsoidea, have been tested for their response to different nitrogen sources, in order to determine its influence in the production of lipids. For S. abundans, autotrophic culture with ammonium nitrate offers the maximum lipid yield, obtaining up to 3.55 mg L(-1) d(-1). For C. ellipsoidea, heterotrophic culture with ammonium nitrate has been shown to be the best condition, reaching a lipid production of 9.27 mg L(-1) d(-1). Moreover, fatty acid composition obtained from these cultures meets international biodiesel standards with an important amount of C18:1, achieving 70% of total fatty acids and thus representing a potential use of these two strains at an industrial scale.