One-pot synthesis of alginate-antimicrobial peptide nanogel.

Nanosized alginate-based particles (NAPs) were obtained in a one-pot solvent-free synthesis procedure, achieving the design of a biocompatible nanocarrier for the encapsulation of IbM6 antimicrobial peptide (IbM6). IbM6 is integrated in the nascent nanosized hydrogel self-assembly guided by electrostatic interactions and by weak interactions, typical of soft matter. The formation of the nanogel is a dynamic and complex process, which presents an interesting temporal evolution. In this work, we optimized the synthesis conditions of IbM6-NAPs based on small-angle X-ray scattering (SAXS) measurements and evaluated its time evolution over several weeks by sensing the IbM6 environment in IbM6-NAPs from photochemical experiments. Fluorescence deactivation experiments revealed that the accessibility of different quenchers to the IbM6 peptide embedded in NAPs is dependent on the aging time of the alginate network. Lifetimes measurements indicate that the deactivation paths of the excited state of the IbM6 in the nanoaggregates are reduced when compared with those exhibited by the peptide in aqueous solution, and are also dependent on the aging time of the nanosized alginate network. Finally, the entrapment of IbM6 in NAPs hinders the degradation of the peptide by trypsin, increasing its antimicrobial activity against Escherichia coli K-12 in simulated operation conditions.

Modulation of TRPV1 on Odontoblast-like Cells Using Capsazepine-Loaded Nanogels.

The modulation of TRPV1 emerges as a promising strategy for dental pain management. This study aimed to assess TRPV1 modulation in a human odontoblast-like cell model using Capsazepine (CZP) loaded in a nanogel delivery system. Gelatin nanogels, synthesized via the emulsification-gelation technique, were characterized and loaded with the TRPV1 antagonist, CZP. HPLC determined a remarkable 67.5 ± 0.04% CZP loading efficiency, with 71.7% of nanogels falling within the 300-950 nm size range, as evidenced by light microscopy. Moreover, CZP-loaded nanogels had a low cytotoxicity. An FTIR analysis showed no adverse chemical interactions, ensuring stability and active release. When examining biological responses, TRPV1 expression and channel activity were assessed in odontoblast-like cells. On the fifth day post-treatment, cells treated with CZP-loaded nanogels exhibited an increased TRPV1 expression and a reduction in calcium fluxes after agonist stimulus (F/F0 ratio 1.18 ± 0.18), resembling the response in free CZP-treated cells (1.28 ± 0.15). A two-way analysis of variance and the Tukey's test were used to determine statistical significance (p < 0.05). This delivery system, proven to be economical and straightforward, holds promise for dental pain management and potential local use. Local administration minimizes systemic adverse effects, making it a practical solution for releasing molecules in the oral cavity.

Wound Healing Effect of Lippia sidoides and Myracrodruon urundeuva Nanogel.

Wound healing is a natural regenerative response to tissue injury and the conventional treatments consists of the use wound dressings with local administration of medicines, but, in some cases, are only partially effective and limited by toxicity or ineffective anti-microbial protection. Medicinal plants such as Lippia sidoides and Myracrodruon urundeuva have shown interesting pharmacological activities, allied to this, the association of these medicinal plants and nanotechnology, could mean an advantage in relation to classical approach. This study investigated the effect of a nanogel loaded with Lippia sidoides essential oil and Myracrodruon urundeuva extract (NAA) in an excisional wound healing model in rats. Animals were anesthetized and skin wounds were made using a metal punch. The groups were treated with vehicle, NAA or collagenase gel, for 7, 14 or 21 days and then sacrificed for tissue analysis. NAA did not show acute dermal irritation, further significantly reduced (p<0.05) the final wound area, accelerated the wound contraction and organization of collagen in the group treated for 14 days. The data presented here demonstrate the therapeutic potential for the use of nanotechnology associated with medicinal plants and provides evidence that corroborate with the use of L. sidoides and M. urundeuva as healing medicinal plants.

Synthesis of poly (N-vinyl pyrrolidone) (PVP) nanogels by gamma irradiation using different saturation atmospheres

Nanogels are internally crosslinked particles of nanometric size used in various fields e.g. as such as carriers in drug delivery systems. They can be produced using ionizing radiation in dilute aqueous solutions. This method is carried out in a pure polymer-solvent system, avoiding the addition of any additives such as monomers, surfactants, catalysts and crosslinking agents and no further purification step is necessary. Poly(N-vinyl pyrrolidone) (PVP K-90) nanogels were prepared by gamma irradiation in an aqueous solution. The samples were prepared in triplicate in multipurpose cobalt-60 gamma irradiator using 1, 10, 25 and 100 mM PVP solutions. Samples were irradiated in argon and nitrous oxide conditions with doses from 1 kGy up to 25 kGy with 10 kGy/h dose rate. The mean particle size (Rh) was determined by Dynamic Light Scattering (DLS) and radius of gyration (Rg) and weight-average molecular weight (Mw) by Static Light Scattering (SLS). These samples were morphologically characterized using Atomic Force Microscopy (AFM) and Transmission Electron Microscopy (TEM). Samples prepared with 100 mM PVP K-90 solution formed macroscopic gels, in the samples obtained with 25 mM PVP K-90 solution there was a prevalence of intermolecular crosslinking. On the other hand, in the samples generated with 10 mM PVP K-90 solution, there was a predominance of intramolecular crosslinking demonstrated in the tendency to: decrease in the radius of gyration (Rg), in the constancy of the weight-average molecular weight (Mw), in the increase in polymer coil density (ρcoil), in the Rg/Rh ratio (shape factor) around 1.0 indicating homogenous, internally cross-linked spheres, in the high relief spherical structures observed in the AFM images and in the spherical particles with high contrast observed in the TEM images. The saturation of the samples with nitrous oxide doubled formation of hydroxyl radicals, favoring the generation of polymeric radicals. Higher average number of radicals in each macromolecule contributed to the higher number of intramolecular crosslinks.

Low nanogel stiffness favors nanogel transcytosis across an in vitro blood-brain barrier.

Transport of therapeutics across the blood-brain barrier (BBB) is a fundamental requirement for effective treatment of numerous brain diseases. However, most therapeutics (>500 Da) are unable to permeate through the BBB and do not achieve therapeutic doses. Nanoparticles (NPs) are being investigated to facilitate drug delivery to the brain. Here, we investigate the effect of nanoparticle stiffness on NP transport across an in vitro BBB model. To this end, fluorescently labeled poly(N-isopropylmethacrylamide) (p(NIPMAM)) nanogels' stiffness was varied by the inclusion of 1.5 mol% (NG1.5), 5 mol% (NG5), and 14 mol% (NG14) N,N'-methylenebis(acrylamide) (BIS) cross-linker and nanogel uptake and transcytosis was quantified. The more densely cross-linked p(NIPMAM) nanogels showed the highest level of uptake by polarized brain endothelial cells, whereas the less densely cross-linked nanogels demonstrated the highest transcytotic potential. These findings suggest that nanogel stiffness has opposing effects on nanogel uptake and transcytosis at the BBB.

Betulinic Acid Nanogels: Rheological, Microstructural Characterization and Evaluation of their Anti-inflammatory Activity.

BACKGROUND: Betulinic Acid (BA) is a lipophilic compound with proven beneficial results in topical inflammation. Nanogels (NG) are carriers of bioactive compounds with properties that make them good candidates to treat skin diseases. OBJECTIVE: The objective of this study was to evaluate the anti-inflammatory activity of BA carried in NG. METHODS: NG were composed of a nanoemulsion and a crosslinking agent (Carbopol 940®) applied at three concentrations (0.5, 1, and 1.5 %) and three activation times (6, 12 and 24 h). In order to select the optimal formulation, the NG were characterized mechanically and micro-structurally followed by evaluation of the BA anti-inflammatory activity in an in vivo model of auricular edema. We determined the edema inhibition activity as percent weight. Additionally, the anti-inflammatory activity of NG was validated through histological analysis. RESULTS: The formulation with the best viscoelastic properties was the one prepared with 0.5% carbopol and 6 h of activation. Microstructural examination of this formulation showed mostly spherical structures with a mean diameter of 65 nm. From the evaluation of edema and the histological analyses, we established that the NG of BA produced 52% inhibition. In contrast, a conventional gel and free BA produced 28% and 19% inhibition, respectively. CONCLUSION: The NG of BA were found to be good vehicles to treat skin inflammation.

Encapsulation of essential oils using cinnamic acid grafted chitosan nanogel: Preparation, characterization and antifungal activity.

Chemical modifications in the chitosan structure may result in obtaining a new material with improved chemical properties, such as an ability to encapsulate lipophilic compounds. This study aimed to synthesize cinnamic acid grafted chitosan nanogel to encapsulate the essential oils of Syzygium aromaticum and Cinnamomum ssp., in order to develop a material to be applied in the control of dermatophytosis caused by the fungus Microsporum canis. The cinnamic acid graft in chitosan was verified by the Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Solid State Nuclear Magnetic Resonance of the 13C Nucleus (13C SSNMR) and Thermal analysis coupled to mass spectrometry (TG-MS) techniques. The nanogel obtained showed affinity for the essential oils of S. aromaticum and Cinnamomum, with encapsulation efficiencies equal to 74% and 89%, respectively. When in an aqueous medium the nanogel with the encapsulated essential oils was able to form stable nanoparticles with average sizes of 176.0 ± 54.3 nm and 263.0 ± 81.4 nm. The cinnamic acid grafted chitosan nanogel showed antifungal activity in vitro against M. canis, inhibiting up to 53.96% of its mycelial growth. Complete inhibition of mycelial growth was achieved by the nanogel with encapsulated essential oils. The results found in this work demonstrated the development of a material with potential application in the control of dermatophytosis caused by the fungus M. canis.

Nanogels as prospective Biomaterial: Radio-induced Synthesis, Characterization, and Biological Assays / Los nanogeles como biomateriales prospectivos: síntesis radioinducida, caracterización y ensayos biológicos

Nowadays, there is a growing interest in biodegradable polymers-based materials due to their diverse application in the biomedical field. Most studied systems involve biocompatible micro and nanodevices, such as liposomes, dendrimer, micelles or polymeric nanogels. The use of Radiation Technology, specifically gamma radiation, to produce micro and nanogels raises the possibility to obtain higher purity products, an important feature for biomedical and pharmaceutical applications. The radio-induced synthesis, characterization, cytotoxicity evaluation, and immunological response of nanogels are described in this study. Nanogel synthesis was performed in the absence of oxygen using aqueous polyvinylpyrrolidone solutions. Crosslinking reactions were carried out at 25 °C in a gamma irradiation chamber with a 60Co source. Nanogels properties were analysed by Scanning Electron Microscopy, Attenuated Total Reflection-Fourier Transform Spectroscopy, Dynamic Light Scattering, and Viscosimetry. The cytotoxicity and immunological response were evaluated by MTT test and analysis of the neutrophil respiratory burst. The results showed that nanogels formation strongly depends on the total absorbed dose. The nanogels have an elliptical shape and their chemical structure is similar to the initial polymer. The nanogels are biocompatible and promote a low-intensity neutrophil activation, similar to the well-characterized biomaterial TiO2, suggesting their potential biomedical uses(AU)

En la actualidad existe un interés creciente en los materiales biodegradables basados en polímeros, debido a sus diversas aplicaciones en la esfera de la biomedicina. En la mayoría de los sistemas estudiados participan micro- y nanodispositivos biocompatibles, tales como liposomas, dendrímeros, micelas o nanogeles poliméricos. El uso de la tecnología de radiaciones, en particular de radiaciones gamma, para producir micro- y nanogeles, eleva la posibilidad de obtener productos de mayor pureza, un rasgo importante con vistas a su aplicación biomédica y farmacéutica. El estudio describe la síntesis radioinducida, caracterización, evaluación de la citotoxicidad y respuesta inmunológica de los nanogeles. La síntesis de los nanogeles se realizó en ausencia de oxígeno, usando soluciones acuosas de polivinilpirrolidona. Las reacciones de entrecruzamiento se realizaron a 25 ºC en cámara de irradiación gamma con una fuente de 60Co. Las propiedades de los nanogeles se analizaron mediante microscopía electrónica de barrido, espectroscopia por transformada de Fourier total atenuada, dispersión dinámica de luz y viscosimetría. La citotoxicidad y la respuesta inmunológica se evaluaron mediante prueba MTT y análisis del estallido respiratorio de neutrófilos. Los resultados muestran que la formación de nanogeles depende en gran medida de la dosis total absorbida. Los nanogeles tienen forma elíptica y su estructura química es similar a la del polímero inicial. Los nanogeles son biocompatibles y promueven una activación de neutrófilos de baja intensidad similar al bien caracterizado material TiO2, lo que sugiere usos biomédicos potenciales(AU)

An Updated Review of Macro, Micro, and Nanostructured Hydrogels for Biomedical and Pharmaceutical Applications.

Hydrogels are materials with wide applications in several fields, including the biomedical and pharmaceutical industries. Their properties such as the capacity of absorbing great amounts of aqueous solutions without losing shape and mechanical properties, as well as loading drugs of different nature, including hydrophobic ones and biomolecules, give an idea of their versatility and promising demand. As they have been explored in a great number of studies for years, many routes of synthesis have been developed, especially for chemical/permanent hydrogels. In the same way, stimuli-responsive hydrogels, also known as intelligent materials, have been explored too, enhancing the regulation of properties such as targeting and drug release. By controlling the particle size, hydrogel on the micro- and nanoscale have been studied likewise and have increased, even more, the possibilities for applications of the so-called XXI century materials. In this paper, we aimed to produce an overview of the recent studies concerning methods of synthesis, biomedical, and pharmaceutical applications of macro-, micro, and nanogels.

Development and optimization of erythromycin-loaded lipid-based gel by Taguchi design: In vitro characterization and antimicrobial evaluation

The foremost aim of the current research was to prolong and sustain the release of erythromycin (ERY) by preparing a solid lipid nanoparticles (SLNs)-based gel formulation for the safe and effective treatment of acne. ERY-loaded SLNs were developed, and various process variables were optimized with respect to particle size, zeta potential, and entrapment efficiency using the Taguchi model. The average particle size, PDI, zeta potential, drug entrapment efficiency, and drug loading of optimized SLN (F4) were found to be 176.2±1.82 nm, 0.275±0.011, -34.0±0.84, 73.56%, and 69.74% respectively. The optimized SLN (F4) was successfully incorporated into the carbopol-based hydrogel. The in vitro release of ERY from the SLN gel and plain gel were compared and found to be 90.94% and 87.94% respectively. In vitro study of ERY-loaded SLN gel showed sustained delivery of drug from formulation thus enhancing the antimicrobial activity after 30 hours when compared to ERY plain gel.

Optimizing Circulating Tumor Cells' Capture Efficiency of Magnetic Nanogels by Transferrin Decoration.

Magnetic nanogels (MNGs) are designed to have all the required features for their use as highly efficient trapping materials in the challenging task of selectively capturing circulating tumor cells (CTCs) from the bloodstream. Advantageously, the discrimination of CTCs from hematological cells, which is a key factor in the capturing process, can be optimized by finely tuning the polymers used to link the targeting moiety to the MNG. We describe herein the relationship between the capturing efficiency of CTCs with overexpressed transferrin receptors and the different strategies on the polymer used as linker to decorate these MNGs with transferrin (Tf). Heterobifunctional polyethylene glycol (PEG) linkers with different molecular weights were coupled to Tf in different ratios. Optimal values over 80% CTC capture efficiency were obtained when 3 PEG linkers with a length of 8 ethylene glycol (EG) units were used, which reveals the important role of the linker in the design of a CTC-sorting system.

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.