Multifunctional Polymeric Biodegradable and Biocompatible Coatings Based on Silver Nanoparticles: A Comparative In Vitro Study on Their Cytotoxicity towards Cancer and Normal Cell Lines of Cytostatic Drugs versus Essential-Oil-Loaded Nanoparticles and on Their Antimicrobial and Antibiofilm Activities.

We report on a comparative in vitro study of selective cytotoxicity against MCF7 tumor cells and normal VERO cells tested on silver-based nanocoatings synthesized by the matrix-assisted pulsed laser evaporation (MAPLE) technique. Silver nanoparticles (AgNPs) were loaded with five representative cytostatic drugs (i.e., doxorubicin, fludarabine, paclitaxel, gemcitabine, and carboplatin) and with five essential oils (EOs) (i.e., oregano, rosemary, ginger, basil, and thyme). The as-obtained coatings were characterized by X-ray diffraction, thermogravimetry coupled with differential scanning calorimetry, Fourier-transform IR spectroscopy, IR mapping, and scanning electron microscopy. A screening of the impact of the prepared nanocoatings on the MCF7 tumor and normal VERO cell lines was achieved by means of cell viability MTT and cytotoxicity LDH assays. While all nanocoatings loaded with antitumor drugs exhibited powerful cytotoxic activity against both the tumor and the normal cells, those embedded with AgNPs loaded with rosemary and thyme EOs showed remarkable and statistically significant selective cytotoxicity against the tested cancercells. The EO-loaded nanocoatings were tested for antimicrobial and antibiofilm activity against Staphylococcus aureus, Escherichia coli, and Candida albicans. For all studied pathogens, the cell viability, assessed by counting the colony-forming units after 2 and 24 h, was significantly decreased by all EO-based nanocoatings, while the best antibiofilm activity was evidenced by the nanocoatings containing ginger and thyme EOs.

Zinc/Cerium-Substituted Magnetite Nanoparticles for Biomedical Applications.

Numerous studies have reported the possibility of enhancing the properties of materials by incorporating foreign elements within their crystal lattice. In this context, while magnetite has widely known properties that have been used for various biomedical applications, the introduction of other metals within its structure could prospectively enhance its effectiveness. Specifically, zinc and cerium have demonstrated their biomedical potential through significant antioxidant, anticancer, and antimicrobial features. Therefore, the aim of the present study was to develop a series of zinc and/or cerium-substituted magnetite nanoparticles that could further be used in the medical sector. The nanostructures were synthesized through the co-precipitation method and their morpho-structural characteristics were evaluated through X-ray diffraction (XRD), inductively coupled plasma mass spectrometry (ICP-MS), X-ray photoelectron spectroscopy (XPS), dynamic light scattering (DLS), zeta potential, scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDX) analyses. Furthermore, the nanostructures were subjected to a ROS-Glo H2O2 assay for assessing their antioxidant potential, MTT assay for determining their anticancer effects, and antimicrobial testing against S. aureus, P. aeruginosa, and C. albicans strains. Results have proven promising for future biomedical applications, as the nanostructures inhibit oxidative stress in normal cells, with between two- and three-fold reduction and cell proliferation in tumor cells; a two-fold decrease in cell viability and microbial growth; an inhibition zone diameter of 4-6 mm and minimum inhibitory concentration (MIC) of 1-2 mg/mL.

Silver/Graphene Oxide Nanostructured Coatings for Modulating the Microbial Susceptibility of Fixation Devices Used in Knee Surgery.

Exploring silver-based and carbon-based nanomaterials' excellent intrinsic antipathogenic effects represents an attractive alternative for fabricating anti-infective formulations. Using chemical synthesis protocols, stearate-conjugated silver (Ag@C18) nanoparticles and graphene oxide nanosheets (nGOs) were herein obtained and investigated in terms of composition and microstructure. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) characterizations revealed the formation of nanomaterials with desirable physical properties, while X-ray diffraction (XRD) analyses confirmed the high purity of synthesized nanomaterials. Further, laser-processed Ag@C18-nGO coatings were developed, optimized, and evaluated in terms of biological and microbiological outcomes. The highly biocompatible Ag@C18-nGO nanostructured coatings proved suitable candidates for the local modulation of biofilm-associated periprosthetic infections.

Dextran-Coated Iron Oxide Nanoparticles Loaded with Curcumin for Antimicrobial Therapies.

The current trend in antimicrobial-agent development focuses on the use of natural compounds that limit the toxicity of conventional drugs and provide a potential solution to the antimicrobial resistance crisis. Curcumin represents a natural bioactive compound with well-known antimicrobial, anticancer, and antioxidant properties. However, its hydrophobicity considerably limits the possibility of body administration. Therefore, dextran-coated iron oxide nanoparticles can be used as efficient drug-delivery supports that could overcome this limitation. The iron oxide nanoparticles were synthesized through the microwave-assisted hydrothermal method by varying the treatment parameters (pressure and reaction time). The nanoparticles were subsequently coated with dextran and used for the loading of curcumin (in various concentrations). The drug-delivery systems were characterized through X-ray diffraction (XRD) coupled with Rietveld refinement, transmission electron microscopy (TEM), high-resolution TEM (HR-TEM), selected area electron diffraction (SAED), dynamic light scattering (DLS) and zeta potential, thermogravimetry and differential scanning calorimetry (TG-DSC), vibrating sample magnetometry (VSM), and UV-Vis spectrophotometry, as well as regarding their antimicrobial efficiency and biocompatibility using the appropriate assays. The results demonstrate a promising antimicrobial efficiency, as well as an increased possibility of controlling the properties of the resulted nanosystems. Thus, the present study represents an important step forward toward the development of highly efficient antimicrobial drug-delivery systems.

Antibacterial Biodegradable Films Based on Alginate with Silver Nanoparticles and Lemongrass Essential Oil-Innovative Packaging for Cheese.

Replacing the petroleum-based materials in the food industry is one of the main objectives of the scientists and decision makers worldwide. Biodegradable packaging will help diminish the environmental impact of human activity. Improving such biodegradable packaging materials by adding antimicrobial activity will not only extend the shelf life of foodstuff, but will also eliminate some health hazards associated with food borne diseases, and by diminishing the food spoilage will decrease the food waste. The objective of this research was to obtain innovative antibacterial films based on a biodegradable polymer, namely alginate. Films were characterized by environmental scanning electron microscopy (ESEM), Fourier-transform infrared spectroscopy (FTIR) and microscopy, complex thermal analysis (TG-DSC-FTIR), UV-Vis and fluorescence spectroscopy. Water vapor permeability and swelling behavior were also determined. As antimicrobial agents, we used silver spherical nanoparticles (Ag NPs) and lemongrass essential oil (LGO), which were found to act in a synergic way. The obtained films exhibited strong antibacterial activity against tested strains, two Gram-positive (Bacillus cereus and Staphylococcus aureus) and two Gram-negative (Escherichia coli and Salmonella Typhi). Best results were obtained against Bacillus cereus. The tests indicate that the antimicrobial films can be used as packaging, preserving the color, surface texture, and softness of cheese for 14 days. At the same time, the color of the films changed (darkened) as a function of temperature and light presence, a feature that can be used to monitor the storage conditions for sensitive food.

Biodegradable Alginate Films with ZnO Nanoparticles and Citronella Essential Oil-A Novel Antimicrobial Structure.

The petroleum-based materials could be replaced, at least partially, by biodegradable packaging. Adding antimicrobial activity to the new packaging materials can also help improve the shelf life of food and diminish the spoilage. The objective of this research was to obtain a novel antibacterial packaging, based on alginate as biodegradable polymer. The antibacterial activity was induced to the alginate films by adding various amounts of ZnO nanoparticles loaded with citronella (lemongrass) essential oil (CEO). The obtained films were characterized, and antibacterial activity was tested against two Gram-negative (Escherichia coli and Salmonella Typhi) and two Gram-positive (Bacillus cereus and Staphylococcus aureus) bacterial strains. The results suggest the existence of synergy between antibacterial activities of ZnO and CEO against all tested bacterial strains. The obtained films have a good antibacterial coverage, being efficient against several pathogens, the best results being obtained against Bacillus cereus. In addition, the films presented better UV light barrier properties and lower water vapor permeability (WVP) when compared with a simple alginate film. The preliminary tests indicate that the alginate films with ZnO nanoparticles and CEO can be used to successfully preserve the cheese. Therefore, our research evidences the feasibility of using alginate/ZnO/CEO films as antibacterial packaging for cheese in order to extend its shelf life.

Electrochemotherapy and Other Clinical Applications of Electroporation for the Targeted Therapy of Metastatic Melanoma.

Electrochemotherapy (ECT) is an effective bioelectrochemical procedure that uses controlled electrical pulses to facilitate the increase of intracellular concentration of certain substances (electropermeabilization/ reversible electroporation). ECT using antitumor drugs such as bleomycin and cisplatin is a minimally invasive targeted therapy that can be used as an alternative for oncologic patients not eligible for surgery or other standard therapies. Even though ECT is mainly applied as palliative care for metastases, it may also be used for primary tumors that are unresectable due to size and location. Skin neoplasms are the main clinical indication of ECT, the procedure reporting good curative results and high efficiency across all tumor types, including melanoma. In daily practice, there are many cases in which the patient's quality of life can be significantly improved by a safe procedure such as ECT. Its popularity must be increased because it has a safe profile and minor local adverse reactions. The method can be used by dermatologists, oncologists, and surgeons. The aim of this paper is to review recent literature concerning electrochemotherapy and other clinical applications of electroporation for the targeted therapy of metastatic melanoma.

MAPLE Coatings Embedded with Essential Oil-Conjugated Magnetite for Anti-Biofilm Applications.

The present study reports on the development and evaluation of nanostructured composite coatings of polylactic acid (PLA) embedded with iron oxide nanoparticles (Fe3O4) modified with Eucalyptus (Eucalyptus globulus) essential oil. The co-precipitation method was employed to synthesize the magnetite particles conjugated with Eucalyptus natural antibiotic (Fe3O4@EG), while their composition and microstructure were investigated using grazing incidence X-ray diffraction (GIXRD), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and dynamic light scattering (DLS). The matrix-assisted pulsed laser evaporation (MAPLE) technique was further employed to obtain PLA/Fe3O4@EG thin films. Optimal experimental conditions for laser processing were established by complementary infrared microscopy (IRM) and scanning electron microscopy (SEM) investigations. The in vitro biocompatibility with eukaryote cells was proven using mesenchymal stem cells, while the anti-biofilm efficiency of composite PLA/Fe3O4@EG coatings was assessed against Gram-negative and Gram-positive pathogens.

Polyphenols of Honeybee Origin with Applications in Dental Medicine.

Honeybee products are a great source of polyphenols with recognized applications in dental medicine. Although their biological mechanisms in oral diseases are not fully understood, numerous in vitro, in vivo and clinical studies have reported promising results in the prevention and treatment of oral diseases. Bioactivities, such as antibacterial, antiviral, antiparasite, anticancer, anti-inflammatory and anti-oxidant properties, recommend their future study in order to develop efficient alternatives in the management of widespread oral conditions, such as dental caries and periodontitis. The most investigated mechanisms of polyphenols in oral health rely on their ability to strengthen the dental enamel, decrease the development of dental plaque formation, inhibit the progression of dental caries and development of dental pathogens and show anti-inflammatory properties. These features recommend them as useful honeybee candidates in the management of emerging oral diseases.

Nanostructured Thin Coatings Containing Anthriscus sylvestris Extract with Dual Bioactivity.

Plant extracts are highly valuable pharmaceutical complexes recognized for their biological properties, including antibacterial, antifungal, antiviral, antioxidant, anticancer, and anti-inflammatory properties. However, their use is limited by their low water solubility and physicochemical stability. In order to overcome these limitations, we aimed to develop nanostructured carriers as delivery systems for plant extracts; in particular, we selected the extract of Anthriscus sylvestris (AN) on the basis of its antimicrobial effect and antitumor activity. In this study, AN-extract-functionalized magnetite (Fe3O4@AN) nanoparticles (NPs) were prepared by the co-precipitation method. The purpose of this study was to synthesize and investigate the physicochemical and biological features of composite coatings based on Fe3O4@AN NPs obtained by matrix-assisted pulsed laser evaporation technique. In this respect, laser fluence and drop-casting studies on coatings were performed. The physical and chemical properties of laser-synthesized coatings were investigated by scanning electron microscopy, while Fourier transform infrared spectroscopy comparative analysis was used for determining the chemical structure and functional integrity. Relevant data regarding the presence of magnetic nanoparticles as the only crystalline phase and the size of nanoparticles were obtained by transmission electron microscopy. The in vitro toxicity assessment of the Fe3O4@AN showed significant cytotoxic activity against human adenocarcinoma HT-29 cells after prolonged exposure. Antimicrobial results demonstrated that Fe3O4@AN coatings inhibit microbial colonization and biofilm formation in clinically relevant bacteria species and yeasts. Such coatings are useful, natural, and multifunctional solutions for the development of tailored medical devices and surfaces.

Nanosystems for Improved Targeted Therapies in Melanoma.

Melanoma is one of the most aggressive forms of skin cancer, with limited therapeutic options. Since its incidence has been rapidly rising in recent years, the study of new targeted therapeutic strategies has increased. The implication of nanoscience in the development of alternative targeted therapies for melanoma has multiple benefits and could significantly improve the outcome of melanoma patients. In this paper, we review the most recent progress in the field of targeted therapies, emphasizing the impact of nanoscale materials on the targeting and controlled release of anti-tumor drugs. The applications of nanomedicine in the management of melanoma are extensive and refer to sentinel lymph node mapping, chemotherapy, and RNA interference; each of these applications harboring the potential to develop efficient and personalized diagnostic techniques and therapies. Further research, especially in clinical trials, is needed to establish whether fighting melanoma on the nanoscale level represents the key to reaching a critical inflection point in mankind's battle with metastatic melanoma.

Novel Therapeutic Strategies Applied to Pseudomonas aeruginosa Infections in Cystic Fibrosis.

Cystic fibrosis (CF) is one of the most prevalent genetic diseases and a total of 1700 different genetic mutations can cause this condition. Patients that suffer this disease have a thickening of the mucus, creating an environment that promotes bacterial infections. Pseudomonas aeruginosa is a ubiquitous bacterium, which is frequently found in the lungs of CF patients. P. aeruginosa is known for its high level of antibiotic resistance as well as its high rate of mutation that allows it to rapidly evolve and adapt to a multitude of conditions. When a CF lung is infected with P. aeruginosa, the decay of the patient is accelerated, but there is little that can be done apart from controlling the infection with antibiotics. Novel strategies to control P. aeruginosa infection are imperative, and nanotechnology provides novel approaches to drug delivery that are more efficient than classic antibiotic treatments. These drug delivery systems are offering new prospects, especially for these patients with special mucus conditions and bacterial characteristics that limit antibiotic use.

Structure-grain size-synthesis route of silver nanoparticles: a correlation with the cytotoxic effect.

Although engineered silver (Ag) nanopowders offer great promise in various fields of biomedical, industrial and ecological applications, insufficient data is known about their cytotoxicity. The purpose of the present study was the synthesis and then the determination of cytotoxicity effect of Ag powders using the pyrosol method, at various temperatures of 600°C, 650°C and 700°C, respectively by sol-gel method and heat treatments at 500°C, 600°C, 700°C and 800°C. From the structural, compositional and morphological point of view, Ag samples were characterized by using X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) coupled with selected area electron diffraction (SAED) techniques. There is an influence of the synthesis route on Ag particles, which is shown as their cytotoxicity, different sizes of micro and nanosilver synthesized powders, which were evaluated in comparison depending on the work methods. Their cytotoxicity was evaluated based on their influence on cellular morphology and proliferation rate, cell cycle and apoptosis of undifferential stem cells, endothelial cells and tumoral cells, assessed through flow cytometry, cloning and MTT assay. The results showed that the cytotoxicity of the obtained Ag nanoparticles (NPs) depends on the synthesis route, the pyrosol synthesized NPs exhibiting a higher cytotoxicity as compared to those obtained by the sol-gel method.

Magnetic Particles for Advanced Molecular Diagnosis.

Molecular diagnosis is the field that aims to develop nucleic-acid-based analytical methods for biological markers and gene expression assessments by combining laboratory medicine and molecular genetics. As it gradually becomes a clinical reality, molecular diagnosis could benefit from improvements resulting from thorough studies that could enhance the accuracy of these methods. The application of magnetic particles in molecular diagnosis tools has led to tremendous breakthroughs in terms of specificity, sensitivity, and discrimination in bioassays. Therefore, the aim of this review is to highlight the principles involved in the implementation of magnetic particles for sample preparation and targeted analyte isolation, purification, and extraction. Furthermore, the most recent advancements in the area of cancer and infectious disease diagnosis are presented, with an emphasis on screening and early stage detection.

Suberin/trans-Cinnamaldehyde Oil Nanoparticles with Antimicrobial Activity and Anticancer Properties When Loaded with Paclitaxel.

Suberin is a biopolyester that can be extracted from bioresources such as cork trees and may find many applications in food packaging, sealing, and biomedical fields. In this work, we describe for the first time the preparation of suberin nanoparticles using the solvent/antisolvent method. Three different suberin extraction methods from cork trees were used to reveal any differences in the production and properties of the nanoparticles. trans-Cinnamaldehyde, a component of cinnamon essential oil, was added to suberin nanoparticles due to its antimicrobial and/or anticancer properties, and its synergistic effect was studied with the inherent biological properties of suberin. Interestingly, both suberin and suberin/trans-cinnamaldehyde nanoparticles showed very good antimicrobial properties against Staphylococcus aureus and Pseudomonas aeruginosa bacteria, and also against Candida albicans (yeast). Suberin nanoparticles containing trans-cinnamaldehyde loaded with 0.1% w/w paclitaxel provided significant anticancer properties against human hepatocellular carcinoma HepG2 cancer cell lines. This study can be used to explore novel biobased polymers as efficient carriers of essential oils and drugs for infection and cancer therapy, by using minimum amount of drugs and therefore reducing their cytotoxicity and side effects.

Cellulose acetate - essential oil nanocapsules with antimicrobial activity for biomedical applications.

This study aimed to obtain bioactive nanosystems by combining cellulose acetate with three selected essential oils (EOs) to create spherical nanocapsules (NCs) using the solvent/anti-solvent technique. The biological activity of the obtained NCs was promoted by the use of some antimicrobial EOs: Peppermint, Cinnamon and lemongrass which were grafted on the cellulose acetate molecules. Due to their chemistry, such as long hydrocarbon tails and heads with functional groups these EOs were playing also the role of surfactant-like substance facilitating the formation of NCs. A dispersion of NCs was obtained in water and various spectroscopy techniques used to examine their size, morphology and chemistry. Dynamic light scattering calculate the size of the NCs whereas scanning electron microscopy showed their morphology. Fluorescent microscopy and Raman spectroscopy proved the attachment of the EOs in the cellulose acetate molecules. The antimicrobial activity of the obtained nanomaterials was tested against four microbial strains (bacteria: Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and a yeast strain of Candida albicans). The obtained results demonstrated that such NCs can be used in a variety of applications including medical, pharmaceutical recipients and in household products for treating or preventing microbial colonization and biofilm development.

Aspects of Gut Microbiota and Immune System Interactions in Infectious Diseases, Immunopathology, and Cancer.

The microbiota consists of a dynamic multispecies community of bacteria, fungi, archaea, and protozoans, bringing to the host organism a dowry of cells and genes more numerous than its own. Among the different non-sterile cavities, the human gut harbors the most complex microbiota, with a strong impact on host homeostasis and immunostasis, being thus essential for maintaining the health condition. In this review, we outline the roles of gut microbiota in immunity, starting with the background information supporting the further presentation of the implications of gut microbiota dysbiosis in host susceptibility to infections, hypersensitivity reactions, autoimmunity, chronic inflammation, and cancer. The role of diet and antibiotics in the occurrence of dysbiosis and its pathological consequences, as well as the potential of probiotics to restore eubiosis is also discussed.

Bioactive mesoporous silica nanostructures with anti-microbial and anti-biofilm properties.

The increasing rate of antibiotic resistant bacteria associated with nosocomial infections in severely ill patients has urged the need for new antibacterial therapies. Nanostructured materials represent emerging innovative approaches to controlled delivery of different antimicrobial drugs. Delivery systems encapsulating natural compounds with antibacterial effects, such as essential oils have shown a great potential. Herein we report the development of SiO2 mesoporous nanosystems loaded with eucalyptus (EUC), orange (ORA), and cinnamon (CIN) essential oils. These systems were characterized with respect to morphology (using scanning electron microscopy, SEM, and transmission electron microscopy, TEM), porosity (by BET and TEM analysis), chemical composition (by X-ray diffraction, XRD, and Fourier transform infrared spectrometry, FTIR) and loading capacity (by thermogravimetric analysis, TGA). The anti-bacterial and anti-adherence effects were tested against clinically relevant microbial species (Staphylococcus aureus ATCC 25923; Escherichia coli ATCC 25922; and Candida albicans ATCC 10231), while the biocompatibility was evaluated by in vitro tests with L929 mouse fibroblast cells.

Phenotypic and Genetic Evaluation of the Influence of Pseudomonas aeruginosa Culture Fractions on the Human Mesenchymal Stem Cells Viability, Apoptotic Pathways and Cytokine Profile.

The objective of this study was to investigate the effects of P. aeruginosa PAO1 cellular and soluble culture fractions on human mesenchymal stem cells (MSCs) death signaling pathways and cytokine profile. The bone marrow isolated MSCs, incubated for different periods of time with one of the three P. aeruginosa PAO1 culture fractions, i.e. low density whole cultures, heat inactivated bacterial cultures sediments and sterile supernatants, were submitted to the following assays: i) fluorescence microscopy evaluation of cellular morphology and viability; ii) bax, caspase 9, relA and bcl-2 genes expression analysis by qRT-PCR; and iii) quantification of the level of IL-1ß, IL-6, IL-8 and IL-10 cytokines released in the MSCs supernatants determined by ELISA. Results were statistically analyzed using the GraphPad In Stat software. The PAO1 whole cultures exhibited the most relevant influences, impacting on MSCs morphology and viability, interfering with apoptotic pathways and significantly stimulating the production of IL-1ß and IL-10, while decreasing the production of IL-6 and IL-8. The culture supernatants increased the production of IL-1ß and reduced the secretion of all other tested cytokines, while heat-inactivated bacterial cells significantly stimulated both IL-1ß and IL-10 production. These data could suggest that in vivo, the fate of P. aeruginosa infection depends on the proportion between different bacterial culture fractions (i.e. the number of viable bacterial cells, the number of dead cells and the amount of bacterial soluble products accumulated locally) that could be influenced by the initial infective dose, by the host defense mechanisms, and also by the administered antimicrobial treatment that may thus interfere with the evolution and magnitude of the induced lesions.

Polylactic Acid-Lemongrass Essential Oil Nanocapsules with Antimicrobial Properties.

Polylactic acid was combined with lemongrass essential oil (EO) to produce functional nanocapsules (NCs). The obtained polylactic acid nanoparticles showed antimicrobial activity both with and without the presence of lemongrass oil; however, the presence of EO improved the activity of the NCs. The presence of lemongrass assisted the formation of well-separated NCs and also provided enhanced antimicrobial properties, since lemongrass is known for its antimicrobial character. Fluorescence microscopy was used to optically observe the nanoparticles and NCs and revealed the attachment of lemongrass oil with the polylactic acid NCs. Dynamic light scattering was used to determine their size. UV absorption was used to determine the exact amount of lemongrass oil found in the polylactic acid-lemongrass oil NCs, which was important for understanding the minimum inhibitory concentration for the antimicrobial experiments. A series of clinically important microbial species were used in the study and the obtained NCs proved to have very good antimicrobial properties against all tested strains. Such NCs can be used for the design of ecological strategies, based on natural alternatives, which may be efficient against severe infections, including those that involve resistant pathogens and biofilms or those with difficult to reach localization.