Ascorbic acid and ascorbyl palmitate-loaded liposomes: Development, characterization, stability evaluation, in vitro security profile, antimicrobial and antioxidant activities.

The objective of this work was to prepare and characterize liposomes containing co-encapsulated ascorbic acid (AA) and ascorbyl palmitate (AP), as well as to evaluate their stability, cytotoxicity, antioxidant, and antimicrobial activity. Through the pre-formulation studies, it was possible to improve the formulation, as leaving it more stable and with a greater antioxidant activity, resulting in a formulation designated LIP-AAP, with 161 nm vesicle size, 0.215 polydispersity index, -31.7 mV zeta potential, and pH of 3.34. Encapsulation efficiencies were 37% for AA and 79% for AP, and the content was 1 mg/mL for each compound. The optimized liposomes demonstrated stability under refrigeration for 60 days, significant antioxidant activity (31.4 µMol of TE/mL), and non-toxicity, but no antimicrobial effects against bacteria and fungi were observed. These findings confirm that the co-encapsulated liposomes are potent, stable antioxidants that maintain their physical and chemical properties under optimal storage conditions.

Eucalyptus oil nanoemulsions against eggs and larvae of Haemonchus contortus.

Haemonchus contortus is a highly pathogenic and prevalent helminth that causes many deaths in sheep herds. Anthelmintics are usually employed to overcome this issue; however, they do not guarantee immediate and lasting efficacy because of the occurrence of drug-resistant parasites. Among substances that are used in scientific studies for parasitic control, essential oils are known to have different pharmacological properties. However, they demonstrate instability owing to several factors, and therefore, nanoemulsification is considered an alternative to control the instability and degradability of these compounds. The objective of this study was to evaluate the cytotoxicity of nanoemulsions containing essential oil of Eucalyptus globulus against the blood of healthy sheep and to verify their activity against the parasite H. contortus in sheep. The results presented adequate nanotechnological characteristics (diameter 72 nm, PDI 0.2, zeta -11 mV, and acidic pH) and adequate morphology. Further, the corona effect and cytotoxic profiles of the free oil and nanoemulsion against blood cells from healthy sheep were evaluated. The tests results did not present a toxicity profile. For evaluating efficacy, we observed an important anthelmintic action of the nanoemulsion containing oil in comparison to the free oil; the results demonstrate a potential role of the nanoemulsion in the inhibition of egg hatchability and the development of larvae L1 to L3 (infective stage). Based on these results, we developed an important and potential anthelmintic alternative for the control of the parasite H. contortus.

Ferulic acid-loaded nanocapsules: Evaluation of mucosal interaction, safety and antioxidant activity in human mononucleated cells.

Ferulic acid (FA) is a phenolic compound that has antioxidant, anti-inflammatory and anticarcinogenic properties besides presenting cytoprotective activity. It has limited oral bioavailability what is a challenge to its therapeutic application. In this way, this investigation aimed to develop FA-loaded nanocapsule suspensions (NC-FA) prepared with ethylcellulose and evaluate their in vitro release profile, mucoadhesion and irritation potential; scavenging capacity, cytotoxicity, cytoprotection and genoprotection against hydrogen peroxide-induced damage in hMNC (human Mononucleated Cells) culture. The nanocapsules presented physicochemical characteristics compatible with colloidal systems (NC-FA: 112 ± 3 nm; NC-B (without FA): 107 ± 3 nm; PdI < 0.2; Span<2.0 and negative zeta potential). In addition, the nanoparticulate system promoted the FA controlled release, increasing the half-life twice through the in vitro dialysis method. NC-FA and NC-B were able to interact with mucin, which is an indicative of mucoadhesive properties and the association of FA with nanocapsules showed decreased irritation by HET-CAM method. Besides, the NC-FA did not present cytotoxicity in hMNC and improved the ATBS radical scavenging capacity. Besides, it prevented, treated and reversed oxidative conditions in a H2O2-induced model in hMNC. Thus, this nanocarrier formulation is promising to perform more preclinical investigations focusing on diseases involving oxidative mechanisms.

Development, characterisation, stability study and antileukemic evaluation of nanoemulsions containing Astrocaryum aculeatum extract.

The objective of this work was to produce and characterise nanoemulsions containing tucumã extract and to evaluate the performance of the nanostructure and the free compound regarding antitumor activity, cytotoxicity, and oxidative metabolism in NB4/APL cells. The nanoemulsions showed adequate physicochemical characteristics (average size approx. 200 nm, polydispersity index less than 0.3, negative zeta potential and acid pH) maintained stable up to 90 days of storage in refrigeration condition. The nanoformulations did not present protein corona formation. Blank nanoemulsion treatments showed moderate toxicity. Furthermore, the nanoemulsion loaded with extract showed better antileukemic results than the free extract. However, nanoemulsions can be promising carriers of natural compounds, emphasising their biological properties and constituting alternatives in treating diseases.

Evaluation of the in vivo safety of tucumã oil nanocapsules in an experimental model of silver catfish Rhamdia quelen.

The aim of this study was to evaluate the toxicity of tucumã oil nanocapsules from the Amazon region in silver catfish, Rhamdia quelen. Fish were exposed to water treated with different concentrations of tucumã nanocapsules, white, solubilized oil and surfactant vehicles. After three days of exposure, fish were euthanized and liver, gills and brain removed for analysis of the dichlorofluorescein, nitric oxide and PicoGreen® assays. Plasma was collected for assay of hepatic transaminases. The nanocapsules had a diameter of 221 ± 1.27 nm, confirmed by atomic force microscopy. The oil nanocapsules were not toxic to this species of fish, but white nanocapsules and surfactant increased the levels of reactive oxygen species. Thus, nanocapsules are promising for the transport of tucumã oil. In view of the anti-inflammatory properties of this oil, it is possible to envisage its application in skin diseases for example, since they present essentially inflammatory conditions.HighlightsThe most abundant carotenoid in tucumã oil was all-trans-beta-carotene.Nanocapsules are good carriers for tucumã oil.Tucumã oil nanocapsules does nothas toxicity effect in catfish.

Vitamin C as a shelf-life extender in liposomes

Abstract The objective of this study was to evaluate the influence of vitamin C (VC) on the stability of stored liposomes under different climatic conditions. Liposomal formulations containing 1 mg/mL of VC (LIP-VC) and blank formulations (LIP-B) were prepared by the reverse-phase evaporation method. After preparation, they were characterized according to their refractive index, average vesicle diameter, polydispersity index (PDI), zeta potential, pH, content, encapsulation efficiency (EE%), morphology, stability and antioxidant activity. For stability, LIP-VC and LIP-B were stored in different climatic conditions (4 °C, 25 °C and 40 °C) for 30 days. The LIP-VC presented 1.3365 refractive index, 161 nm of mean diameter, 0.231 PDI, -7.3 mV zeta potential, 3.2 pH, 19.4% EE%, spherical morphology, 1 mg/mL of VC content, and antioxidant activity of 12 and 11.4 μmol of TE/mL for the radical DPPH and ABTS+, respectively. During stability, the LIP-B stored in 40 °C showed an instability in the parameters: PDI, vesicle size and zeta potential after 15 days, while the LIP-VC remained stable in its size and PDI for 30 days. After that, it is shown that VC can be used as an antioxidant and stabilizer in liposomes to increase the stability and shelf-life of vesicles.

Grain filling in barley relies on developmentally controlled programmed cell death.

Cereal grains contribute substantially to the human diet. The maternal plant provides the carbohydrate and nitrogen sources deposited in the endosperm, but the basis for their spatial allocation during the grain filling process is obscure. Here, vacuolar processing enzymes have been shown to both mediate programmed cell death (PCD) in the maternal tissues of a barley grain and influence the delivery of assimilate to the endosperm. The proposed centrality of PCD has implications for cereal crop improvement.

Low-dose curcumin-loaded Eudragit L-100-nanocapsules in the diet of dairy sheep increases antioxidant levels and reduces lipid peroxidation in milk.

The main objective of this study was to evaluate whether the addition of curcumin-loaded nanocapsules (prepared and characterized) in the diets of dairy sheep improved milk quality. The nanocapsules were prepared using two polymers: poly-ε-caprolactone (PCL) and Eudragit L-100. The nanocapsules contained 0.25 mg/ml (Nano-Eudragit L-100) and 2 mg/ml (Nano-PCL) of curcumin. Dairy sheep were divided into four groups: A (control), B (30 mg free curcumin/kg concentrate), C (3 mg Nano-PCL/kg concentrate), and D (3 mg Nano-Eudragit/kg concentrate). We observed that the number of total leukocytes and serum globulin levels were lower in Group D than in the control (Group A) (p < 0.05). Antioxidant capacity against peroxyl radicals (ACAP) and catalase enzymes was elevated in Group D, with consequently reduced lipid peroxidation (LPO; p < 0.05). In milk, there were no differences in production and composition between groups during the experimental period (p > 0.05); however, ACAP increased and LPO decreased in milk. PRACTICAL APPLICATIONS: Curcumin is a functional molecule with potent antioxidant, anti-inflammatory, and antimicrobial actions, used frequently and with medical indications in human food. Free curcumin in sheep diets improves milk quality and increases its shelf life. This study showed that curcumin nanocapsules produced from the Eudragit L-100 polymer potentiated the anti-inflammatory and antioxidant actions of dairy sheep when used in the diet daily, at doses 10 times lower than that of free curcumin. These positive effects were reflected in higher total antioxidant capacity and lower lipid peroxidation in milk in sheep-fed curcumin-loaded Eudragit L-100 nanocapsules, generating desirable milk properties. In practice, the use of nanotechnology enhances the beneficial effects of curcumin in milk, possibly creating a nutraceutical food desirable to consumers.

Characterisation and anti-biofilm activity of glycerol monolaurate nanocapsules against Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a ubiquitous microorganism that commonly causes hospital-acquired infections, including pneumonia, bloodstream and urinary tract infections and it is well known for chronically colonising the respiratory tract of patients with cystic fibrosis, causing severe intermittent exacerbation of the condition. P. aeruginosa may appear in the free form cell but also grows in biofilm communities adhered to a surface. An alternative to conventional antimicrobial agents are nanoparticles that can act as carriers for antibiotics and other drugs. In this context, the study aimed to characterise and verify the anti-biofilm potential of GML Nanocapsules against P. aeruginosa. The nanocapsules showed a mean diameter of 190.7 nm, polydispersion index of 0.069, the zeta potential of -23.3 mV. The microdilution test showed a MIC of 62.5 µg/mL to GML and 15.62 µg/mL to GML Nanocapsules. The anti-biofilm experiments demonstrated the significant reduction of biomass, proteins, polysaccharide and viable P. aeruginosa in biofilm treated with GML Nanocapsules while the free GML did not cause an effect. The AFM images showed a decrease in a biofilm which received GML. The positive results suggest an alternative for the public health trouble related to infections associated with biofilm.

Adhesion to a lithium disilicate glass ceramic etched with hydrofluoric acid at distinct concentrations

Abstract This study evaluated the effect of different hydrofluoric acid (HF) concentrations on the bond strength between a lithium disilicate-based glass ceramic and a resin cement. Eighty ceramic-blocks (12×7×2 mm) of IPS e.Max CAD (Ivoclar Vivadent) were produced and randomly assigned to 8 groups, considering 2 study factors: HF concentration in 4 levels, i.e., 1% (HF1), 3% (HF3), 5% (HF5), and 10% (HF10), and storage in 2 levels, i.e., baseline (tests were performed 24 h after cementation), and aged (storage for 150 days + 12,000 thermal-cycles at 5°C and 55°C). Acid etching (20 s) was performed, followed by washing, drying, and silanization. Four resin cement cylinders (ϕ= 0.96 mm) were built-up from starch matrices on each ceramic sample (n= 40). Additional ceramic samples were etched and analyzed for contact angle, micro-morphology, and roughness. In baseline condition (without aging), the HF3, HF5, and HF10 groups showed similar bond strength values (13.9 - 15.9 MPa), and HF1 (11.2 MPa) presented lower values than HF5, being that statistically different (p= 0.012). After aging, all the mean bond strengths statistically decreased, being that HF3, HF5, and HF10 (7.8 - 11 MPa) were similar and higher than HF1 (1.8 MPa) (p= 0.0001). For contact angle, HF3, HF5, and HF10 presented similar values (7.8 - 10.4°), lower than HF1 and CTRL groups. HF5 and HF10 presented rougher surfaces than other conditions. For better bond strength results, the tested ceramic may be etched by HF acid in concentrations of 3%, 5%, and 10%.

Resumo Este estudo avaliou o efeito de diferentes concentrações de ácido fluorídrico (HF) na resistência de união entre uma cerâmica vítrea à base de dissilicato de lítio e um cimento resinoso. Oitenta blocos cerâmicos (12×7×2 mm) de IPS e.Max CAD (Ivoclar Vivadent) foram produzidos e distribuídos aleatoriamente em 8 grupos, considerando 2 fatores de estudo: concentração de HF em 4 níveis, isto é, 1% (HF1), 3% (HF3), 5% (HF5), e 10% (HF10), e armazenamento em 2 níveis, isto é, condição inicial (testes foram realizados 24 h após a cimentação), e envelhecidos (150 dias de armazenamento + 12.000 ciclos térmicos a 5°C e 55°C). Condicionamento ácido (20 s) foi realizado, seguido por lavagem, secagem e silanização. Quatro cilindros de cimento resinoso (ϕ= 0.96 mm) foram construídos a partir de matrizes de amido em cada amostra cerâmica (n= 40). Amostras cerâmicas adicionais foram condicionadas e analisadas quanto ao ângulo de contato, micro-morfologia e rugosidade. Na condição inicial (sem envelhecimento), os grupos HF3, HF5, e HF10 mostraram valores de resistência de união similares (13.9 - 15.9 MPa), e HF1 apresentou valores menores que HF5, sendo estatisticamente diferente (p= 0.012). Após o envelhecimento, todas as médias de resistência de união diminuíram estatisticamente, sendo que HF3, HF5 e HF10 foram similares e maiores que HF1 (p= 0.0001). Para o ângulo de contato, HF3, HF5 e HF10 apresentaram valores similares (7.8 - 10.4°), menores que os grupos HF1 e CTRL. HF5 e HF10 apresentaram superfícies mais rugosas que as outras condições. Para melhores resultados de resistência de união, a cerâmica testada pode ser condicionada com ácido fluorídrico nas concentrações de 3%, 5% e 10%.

Nanoemulsions containing Cymbopogon flexuosus essential oil: Development, characterization, stability study and evaluation of antimicrobial and antibiofilm activities.

The increase of microbial resistance generates the search for new substances with antimicrobial potential. The essential oil of Cymbopogon flexuosus (Lemongrass) stands out in the literature for its antimicrobial, insecticide and antioxidant properties, but it has high volatilization and low stability, and the nanoencapsulation of this oil could be an alternative to overcome these limitations. Thus, the objective of this study was to develop, for the first time, nanoemulsions containing the essential oil of C. flexuosus, through a method that does not use organic solvent and with temperature control to avoid the volatilization of the oil, characterize and evaluate of stability and the antimicrobial and antibiofilm activities of these nanoemulsions. Nanoemulsions presented adequate physicochemical characteristics (average size less than 200 nm, polydispersity index less than 0.3, negative zeta potential and acid pH) which were maintained during 90 days of storage, and the nanoencapsulation of the C. flexuosus oil enhanced its therapeutic efficacy against the microorganisms evaluated in this study compared to the free oil. These results are very promising because among the microorganisms that the nanoemulsion containing C. flexuosus was able to inhibit the formation of biofilm are the bacteria Pseudomonas aeruginosa and Staphylococcus aureus, which were recently listed by the World Health Organization as priority pathogens for development of new antibiotics.

Optimization of Curcuma Oil/Quinine-Loaded Nanocapsules for Malaria Treatment.

Quinine, a treatment used in chloroquine-resistant falciparum malaria, was loaded into poly(ɛ-caprolactone) or Eudragit® RS100 nanocapsules using Curcuma oil as the oil-based core. Until now, the effect of cationic nanocapsules on malaria has not been reported. A 24 factorial design was adopted using, as independent variables, the concentration of Curcuma oil, presence of quinine, type of polymer, and aqueous surfactant. Diameter, zeta potential, and pH were the responses studied. The formulations were also evaluated for drug content, encapsulation efficiency, photostability, and antimalarial activity against Plasmodium berghei-infected mice. The type of polymer influenced all of the responses studied. Quinine-loaded Eudragit® RS100 (F13) and PCL nanocapsules (F9), both with polysorbate 80 coating, showed nanometric particle size, positive zeta potential, neutral pH, high drug content, and quinine photoprotection ability; thus, these nanocapsules were selected for in vivo tests. Both formulations showed lower levels of parasitemia from the beginning of the experiment (5.78 ± 3.60 and 4.76 ± 3.46% for F9 and F13, respectively) and highest survival mean time (15.3 ± 2.0 and 14.9 ± 5.6 days for F9 and F13, respectively). F9 and F13 showed significant survival curve compared to saline, thus demonstrating that nanoencapsulation improved bioefficacy of QN and co-encapsulated curcuminoids, regardless of the surface charge.

Antimicrobial and antibiofilm activities of nanoemulsions containing Eucalyptus globulus oil against Pseudomonas aeruginosa and Candida spp.

Candida species are the main responsible microorganisms for causing fungal infections worldwide, and Candida albicans is most frequently associated with infectious processes. Pseudomonas aeruginosa is a gram-negative bacterium commonly found in immunocompromised patients. The infection persistence caused by these microorganisms is often related to antimicrobial resistance and biofilm formation. In this context, the objective of the present study was to prepare and characterize nanoemulsions containing Eucalyptus globulus oil and to verify its antimicrobial and antibiofilm activities against P. aeruginosa and Candida spp. The nanoemulsions had a size of approximately 76 nm, a polydispersity index of 0.22, a zeta potential of - 9,42 mV and a pH of approximately 5.0. The E. globulus oil was characterized by gas chromatography, being possible to observe its main components, such as 1-8-Cineol (75.8%), p- Cymene (7.5%), α-Pinene (7.4%) and Limonene (6.4%). The antimicrobial activity of the nanoemulsion was determined from the macrodilution tests and the cell viability curve, where the minimum fungicidal concentration of 0.7 mg/mL for C. albicans and 1.4 mg/mL for C. tropicalis and C. glabrata were obtained. However, the nanoemulsions did not present antimicrobial activity against P. aeruginosa, since it contains only 5% of the oil, being ineffective for this microorganism. The nanoencapsulated oil action against the formed biofilm was evaluated by atomic force microscopy and calcofluor staining, and the nanoemulsion was more efficient for two of the three Candida species when compared to free oil.

Anti-Candida activity assessment of Pelargonium graveolens oil free and nanoemulsion in biofilm formation in hospital medical supplies.

Infections due to microbial biofilm formation on the surface of catheters and other medical devices are constantly reported as a major cause of morbidity and mortality in patients admitted to hospitals. Furthermore, sessile cells are more resistant to phagocytosis and most antimicrobial, which complicates the treatment of such infections. Researches aimed at new antimicrobial originating mainly from plants have increased in recent years and the development of new strategies for their release is critical in combating the formation of biofilms. Geranium oil (GO) has proven antimicrobial activity. Because of this, the aim of this study was to develop nanoemulsions containing this oil (NEG) and evaluate its activity after the biofilm formation of Candida albicans, Candida tropicalis, Candida glabrata, and Candida krusei in hospital medical supplies. For quantification of the biofilm, crystal violet, total protein, and ATP-bioluminescence assays were used. The results revealed that GO and NEG showed lower MIC for C. albicans and C. tropicalis. The biofilms formed by different species of Candida on the surfaces of polyethylene and polyurethane were quantified. GO and NEG significantly inhibited the formation of biofilms in all species tested on the surfaces of polyethylene. However, NEG antibiofilm has had better activity than GO for C. albicans, C. tropicalis and C. glabrata, according to the surface potential analysis by atomic force microscopy (AFM). The analysis of the biofilm formation on the polyethylene surface by ATP-bioluminescence and CFU showed similar results. In both methods the formation of biofilm in the catheter occurred in greater quantity for C. albicans and C. tropicalis. GO did not significantly inhibit the formation of biofilms only in C. krusei, although NEG significantly increased this activity GO in all species tested when compared to the control training biofilm. The following study shows that the development of NEG may become an effective alternative to reduce the adhesion of microorganisms and prevent infections resulting from the use of some hospital medical materials.

Effect of etching with different hydrofluoric acid concentrations on bond strength between glazed feldspathic ceramic and metal brackets / Efeito de diferentes concentrações de ácido fluorídrico na resistência adesiva entre uma cerâmica feldspatica glazeada e bráquetes metálicos

Objetivo: O objetivo deste estudo foi avaliar o efeito do condicionamento com diferentes concentrações de ácido fluorídrico (AF) na resistência de união ao cisalhamento entre cerâmica feldspática glazeada e bráquetes metálicos. Material e Métodos: Setenta e cinco blocos de cerâmica feldspática glazeada foram produzidos e distribuídos aleatoriamente em 5 grupos: Ctrl- silano; HF1- AF 1% + silano; HF3- AF 3% + silano; HF5- AF 5% + silano; HF10- AF 10% + silano (procedimento padrão: condicionamento por 1 min + lavagem + secagem + silanização). Bráquetes metálicos de incisivo central superior (Edgewise Standard) foram colados sobre a superfície da cerâmica com o uso de um sistema adesivo e resina composta fotopolimerizável (Transbond TM XT, 3M). Os espécimes foram envelhecidos por 60 dias (termociclagem: 10000x a 5-55°C; estocados em água destilada a 37°C). O teste de cisalhamento foi realizado e espécimes foram classificados quanto ao seu Índice de Remanescente Adesivo (IRA). Inspeção topográfica e análise do ângulo de contato da superfície da cerâmica condicionada foram realizados. Dados de resistência adesiva foram analisados estatisticamente usando o teste de Kruskal-Wallis, enquanto os dados de ângulo de contato foram analisados com ANOVA 1-fator e teste de Tukey (p<0.05). Resultados: Nenhuma diferença significativa foi detectada para a resistência de união entre os grupos; mas o condicionamento da superfície teve uma influência significativa sobre os resultados de ângulo de contato (p<0.00001). O grupo controle apresentou a mais alta média de ângulo de contato (61,8± 17,2°). Todos os espécimes apresentaram falhas adesivas na interface cerâmica-resina. Conclusão: As concentrações de AF testadas não influenciaram significativamente a resistência de união de bráquetes metálicos aderidos na superfície de cerâmica feldspática glazeada

Objective: The aim of this study was evaluate the effect of etching with different hydrofluoric acid (HF) concentrations on the shear bond strength between glazed feldspathic ceramic and metal brackets. Material and Methods: –Seventyfive blocks of glazed feldspathic ceramic were produced and randomly allocated to 5 groups: Ctrl- silane application only; HF1- HF1%+silane; HF3- HF3%+silane; HF5- HF5%+silane; HF10 HF10%+silane (standard procedures: etching for 1 min + washing + drying + silanization). Metal brackets for upper central incisors (Edgewise Standard) were bonded on the ceramic surface with the use of an adhesive system and light-cured composite resin (TransbondTM XT, 3M). The specimens were aged for 60 days (thermocycling: 10000x at 5-55 °C; stored in distilled water at 37°C). Shear testing was performed, and specimens were classified for their AdhesiveRemnant-Index (ARI). Topographical inspectionand contact angle analysis of the etched ceramic surfaces were performed. Data were statistically analyzed using the non-parametric Kruskal-Wallis test. One-way ANOVA and post-hoc Tukey’s tests were applied to the contact angle data (p<0.05). Results: No significant difference was detected between the shear bond strength of the groups; but surface etching had a significant influence on the contact angle results (p<0.00001). The control group presented the highest mean contact angle (61.8±17.2°). All specimens showed adhesive failure at the resin ceramic interface. Conclusion The tested HF concentrations did not significantly influence the obtained shear bond strength of metal brackets adhered to glazed feldspathic ceramic surfaces.