Nanoemulsões de uso tópico contendo extrato de achyrocline satureioides : estudos de formulação, permeação e atividade anti-herpética

Achyrocline satureioides (AS) - Asteraceae, conhecida como marcela, é uma planta medicinal amplamente utilizada na América do Sul. Estudos recentes demonstraram a atividade anti-herpética dos extratos hidroalcoólicos de AS. Tal atividade tem sido relacionada à presença de constituintes flavonoídicos (i.e. quercetina, luteolina e 3-O- metilquercetina) nesses extratos. Neste contexto, o objetivo do presente estudo foi desenvolver nanoemulsões de uso tópico contendo o extrato de AS visando o tratamento do herpes simples. Em uma primeira etapa, o extrato hidroetanólico líquido ou seco (aspersão ou liofilização) de AS foi incorporado em nanoemulsões constituídas de um núcleo oleoso de triglicerídeos de cadeia média estabilizado por lecitina de gema de ovo e polissorbato 80. As nanoemulsões foram obtidas por emulsificação espontânea. As formulações foram adicionalmente caracterizadas por medidas de tamanho, potencial zeta, viscosidade e morfologia. A incorporação dos flavonoides nas nanoemulsões foi total e mostrou-se estável por quatro meses. A liberação da 3-O-metilquercetina a partir das formulações contendo extratos ocorreu dentro de 8 horas seguindo uma cinética de 1ª ordem, utilizando células de difusão de Franz. Quando a membrana utilizada foi pele de orelha suína, não foram detectados flavonoides no fluido receptor. Os flavonoides foram detectados nas camadas da pele, especialmente quando essa foi lesionada, o que foi confirmado por microscopia confocal usando Vermelho do Nilo como marcador fluorescente. A utilização de mucosa esofágica como membrana conduziu a uma maior retenção dos flavonóides em comparação à pele, e a remoção da camada superficial permitiu a permeação dos flavonóides. A determinação dos flavonoides tanto nas formulações como na pele e mucosa suína foi realizada por cromatografia líquida em condições analíticas validadas ao longo do estudo. A avaliação da atividade anti-herpética in vitro frente ao HSV (tipo 1) demonstrou que a incorporação de extrato de AS em nanoemulsões conduz à redução da concentração inibitória 50% e ao aumento do índice de seletividade. O conjunto dos resultados demonstra que nanoemulsões são potenciais carreadores de uso tópico para o extrato de AS.Achyrocline satureioides (AS) - Asteraceae, known as marcela, is a medicinal plant widely used in South America. Recent studies have demonstrated the antiherpetic activity of hydroalcoholic extracts of AS. Such activity has been related to the presence of flavonoids (i.e. quercetin, luteolin, and 3-O-methylquercetin) in these extracts. In this context, the objective of this study was to develop topical nanoemulsions containing extract of AS intended for the treatment of herpes simples. First, the liquid or dry (spray- or freeze-dried) hydroethanolic extract was incorporated in nanoemulsions, composed of an oil core of medium chain triglycerides stabilized by egg-lecithin and polysorbate 80. The extracts were incorporated during the preparation of the nanoemulsion by spontaneous emulsification. The formulations were further characterized by evaluating size, zeta potential, viscosity, and morphology. The incorporation of the flavonoids in the nanoemulsions was total and the formulation was stable for four months. The release of 3-O-methylquercetin from nanoemulsions containing extracts occurred within 8 hours following 1st-order kinetics, using the Franz diffusion cells. When the membrane used was pig ear skin, flavonoids were not detected in the receptor fluid. The flavonoids were detected in the skin layers, especially when this was injured, which was confirmed by confocal microscopy using Nile Red as a fluorescent marker. The use of esophageal mucosa as membrane led to greater retention of flavonoids in comparison to skin, and the removal the superficial layer of epithelium allowed the permeation of the flavonoids.The determination of the flavonoids in the formulations and the pig ear skin layers was assayed by liquid chromatography in analytical conditions validated throughout the study. The evaluation of the in vitro antiherpetic activity over HSV (Type 1) demonstrated that the incorporation of AS extract into nanoemulsions led to a reduction of the inhibitory concentration and an increase in the selectivity index. The overall results show that nanoemulsions are potential carriers for the topical use of AS extract

Desenvolvimento de microesferas a partir do poli-(3-hidroxibutirato) e diferentes adjuvantes de formulação visando o prolongamento da liberação do ibuprofeno para o tratamento localizado da artrite

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências da Saúde. Programa de Pós-Graduação em Farmácia.Polihidroxialcanoatos (PHAs) são poliésters biodegradáveis e biocompátiveis, produzidos por uma ampla variedade de microorganismos e estocados como fonte de carbono e energia. Poli-(3-hidroxibutirato) (P(3HB)) é o membro mais comum desta família e tem sido usado na obtenção de carreadores de fármacos. Na maior parte dos casos, a liberação de fármacos a partir de microesferas de P(3HB) ocorre em taxas excessivas e tem sido assumido que o fenômeno de liberação é mais dependente da velocidade de dissolução do fármaco do que da degradação da matriz. A estratégia de misturar diferentes materiais em uma formulação pode ser empregada para modificar as propriedades físico-químicas de microesferas e controlar a liberação. Neste contexto, este trabalho teve como objetivo avaliar o efeito da formação de blendas de P(3HB) com trimiristato de gricerila (TMG) ou poli (D,L-ácido lático)-b-poli (etilenoglicol) (PLA-PEG) e da obtenção de partículas compostas de P(3HB) e gelatina (GEL) sobre as características físico-químicas das microesferas e o perfil de liberação do ibuprofeno. O método de emulsão/evaporação do solvente foi empregado no preparo das microesferas de P(3HB):TMG (9:1, 4:1, 3:1 e 1:1) e P(3HB):PLA-PEG (3:1 e 1:1), usando clorofórmio ou diclorometano/ álcool isopropílico como solventes. As microesferas de P(3HB):GEL 10:1 foram preparadas usando o procedimento de dupla emulsão/evaporação do solvente. Elevados valores de eficiência de encapsulação foram verificados em todas as formulações testadas. Partículas esféricas e de superfície rugosa, apresentando diâmetro médio entre 21,93 e 41,55 mm, foram obtidas. A utilização de diclorometano e o aumento da proporção de TMG na blenda conduziram à redução da porosidade das partículas. Entretanto, os estudos de liberação em tampão fosfato 0,02 M pH 7,4 mostraram que a associação de P(3HB) e TMG não conduz ao controle de liberação do IBF, a qual foi bastante pronunciada na primeira hora de ensaio. Elevados valores de liberação inicial (efeito burst) também foram observados nas microesferas preparadas a partir das blendas de P(3HB) e PLA-PEG 3:1 e P(3HB):GEL 10:1. Neste último caso, a adição de etanol acelerou a liberação do IBF, indicando que a adição do solvente afeta a localização do fármaco na matriz. As análises por calorimetria exploratória diferencial (DSC) e difração de raios-x indicaram a redução da cristalinidade do P(3HB) somente quando a blenda P(3HB):PLA-PEG 1:1 foi testada. Entretanto, independente das condições de preparo das microesferas, o IBF pareceu estar parcialmente disperso na sua forma amorfa ou molecular na matriz. Apesar da utilização da blenda de P(3HB) e PLA-PEG 1:1 ter conduzido à redução da cristalinidade, a velocidade de liberação foi mais prolongada quando as microesferas de P(3HB):PLA-PEG 3:1 foram testadas, sugerindo que o grau de hidratação da matriz pode ter interferido na cinética de liberação. A avaliação in vivo do ibuprofeno encapsulado nas microesferas de P(3HB):PLA-PEG 3:1 foi conduzida em ratos artríticos (modelo CFA). Os resultados obtidos indicaram redução do edema articular e da migração celular após administração das microesferas contendo IBF, quando comparado aos grupos tratados com IBF livre e microesferas brancas, respectivamente

Curcumin and Quercetin-Loaded Nanoemulsions: Physicochemical Compatibility Study and Validation of a Simultaneous Quantification Method

Biphasic oil/water nanoemulsions have been proposed as delivery systems for the intranasal administration of curcumin (CUR) and quercetin (QU), due to their high drug entrapment efficiency, the possibility of simultaneous drug administration and protection of the encapsulated compounds from degradation. To better understand the physicochemical and biological performance of the selected formulation simultaneously co-encapsulating CUR and QU, a stability test of the compound mixture was firstly carried out using X-ray powder diffraction and thermal analyses, such as differential scanning calorimetry (DSC) and thermogravimetric analyses (TGA). The determination and quantification of the encapsulated active compounds were then carried out being an essential parameter for the development of innovative nanomedicines. Thus, a new HPLC-UV/Vis method for the simultaneous determination of CUR and QU in the nanoemulsions was developed and validated. The X-ray diffraction analyses demonstrated that no interaction between the mixture of active ingredients, if any, is strong enough to take place in the solid state. Moreover, the thermal analysis demonstrated that the CUR and QU are stable in the nanoemulsion production temperature range. The proposed analytical method for the simultaneous quantification of the two actives was selective and linear for both compounds in the range of 0.5-12.5 µg/mL (R2 > 0.9997), precise (RSD below 3%), robust and accurate (recovery 100 ± 5 %). The method was validated in accordance with ICH Q2 R1 "Validation of Analytical Procedures" and CDER-FDA "Validation of chromatographic methods" guideline. Furthermore, the low limit of detection (LOD 0.005 µg/mL for CUR and 0.14 µg/mL for QU) and the low limit of quantification (LOQ 0.017 µg/mL for CUR and 0.48 µg/mL for QU) of the method were suitable for the application to drug release and permeation studies planned for the development of the nanoemulsions. The method was then applied for the determination of nanoemulsions CUR and QU encapsulation efficiencies (> 99%), as well as for the stability studies of the two compounds in simulated biological fluids over time. The proposed method represents, to our knowledge, the only method for the simultaneous quantification of CUR and QU in nanoemulsions

Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model

Mucopolysaccharidosis type I (MPS I) is an autosomal disease caused by alpha-l-iduronidase (IDUA) deficiency. This study used IDUA knockout mice as a model to evaluate whether parameters such as dose of plasmid and time of treatment could influence the transfection efficiency of complexes formed with PEGylated cationic nanoemulsions and plasmid (pIDUA), which contains the gene that encodes for IDUA. Formulations were composed of medium chain triglycerides, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(amino[polyethylene glycol]-2000), 1,2-dioleoyl-sn-glycero-3-trimethylammonium propane (DOTAP), glycerol, and water and were prepared by the adsorption or encapsulation of preformed pIDUA–DOTAP complexes by high-pressure homogenization. A progressive increase in IDUA expression was observed with an increase in the dose and time of transfection for mice treated with both complexes (adsorbed and encapsulated), especially in the liver. Regardless of the complex administered, a significant increase in IDUA activity was detected in lungs and liver compared with nontreated MPS I when a dose of 60 μg was administered and IDUA activity was measured 7 days postadministration. Tissue sections of major organs showed no presence of cell necrosis, inflammatory infiltrate, or an increase in apoptosis. Furthermore, immunohistochemistry for CD68 showed no difference in the number of macrophage cells in treated and nontreated animals, indicating the absence of inflammatory reaction caused by the treatment. The data set obtained in this study allowed establishing that factors such as dose and time can influence transfection efficiency in different degrees and that these complexes did not lead to any lethal effect in the MPS I murine model used

Composição para terapia gênica do sistema nervoso central, processo de obtenção e uso da mesma

Universidade Federal do Rio Grande do SulCiências Básicas da SaúdeCiências BiológicasFarmáciaDepositad

Achyrocline satureioides (Lam.) DC (Asteraceae) extract-loaded nanoemulsions as a promising topical wound Healing delivery system : in vitro assessments in human keratinocytes (HaCaT) and HET-CAM irritant potential

Achyrocline satureioides (Lam.) DC Asteraceae extracts (ASEs) have been investigated for the treatment of various skin disorders. This study reports the effects of ASE-loaded nanoemulsions (NEASE) on the cellular viability, death by necrosis, and migration of immortalized human keratinocytes (HaCaT cell line), as well as the irritant potential through the hen’s egg chorioallantoic membrane test (HET-CAM). NEASE exhibited a polydispersity index above 0.12, with a droplet size of 300 nm, -potential of 40 mV, and content of flavonoids close to 1 mg/mL. No cytotoxicity of the ASE was observed on HaCaT by MTT assay (up to 10 g/mL). A significant increase of HaCaT viability was observed to NEASE (up to 5 g/mL of flavonoids), compared to treatment with the ASE. The necrosis death evaluation demonstrated that only NEASE did not lead to cell death at all the tested concentrations. The scratch assay demonstrated that NEASE was able to increase the cell migration at low flavonoid concentrations. Finally, the HET-CAM test proved the non-irritative potential of NEASE. Overall, the results indicate the potential of the proposed formulations for topical use in wound healing, in view of their promising effects on proliferation and migration in keratinocytes, combined with an indication of the absence of cytotoxicity and non-irritating potential

Curcumin and quercetin-loaded lipid nanocarriers : development of omega-3 mucoadhesive nanoemulsions for intranasal administration

Curcumin (CUR) and quercetin (QU) are potential compounds for treatment of brain diseases such as neurodegenerative diseases (ND) because of their anti-inflammatory and antioxidant properties. However, low water solubility and poor bioavailability hinder their clinical use. In this context, nanotechnology arises as a strategy to overcome biopharmaceutical issues. In this work, we develop, characterize, compare, and optimize three different omega-3 (!-3) fatty acids nanoemulsions (NEs) loaded with CUR and QU (negative, cationic, gelling) prepared by two different methods for administration by intranasal route (IN). The results showed that formulations prepared with the two proposed methods exhibited good stability and were able to incorporate a similar amount of CUR and QU. On the other side, differences in size, zeta potential, in vitro release kinetics, and permeation/retention test were observed. Considering the two preparation methods tested, high-pressure homogenization (HPH) shows advantages, and the CQ NE- obtained demonstrated potential for sustained release. Toxicity studies demonstrated that the formulations were not toxic for Caenorhabditis elegans. The developed !-3 fatty acid NEs have shown a range of interesting properties for the treatment of brain diseases, since they have the potential to increase the nose-tobrain permeation of CUR and QU, enabling enhanced treatments efficiency

Toward a platform for the treatment of burns: An assessment of nanoemulsions vs. nanostructured lipid carriers loaded with curcumin

Curcumin is a highly promising substance for treating burns, owing to its anti-inflammatory, antioxidant, antimicrobial, and wound-healing properties. However, its therapeutic use is restricted due to its hydrophobic nature and low bioavailability. This study was conducted to address these limitations; it developed and tested two types of lipid nanocarriers, namely nanoemulsions (NE-CUR) and nanostructured lipid carriers (NLC-CUR) loaded with curcumin, and aimed to identify the most suitable nanocarrier for skin burn treatment. The study evaluated various parameters, including physicochemical characteristics, stability, encapsulation efficiency, release, skin permeation, retention, cell viability, and antimicrobial activity. The results showed that both nanocarriers showed adequate size (~200 nm), polydispersity index (~0.25), and zeta potential (~>20 mV). They also showed good encapsulation efficiency (>90%) and remained stable for 120 days at different temperatures. In the release test, NE-CUR and NCL-CUR released 57.14% and 51.64% of curcumin, respectively, in 72 h. NE-CUR demonstrated better cutaneous permeation/retention in intact or scalded skin epidermis and dermis than NLC-CUR. The cell viability test showed no toxicity after treatment with NE-CUR and NLC-CUR up to 125 g/mL. Regarding microbial activity assays, free curcumin has activity against P. aeruginosa, reducing bacterial growth by 75% in 3 h. NE-CUR inhibited bacterial growth by 65% after 24 h, and the association with gentamicin had favorable results, while NLC-CUR showed a lower inhibition. The results demonstrated that NE-CUR is probably the most promising nanocarrier for treating burns.FCT -Fundação para a Ciência e a Tecnologia(2022.13494)info:eu-repo/semantics/publishedVersio

Processo de extração de compostos fenólicos a partir da microalga Pediastrum boryanum, formulação farmacêutica, processo de preparo das formulações farmacêuticas e uso da formulação

Universidade Federal de Rio GrandeUniversidade Federal do Rio Grande do SulFarmáciaDepositad