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This study aimed to develop a film dressing prepared by incorporating a complex of cannabidiol and 2-hydroxypropyl-ß-cyclodextrin (CBD/HP-ß-CD) into a fibroin-based film and to investigate its wound healing capabilities. The fibroin from silkworm cocoons exhibited a total protein content of 96.34 ± 0.14% w/w and a molecular weight range of 25 to 245 kDa. Fourier-transform infrared spectroscopy (FTIR) revealed the presence of characteristic amide peaks (I, II, and III) in the isolated fibroin. The CBD/HP-ß-CD complex, prepared with a molar ratio of 1:2 (CBD to HP-ß-CD), had 81.5 ± 1.2% w/w CBD content, as determined by high-performance liquid chromatography (HPLC). X-ray diffraction (XRD) and FTIR analyses demonstrated successful encapsulation of CBD's hydrophobic aromatic rings by HP-ß-CD. Blending the fibroin solution with the CBD/HP-ß-CD complex produced a transparent, slightly yellowish film. Mechanical testing revealed a tensile strength of 48.67 ± 2.57 MPa and a % elongation at a break of 1.71 ± 0.21%. XRD and FTIR analyses showed distinctive crystalline and chemical structures of the film. In subsequent in vitro experiments with normal human dermal fibroblasts, the film demonstrated potential for wound healing. An increase in cell division (G2/M phase) was observed compared to the fibroin film without the CBD/HP-ß-CD complex. Additionally, fibroblasts treated with the film exhibited enhanced cell migration in a scratch assay and increased expression of vascular endothelial growth factor protein compared to the control group. Overall, these findings underscore the film's potential for enhancing wound healing outcomes.
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Tailored porous structures of poly(2-hydroxyethyl methacrylate) (PHEMA) and silk sericin (SS) were used to create porous hydrogel scaffolds using two distinct crosslinking systems. These structures were designed to closely mimic the porous nature of the native extracellular matrix. Conventional free radical polymerization of 2-hydroxyethyl methacrylate (HEMA) was performed in the presence of different concentrations of SS (1.25, 2.50, 5.00% w/v) with two crosslinking systems. A chemical crosslinking system with N'N-methylene bisacrylamide (MBAAm) and a physical crosslinking system with dimethylurea (DMU) were used: C-PHEMA/SS (crosslinked using MBAAm) and C-PHEMA/pC-SS (crosslinked using MBAAm and DMU). The focus of this study was on investigating the impact of these crosslinking methods on various properties of the scaffolds, including pore size, pore characteristics, polymerization time, morphology, molecular interaction, in vitro degradation, thermal properties, and in vitro cytotoxicity. The various crosslinked networks were found to appreciably influence the properties of the scaffolds, especially the pore sizes, in which smaller sizes and higher numbers of pores with high regularity were seen in C-PHEMA/1.25 pC-SS (17 ± 2 µm) than in C-PHEMA/1.25 SS (34 ± 3 µm). Semi-interpenetrating networks were created by crosslinking PHEMA-MBAAm-PHEMA while incorporating free protein molecules of SS within the networks. The additional crosslinking step involving DMU occurred through hydrogen bonding of the -C=O and -N-H groups with the SS, resulting in the simultaneous incorporation of DMU and SS within the PHEMA networks. As a consequence of this process, the scaffold C-PHEMA/pC-SS exhibited smaller pore sizes compared to scaffolds without DMU crosslinking. Moreover, the incorporation of higher loadings of SS led to even smaller pore sizes. Additionally, the gelation time of C-PHEMA/pC-SS was delayed due to the presence of DMU in the crosslinking system. Both porous hydrogel scaffolds, C-PHEMA/pC-SS and PHEMA, were found to be non-cytotoxic to the normal human skin dermal fibroblast cell line (NHDF cells). This promising result indicates that these hydrogel scaffolds have potential for use in tissue engineering applications.
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Recently, there has been a growing concern among consumers regarding the safety of packaging products, particularly due to the presence of potentially harmful substances like synthetic pigments and inorganic dyes. These substances, which are often used to attract consumer attention, can migrate and contaminate products over extended shelf storage periods. To address this issue, the focus of this research was the development of a biodegradable packaging film using poly(butylene succinate) (PBS) incorporated with natural colorants extracted from roselle (RS) and sappan heartwood (SP). RS and SP serve as non-toxic and alternative pigments when compared to synthetic colorants. The biodegradable packaging films were prepared using blown film extrusion, encompassing different weight percentages of RS and SP (0.1%, 0.2%, and 0.3%). The films exhibited distinct colors, with RS films appearing pink to purple and SP films exhibiting an orange hue. The water vapor transmission rate slightly decreased with an increasing content of RS and SP extracts, indicating improved barrier properties. Additionally, the films showed reduced light transmittance, as evidenced by the UV-Vis light barrier results. The degree of crystallinity in the films was enhanced, as confirmed by X-ray diffraction and differential scanning calorimetry techniques. Regarding mechanical properties, the PBS/RS and PBS/SP films exhibited slight increases in tensile strength and elongation compared to neat PBS films. Moreover, the blended films demonstrated higher stability after undergoing an aging test, further highlighting their potential for use in biodegradable packaging applications. The key advantages of these films lie in their non-toxicity, biodegradability, and overall environmental friendliness.
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Hydroquinine has antimicrobial potential with demonstrated activity against several bacteria, including multidrug-resistant (MDR) P. aeruginosa reference strains. Despite this, there is limited evidence confirming the antibacterial activity of hydroquinine against clinical isolates and the underlying mechanism of action. Here, we aimed to investigate the antibacterial effect of hydroquinine in clinical P. aeruginosa strains using phenotypic antimicrobial susceptibility testing and synergistic testing. In addition, we examined the potential inhibitory mechanisms against MDR P. aeruginosa isolates using informatic-driven molecular docking analysis in combination with RT-qPCR. We uncovered that hydroquinine inhibits and kills clinical P. aeruginosa at 2.50 mg/mL (MIC) and 5.00 mg/mL (MBC), respectively. Hydroquinine also showed partial synergistic effects with ceftazidime against clinical MDR P. aeruginosa strains. Using SwissDock, we identified potential interactions between arginine deiminase (ADI)-pathway-related proteins and hydroquinine. Furthermore, using RT-qPCR, we found that hydroquinine directly affects the mRNA expression of arc operon. We demonstrated that the ADI-related genes, including the arginine/ornithine antiporter (arcD) and the three enzymes (arginine deiminase (arcA), ornithine transcarbamylase (arcB), and carbamate kinase (arcC)), were significantly downregulated at a half MIC of hydroquinine. This study is the first report that the ADI-related proteins are potential molecular targets for the inhibitory effect of hydroquinine against clinically isolated MDR P. aeruginosa strains.
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Antiinfecciosos , Pseudomonas aeruginosa , Pseudomonas aeruginosa/metabolismo , Simulación del Acoplamiento Molecular , Genes Bacterianos , Antiinfecciosos/metabolismo , Antibacterianos/farmacología , Antibacterianos/metabolismo , Pruebas de Sensibilidad Microbiana , Arginina/metabolismoRESUMEN
Natural substances have gained considerable attention for skin protection against UV light reactions. Artocarpus altilis plant's heartwood extract is comprised of artocarpin as a major substance, already known for its interesting biological attributes as an antimicrobial, an anti-inflammatory, an antioxidant, and a melanogenesis inhibitor. The present work clarified the mechanism of natural artocarpin (NAR) with a purity of approximately 99% against the effects of UVB-induced HaCaT keratinocyte apoptosis. The indicated results showed that NAR suppresses free radical production (ROS and nitrite) and apoptosis-related molecule activation (caspase-3, p-p53, p-p38, and NF-κB p65) and secretion (TNF-α). Additionally, NAR prevented structural damages (nuclei condensation and fragmentation, apoptotic body formation, impaired cell adherence and round cell shape, disruption of F-actin filament, and clustering of cell death receptor CD95/Fas) and biophysical changes (plasma membrane rigidification). Thus, NAR acts directly from scavenging free radicals generated by UV and indirectly by suppressing morphological and biochemical UV-induced cell damages. Its biological effects are mainly attributed to antioxidant and antiapoptotic properties. Taken together, NAR could be considered as an effective natural product for photoprotective formulations.
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Artocarpus/efectos de los fármacos , Células HaCaT/efectos de los fármacos , Células HaCaT/patología , Lectinas de Unión a Manosa/farmacología , Lectinas de Plantas/farmacología , Rayos Ultravioleta/efectos adversos , Antioxidantes/metabolismo , Artocarpus/metabolismo , Caspasa 3/efectos de los fármacos , Humanos , Queratinocitos/efectos de los fármacos , Queratinocitos/metabolismo , Queratinocitos/patología , Protectores contra Radiación/farmacología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Apoptosis, a well-known pattern of programmed cell death, occurs in multicellular organisms not only for controlling tissue homeostasis but also for getting rid of severely damaged cells in order to protect the redundant growth of abnormal cells undergoing cancerous cells. The epidermis of the human skin, composed largely of keratinocytes (KCs), is renewed continuously. Therefore, KCs apoptosis plays a critical role in the maintenance of epidermis structure and function. However, regulated cell death can be disturbed by environmental factors especially ultraviolet radiation (UV) B, leading to the formation of sunburn cells (KCs undergoing UVB-induced apoptosis) and impairing the skin integrity. In the present study, we firstly reported the potential of the natural artocarpin (NAR) to regulate UVB-induced human KCs apoptosis. The NAR showed antilipid peroxidation with an IC50 value of 18.2 ± 1.6 µg/mL, according to TBARS assay while the IC50 value of trolox, a well-known antioxidant, was 7.3 ± 0.8 µg/mL. For cell-based studies, KCs were pretreated with 3.1 µg/mL of the NAR for 24 hr and then exposed to UVB at 55 mJ/cm2. Our data indicated that the NAR pretreatment reduces UVB-induced oxidative stress by scavenging free radicals and nitric oxide and therefore prevents reactive oxygen species (ROS) and reactive nitrogen species- (RNS-) mediated apoptosis. The NAR pretreatment has been shown also to reduce the UVB-induced cyclobutane pyrimidine dimer (CPD) lesions by absorbing UVB radiation and regulating the cell cycle phase. Additionally, the NAR pretreatment was found to modulate the expression of cleaved caspases-3 and 8 that trigger different signalling cascades leading to apoptosis. Thus, these results provide a basis for the investigation of the photoprotective effect of the NAR isolated from A. altilis heartwood and suggest that it can be potentially used as an agent against UVB-induced skin damages.
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Apoptosis/efectos de los fármacos , Lectinas de Unión a Manosa/química , Lectinas de Plantas/química , Protectores contra Radiación/farmacología , Rayos Ultravioleta , Antioxidantes/química , Apoptosis/efectos de la radiación , Artocarpus/química , Artocarpus/metabolismo , Caspasa 3/metabolismo , Puntos de Control del Ciclo Celular/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de la radiación , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Cromatografía Líquida de Alta Presión , Humanos , Queratinocitos/citología , Queratinocitos/metabolismo , Lectinas de Unión a Manosa/aislamiento & purificación , Lectinas de Unión a Manosa/farmacología , Óxido Nítrico/metabolismo , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/química , Lectinas de Plantas/aislamiento & purificación , Lectinas de Plantas/farmacología , Protectores contra Radiación/química , Protectores contra Radiación/aislamiento & purificación , Especies de Nitrógeno Reactivo/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
OBJECTIVE: The moisturizing and irritation effects of sacha inchi oil were evaluated. STUDY DESIGN: The moisturizing effect on the skin was clinically assessed using a regression study design. Sacha inchi oil or olive oil (benchmark) was applied on the left or right lower leg of the subjects for 14 days followed by application discontinuation for 2 days. The TEWL, skin moisture content and dryness appearance were observed. METHODS: The fatty acid composition and characteristics of cold-pressed sacha inchi seed oil were determined. Skin tissues cultured ex vivo were used to assess primary irritation induced by the oil by examining keratin 1 expression and TNF-α and IL-1α release from the oil-applied tissues. RESULTS: The sacha inchi oil contained 42.3% linolenic acid and 39.5% linoleic acid. This oil's saponification, iodine, acid and peroxide values were 168.58 ± 1.55 mg KOH/g, 203.00 ± 0.04 g I2 /100 g, 1.68 ± 0.03 mg KOH/g, and 1.95 ± 0.26 mEq peroxide/kg, respectively. Compared with nontreated skin tissues, induced secretion of TNF-α and IL-1α and disruption of keratin 1 integrity in the stratum corneum layer were not found in the sacha inchi oil-treated tissues. In a clinical study with 13 volunteers, the improvement in moisture content and skin dryness appearance at the sacha inchi oil-applied site was comparable with that observed at the olive oil-applied site. CONCLUSIONS: The sacha inchi oil was mild to the skin and benefited dry skin.
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Cosmecéuticos/administración & dosificación , Epidermis/efectos de los fármacos , Euphorbiaceae/química , Aceites de Plantas/administración & dosificación , Semillas/química , Adulto , Biopsia , Cosmecéuticos/efectos adversos , Cosmecéuticos/química , Elasticidad/efectos de los fármacos , Epidermis/metabolismo , Epidermis/patología , Femenino , Voluntarios Sanos , Humanos , Interleucina-1alfa/metabolismo , Ácido Linoleico/análisis , Persona de Mediana Edad , Aceites de Plantas/efectos adversos , Aceites de Plantas/química , Pruebas de Irritación de la Piel , Resultado del Tratamiento , Factor de Necrosis Tumoral alfa/metabolismo , Pérdida Insensible de Agua/efectos de los fármacos , Adulto Joven , Ácido alfa-Linolénico/análisisRESUMEN
The purpose of this study was to investigate the cytokine production and liver injury induced by lipoplexes prepared with DOTMA/cholesterol and DOTAP/cholesterol liposomes with various mixing ratios in mice. Lipoplexes were prepared with pCMV-Luc and DOTMA/cholesterol or DOTAP/cholesterol liposomes. After intravenous administration into the mice, organ luciferase activity and serum TNFalpha and ALT were measured to evaluate the transfection efficacy, cytokine production and liver injury. After intravenous administration of these lipoplexes, basically the serum TNFalpha and ALT levels were in agreement with the transfection efficacy of the lipoplexes. The cytokine production and liver injury were markedly suppressed by reducing the pDNA dose, and achieved normal levels at a pDNA dose of 0.47 mg/kg. As far as the effects of the charge ratio at this low pDNA dose are concerned, the charge ratios of the lipoplexes affected the transfection efficacy, but not the cytokine production and liver injury. After intravenous administration of either DOTAP/cholesterol or DOTMA/cholesterol liposomes, serum TNFalpha and ALT levels were normal, suggesting that liver injury as well as cytokine production was caused by lipoplexes, but not by cationic liposomes. This information will be valuable for the future optimization of the preparation conditions of lipoplexes for use in clinical gene therapy.
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ADN/administración & dosificación , Técnicas de Transferencia de Gen , Hígado/patología , Alanina Transaminasa/sangre , Animales , Cationes , Colesterol/química , Citocinas/metabolismo , ADN/toxicidad , Ácidos Grasos Monoinsaturados/química , Femenino , Dosificación de Gen , Mediadores de Inflamación/metabolismo , Inyecciones Intravenosas , Liposomas , Luciferasas/metabolismo , Ratones , Ratones Endogámicos ICR , Plásmidos/administración & dosificación , Plásmidos/toxicidad , Compuestos de Amonio Cuaternario/química , Transfección/métodos , Factor de Necrosis Tumoral alfa/sangreRESUMEN
PURPOSE: All-trans retinoic acid (ATRA) polymeric micelles were developed for parenteral administration. The distribution characteristics and antitumor activities of ATRA polymeric micelles were evaluated after intravenous administration to mice bearing CT26 solid tumors. METHODS: ATRA incorporated in poly(ethylene glycol)-poly(benzyl aspartate) block copolymer was prepared by the evaporation method. The levels of [3H]ATRA in blood and tissue including tumor were determined by measuring the radioactivity after injection into mice. The tumor volume and the survival of the mice were determined to assess the anticancer activity. RESULTS: The delivery of ATRA by polymeric micelles prolonged the blood circulation and enhanced the accumulation of ATRA in the tumor tissue compared with the administration of free ATRA. Tumor growth was significantly delayed and the survival time of mice was prolonged following the treatment by ATRA polymeric micelles demonstrating the improved anticancer activity of ATRA. CONCLUSION: Polymeric micelles are a promising and effective carrier of ATRA in order to enhance tumor delivery and they have a promising potential application in the treatment of solid tumors.
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Micelas , Neoplasias Experimentales/tratamiento farmacológico , Tretinoina/administración & dosificación , Animales , Masculino , Ratones , Neoplasias Experimentales/mortalidad , Distribución Tisular , Tretinoina/farmacocinéticaRESUMEN
BACKGROUND: All-trans retinoic acid (ATRA) is a natural derivative of vitamin A, which is well known to suppress inflammatory cytokine production. To date, there have been few reports about the systemic use of ATRA for inflammation because of acute resistance and the highly lipophilic nature of ATRA. METHODS: ATRA-lipoplexes were prepared by mixing CMV-Luc plasmid DNA with ATRA-incorporated 1,2-dioleoyl-3-trimethylammoniopropane (DOTAP)/cholesterol liposome. After intravenous injection, tissue accumulation, transfection efficacy, NFkappaB activation, cytokine production, and hepatic toxicity of ATRA-lipoplexes were evaluated and compared with lipoplexes lacking ATRA. RESULTS: The particle size and zeta potential of ATRA-lipoplexes were similar to those of lipoplexes. After intravenous injection of ATRA-lipoplexes, tissue accumulation in liver and gene expression in liver and lung were similar to those of lipoplexes, supporting the hypothesis that ATRA incorporation did not affect the delivery and gene transfection efficacy. In addition, ATRA incorporated in ATRA-lipoplexes was delivered to liver in a manner similar to that for ATRA incorporated in liposomes. In addition, intravenous injection of ATRA-lipoplexes inhibited the activation of NFkappaB in liver, and subsequently suppressed the serum levels of tumor necrosis factor-alpha (TNF-alpha) and alanine aminotransferase (ALT) compared with lipoplexes. Liver histology data also demonstrated a low degree of liver injury produced by ATRA-lipoplexes compared with lipoplexes. CONCLUSIONS: ATRA-incorporated lipoplexes effectively suppress NFkappaB activation, cytokine response and liver injury induced by lipoplexes without affecting gene delivery and transfection efficacy in vivo.
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Hepatopatías/patología , Hepatopatías/prevención & control , FN-kappa B/metabolismo , Tretinoina/metabolismo , Alanina Transaminasa/sangre , Animales , ADN/metabolismo , Femenino , Perfilación de la Expresión Génica , Inyecciones Intravenosas , Interleucina-6/sangre , Liposomas , Ratones , Ratones Endogámicos ICR , Especificidad de Órganos/efectos de los fármacos , Tamaño de la Partícula , Radioisótopos de Fósforo , Tretinoina/administración & dosificación , Tretinoina/farmacología , Tritio , Factor de Necrosis Tumoral alfa/sangreRESUMEN
The purpose of this study was to evaluate the cytokine response induced by linear and branched polyethylenimine (PEI)/plasmid DNA (pDNA) complex (polyplex) in relation to the ratio of PEI nitrogen and DNA phosphate (N/P ratio) of the polyplex, dose of pDNA, and structure and molecular weight of PEI, which are important for transfection efficacy of PEI polyplex. As a control, a N-[1-(2, 3-dioleyloxy) propyl]-n,n,n-trimethylammonium chloride/cholesterol liposome/pDNA complex (lipoplex) was selected for its high transfection efficacy in vivo. The concentration of proinflammatory cytokines such as tumor necrosis factor (TNF)-alpha were much lower after the administration of polyplex than lipoplex irrespective of the N/P ratio, dose of pDNA, or structure and molecular weight of PEI, although these factors affected the transfection efficacy in vivo. We demonstrated that the amount of activated nuclear factor-kappaB, which contributes substantially to the production of cytokines, was comparable with the control (no treatment) level, and significantly less than that obtained with lipoplex. Although the production of proinflammatory cytokines (TNF-alpha, interferon-gamma, and interleukin-12) was reduced on the administration of the linear PEI polyplex, serum alanine aminotransferase levels were significantly enhanced by pDNA in a dose-dependent manner, suggesting that such hepatic damage is not induced by proinflammatory cytokines.