RESUMEN

PURPOSE: Sonodynamic therapy (SDT) is a new therapeutic modality for the noninvasive cancer treatment based on the association of ultrasound and sonosensitizer drugs. Topical SDT requires the development of delivery systems to properly transport the sonosensitizer, such as zinc phthalocyanine (ZnPc), to the skin. In addition, the delivery system itself can participate in sonodynamic events and influence the therapeutic response. This study aimed to develop ZnPc-loaded micelle to evaluate its potential as a topical delivery system and as a cavitational agent for low-frequency ultrasound (LFU) application with the dual purpose of promoting ZnPc skin penetration and generating reactive oxygen species (ROS) for SDT. METHODS: ZnPc-loaded micelles were developed by the thin-film hydration method and optimized using the Quality by Design approach. Micelles' influence on LFU-induced cavitation activity was measured by potassium iodide dosimeter and aluminum foil pits experiments. In vitro skin penetration of ZnPc was assessed after pretreatment of the skin with LFU and simultaneous LFU treatment using ZnPc-loaded micelles as coupling media followed by 6 h of passive permeation of ZnPc-loaded micelles. The singlet oxygen generation by LFU irradiation of the micelles was evaluated using two different hydrophilic probes. The lipid peroxidation of the skin was estimated using the malondialdehyde assay after skin treatment with simultaneous LFU using ZnPc-loaded micelles. The viability of the B16F10 melanoma cell line was evaluated using resazurin after treatment with different concentrations of ZnPc-loaded micelles irradiated or not with LFU. RESULTS: The micelles increased the solubility of ZnPc and augmented the LFU-induced cavitation activity in two times compared to water. After 6 h ZnPc-loaded micelles skin permeation, simultaneous LFU treatment increased the amount of ZnPc in the dermis by more than 40 times, when compared to non-LFU-mediated treatment, and by almost 5 times, when compared to LFU pretreatment protocol. The LFU irradiation of micelles induced the generation of singlet oxygen, and the lipoperoxidation of the skin treated with the simultaneous LFU was enhanced in three times in comparison to the non-LFU-treated skin. A significant reduction in cell viability following treatment with ZnPc-loaded micelles and LFU was observed compared to blank micelles and non-LFU-treated control groups. CONCLUSION: LFU-irradiated mice can be a potential approach to skin cancer treatment by combining the functions of increasing drug penetration and ROS generation required for SDT.

Asunto(s)

Indoles/farmacología , Micelas , Compuestos Organometálicos/farmacología , Ultrasonido , Aluminio/química , Animales , Supervivencia Celular/efectos de los fármacos , Humanos , Isoindoles , Melanoma/tratamiento farmacológico , Melanoma/patología , Melanoma Experimental/patología , Fosfatidiletanolaminas/química , Fármacos Fotosensibilizantes/farmacología , Polietilenglicoles/química , Yoduro de Potasio/química , Oxígeno Singlete/química , Neoplasias Cutáneas/tratamiento farmacológico , Neoplasias Cutáneas/patología , Porcinos , Compuestos de Zinc

RESUMEN

OBJECTIVES: To evaluate the effects of a topical emulsion containing pyrrolidine dithiocarbamate (PDTC) (EcPDTC) in skin oxidative stress and inflammation triggered by ultraviolet B (UVB) irradiation (dose of 4.14 J/cm2 ). METHODS: Hairless mouse received treatment with 0.5 g of EcPDTC or control emulsion (CTRLE) on the dorsal surface skin 12 h, 6 h and 5 min before and 6 h after the irradiation. Oxidative stress was evaluated by ferric reducing antioxidant power (FRAP), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS) scavenging capacity, reduced glutathione quantitation, catalase activity, superoxide anion production and lipid peroxidation products. Inflammation parameters were as follows: skin oedema, myeloperoxidase activity (neutrophil marker), matrix metalloproteinase-9 activity, collagen fibre damage, mast cell and sunburn cell counts, and cytokine production. KEY FINDINGS: Topical treatment with EcPDTC protected from UVB-induced skin injury by maintaining the antioxidant capacity levels similar to non-irradiated control group. Furthermore, EcPDTC inhibited UVB irradiation-induced superoxide anion production, lipid peroxidation and reduced skin inflammation by inhibiting skin oedema, neutrophil recruitment, metalloproteinase-9 activity, collagen fibre damage, mast cell and sunburn cell counts, and cytokine (TNF-α and IL-1ß) production. CONCLUSIONS: Topical treatment with EcPDTC improves antioxidant systems and inhibits inflammation, protecting the skin from the damaging effects of UVB irradiation.

Asunto(s)

Antiinflamatorios/administración & dosificación , Antioxidantes/administración & dosificación , Pirrolidinas/administración & dosificación , Piel/efectos de los fármacos , Quemadura Solar/prevención & control , Tiocarbamatos/administración & dosificación , Rayos Ultravioleta , Administración Cutánea , Animales , Antiinflamatorios/química , Antioxidantes/química , Citocinas/metabolismo , Modelos Animales de Enfermedad , Composición de Medicamentos , Emulsiones , Mediadores de Inflamación/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Ratones Pelados , Estrés Oxidativo/efectos de los fármacos , Pirrolidinas/química , Piel/metabolismo , Piel/patología , Quemadura Solar/metabolismo , Quemadura Solar/patología , Tiocarbamatos/química

RESUMEN

trans-Chalcone is a plant flavonoid precursor, which lacks broad investigation on its biological activity in inflammatory processes. In the present study, anti-inflammatory and antioxidant mechanisms of systemic administration with trans-chalcone, a flavonoid precursor, on ultraviolet (UV) irradiation-induced skin inflammation and oxidative stress in hairless mice were investigated by the following parameters: skin edema, myeloperoxidase activity (neutrophil marker), matrix metalloproteinase-9 activity, reduced glutathione levels, catalase activity, lipid peroxidation products, superoxide anion production, gp91phox (NADPH oxidase subunit) mRNA expression by quantitative PCR and cytokine production by ELISA. Systemic treatment with trans-chalcone inhibited skin inflammation by reducing skin edema and neutrophil recruitment, and also inhibited matrix metalloproteinase-9 activity. trans-Chalcone also inhibited oxidative stress, gp91phox mRNA expression, and the production of a wide range of pro-inflammatory cytokines, while it did not affect anti-inflammatory cytokines induced by UV irradiation. However, trans-chalcone did not prevent oxidative stress in vitro, suggesting that its in vivo effect is more related to anti-inflammatory properties rather than a direct antioxidant effect. In conclusion, treatment with trans-chalcone inhibited UV-induced skin inflammation resulting in oxidative stress inhibition in vivo. Therefore, systemic supplementation with this compound may represent an important therapeutic approach in inflammatory skin diseases induced by UV irradiation.

Asunto(s)

Chalcona/farmacología , Citocinas/biosíntesis , Inflamación/prevención & control , NADPH Oxidasas/metabolismo , Estrés Oxidativo/efectos de los fármacos , Piel/efectos de los fármacos , Piel/efectos de la radiación , Rayos Ultravioleta , Animales , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/farmacología , Chalcona/química , Relación Dosis-Respuesta a Droga , Edema/tratamiento farmacológico , Ensayo de Inmunoadsorción Enzimática , Inflamación/metabolismo , Masculino , Ratones , Ratones Pelados , Estructura Molecular , Estrés Oxidativo/efectos de la radiación , Piel/metabolismo , Piel/patología , Relación Estructura-Actividad

RESUMEN

Evidence shows that sunscreens undergo degradation processes induced by UV irradiation forming free radicals, which reduces skin protection. In this regard, the biological effects of three commercial sunscreen formulations upon UVB irradiation in the skin were investigated. The three formulations had in common the presence of benzophenone-3 added with octyl methoxycinnamate or octyl salycilate or both, which are regular UV filters in sunscreens. The results show that formulations F1 and F2 presented partial degradation upon UVB irradiation. Formulations F1 and F2 presented higher skin penetration profiles than F3. None of the formulations avoided UVB irradiation-induced GSH depletion, but inhibited reduction of SOD activity, suggesting the tested formulations did not present as a major mechanism inhibiting all UVB irradiation-triggered oxidative stress pathways. The formulations avoided the increase of myeloperoxidase activity and cytokine production (IL-1ß and TNF-α), but with different levels of protection in relation to the IL-1ß release. Concluding, UVB irradiation can reduce the stability of sunscreens, which in turn, present the undesirable properties of reaching viable skin. Additionally, the same SPF does not mean that different sunscreens will present the same biological effects as SPF is solely based on a skin erythema response. This found opens up perspectives to consider additional studies to reach highly safe sunscreens.

Asunto(s)

Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/efectos de la radiación , Piel/efectos de los fármacos , Piel/efectos de la radiación , Protectores Solares/química , Protectores Solares/farmacología , Rayos Ultravioleta/efectos adversos , Animales , Composición de Medicamentos , Estabilidad de Medicamentos , Femenino , Glutatión/metabolismo , Inflamación/tratamiento farmacológico , Inflamación/etiología , Interleucina-1beta/metabolismo , Masculino , Ratones , Peroxidasa/metabolismo , Piel/metabolismo , Factor de Protección Solar , Protectores Solares/uso terapéutico , Superóxido Dismutasa/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

Naringenin (NGN) exhibits anti-inflammatory and antioxidant activities, but it remains undetermined its topical actions against ultraviolet B (UVB)-induced inflammation and oxidative stress in vivo. The purpose of this study was to evaluate the physicochemical and functional antioxidant stability of NGN containing formulations, and the effects of selected NGN containing formulation on UVB irradiation-induced skin inflammation and oxidative damage in hairless mice. NGN presented ferric reducing power, ability to scavenge 2,2'-azinobis (3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS) and hydroxyl radical, and inhibited iron-independent and dependent lipid peroxidation. Among the three formulations containing NGN, only the F3 kept its physicochemical and functional stability over 180 days. Topical application of F3 in mice protected from UVB-induced skin damage by inhibiting edema and cytokine production (TNF-α, IL-1ß, IL-6, and IL-10). Furthermore, F3 inhibited superoxide anion and lipid hydroperoxides production and maintained ferric reducing and ABTS scavenging abilities, catalase activity, and reduced glutathione levels. In addition, F3 maintained mRNA expression of cellular antioxidants glutathione peroxidase 1, glutathione reductase and transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2), and induced mRNA expression of heme oxygenase-1. In conclusion, a formulation containing NGN may be a promising approach to protecting the skin from the deleterious effects of UVB irradiation.

Asunto(s)

Antioxidantes/farmacología , Edema/prevención & control , Flavanonas/farmacología , Estrés Oxidativo/efectos de los fármacos , Piel/efectos de los fármacos , Rayos Ultravioleta/efectos adversos , Administración Cutánea , Animales , Benzotiazoles/antagonistas & inhibidores , Benzotiazoles/metabolismo , Catalasa/genética , Catalasa/metabolismo , Edema/etiología , Edema/genética , Edema/patología , Expresión Génica/efectos de los fármacos , Glutatión/metabolismo , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/metabolismo , Glutatión Reductasa/genética , Glutatión Reductasa/metabolismo , Hemo-Oxigenasa 1/genética , Hemo-Oxigenasa 1/metabolismo , Radical Hidroxilo/antagonistas & inhibidores , Radical Hidroxilo/metabolismo , Inflamación , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Ratones , Ratones Pelados , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Piel/metabolismo , Piel/patología , Piel/efectos de la radiación , Ácidos Sulfónicos/antagonistas & inhibidores , Ácidos Sulfónicos/metabolismo , Superóxidos/antagonistas & inhibidores , Superóxidos/metabolismo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Glutatión Peroxidasa GPX1

RESUMEN

Ultraviolet B (UVB) irradiation may cause oxidative stress- and inflammation-dependent skin cancer and premature aging. Pyrrolidine dithiocarbamate (PDTC) is an antioxidant and inhibits nuclear factor-κB (NF-κB) activation. In the present study, the mechanisms of PDTC were investigated in cell free oxidant/antioxidant assays, in vivo UVB irradiation in hairless mice and UVB-induced NFκB activation in keratinocytes. PDTC presented the ability to scavenge 2,2'-azinobis-(3-ethyl benzothiazoline-6-sulfonic acid) radical (ABTS), 2,2-diphenyl-1-picryl-hydrazyl radical (DPPH) and hydroxyl radical (OH); and also efficiently inhibited iron-dependent and -independent lipid peroxidation as well as chelated iron. In vivo, PDTC treatment significantly decreased UVB-induced skin edema, myeloperoxidase (MPO) activity, production of the proinflammatory cytokine interleukin-1ß (IL-1ß), matrix metalloproteinase-9 (MMP-9), increase of reduced glutathione (GSH) levels and antioxidant capacity of the skin tested by the ferric reducing antioxidant power (FRAP) and ABTS assays. PDTC also reduced UVB-induced IκB degradation in keratinocytes. These results demonstrate that PDTC presents antioxidant and anti-inflammatory effects in vitro, which line up well with the PDTC inhibition of UVB irradiation-induced skin inflammation and oxidative stress in mice. These data suggest that treatment with PDTC may be a promising approach to reduce UVB irradiation-induced skin damages and merits further pre-clinical and clinical studies.

Asunto(s)

Antioxidantes/farmacología , Edema/etiología , Estrés Oxidativo/efectos de los fármacos , Pirrolidinas/farmacología , Piel/efectos de la radiación , Tiocarbamatos/farmacología , Rayos Ultravioleta , Animales , Antioxidantes/química , Línea Celular , Femenino , Glutatión/metabolismo , Humanos , Proteínas I-kappa B/metabolismo , Interleucina-1beta/metabolismo , Masculino , Metaloproteinasa 9 de la Matriz/metabolismo , Ratones , Ratones Pelados , Estrés Oxidativo/efectos de la radiación , Peroxidasa/metabolismo , Pirrolidinas/química , Tiocarbamatos/química

RESUMEN

This study evaluated the chemical properties (polyphenol and genistein contents) of soybean extracts obtained by biotransformation and dried by spray dryer at different conditions and their in vivo ability to inhibit 12-O-tetradecanoylphorbol-13-acetate- (TPA-) induced biochemical alterations in the skin of hairless mice. By comparing the obtained data with that of the well-known active soybean extract Isoflavin beta, we evaluated the influence of the fermentation and drying process in the extracts efficacy. The results demonstrated that inlet gas temperature and adjuvant concentration for the extract drying process have significantly affected the total polyphenol contents and, to a minor degree, the genistein contents. However, the effect of topical stimulus with TPA, an oxidative stress inducer, which caused significant depletion of reduced glutathione (GSH) and catalase, with increased levels of H2O2 and lipid peroxidation (MDA) in the skin of hairless mice, was significantly prevented by the soybean extracts treatment. These results indicate that the spray drying processing resulted in a product capable of limiting the oxidative stress with possible therapeutic applicability as an antioxidant in pharmaceutical forms.

Asunto(s)

Fermentación/efectos de los fármacos , Glycine max/química , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/farmacología , Sustancias Protectoras/farmacología , Piel/efectos de los fármacos , Piel/patología , Animales , Catalasa/metabolismo , Desecación , Femenino , Genisteína/farmacología , Glutatión/metabolismo , Peróxido de Hidrógeno/farmacología , Masculino , Ratones , Ratones Pelados , Polifenoles/farmacología , Acetato de Tetradecanoilforbol

RESUMEN

Plants rich in antioxidant substances may be a promising strategy for preventing UV-induced oxidative and inflammatory damage of the skin. Pimenta pseudocaryophyllus is native to Brazil and presents flavonoids and other polyphenolic compounds in high concentration. Thus, the present study evaluated the possible effects of topical formulations containing P. pseudocaryophyllus ethanolic extract (PPE) at inhibiting UV-B irradiation-induced oxidative stress and inflammation. PPE was administered on the dorsal skin of hairless mice using two formulations: F1 (non-ionic emulsion with high lipid content) and F2 (anionic emulsion with low lipid content) before and after UV-B irradiation. The following parameters were evaluated in skin samples: edema, myeloperoxidase activity, cytokines levels, matrix metalloprotease-9 (MMP-9) secretion/activity, reduced glutathione (GSH), superoxide anion and lipid peroxidation levels, and mRNA expression for glutathione reductase and gp91phox. The UV-B irradiation increased all parameters, except for IL-10 levels and glutathione reductase mRNA expression, which were not altered, and GSH levels, which were reduced by exposure to UV-B light. Treatments with F1 and F2 containing PPE inhibited UV-B-induced edema formation (89% and 86%), myeloperoxidase activity (85% and 81%), IL-1ß production (62% and 82%), MMP-9 activity (71% and 74%), GSH depletion (73% and 85%), superoxide anion (83% and 66%) and TBARS (100% and 100%) levels, increased glutathione reductase (2.54 and 2.55-fold) and reduced gp91phox (67% and 100%) mRNA expression, respectively. F2 containing PPE also increased IL-10 levels. Therefore, this study demonstrates the effectiveness of topical formulations containing PPE in inhibiting UV-B irradiation-induced inflammation and oxidative stress of the skin.

Asunto(s)

Estrés Oxidativo/efectos de los fármacos , Pimenta/química , Extractos Vegetales/farmacología , Rayos Ultravioleta/efectos adversos , Administración Tópica , Animales , Química Farmacéutica , Citocinas/metabolismo , Edema/etiología , Edema/prevención & control , Etanol/química , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/efectos de la radiación , Glutatión Reductasa/genética , Inflamación/etiología , Inflamación/prevención & control , Masculino , Metaloproteinasa 9 de la Matriz/metabolismo , Glicoproteínas de Membrana/genética , Ratones , Ratones Pelados , NADPH Oxidasa 2 , NADPH Oxidasas/genética , Estrés Oxidativo/efectos de la radiación , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , ARN Mensajero/genética , ARN Mensajero/metabolismo , Piel/efectos de los fármacos , Piel/metabolismo

RESUMEN

Tephrosia toxicaria (Sw.) Pers., which is currently known as T. sinapou (Buc'hoz) A. Chev., Fabaceae, is a source of compounds such as flavonoids, however, few studies addressed the anti-inflammatory and antioxidant effects of T. sinapou. Therefore, we evaluated the antioxidant mechanisms of the T. sinapou ethyl acetate extract in vitro, and whether the extract affects leukocyte recruitment in four models of inflammation and the involvement of nitric oxide and cytokines in its mechanism. In vitro, it was observed that the extract presented hydrogen donating ability to 2,2-diphenyl-1-picryl-hydrazyl radical (DPPH), 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulphonic acid) radical (ABTS+), and also efficiently inhibited iron-dependent and independent lipid peroxidation and iron chelation assays. In vivo, it inhibited the recruitment of total leukocytes and neutrophil induced by carrageenin, zymosan, glycogen and lipopolysaccharide in the peritoneal cavity of mice. Two mechanisms were detected: 1) T. sinapou effect on leukocyte recruitment depends on nitric oxide since was dose-dependently inhibited by treatment with L-NAME (nitric oxide synthase inhibitor), and 2) the extract also inhibited the production of crucial cytokines for the leukocyte recruitment; tumor necrosis factor α and interleukin-1β. Concluding, T. sinapou ethyl acetate extract reduces oxidative stress in vitro, and inflammatory leukocyte recruitment by a mechanism related to inhibition of cytokine production, and in a nitric oxide dependent manner in vivo.

RESUMEN

BACKGROUND: Topical flavonoids, such as quercetin, have been shown to reduce ultraviolet (UV) irradiation-mediated skin damage. However, the mechanisms and signaling pathways involved in this protective effect are not clear. UV irradiation leads to activation of two major signaling pathways, namely nuclear factor kappa B (NF-κB) and activator protein-1 (AP-1) pathways. Activation of NF-κB pathway by UV irradiation stimulates inflammatory cytokine expression, whereas activation of AP-1 pathway by UV irradiation promotes matrix metalloproteinase (MMP) production. Both pathways contribute to UV irradiation-induced skin damage, such as photoaging and skin tumor formation. OBJECTIVE: To elucidate the underlying mechanism, we examined the effect of quercetin on UV irradiation induced activation of NF-κB and AP-1 pathways. METHODS: Primary human keratinocytes, the major skin cell type subjected to physiological solar UV irradiation, were used to study the effects of quercetin on UV irradiation-induced signal transduction pathways. RESULTS: Quercetin decreased UV irradiation-induced NF-κB DNA-binding by 80%. Consequently, quercetin suppressed UV irradiation-induced expression of inflammatory cytokines IL-1ß (∼60%), IL-6 (∼80%), IL-8 (∼76%) and TNF-α (∼69%). In contrast, quercetin had no effect on UV irradiation activation of three MAP kinases, ERK, JNK, or p38. Accordingly, induction of AP-1 target genes such as MMP-1 and MMP-3 by UV irradiation was not suppressed by quercetin. CONCLUSION: Our data indicate that the ability of quercetin to block UV irradiation-induced skin inflammation is mediated, at least in part, by its inhibitory effect on NF-κB activation and inflammatory cytokine production.

Asunto(s)

Citocinas/metabolismo , Queratinocitos/efectos de los fármacos , Queratinocitos/efectos de la radiación , FN-kappa B/metabolismo , Quercetina/farmacología , Transducción de Señal/efectos de los fármacos , Rayos Ultravioleta , Antioxidantes/farmacología , Células Cultivadas , Citocinas/antagonistas & inhibidores , Humanos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Queratinocitos/metabolismo , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 3 de la Matriz/metabolismo , FN-kappa B/antagonistas & inhibidores , Factor de Transcripción AP-1/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

Recent in vitro data have suggested that the flavonoid quercetin (1) does not affect the functioning of neutrophils. Therefore, we evaluated in vivo and in vitro whether or not 1 affects neutrophil function, focusing on recruitment. The in vivo treatment with 1 inhibited in a dose-dependent manner the recruitment of neutrophils to the peritoneal cavity of mice induced by known chemotatic factors such as CXCL1, CXCL5, LTB(4), and fMLP. Furthermore, 1 also inhibited in a concentration-dependent manner the chemoattraction of human neutrophils induced by CXCL8, LTB(4), and fMLP in a Boyden chamber. In vitro treatment with 1 did not affect human neutrophil surface expression of CXCR1, CXCR2, BLT1, or FLPR1, but rather reduced actin polymerization. These results suggest that 1 inhibits actin polymerization, hence, explaining the inhibition of neutrophil recruitment in vivo and in vitro and highlighting its possible usefulness to diminish excessive neutrophil migration during inflammation.

Asunto(s)

Actinas/metabolismo , Quimiocina CXCL5/inmunología , Interleucina-8/efectos de los fármacos , Leucotrieno B4/inmunología , N-Formilmetionina Leucil-Fenilalanina/inmunología , Infiltración Neutrófila/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Quercetina/farmacología , Actinas/efectos de los fármacos , Animales , Factores Quimiotácticos/inmunología , Quimiotaxis/inmunología , Relación Dosis-Respuesta a Droga , Humanos , Inflamación/inmunología , Interleucina-8/inmunología , Masculino , Ratones , Estructura Molecular , Infiltración Neutrófila/inmunología , Neutrófilos/inmunología , Quercetina/química , Quercetina/inmunología

RESUMEN

BACKGROUND: The effectiveness of a water/oil (w/o) microemulsion containing quercetin against ultraviolet B radiation (UVB) induced damage was recently demonstrated by our group. However, during the development of new pharmaceutical products, the evaluation of percutaneous absorption and in vivo effectiveness should be accompanied by evaluation of stability parameters as an integral part of the process. OBJECTIVE: The aim was to investigate the stability of the final microemulsion formulation considering the temperature ranges of storage and application. METHODS: The physical, chemical, and functional stability of this formulation under different conditions of storage during 12 months and the photostability of quercetin incorporated into this system over UVB exposure for 7 days were evaluated. RESULTS: Although the results indicated a notable physical stability of the w/o microemulsions during the experimental period under all employed conditions, in both, the chemical and functional studies, a significant loss of quercetin content and antioxidant activity was found after 6 months of storage at 30°C/70% relative humidity and after 2 months at 40°C/70% relative humidity. The photostability study results demonstrated that the incorporation of quercetin into the w/o microemulsion maintained the previously demonstrated photostability of this flavonoid under forced exposure to UVB irradiation. CONCLUSION: Thus, this work demonstrates that special storage conditions (at 4 ± 2°C) are necessary to maintain the functionality of the w/o microemulsion containing quercetin and mainly emphasizes the importance of studying physical, chemical, and functional parameters at the same time during stability evaluation of active principles.

Asunto(s)

Antioxidantes/química , Aceites/química , Quercetina/química , Agua/química , Antioxidantes/farmacología , Química Farmacéutica , Formas de Dosificación , Estabilidad de Medicamentos , Almacenaje de Medicamentos , Emulsiones/química , Emulsiones/efectos de la radiación , Humedad , Tamaño de la Partícula , Quercetina/farmacología , Temperatura , Rayos Ultravioleta

RESUMEN

PURPOSE: In the present study, histological aspects were considered in order to evaluate the in vivo photoprotective effect of a w/o microemulsion containing quercetin against UVB irradiation-induced dermal damages. The toxicity in cell culture and the potential skin irritation resulting from topical application of this formulation were also investigated. METHODS: Mouse dorsal surfaces were treated topically with 300 mg of the unloaded and quercetin-loaded (0.3%, w/w) microemulsions before and after exposure to UVB (2.87 J/cm2) irradiation. The untreated control groups irradiated and non-irradiated were also evaluated. UVB-induced histopathological changes as well as the photoprotective effect of this formulation were evaluated considering the parameters of infiltration of inflammatory cells, epidermis thickening (basale and spinosum layers) and collagen and elastic fiber contents. The cytotoxicity of the reported formulation was evaluated in L929 mice fibroblasts by MTT assay and the skin irritation was investigated after topical application of both unloaded and quercetin-loaded microemulsions once a day for 15 days. RESULTS: The results demonstrated that the w/o microemulsion containing quercetin reduced the incidence of histological skin alterations, mainly the connective-tissue damage, induced by exposure to UVB irradiation, this allows the suggestion that protective effects of this formulation against UV-induced responses are not secondary to the interference of UV transmission (i.e., blocking the UVB radiation from being absorbed by the skin), as is usually done with UVB absorbers and sunscreens, but is instead due to different biological effects of this flavonoid. Furthermore, by evaluating the cytotoxic effect on L929 cells and histological aspects such as infiltration of inflammatory cells and epidermis thickness of hairless mice, the present study also demonstrated no toxicity of the proposed system. CONCLUSIONS: Therefore, based on these mouse models, a detailed characterization of the w/o microemulsion incorporating quercetin effects as a photochemoprotective agent on human skin is thus indicated.

Asunto(s)

Antioxidantes/farmacología , Quercetina/farmacología , Rayos Ultravioleta/efectos adversos , Animales , Antioxidantes/administración & dosificación , Antioxidantes/toxicidad , Células Cultivadas , Colágeno/metabolismo , Elastina/metabolismo , Emulsiones , Femenino , Fibroblastos/metabolismo , Inflamación/patología , Masculino , Ratones , Ratones Pelados , Quercetina/administración & dosificación , Quercetina/toxicidad , Piel/efectos de los fármacos , Piel/efectos de la radiación

RESUMEN

Quercetin (1) is known to have both antioxidant and antinociceptive effects. However, the mechanism involved in its antinociceptive effect is not fully elucidated. Cytokines and reactive oxygen species have been implicated in the cascade of events resulting in inflammatory pain. Therefore, we evaluated the antinociceptive mechanism of 1 focusing on the role of cytokines and oxidative stress. Intraperitoneal and oral treatments with 1 dose-dependently inhibited inflammatory nociception induced by acetic acid and phenyl-p-benzoquinone and also the second phase of formalin- and carrageenin-induced mechanical hypernociception. Compound 1 also inhibited the hypernociception induced by cytokines (e.g., TNFalpha and CXCL1), but not by inflammatory mediators that directly sensitize the nociceptor such as PGE2 and dopamine. On the other hand, 1 reduced carrageenin-induced IL-1beta production as well as carrageenin-induced decrease of reduced glutathione (GSH) levels. These results suggest that 1 exerts its analgesic effect by inhibiting pro-nociceptive cytokine production and the oxidative imbalance mediation of inflammatory pain.

Asunto(s)

Analgésicos/farmacología , Antioxidantes/farmacología , Productos Biológicos/farmacología , Citocinas/efectos de los fármacos , Inflamación/fisiopatología , Estrés Oxidativo , Dolor/tratamiento farmacológico , Quercetina/farmacología , Analgésicos/química , Antioxidantes/química , Productos Biológicos/química , Citocinas/metabolismo , Estructura Molecular , Quercetina/química

RESUMEN

The purpose of this study was to develop a lyotropic liquid crystalline formulation using the emulsifier vitamin E TPGS and evaluate its behavior after incorporation of a flavonoid, quercetin. The physical (macro and microscopic), chemical (determination of quercetin content by the HPLC method) and functional (determination of quercetin antioxidant activity by DPPH(*) assay) stability of the lamellar liquid crystalline formulation containing flavonoid was evaluated when stored at 4 +/- 2 degrees C; 30 +/- 2 degrees C/70 +/- 5% RH (relative humidity) and 40 +/- 2 degrees C/70 +/- 5% RH during 12 months. The lamellar liquid crystalline structure of the formulation was maintained during the experiment, however chemical and functional stability results showed a great influence of the storage period in all conditions tested. A significant decrease in quercetin content (approximately 40%) was detected during the first month of storage and a similar significant loss in antioxidant activity was detected after 6 months. The remaining flavonoid content was unchanged during the final 6 months of the experimental period. The results suggest possible interactions between quercetin and the liquid crystalline formulation, which could inhibit or reduce the quercetin activity incorporated in the system. In conclusion, the present study demonstrated that incorporation of quercetin (1%) did not affect the liquid crystalline structure composed of vitamin E TPGS/IPM/PG-H2O (1:1) at 63.75/21.25/15 (w/w/w). Nevertheless, of the total quercetin incorporated in the system only 60% was free to act as an antioxidant.

Asunto(s)

Depuradores de Radicales Libres/química , Cristales Líquidos , Quercetina/química , Química Farmacéutica , Cromatografía Líquida de Alta Presión , Formas de Dosificación , Composición de Medicamentos , Estabilidad de Medicamentos , Emulsionantes/química , Humedad , Miristatos/química , Polietilenglicoles/química , Propilenglicol/química , Tecnología Farmacéutica/métodos , Temperatura , Factores de Tiempo , Vitamina E/análogos & derivados , Vitamina E/química

RESUMEN

The present study evaluated the potential of a w/o microemulsion as a topical carrier system for delivery of the antioxidant quercetin. Topical and transdermal delivery of quercetin were evaluated in vitro using porcine ear skin mounted on a Franz diffusion cell and in vivo on hairless-skin mice. Skin irritation by topical application of the microemulsion containing quercetin, and the protective effect of the formulation on UVB-induced decrease of endogenous reduced glutathione levels and increase of cutaneous proteinase secretion/activity were also investigated. The w/o microemulsion increased the penetration of quercetin into the stratum corneum and epidermis plus dermis at 3, 6, 9 and 12h post-application in vitro and in vivo at 6h post-application. No transdermal delivery of quercetin occurred. By evaluating established endpoints of skin irritation (erythema formation, epidermis thickening and infiltration of inflammatory cells), the study demonstrated that the daily application of the w/o microemulsion for up to 2 days did not cause skin irritation. W/o microemulsion containing quercetin significantly prevented the UVB irradiation-induced GSH depletion and secretion/activity of metalloproteinases.

Asunto(s)

Antioxidantes/farmacología , Antioxidantes/farmacocinética , Quercetina/farmacología , Quercetina/farmacocinética , Protectores contra Radiación/farmacología , Protectores contra Radiación/farmacocinética , Animales , Antioxidantes/administración & dosificación , Química Farmacéutica , Cromatografía Líquida de Alta Presión , Portadores de Fármacos , Sistemas de Liberación de Medicamentos , Electroforesis en Gel de Poliacrilamida , Emulsiones , Glutatión/metabolismo , Irritantes/toxicidad , Ratones , Ratones Pelados , Aceites , Péptido Hidrolasas/química , Péptido Hidrolasas/metabolismo , Quercetina/administración & dosificación , Protectores contra Radiación/administración & dosificación , Piel/efectos de los fármacos , Piel/enzimología , Absorción Cutánea , Rayos Ultravioleta , Agua

RESUMEN

In the present study it was evaluated the: (i) functional stability of the soybean extract as a raw material and dispersed in two different topical formulations, (ii) skin retention using modified Franz diffusion cells, and (iii) in vivo activity of these formulations to inhibit 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced hydrogen peroxide (H(2)O(2)) and malondialdehyde (MDA) increases in the skin of hairless mice. The physico-chemical stability was evaluated by pH, globule size and centrifugation test. Furthermore, functional stability was also evaluated by antilipoperoxidative activity. The two topical formulations were stored at 4 degrees C, 30 degrees C/60% RH and 40 degrees C/70% RH for 6 months. The evaluation of the antiperoxidative stability of soybean extract itself and incorporated in formulations did not demonstrate loss of activity by storage at 4 degrees C/6 months. During 6 months of the study in different storage conditions the formulations 1 and 2 added or not with soybean extract were stable to physico-chemical tests. The effect of antioxidant compounds detected by the inhibition of MDA formation was time-dependent for formulation 2 as detected in the skin retention study. Pretreatment with formulation 1 or 2 significantly diminished TPA-induced H(2)O(2) and MDA generation. In conclusion, the present results suggest for the first time that formulations containing soybean extract may be a topical source of antioxidant compounds that decrease oxidative damages of the skin.

Asunto(s)

Antioxidantes/farmacología , Fármacos Dermatológicos/farmacología , Glycine max , Estrés Oxidativo/efectos de los fármacos , Piel/efectos de los fármacos , Administración Cutánea , Animales , Antioxidantes/administración & dosificación , Antioxidantes/química , Antioxidantes/metabolismo , Química Farmacéutica , Fármacos Dermatológicos/administración & dosificación , Fármacos Dermatológicos/química , Fármacos Dermatológicos/metabolismo , Cámaras de Difusión de Cultivos , Estabilidad de Medicamentos , Almacenaje de Medicamentos , Humedad , Peróxido de Hidrógeno/metabolismo , Concentración de Iones de Hidrógeno , Peroxidación de Lípido/efectos de los fármacos , Masculino , Malondialdehído/metabolismo , Ratones , Ratones Pelados , Tamaño de la Partícula , Extractos Vegetales/farmacología , Piel/metabolismo , Absorción Cutánea , Glycine max/química , Porcinos , Temperatura , Acetato de Tetradecanoilforbol/farmacología , Factores de Tiempo

RESUMEN

Recently, it was demonstrated that two different formulations containing quercetin inhibit the UVB-induced cutaneous oxidative stress and inflammation. Therefore, in the present study it was evaluated the functional stability of those formulations by the antioxidant activity, the release of quercetin from the formulations, and its skin retention using modified Franz diffusion cells. Both formulations tested ((1) non-ionic emulsion with high lipid content and (2) anionic emulsion with low lipid content) remained functionally (hydrogen-donating ability) stable during 180 days. Furthermore, quercetin was released from both formulations as determined using nitrocellulose membrane. In vitro antioxidant activity of retained quercetin into the skin was observed for both formulations as detected by the inhibition of malondialdehyde formation. The effect of quercetin retention was time-dependent for formulation 1. Concluding, this study demonstrates that quercetin remains functionally stable in formulations, and measuring the antioxidant activity is an efficient approach to evaluate quercetin skin retention with minimal interference of the tissue products. Furthermore, the present results on skin retention explain the previous study on quercetin in vivo activities, and together, these data suggest that formulations containing quercetin may be used as topical active products to control UVB-mediated oxidative damage of the skin.

Asunto(s)

Antioxidantes/química , Antioxidantes/metabolismo , Quercetina/química , Quercetina/metabolismo , Piel/metabolismo , Administración Cutánea , Animales , Compuestos de Bifenilo , Estabilidad de Medicamentos , Excipientes/química , Radicales Libres/química , Técnicas In Vitro , Picratos/química , Polisorbatos/química , Absorción Cutánea , Solubilidad , Porcinos

RESUMEN

The present study evaluates the antioxidant activity of the flavonol quercetin, and its functional stability as a raw material and when added in formulations. The iron-chelating activity was determined using the bathophenanthroline assay, and the functional stability was evaluated with the antilipoperoxidative assay. Raw material presented concentration-dependent antilipoperoxidative and iron-chelating activities. The initial antilipoperoxidative activity of the raw material, cream and gel-cream were 63%, 78%, and 69%, respectively. There was no detectable loss of activity during 182 days (6 months) of storage at all tested temperatures (4 degrees C, room temperature [RT], 37 degrees C, and 45 degrees C) for the raw material. Considering the method variability of 10%, activity loss greater than 10% for nonionic cream was detected after 126 days at 4 degrees C (20.1%), decreasing thereafter to 22.2% after 182 days. At 45 degrees C, the loss of activity started after 182 days (13.2%). For the anionic gel-cream, activity loss started after 84 days (28.4%, 45 degrees C), decreasing after 182 days to 40.3% at 45 degrees C. At 37 degrees C, activity loss was detected after 182 days (12%). In conclusion, the results suggest that the activity of quercetin depends on iron chelation, and its possible usefulness as a topical antioxidant to prevent oxidative stress-induced skin damage depends on maintaining its antilipoperoxidative activity stored at RT, which avoids special storage conditions.

Asunto(s)

Antioxidantes/administración & dosificación , Peroxidación de Lípido/efectos de los fármacos , Quercetina/administración & dosificación , Administración Tópica , Química Farmacéutica , Relación Dosis-Respuesta a Droga , Estabilidad de Medicamentos , Quelantes del Hierro/farmacología , Quercetina/química , Quercetina/farmacología , Temperatura

RESUMEN

UV radiation-induced skin damages may result in pre-cancerous and cancerous skin lesions, and acceleration of skin aging. It involves an imbalance of the endogenous antioxidant system that leads to the increase of free radical levels and inflammation. Therefore, antioxidant supplementation might inhibit such imbalance. In this regard, quercetin is a promising drug, this plant derived lipophilic flavonoid presents the higher antioxidant activity among flavonoids and multiple antioxidant mechanisms. Thus, the present study investigated the possible beneficial effects of topical formulations containing quercetin to inhibit UVB irradiation-induced oxidative damages. Quercetin was administered on the dorsal skin of hairless mice using two formulations, formulation 1 (non-ionic emulsion with high lipid content) and formulation 2 (anionic emulsion with low lipid content). The UVB irradiation (0.31-3.69 J/cm(2)) induced a dose-dependent increase in the myeloperoxidase (MPO) activity (4-2708%) and depletion of reduced glutathione (GSH) (22-68%) in the skin of hairless mice after 6h. These results demonstrated that the UVB doses are not excessive, and additionally, they are lower than the doses used in other similar studies. Proteinases secretion/activity, detected by the qualitative sodium dodecyl sulphate polyacrylamide gel electrophoresis substrate-embedded enzymography (zymography), was also enhanced in the same manner as MPO activity using the UVB dose of 1.23J /cm(2). Formulations 1 and 2 inhibited the MPO activity increase (62% and 59%, respectively), GSH depletion (119% and 53%, respectively) and proteinases secretion/activity. To our knowledge, this is the first study to demonstrate the effectiveness of topical formulations containing quercetin to inhibit the UVB irradiation-induced skin damages. Thus, these data suggest the possible usefulness of topical formulations containing quercetin to prevent UVB radiation skin damages.

Asunto(s)

Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/efectos de la radiación , Quercetina/farmacología , Piel/efectos de los fármacos , Piel/efectos de la radiación , Rayos Ultravioleta , Administración Tópica , Animales , Glutatión/efectos de los fármacos , Glutatión/metabolismo , Ratones , Ratones Pelados , Péptido Hidrolasas/efectos de los fármacos , Péptido Hidrolasas/metabolismo , Peroxidasa/efectos de los fármacos , Peroxidasa/metabolismo , Quercetina/administración & dosificación , Protectores contra Radiación/administración & dosificación , Protectores contra Radiación/farmacología , Piel/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/efectos de la radiación