Effects of an aglycone-rich biotransformed soybean extract in human endothelial cells.

Estrogen replacement therapy is not thought to be a safe treatment for prevention of cardiovascular disease in menopausal women; isoflavones are a possible alternative. Estrogen produces beneficial effects on the cardiovascular system by enhancing production of nitric oxide, a vasoprotective and antiatherosclerotic agent. Estrogen-like compounds such as isoflavones are also suggested for increasing nitric oxide production. Isoflavones are present mainly in soy foods as glucosides, but soy isoflavone aglycones, the biologically active estrogen-like compounds, are absorbed faster and in higher amounts than their glucoside derivatives and show higher biological activity, implying that they may be more effective in preventing chronic diseases such as coronary heart disease. We evaluated an extract of soybeans fermented by Aspergillus awamori on which polyphenol glucosides were biotransformed to aglycone forms on production of nitric oxide, prostaglandin E2 and endothelin-1 in vitro in human endothelial cells, comparing it with a non-fermented extract. Bioconverted soybean extracts enhanced endothelin-1, nitric oxide and prostaglandin E2 production, while the unfermented extract only enhanced endothelin-1 production. Thus, only the aglycone-rich forms of soybean extracts were able to increase nitric oxide and prostaglandin E2 production, demonstrating that, in endothelial cells in vitro, they may be usable as therapeutic agents against the development of atherosclerosis.

Sunscreen protection against ultraviolet-induced oxidative stress: evaluation of reduced glutathione levels, metalloproteinase secretion, and myeloperoxidase activity.

Several studies have demonstrated the skin protection by sunscreens considering the aspects skin penetration, photostability, and protection against erythema and sunburn. However, little is known about the effect of topically applied sunscreen formulations on the antioxidant defense, metalloproteinases, and inflammatory processes of skin in response to UVR exposure. Therefore, this study aimed to investigate the use of a cream gel formulation containing the UV filters benzophenone-3, octyl methoxycinnamate, and octyl salicylate to prevent skin damage from a single dose of UVR (2.87 J/cm2). This protective effect was evaluated in vivo by measuring the following biochemical parameters: reduced glutathione levels, secretion of matrix metalloproteinases, and myeloperoxidase activity. The results showed that the sunscreen formulation, despite having sun protection factor (SPF) 15, was not completely effective to protect the skin against GSH depletion, MMP-9 secretion and the inflammatory process induced by UVR. These results demonstrate the importance of analyzing UV-altered biochemical parameters of skin in order to propose new sunscreen formulations that can completely protect skin against UVR-induced damage.

Biotransformed soybean cream as a new nutraceutical for skin care: collagen stimulation in vitro and ex vivo.

Treatments that attenuate the effects of hypoestrogenism in menopausal women have been gaining visibility. This study investigated the skin response to a phytoestrogen-enriched cosmetic formulation created by incorporating a biotransformed soybean extract (BE) into a cream-like matrix. Collagen-I expression was analyzed both in vitro (fibroblast cells) and ex vivo (skin explants). The results revealed an increased amount of collagen-I both in fibroblasts and human skin when treated with BE and BE-incorporated cream. Also, this collagen-I overexpression was inhibited by PHTPP, indicating a dependence on estrogen hormone receptor beta (ERß) signaling. Moreover, BE was not harmful to skin microbiota, showing a promising nutricosmetic potential. Thus, this work presented a fully functional cream-like formulation that was shown to be safe and effectively increase collagen-I levels both in vitro and ex vivo.

In vivo evaluation of a metronidazole-containing gel for the adjuvant treatment of chronic periodontitis: preliminary results.

The present study developed an experimental metronidazole-based gel and evaluated its efficacy for the adjuvant treatment of chronic periodontitis. Sixteen patients were randomly allocated into two groups of eight subjects according to the following proposed treatments: (1) scaling and root planing (active control) or (2) scaling and root planing and direct periodontal intrapocket application of 15% metronidazole-based gel in two sites (≥5 mm in depth) (experimental group). Potential changes in the subgingival microbiota were assessed using a DNA Checkerboard method at three proposed times: baseline and following 7 or 30 days of drug administration. High-performance liquid chromatography (HPLC) monitored metronidazole concentrations in the crevicular fluid during treatment. The metronidazole experimental group presented lower bacterial counts than the control group at the three evaluated times (p<0.01 for baseline, p<0.001 for 7 or 30 days) when the target species were analyzed as a pool of bacteria. Samples revealed significantly lower counts 7 days after drug administration compared with baseline or after 30 days (p<0.05). HPLC analysis detected gel 1 h after application. The metronidazole-based gel significantly decreased the total bacterial count at the three evaluated times. Periodontopathogenic species were not different after gel administration.

Enhanced in vitro and in vivo antioxidant activity and mobilization of free phenolic compounds of soybean flour fermented with different beta-glucosidase-producing fungi.

AIMS: To evaluate the soybean polyphenol glucosides bioconversion to aglycone forms by different beta-glucosidases-producing filamentous fungi to enhance their antioxidant activity. METHODS AND RESULTS: Soybean defatted flour was submitted to solid-state fermentation with Aspergillus niger, Aspergillus niveus and Aspergillus awamori. The fungi studied produced approximately the same beta-glucosidase activity units amount when p-nitrophenyl-beta-d-glucopyranoside was used as substrate for the assay. However, electrophoretic analysis, using 4-methylumbellipheryl-beta-d-glucopyranoside as substrate, showed that beta-glucosidase produced by A. niveus was more active. Fermented methanolic extracts showed an increase in polyphenol and genistein contents and antioxidant activities. The highest genistein content was found in soybean fermented by A. niveus. Methanolic extracts of the soybean fermented by the different fungi showed a similar capacity of scavenging H(2)O(2) generated in vivo by the tumour promoter 12-O-tetradecanoyl phorbol-13-acetate. CONCLUSIONS: A. niveus synthesized a beta-glucosidase with higher specificity to hydrolyse genistin beta-glycosidic bond than those produced by A. awamori and A. niger. SIGNIFICANCE AND IMPACT OF THE STUDY: The utilization of these beta-glucosidases-producing fungi in soybean fermentation processes resulted in the obtaining of methanolic extracts with different antioxidant potentials that could be used either therapeutically or as an antioxidant in nonphysiological oxidative stress conditions, as the one induced in skin by UV radiation.

Biotransformed soybean cream as a new nutraceutical for skin care: collagen stimulation in vitro and ex vivo

Treatments that attenuate the effects of hypoestrogenism in menopausal women have been gaining visibility. This study investigated the skin response to a phytoestrogen-enriched cosmetic formulation created by incorporating a biotransformed soybean extract (BE) into a cream-like matrix. Collagen-I expression was analyzed both in vitro (fibroblast cells) and ex vivo (skin explants). The results revealed an increased amount of collagen-I both in fibroblasts and human skin when treated with BE and BE-incorporated cream. Also, this collagen-I overexpression was inhibited by PHTPP, indicating a dependence on estrogen hormone receptor beta (ERβ) signaling. Moreover, BE was not harmful to skin microbiota, showing a promising nutricosmetic potential. Thus, this work presented a fully functional cream-like formulation that was shown to be safe and effectively increase collagen-I levels both in vitro and ex vivo.

Development of topical functionalized formulations added with propolis extract: stability, cutaneous absorption and in vivo studies.

Propolis, which is a natural product widely consumed in the folk medicine, is a serious candidate to be applied topically due to its outstanding antioxidant properties. So, the purpose of this study was to develop stable topical formulations added with propolis extract in an attempt to prevent and/or treat the diseases occurring in skin caused by UV radiation. The antioxidant activity using a chemiluminescent method was used to evaluate the functional stability and the permeation/retention in skin of these formulations. In the long-term stability study, the formulations were stored at 25+/-2 degrees C/AH and at 40+/-2 degrees C/70% RH for 360 days. It was found in this study, that the formulations prepared with Polawax showed functional and physical stability in the period of study. In addition, this formulation presented good results in the percutaneous study, allowing the antioxidant compounds present in the propolis extract to reach lower layers in pig ear skin and in the whole hairless mice skin (retention=0.12 and 0.13 microL of propolis/g of skin, respectively). In the in vivo study, it was also suggested that this formulation may be effective in protecting skin from UVB photodamage, nevertheless other assays need to be done in order to have a complete understanding of the protective effect of formulations added with propolis extract.

Spray-dried propolis extract. I: physicochemical and antioxidant properties.

The effect of spray drying on the chemical and biological properties of alcoholic extract of green propolis was investigated. The total polyphenol and flavonoid contents in spray-dried propolis extract were determined by the Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively. The antioxidant activity of the dry extract was assessed by the membrane lipid peroxidation inhibition method, using quercetin as reference. The polyphenol content was shown to depend on the drying air outlet temperature and its square at the 0.5% significance level, while the flavonoid content depended only on the square of the outlet temperature at the 5% significance level. Polyphenol and flavonoid recovery after spray-drying ranged from 45.1 to 54.9% and 30.6 to 40.8%, respectively. The antioxidant activity of the spray-dried propolis was shown to be affected by the extract feed rate and air outlet temperature at a significance level of 0.1%. The spray-dried propolis extract showed significant antioxidant activity, with 50% lipid peroxidation inhibition at concentrations ranging from 2.5 to 5.0 microg/ml.

Liquid Crystalline Nanodispersions Functionalized with Cell-Penetrating Peptides for Topical Delivery of Short-Interfering RNAs: A Proposal for Silencing a Pro-Inflammatory Cytokine in Cutaneous Diseases.

Short-interfering RNAs (siRNAs) are a potential strategy for the treatment of cutaneous diseases. In this context, liquid crystalline nanoparticles functionalized with specific proteins and peptide-transduction domains (PTDs), which act as penetration enhancers, are a promising carrier for siRNA delivery through the skin. Herein, hexagonal phase liquid crystal nanoparticles based on monoolein (MO) and/or oleic acid (OA) containing (or lacking) the cationic polymer polyethylenimine (PEI) and the cationic lipid oleylamine (OAM) were functionalized with the membrane transduction peptides transcriptional activator (TAT) or penetratin (PNT). These nanoparticles were complexed with siRNA and characterized by particle size, polydispersity, zeta potential, complexation efficiency and siRNA release. The formulations containing cationic agents presented positive zeta potentials, sizes on the nanometer scale, and complexed siRNAs at concentrations of 10 µM; these agents were successfully released in a heparin competition assay. Cell culture studies demonstrated that nanoparticles composed of MO:OA:PEI functionalized with TAT were the most efficient at transfecting L929 cells, and the uptake efficiency was enhanced by TAT peptide functionalization. Thereafter, the selected formulations were evaluated for in vivo skin irritation, penetration and in vivo efficacy using a chemically induced inflammatory animal model. These nanoparticles did not irritate the skin and provided higher siRNA penetration and delivery into the skin than control formulations. Additionally, efficacy studies in the animal model showed that the association of TAT with the nanodispersion provided higher suppression of tumor necrosis factor (TNF)-α. Thus, the development of liquid crystalline nanodispersions containing TAT may lead to improved topical siRNA delivery for the treatment of inflammatory skin diseases.

Physical and chemical stability of different formulations with superoxide dismutase.

Topical formulations with superoxide dismutase (SOD), a scavenger of superoxide radicals, have proved to be effective against some skin diseases. Nevertheless, formulations with proteins are susceptible to both chemical and physical instability. Three different formulations (anionic and non-ionic gel and emulsion) were developed and supplemented with SOD in order to determine the most stable formulation that would maintain SOD activity. Physical stability was evaluated by assessing the rheological behavior of the formulations stored at room temperature, 37 and 45 degrees C. Chemical stability was evaluated by the measurement of enzymatic activity in the formulations stored at room temperature and at 45 degrees C. Formulations showed a flow index less than one, characterizing pseudoplastic behavior. There was no significant difference in initial values of flow index, tixotropy or minimum apparent viscosity. Neither gel showed significant changes in minimum apparent viscosity concerning storage time or temperature, as well, SOD presence and its activity. The emulsion showed decreased viscosity by the 28th day, but no significant changes concerning storage temperature or SOD presence, although it showed a decreased activity. The addition of SOD to the formulations studied did not affect their physical stability but gel formulations seem to be better bases for enzyme addition.