Quercetin and 3-O-methylquercetin in vitro skin layers permeation/retention from hydrogels: why only a methoxy group difference determines different behaviors?

OBJECTIVES: The present study was designed to verify if quercetin (QCT), a flavonoid with antioxidant and antiviral activity, and 3-O-methylquercetin (3OMQ), a quercetin C3-methoxylated derivative, present differences in their behavior against complexation with ß-cyclodextrin (ß-CD) and the corresponding permeation/retention trhough porcine ear skin, when incorporated into hydroxypropyl methylcellulose (HPMC) or chitosan (CS) hydrogels. METHODS: The influence of ß-CD on the skin permeation/retention of QCT and 3OMQ from hydrogels is comparatively evaluated for both flavonoids using porcine ear skin in Franz cells model. The properties of the two flavonoids using the semi-empirical method Recife Model was studied. KEY FINDINGS: Quercetin presented higher skin retention compared with its C3-methoxy derivative 3OMQ. The best permeation/retention of QCT was observed when it was incorporated into CS hydrogel containing 5% ß-CD, whereas, for 3OMQ, the HPMC hydrogel containing 5% ß-CD was the best formulation. The flavonoids complexation with ß-CD in water occurred preferentially with the insertion of the B ring through the secondary OH rim. CONCLUSIONS: The dynamic molecular modeling revealed that the methyl group at C3 in 3OMQ molecule determined significant difference in its complexation with ß-CD, in comparison to its analogous QCT and that difference is coincident with the permeation behavior of these flavonoids, denoting a possible relationship with their molecular dynamics.

Comparative evaluation of rivastigmine permeation from a transdermal system in the Franz cell using synthetic membranes and pig ear skin with in vivo-in vitro correlation.

In the present study, in vitro permeation experiments in a Franz diffusion cell were performed using different synthetic polymeric membranes and pig ear skin to evaluate a rivastigmine (RV) transdermal drug delivery system. In vitro-in vivo correlations (IVIVC) were examined to determine the best model membrane. In vitro permeation studies across different synthetic membranes and skin were performed for the Exelon(®) Patch (which contains RV), and the results were compared. Deconvolution of bioavailability data using the Wagner-Nelson method enabled the fraction of RV absorbed to be determined and a point-to-point IVIVC to be established. The synthetic membrane, Strat-M™, showed a RV permeation profile similar to that obtained with pig ear skin (R(2)=0.920). Studies with Strat-M™ resulted in a good and linear IVIVC (R(2)=0.991) when compared with other synthetic membranes that showed R(2) values less than 0.90. The R(2) for pig ear skin was 0.982. Strat-M™ membrane was the only synthetic membrane that adequately simulated skin barrier performance and therefore it can be considered to be a suitable alternative to human or animal skin in evaluating transdermal drug transport, potentially reducing the number of studies requiring human or animal samples.

Simultaneous quantification of flavonoids from Achyrocline satureioides by a polar-reversed phase LC method--application to skin permeation/retention studies.

A selective and sensitive polar-reversed phase LC method was validated for simultaneous quantification of the main Achyrocline satureioides flavonoids (quercetin, luteolin, and 3-O-methylquercetin) in skin samples after permeation/retention studies from topical nanoemulsions. The method was linear in a range of 0.25 to 10.0 microg/mL exhibiting a coefficient of determination higher than 0.999 for all flavonoids. No interference of the nanoemulsion excipients or skin components was observed in the retention times of all flavonoids. The R.S.D. values for intra- and inter-day precision experiments were lower than 6.73%. Flavonoids recovery from nanoemulsions and skin matrices was between 90.05 and 109.88%. In a permeation/retention study with porcine ear high amount of 3-O-methylquercetin was found in the skin sample (0.92 +/- 0.22 microg/g) after two hours. The proposed method was suitable to quantify the main flavonoids of A. satureioides in skin permeation/retention studies from topical nanoemulsions.

Improving the topical delivery of zinc phthalocyanine using oleic acid as a penetration enhancer: in vitro permeation and retention.

Topical photodynamic therapy with zinc phthalocyanine (ZnPc), second-generation photosensitizer, can be an alternative method for the treatment of skin cancer. However, ZnPc has poor penetration in the skin. This study was aimed at investigating whether the presence of oleic acid (chemical enhancer) in propylene glycol can improve the topical delivery of ZnPc. The topical (to the skin) and transdermal (across the skin) delivery of ZnPc were evaluated in vitro using suine ear skin mounted in Franz diffusion cell. Photosensitizer was quantified by fluorescence emission, which is a sensitive and selective method. At 5 and 10%, oleic acid increased the topical and transdermal delivery significantly. When the concentration of oleic acid was further increased (20-60% w/w), the topical delivery of ZnPc was still elevated, but its transdermal delivery was substantially reduced. It was concluded that oleic acid (in propylene glycol formulations) can promote the topical delivery of ZnPc, with reduced transdermal delivery. This approach can be effective for the treatment of skin cancer by topical photodynamic therapy.

Nitric oxide photorelease from hydrogels and from skin containing a nitro-ruthenium complex.

Nitric oxide (NO) is a gaseous molecule that has specific functions dictated by its localization and its kinetics of release. As NO-donors have a range of potential uses in the skin, much attention has been paid to the development of topical NO delivery systems. The aim of this work was to study the release rate and the skin penetration of the NO-donor cis-[Ru(NO(2))(bpy)(2)(4-pic)](+) from different gel formulations and their potential as topical NO delivery systems under light stimuli. Among the formulations developed, the anionic gel retarded the nitro-ruthenium complex diffusion and also obstructed NO release after light irradiation. On the other hand, NO release before light irradiation was observed when the complex was dispersed in the cationic chitosan gel, possibly due to oxi-redox reactions between the amino groups of the polymer and the drug molecule. Finally, the non-ionic gel released the NO after light irradiation to the same extent as a drug aqueous solution at the same pH. The drug dispersed in this gel also penetrated into the stratum corneum skin layer, and the nitro-ruthenium complex present in the skin was able to release the NO after light stimuli, suggesting the potential use of this formulation as a topical NO delivery system.

Evaluation of the transdermal permeation of different paraben combinations through a pig ear skin model.

Although parabens have several features of ideal preservatives, different studies have shown that they may affect human health due to their estrogenic activity. Therefore, various strategies have been applied to reduce their skin penetration. However, the effect of paraben combinations on transdermal permeation has not yet been investigated. Thus, the objective of this study was to evaluate paraben permeation in pig ear skin using a Franz diffusion cell system with capillary electrophoresis detection, in order to identify which paraben combinations (defined by a factorial design) have the lowest skin permeation. The permeation of isolated parabens was also evaluated and the permeation characteristics, obtained by the Moser model, confirmed that lipophilicity and molecular weight may influence the systemic absorption of these compounds. In previous tests using isolated parabens, methyl and ethyl parabens presented greater retention in the epidermis compared to the dermis, while propyl and butyl parabens had similar retention profiles in these layers. An increase in ethanol concentration and experimental time promoted greater parabens retention in the dermis compared to the epidermis. The binary combinations of methyl and ethyl parabens as well as of methyl and propyl parabens (added to several cosmetic products in order to increase the antimicrobial spectrum) reduced significantly their permeation rates through pig ear skin (with the exception of EP), probably due to the high retention of these parabens in the epidermis and dermis.

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.

Effects of phycocyanin extract on prostaglandin E2 levels in mouse ear inflammation test.

Recently it was demonstrated that phycocyanin, a biliprotein isolated from microalgae Spirulina, exerts anti-inflammatory activity in several animal models of inflammation. In this report, the effects of phycocyamin on prostaglandin E2 (PGE2) concentrations and phospholipase A2 (PLA2) activity were determined in arachidonic acid (AA) and 12-O-tetradecanoyl phorbol 13-acetate (TPA)-induced mouse ear oedema, respectively. Phycocyanin (50-200 mg/kg p.o.) inhibited in a dose-dependent manner PGE2 levels in mouse ear treated with AA. Also, phycocyanin (100-400 mg/kg p.o.) moderately reduced PLA2 activity in TPA-induced mouse ear inflammation test. In this model triamcinolone (10 mg/kg p.o.) used as reference drug exerted a remarkable inhibitory effect on PLA2 activity. These results provide the first evidence that the anti-inflammatory effects of phycocyanin may result, at least partially, from inhibition of PGE2 production and a moderate inhibition of PLA2 activity.

Evaluation of some Samoan and Peruvian medicinal plants by prostaglandin biosynthesis and rat ear oedema assays.

In our ongoing program to find new anti-inflammatory compounds, 58 extracts from 46 different medicinal plant species, used in treatment of inflammatory disorders-38 plants from the traditional medicine of Western Samoa and eight originating from the indigenous medicine of the Shipibo-Conibo tribe of Peruvian Amazonia-ere evaluated. The ability of all extracts to inhibit cyclooxygenase-1 catalysed prostaglandin biosynthesis in vitro was examined. Of the plant species tested 14 showed moderate to strong inhibition; including 11 Samoan and three Peruvian species. Further, 12 Samoan and all eight Peruvian species were investigated on their inhibitory activity of ethyl phenylpropiolate induced rat ear oedema in vivo. Significant activity was shown by 10 of the Samoan and by all eight Peruvian species. An additional evaluation of the most active species was provided through a compilation of existing literature documenting traditional medicinal uses, pharmacological activity and chemical constituents. Several known cyclooxygenase-1 inhibitors were reported to which the observed pharmacological activity can be attributed at least partly. The combination of chemical and pharmacological literature data and our experimental data may help to explain the anti-inflammatory use of these species in indigenous medicine.

Diffusion versus iontophoresis in the transport of gentamicin in the burned rabbit ear model.

This experimental study demonstrates that iontophoresis can be used to transport biologically active gentamicin into auricular cartilage. Fifty female New Zealand White rabbits were divided into three groups: live controls with unburned ears, live animals with burn-injured ears (deep or partial-thickness), and euthanized animals with unburned ears. Each group was then divided into subgroups that received gentamicin transported by iontophoresis or diffusion. In ears subjected to iontophoresis, adequate gentamicin activity could be demonstrated only in the cartilage of euthanized animals with unburned ears and live animals with full-thickness burned ears. Diffusion did not transport a detectable level of gentamicin in any instance. Diffusion is ineffective in the transport of gentamicin and should not be used as a primary method of treatment for ear chondritis. Iontophoresis did not transport active gentamicin across partial-thickness burns, presumably because the migrating antibiotic molecules were dispersed throughout the body by the circulation. However, if the circulation is destroyed, as in a full-thickness burn, iontophoresis can move the biologically active gentamicin into the cartilage.