Skin adhesive hydrogels for the topical delivery of active agents

This thesis illustrates the development of tailor-made, partially hydrated skin adhesive hydrogels as a vehicle for the topical delivery of moisturising agents. Maintaining an optimum hydration level of the stratum corneum ensures that the barrier properties of the skin are preserved. An unsaturated ionic monomer 2-acrylamido-2-methylpropanesulfonic acid sodium salt, glycerol, water, a photoinitiator Irgacure 184 and crosslinker Ebacryl II facilitated the production of monophasic sheet skin adhesives using photopolymerisation. The exploration and modification of the hydrogel components coupled with their influence on the adhesive and dynamic mechanical behaviour led to the development of novel monophasic and biphasic hydrogels. Biphasic pregels comprising of a hydrophobic monomer (epoxidised soybean oil acrylate, lauryl acrylate or stearyl acrylate) micellised with a non ionic surfactant Tween 60 allowed a homogeneous distribution throughout a predominantly hydrophilic phase (2-acrylamido-2-methylpropanesulfonic acid sodium salt, 4-acryloylmorpholine, glycerol and water). Further development of biphasic hydrogel technology led to the incorporation of preformed commercial O/W emulsions (Acronal, Flexbond 150, DM137 or Texicryl 13056WB) allowing the hydrophobic component to be added without prior stabilisation. The topical release of moisturising agents 2-pyrrolidone-5-carboxylic acid, lactobionic acid and d-calcium pantothenate results in the deposition onto the skin by an initial burst mechanism. The hydration level of the stratum corneum was measured using a Comeometer CM 825, Skin Reader MY810 or FT-ATR. The use of hydrophilic actives in conjunction with lipophilic agents for example Vitamin E or Jojoba oil provided an occlusive barrier, which reduced the rate of transepidermal water loss. The partition coefficients of the release agents provided invaluable information which enabled the appropriate gel technology to be selected. In summary the synthetic studies led to the understanding and generation of transferable technology. This enabled the synthesis of novel vehicles allowing an array of actives with a range of solubilities to be incorporated.

Desenvolvimento e caracterização de nanoemulsões cosméticas contendo extrato de Opuntia ficus-indica L. Mill e avaliação in vivo da eficácia hidratante

Nanoemulsions are emulsified systems, characterized for reduced droplet size (50- 500nm), which the main characteristic are kinect stability and thermodynamic instability. These are promising systems on cosmetic area due to their droplet size that provide different advantages when compared to conventional systems, among others, larger surface area and better permeability. The Opuntia ficus-indica (L.) Mill is a plant cultivated on Caatinga Brazilian biome, which has great socioeconomic importance to region. This plant shows carbohydrates utilized for cosmetic industry as moisturizing active in their chemical composition. The aim of study was to develop, characterize, evaluate stability and moisturizing efficacy of cosmetic nanoemulsions added to Opuntia ficus-indica (L.) Mill extract. Nanoemulsions preparation was made using a low energy method. Different nanoemulsions were formulated varying the ratio of oil, water and surfactant phases beyond xanthan gum (0.5% e 1%) and Opuntia ficus-indica (L.) Mill hydroglycolic extract addition on 1% and 3%. Obtained nanoemulsions were submitted to preliminary and accelerated stability tests. The evaluated parameters monitored were: macroscopic aspect, pH value, droplet size, zeta potential and polydispersion index, during 60 days on different temperatures. Stable formulations were submitted to moisturizing efficacy assessment by capacitance and transepidermal water loss methodologies during 5 hours. Stable samples were white and showed homogeneous and fluid aspect, pH value was inside ideal range (4,5-6,0) to topical application and droplet size under 200nm characterizing these system as nanoemulsions. Developed nanoemulsions did not decrease transepidermal water loss, however increased the water content on stratum corneum, highlighting the nanoemulsions containing 0.5% of xanthan gum and 1% of hydroglycolic extract. This work presents cosmetic moisturizing nanoemulsions composed to vegetal raw material from Brazilian Caatinga with potential to be used on cosmetic area.

Development of a dissolvable microneedle influenza vaccine patch

Delivery of large molecular weight biological molecules to the epidermis and dermis is constrained by the tough outer layer of the epidermis, the stratum corneum (sc). Microneedle technologies attempt to overcome this physical barrier using sharp micron-size projections to penetrate the sc. Dissolvable microneedles (DMN), are a particular microneedle design whereby the needle structure is composed of a soluble matrix that upon application to the skin, dissolves releasing the vaccine load into skin. This thesis examines (1) the formulation and processing considerations around DMN fabrication, (2) the immunogenicity of DMN containing trivalent influenza vaccine (TIV) in pre-clinical mouse and pig models and (3) the thermostability of these DMN formulations during storage. The results demonstrate the importance of formulation for microneedle formation and mechanical strength. Trehalose and polyvinylalcohol based formulations produced optimal microneedle structures and were amenable to piezoelectric dispensing; allowing for precise multi-layered DMN to be fabricated. The effect of drying conditions was assessed and found to be critical for DMN mechanical strength and skin penetration. The antibody responses to TIV generated by DMN-mediated vaccination were comparable or greater to those induced by immunization with a commercial TIV via the IM route in mice. DMN mediated immunisation resulted in a significantly broader humoral response to heterotypic influenza viruses compared to IM delivery. Stored at 40°C, a licensed seasonal influenza vaccine incorporated into DMN array was thermostable for at least 6 month as determined by Single Radial Immunodiffusion and immunogenicity in mice. The thesis advances the field of DMN influenza vaccination by elucidating important processing and formulation considerations in the fabrication of highly reproducible DMN. It also demonstrated that DMN can induce broader, larger humoral responses than conventional IM administration while demonstrating enhanced accelerated stability. Crucially, this works advances an automated fabrication system that will allow for clinical translation of DMN.

Contact dermatitis from shoes: an update

Contact dermatitis is a common inflammatory skin condition characterized by erythematous and pruritic skin lesions that occur after contact with a foreign substance. There are two forms of contact dermatitis: irritant and allergic. Irritant contact dermatitis is caused by the non–immune-modulated irritation of the skin by a substance, leading to skin changes. Allergic contact dermatitis is a delayed hypersensitivity reaction in which a foreign substance comes into contact with the skin; skin changes occur after reexposure to the substance. A medical condition referred to as “shoe dermatitis” is a form of contact dermatitis caused by the contact of the foot with parts of the shoe due to these materials. Shoe dermatitis is a diagnostic and therapeutic challenge and is a common type of contact dermatitis. It is imperative the foot and ankle physician become familiar with recognizing signs and symptoms of shoe dermatitis so that their patients can be accurately diagnosis and appropriately treated to avoid secondary infections and disability. This review will first present causative factors for the etiology of shoe contact dermatitis supported by clinical-based evidence as found in the medical literature. Secondly, a description of the signs and symptoms of shoe contact dermatitis will be presented in a narrative fashion. Finally, both treatment options and preventative measures to avoid shoe.

Avaliação da eficácia in vivo da L-arginina na elasticidade da pele

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ciências da Saúde, Programa de Pós-Graduação em Ciências da Saúde, 2012.

Protective effect of (-)-epigallocatechin gallate on ultraviolet b-induced skin damage in hairless mice

Purpose: To investigate the protective effect of green tea (-)-epigallocatechin gallate (EGCg) on ultraviolet B (UV-B)-induced skin damages in hairless mice in order to develop a natural sunscreen compound for use in skin care products and cosmetics. Methods: EGCg was dissolved in acetone at concentrations of 1.0, 10.0 and 50.0 mg/mL, and topically applied to the skin of hairless mice at doses of 0.2 mL/cm2, with acetone as control. The mice were then irradiatd m2 UV-B for 30 min daily. EGCg treatment and UV-B irradiation were carried out daily for 28 consecutive days. The mice were then sacrificed and their dorsal skin examined by transmission electron microscopy (TEM) on the 28th day. Results: UV-B irradiation induced severe macroscopic skin damage including chapping, cracking and abnormal desquamation in the treated hairless mice. EGCg showed dose-dependent protective effects against UV-B induced damage on the skin. Treatments with 10.0 and 50.0 mg/mL EGCg alleviated UVB-induced skin damage by suppressing both keratinocyte apoptosis and mitochondrial dysfunction, along with inhibiting the production of melanin pigment. Conclusion: Topical application of green tea EGCg shows dose-dependent protective effect against UV-B-induced damage on hairless mouse skin. Thus, the plant compound can potentially be used as an alternative agent for photoprotection against UV-B exposure.

Estudio molecular en dos hermanas afectadas por ictiosis congénita autosómica recesiva : descripción de una nueva mutación causal en TGM1

Se describe la variante homocigota c.320-2A>G de TGM1 en dos hermanas con ictiosis congénita autosómica recesiva. El clonaje de los transcritos generados por esta variante permitió identificar tres mecanismos moleculares de splicing alternativos.

Plasma hormonal profiles and dendritic spine density and morphology in the hippocampal CA1 stratum radiatum, evidenced by light microscopy, of virgin and postpartum female rats

Successful reproduction requires that changes in plasma follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), oxytocin (OT), estrogen (E-2) and progesterone (P-4) occur together with the display of maternal behaviors. Ovarian steroids and environmental stimuli can affect the dendritic spines in the rat hippocampus. Here, studying Wistar rats, it is described: (a) the sequential and concomitant changes in the hormonal profile of females at postpartum days (PP) 4, 8, 12, 16, 20 and 24, comparing to estrous cycle referential values; (b) the dendritic spine density in the stratum radiatum of CA1 (CA1-SR) Golgi-impregnated neurons in virgin females across the estrous cycle and in multiparous age-matched ones; and (c) the proportion of different types of spines in the CAI-SR of virgin and postpartum females, both in diestrus. Plasma levels of gonadotrophins and ovarian hormones remained low along PP while LH increased and PRL decreased near the end of the lactating period. The lowest dendritic spine density was found in virgin females in estrus when compared to diestrus and proestrus phases or to postpartum females in diestrus (p < 0.03). Other comparisons among groups were not statistically significant (p > 0.4). There were no differences in the proportions of the different spine types in nulliparous and postpartum females (p > 0.2). Results suggest that medium layer CA1-SR spines undergo rapid modifications in Wistar females across the estrous cycle (not quite comparable to Sprague-Dawley data or to hormonal substitutive therapy following ovariectomy), but persistent effects of motherhood on dendritic spine density and morphology were not found in this area. (c) 2008 Elsevier Ireland Ltd. All rights reserved.

Diameter structure of stratum and ecological groups of a Semideciduous Forest area in Dionísio-MG

The objective of this study was to obtain homogeneous groups of species and information on their density, dominance and volume, in terms of ecological group and diameter structure of an area of Submontane Semideciduous forest (Mata do Mumbaça) in Dionísio, MG. This work was conducted with data of the diameter distribution per species from floristic and phytosociological (Mata do Mumbaça) survey of 120 plots with 10 x 10 m each one. The 120 plots were contiguous and corresponding to a total sample area of 12,000 m² distributed over the topographic units (Low Ramp, Lower Slope, Upper Slope and Hill Top). The topographic units Low Ramp, Lower Slope and Upper Slope were in the middle stage of succession as they presented incipient stratification into two strata (canopy and understory) i.e. canopy ranging from 5 to 12 m high. However, the stratum Hill Top was classified as intermediate/advanced succession because it had a total height equal to or greater than 12 m. The distribution of individual trees of the four strata on diameter classes showed a typical J-inverted pattern that is, high concentration of individuals in smaller diameter classes and a sharp reduction towards the larger classes. In relation to absolute dominance and total volume of species, the ecological group that stood out in the four strata (Low Ramp, Lower Slope, Upper Slope and Hill Top) was the initial secondary, which were in the intermediate stage of secondary, rapidly developing into the mature phase.

NATURAL REGENERATION STRATUM AS AN INDICATOR OF RESTORATION IN AREA OF ENVIRONMENTAL COMPENSATION FOR MINING LIMESTONE, MUNICIPALITY OF BARROSO, MG, BRAZIL

ABSTRACT This study was conducted in a forest under restoration process, which belongs to the company Holcim Brasil S/A, in the municipality of Barroso, state of Minas Gerais (21º00'to 22º00'S and 43º00' to 44º00'W), where 40 plots (2 x 2 m) were set, spaced at 10 m, forming eight strata parallel to the watercourse present in the area. Floristic composition and natural regeneration stratum were characterized, and the formed strata allowed evaluating whether the riparian vegetation and watercourse influence on the local regeneration. It was found 162 individuals of 13 families, 18 genera and 22 species, and 10,125 individuals/ha were estimated. Successional classes from pioneer and early secondary and zoochory dispersion syndrome prevailed among species and individuals. The watercourse and riparian vegetation did not exercise significant influence (p> 0.05) on the number of species and regenerating individuals among the different strata of the forest. The diversity index of Shannon-Wiener (H') and equability of Pielou (J') were 2.691 and 0.870, respectively. The species Psidium guajava and Myrtaceae families presented the highest VI (value of importance). Natural regeneration analysis showed the low floristic diversity in the area, suggesting that corrective management actions should be adopted.

Maintenance of the corneal epithelium is carried out by germinative cells of its basal stratum and not by presumed stem cells of the limbus

The purpose of this investigation was to analyze the proliferative behavior of rabbit corneal epithelium and establish if any particular region was preferentially involved in epithelial maintenance. [3H]-thymidine was injected intravitreally into both normal eyes and eyes with partially scraped corneal epithelium. Semithin sections of the anterior segment were evaluated by quantitative autoradiography. Segments with active replication (on) and those with no cell division (off) were intermingled in all regions of the tissue, suggesting that the renewal of the epithelial surface of the cornea followed an on/off alternating pattern. In the limbus, heavy labeling of the outermost layers was observed, coupled with a few or no labeled nuclei in the basal stratum. This suggests that this region is a site of rapid cell differentiation and does not contain many slow-cycling cells. The conspicuous and protracted labeling of the basal layer of the corneal epithelium suggests that its cells undergo repeated cycles of replication before being sent to the suprabasal strata. This replication model is prone to generate label-retaining cells. Thus, if these are adult stem cells, one must conclude that they reside in the corneal basal layer and not the limbal basal layer. One may also infer that the basal cells of the cornea and not of the limbus are the ones with the main burden of renewing the corneal epithelium. No particular role in this process could be assigned to the cells of the basal layer of the limbal epithelium.

Modelling 2,4-dichlorophenol bioavailability and bioaccumulation by the freshwater fingernail clam Sphaerium corneum using artificial particles and humic acids

The complex and variable composition of natural sediments makes it very difficult to predict the bioavailability and bioaccumulation of sediment-bound contaminants. Several approaches have been proposed to overcome this problem, including an experimental model using artificial particles with or without humic acids as a source of organic matter. For this work, we have applied this experimental model, and also a sample of a natural sediment, to investigate the uptake and bioaccumulation of 2,4-dichlorophenol (2,4-DCP) by Sphaerium corneum. Additionally, the particle-water partition coefficients (K-d) were calculated. The results showed that the bioaccumulation of 2,4-DCP by clams did not depend solely on the levels of chemical dissolved, but also on the amount sorbed onto the particles and the characteristics and the strength of that binding. This study confirms the value of using artificial particles as a suitable experimental model for assessing the fate of sediment-bound contaminants. (c) 2006 Elsevier Ltd. All rights reserved.

Floristic composition of the tree stratum of a semideciduous mesophytic forest in the municipality of Jahu, SP, Brazil

A floristic study was carried out in an area of semideciduous mesophytic forest, lied in the municipality of Jahu, São Paulo State, Brazil. Due to the fact of the forest be crossed by three small rivers, for the sampling process, the forest was divided in two sampling areas: Area I (under the influence of the rivers) and Area II (without this influence). Differences in the abundance of some species between the two sampling areas were observed.

Phytosociological study on the tree stratum of a semideciduous mesophytic forest in the municipality of Jau, SP, Brazil

Through the point-centered quarter method, a phytosociological study was carried out in an area of semideciduous mesophytic forest with 190 ha, in the municipality of Jahu,SP. Due to the fact of the forest be crossed by three small rivers, for the sampling process, the Total Area was divided in two sampling areas: the Area I (under the influence of the rivers) and the Area II (without this influence). Some differences between the two regions were observed.

Floristic composition of the tree stratum of a semideciduous mesophytic forest in the municipality of Botucatu, São Paulo state

The floristic composition was studied in an area of semideciduous mesophytic forest, with 120 ha, in the municipality of Botucatu, SP, Brazil. This forest lies on the ascent of the Cuesta of Botucatu. Due to the heterogeneity of the relief, for the sampling process, the forest was divided in three regions: the upper one (comprising the strip of forest on the plateau), middle on (the forestal area lied on the ascent properly said) and the lower one (the strip of forest on the plain). The tree sampling regions showed some differences among them and this must be connected to the differences in the soil.