RESUMO
The main barrier of the human skin is the stratum corneum (SC). Its properties (also depending on the health and age of the individual) and its influence on improved penetration of active ingredients into the skin are the subject of many research projects. Since the availability of human skin, as the ideal model membrane, is limited, the aim of this study was to find a suitable alternative model membrane from the animal kingdom. The alternative model membrane should be used in subsequent permeation experiments with the Teflon diffusion cell instead of human SC. Previous studies have already investigated the permeation properties of pig, snake, and human skin, but not in a Teflon diffusion cell using ATR. Therefore, it first had to be proven that comparable results can be achieved with animal membranes even under these measurement conditions. This is the precondition for meaningful future permeation experiments with potential enhancers. For this purpose, permeation experiments on various model membranes (human isolated SC, sunburned SC, pig isolated SC, and shed snake skin) by means of FTIR-ATR in a Teflon diffusion cell containing the acceptor and the donor compartment as well as the model membrane were conducted and concentration-time courses of the model peptide PKEK determined. These concentration-time courses were used to calculate and compare the pharmacokinetic parameters (permeation coefficients, lag time, and flux). The starting point was a 10% PKEK solution in D2O. It turned out that snake skin is the appropriate alternative model membrane for this type of permeation test.
Assuntos
Oligopeptídeos/administração & dosagem , Absorção Cutânea , Pele/metabolismo , Serpentes , Administração Cutânea , Animais , Coloides , Difusão , Humanos , Espectroscopia de Infravermelho com Transformada de Fourier , Queimadura Solar , SuínosRESUMO
The aim of this work is to develop a curcumin nanoemulsion for transdermal delivery. The incorporation of curcumin inside a nanoglobul should improve curcumin stability and permeability. A nanoemulsion was prepared by the self-nanoemulsification method, using an oil phase of glyceryl monooleate, Cremophor RH40 and polyethylene glycol 400. Evaluation of the nanoemulsion included analysis of particle size, polydispersity index, zeta potential, physical stability, Raman spectrum and morphology. In addition, the physical performance of the nanoemulsion in Viscolam AT 100P gel was studied. A modified vertical diffusion cell and shed snake skin of Python reticulatus were used to study the in vitro permeation of curcumin. A spontaneously formed stable nanoemulsion has a loading capacity of 350 mg curcumin/10 g of oil phase. The mean droplet diameter, polydispersity index and zeta potential of optimized nanoemulsion were 85.0 ± 1.5 nm, 0.18 ± 0.0 and -5.9 ± 0.3 mV, respectively. Curcumin in a nanoemulsion was more stable than unencapsulated curcumin. Furthermore, nanoemulsification significantly improved the permeation flux of curcumin from the hydrophilic matrix gel; the release kinetic of curcumin changed from zero order to a Higuchi release profile. Overall, the developed nanoemulsion system not only improved curcumin permeability but also protected the curcumin from chemical degradation.
Assuntos
Anti-Inflamatórios não Esteroides/metabolismo , Curcumina/metabolismo , Portadores de Fármacos/metabolismo , Modelos Biológicos , Nanoestruturas/química , Absorção Cutânea , Pele/metabolismo , Administração Cutânea , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/análise , Anti-Inflamatórios não Esteroides/química , Boidae , Curcumina/administração & dosagem , Curcumina/análise , Curcumina/química , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/análise , Portadores de Fármacos/química , Composição de Medicamentos , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Emulsões , Excipientes/química , Glicerídeos/química , Nanoestruturas/ultraestrutura , Tamanho da Partícula , Polietilenoglicóis/química , Propriedades de SuperfícieRESUMO
The role of lipids in controlling water exchange is fundamentally a matter of molecular organization. In the present study we have observed that in snake molt the water permeability drastically varies among species living in different climates and habitats. The analysis of molts from four snake species: tiger snake, Notechis scutatus, gabon viper, Bitis gabonica, rattle snake, Crotalus atrox, and grass snake, Natrix natrix, revealed correlations between the molecular composition and the structural organization of the lipid-rich mesos layer with control in water exchange as a function of temperature. It was discovered, merging data from micro-diffraction and micro-spectroscopy with those from thermal, NMR and chromatographic analyses, that this control is generated from a sophisticated structural organization that changes size and phase distribution of crystalline domains of specific lipid molecules as a function of temperature. Thus, the results of this research on four snake species suggest that in snake skins different structured lipid layers have evolved and adapted to different climates. Moreover, these lipid structures can protect, "safety", the snakes from water lost even at temperatures higher than those of their usual habitat.
Assuntos
Lipídeos/fisiologia , Muda/fisiologia , Pele/metabolismo , Pele/fisiopatologia , Serpentes/metabolismo , Serpentes/fisiologia , Água/metabolismo , Animais , Ecossistema , Permeabilidade , TemperaturaRESUMO
Introduction: Shed snake skin (SSS) is commonly used empirically in ethnomedicine to treat psoriasis, acne, warts, eczema, scabies, open wounds, hemorrhoids, and glaucoma. Although a few studies exist, SSS extracts' in vitro immunological effects have yet to be well described. Therefore, we aimed to investigate the immunomodulatory effects of SSS extract on murine lymphocytes and T cells. Methods: Hexane, methanol, and chloroform extractions were conducted in collected SSS samples. Protein concentrations in the SSS extract were measured. The cytotoxic and anticancer activities were measured using L929 Fibroblast and SK MEL 30 Cell Lines via MTT assay as described in TS EN ISO 10993-5. Immunomodulatory activities of SSS extract on total lymphocytes or enriched CD4+ T cell cultures, their cell-specific pro-inflammatory cytokines (IL-6, IL-1ß. IL-12p40, IL-23p19, TNF-α, IL-17A, IFN-γ, IL-10, TGFß1) levels were measured via FACS ARIA III analysis and related gene expression with Real-Time Quantitative Polymerase Chain Reaction (Rt-qPCR). Results: Hexane, methanol, and chloroform-extracted SSS were tested on SK-MEL-30 cells via MTT and revealed a superior anti-proliferative effect for hexane extract of SSS at low concentrations. SSS treatment of murine lymphocytes augmented Tnf-α and IFN-γ levels produced by CD3+ T cells when lymphocytes were activated with anti-CD3/CD28 or LPS stimulation. This effect required the presence of non-T cells, possibly antigen-presenting cells, and was not observed on purified CD4+ T cells. Additionally, SSS significantly blocked suppressive cytokine Tgfb gene expression (but not Il10) without altering in vitro Treg generation/or expansion. Discussion: This is the first in vitro study investigating SSS's anti-tumor and immunomodulatory effects. Our data provide evidence for SSS's anti-proliferative activity on SK-MEL-30 cells and its pro-inflammatory role on murine lymphocytes, which warrants further investigation of the potential use of SSS extract with in vivo disease models.
RESUMO
Advancements in developing antipathogenic interfaces are critical in mitigating the risk of infection spread amid the practical limitations of hygienic control in crowded and resource-limited settings. Such requirements are also extremely compelling in busy patient care centers including intensive care units where the statutory maintenance of environmental standards often appears to be impractical because of the overflooded patient loads. While advances in surface engineering have emerged with great promises to cater these needs, the underlying technological complexities appear to be prohibitive against practicable applications amid constrained technological resources. Here, we harnessed the role of unique topographical features of the skin of Ptyas mucosa (oriental rat snake), a commonly found snake species in south and southeast Asia, in terms of exhibiting supreme antifouling properties via natural inheritance, leading to pathogenic resistance. Our characterization studies unveiled that unlike the previously reported vertical pillars, hairs, and needles, arrays of horizontal denticulation, offering favorable topographical characteristics of structured roughness and hierarchical features, emerged to be responsible for exhibiting the desired functionalities. We subsequently adapted these structures with certain simplifications by biomimicking artificially engineered topologies on a polydimethylsiloxane (PDMS) surface. The resulting surfaces were proven to offer dual antimicrobial mechanisms such as resistances to adhesion or colonization of different bacteria (Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus mutans) and facilitation for cell wall deformation and programmed cell death as evidenced by an abundance of oxidative stresses. These results opened up strategies of producing biomimetic surface textures and their effective implementation against pathogenic invasion in a plethora of applications ranging from medical implants to marine propulsion.
Assuntos
Pseudomonas aeruginosa , Staphylococcus aureus , Animais , Escherichia coli , Klebsiella pneumoniae , SerpentesRESUMO
The understanding of the tribological behavior of natural structures has been used as inspiration to design and optimize surfaces for diverse applications in engineering. In the present work, morphological, microstructural, mechanical and tribological characterization of the shed skin of two snake species, namely Boa Red Tail and Python Regius was carried out. Atomic Force Microscopy (AFM) and Scanning Electron Microscopy (SEM) analyses showed the existence of deterministic patterns, i.e., ordered arrays of geometrical features at the surface, while Transmission Electron Microscopy (TEM) allowed studying the internal structure and chemical composition of the skin sheds. Nanoindentation measurements showed significant variations in hardness and elastic modulus from the surface to the inner layers of the skin, and pin-on-disc tests revealed anisotropic behavior of the friction coefficient (COF) as a function of the sliding direction against balsa wood in dry conditions. Correlations between the friction data, nano-indentation mechanical properties and subsurface skin structure were established for both species taking into account the ways in which the skins' deterministic patterns influence the tribological performance.
Assuntos
Boidae , Animais , Módulo de Elasticidade , Fricção , Dureza , Microscopia de Força Atômica , Propriedades de SuperfícieRESUMO
BACKGROUND: Tabernaemontana catharinensis, popularly known as snake skin, has been empirically used as an anti-inflammatory to treat cutaneous skin disorders. However, no study proves its effectiveness as a topical anti-inflammatory. STUDY DESIGN: We investigated the topical anti-inflammatory effect of T.catharinensis leaves crude extract (TcE) in irritant contact dermatitis models in mice and its preliminary toxicity profile. METHODS: The topical anti-inflammatory effect was evaluated by ear thickness measurement, inflammatory cell infiltration (MPO activity measurement and histological procedure) and cytokines levels. TcE qualitative phytochemical analysis was performed by UHPLC-ESI-HRMS and the TcE effect (therapeutic dose; 10⯵g/ear) on preliminary toxicological parameters was also evaluated (on the 14°â¯day of experiment). RESULTS: TcE (10⯵g/ear) prevented the development of ear edema induced by cinnamaldehyde, capsaicin, arachidonic acid, phenol, and croton oil with maximum inhibition of 100% to cinnamaldehyde, arachidonic acid, phenol, and croton oil and 75⯱â¯6% to capsaicin. Besides, the TcE (10⯵g/ear) also prevented the increase of MPO activity by 96⯱â¯2%, 48⯱â¯7%, 100%, 87⯱â¯8%, and 93⯱â¯4%, respectively, to the same irritant agents. The positive controls also prevented both ear edema and the increased of MPO activity by 100% and 42⯱â¯8% (HC-030031), 54⯱â¯6% and 80⯱â¯4% (SB-366791), 100% and 54⯱â¯5% (indomethacin), 100% and 80⯱â¯4% (dexamethasone in skin inflammation model induced by phenol) and 100% and 97⯱â¯3% (dexamethasone in inflammation model induced by croton oil), respectively. TcE also prevented the inflammatory cells infiltration and the increase of MIP-2, IL-1ß and TNF-α levels irritant agents-induced. TcE topical anti-inflammatory effect may be attributed to the combined effect of indole alkaloids, terpenes, and phenolic compounds found in the extract and identified by dereplication method. The TcE' therapeutic dose proved to be safe in preliminary toxicological tests. CONCLUSION: Our results suggest that TcE could be an interesting strategy for the treatment of inflammatory diseases.
Assuntos
Anti-Inflamatórios/uso terapêutico , Edema/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Tabernaemontana , Animais , Citocinas/imunologia , Edema/induzido quimicamente , Edema/imunologia , Irritantes , Masculino , Camundongos , Fitoterapia , Folhas de PlantaRESUMO
Snakes are limbless tetrapods highly specialized for sliding locomotion on various substrates. Their skin is constantly exposed to high friction forces, which promotes abrasion. Snake skin has material and surface specializations, presumably optimized for friction and abrasion resistance. We found that different snake species living in different habitats have different abrasion patterns and hypothesized that this correlates with specific epidermal architecture and surface topography. To test this hypothesis artificial scratches, under controlled load conditions, were created on the ventral skin material (epidermis) of four snake species adapted to different habitats: Lampropeltis getula californiae (stony and sandy soil substrates), Epicrates cenchria cenchria (trees, soil and water), Morelia viridis (trees), and Gongylophis colubrinus (burrowing in sand). Abrasion appearance on the skin surface was examined using scanning electron microscopy and white light interferometry. The material failure was different between the species, which we attribute to differences in the epidermis' response to the same abrasive challenge. We also discuss abrasion resistance mechanisms and the correlation with the different ultrastructure and surface microstructure.
Assuntos
Boidae/fisiologia , Epiderme/lesões , Epiderme/fisiologia , Animais , Ecossistema , Feminino , Especificidade da EspécieRESUMO
The microstructure investigated in this study was inspired by the anisotropic microornamentation of scales from the ventral body side of the California King Snake (Lampropeltis getula californiae). Frictional properties of snake-inspired microstructured polymer surface (SIMPS) made of epoxy resin were characterised in contact with a smooth glass ball by a microtribometer in two perpendicular directions. The SIMPS exhibited a considerable frictional anisotropy: Frictional coefficients measured along the microstructure were about 33% lower than those measured in the opposite direction. Frictional coefficients were compared to those obtained on other types of surface microstructure: (i) smooth ones, (ii) rough ones, and (iii) ones with periodic groove-like microstructures of different dimensions. The results demonstrate the existence of a common pattern of interaction between two general effects that influence friction: (1) molecular interaction depending on real contact area and (2) the mechanical interlocking of both contacting surfaces. The strongest reduction of the frictional coefficient, compared to the smooth reference surface, was observed at a medium range of surface structure dimensions suggesting a trade-off between these two effects.
RESUMO
A rutina é empregada como antioxidante e na prevenção da fragilidade capilar. Estudos de penetração in vitro através da pele humana seria a situação ideal, entretanto, há dificuldades de sua obtenção e manutenção de sua viabilidade. Entre os demais modelos de membrana, a muda de pele de cobra se apresenta como estrato córneo puro, fornecendo barreira similar ao humano e é obtida sem a morte do animal. Os objetivos desta pesquisa foram desenvolver e avaliar a estabilidade de uma emulsão cosmética contendo rutina e, como promotor de penetração cutânea, o propilenoglicol; e avaliar a penetração e a retenção cutânea in vitro da referida substância ativa da formulação, empregando um modelo de biomembrana alternativo. A emulsão foi desenvolvida com rutina e propilenoglicol, ambos a 5,0 por cento p/p. Quantificou-se a rutina das emulsões por espectrofotometria a 361,0 nm, método previamente validado. A penetração e retenção cutânea in vitro foram realizadas em células de difusão vertical com muda de pele de cobra de Crotalus durissus, como modelo de biomembrana alternativo, e água destilada e álcool etílico absoluto 99,5 por cento (1:1), como fluido receptor. O experimento foi conduzido em um período de seis horas, a 37,0 ± 0,5 ºC e agitação constante de 300 rpm. Empregou-se o método espectrofotométrico validado a 410,0 nm para a quantificação da rutina após penetração e retenção cutânea. A emulsão não promoveu a penetração cutânea da rutina através da muda de pele de C. durissus, retendo 0,931 ± 0,0391 mg de rutina/mg de muda de pele de cobra. Nas condições de armazenamento a 25,0 ± 2,0 ºC; 5,0 ± 0,5 ºC e 45,0 ± 0,5 ºC, a emulsão apresentou-se quimicamente estável durante 30 dias. De acordo com os resultados, a emulsão não favoreceu a penetração cutânea da rutina, mas apenas sua retenção no estrato córneo de C. durissus, condição considerada estável no período de 30 dias.
Rutin is employed as antioxidant and to prevent the capillary fragility and, when incorporated in cosmetic emulsions, it must target the action site. In vitro cutaneous penetration studies through human skin is the ideal situation, however, there are difficulties to obtain and to maintain this tissue viability. Among the membrane models, shed snake skin presents itself as pure stratum corneum, providing barrier function similar to human and it is obtained without the animal sacrifice. The objectives of this research were the development and stability evaluation of a cosmetic emulsion containing rutin and propylene glycol (penetration enhancer) and the evaluation of rutin in vitro cutaneous penetration and retention from the emulsion, employing an alternative model biomembrane. Emulsion was developed with rutin and propylene glycol, both at 5.0 percent w/w. Active substance presented on the formulation was quantified by a validated spectrophotometric method at 361.0 nm. Rutin cutaneous penetration and retention was performed in vertical diffusion cells with shed snake skin of Crotalus durissus, as alternative model biomembrane, and distilled water and ethanol 99.5 percent (1:1), as receptor fluid. The experiment was conducted for six hours, at 37.0 ± 0.5 ºC with constant stirring of 300 rpm. Spectrophotometry at 410.0 nm, previously validated, determined the active substance after cutaneous penetration/retention. Emulsion did not promote rutin cutaneous penetration through C. durissus skin, retaining 0.931 ± 0.0391 mg rutin/mg shed snake skin. The referred formulation was chemically stable for 30 days after stored at 25.0 ± 2.0 ºC, 5.0 ± 0.5 ºC and 45.0 ± 0.5 ºC. In conclusion, it has not been verified the active cutaneous penetration through the model biomembrane, but only its retention on the Crotalus durissus stratum corneum, condition considered stable for 30 days.