Transepidermal Transport Research Articles

Cellular response of keratinocytes to the entry and accumulation of nanoplastic particles

Plastic accumulation in the environment is rapidly increasing, and nanoplastics (NP), byproducts of environmental weathering of bulk plastic waste, pose a significant public health risk. Particles may enter the human body through many possible routes such as ingestion, inhalation, and skin absorption. However, studies on NP penetration and accumulation in human skin are limited. Loss or reduction of the keratinized skin barrier may enhance the skin penetration of NPs. The present study investigated the entry of NPs into a human skin system modeling skin with compromised barrier functions and cellular responses to the intracellular accumulations of NPs. Two in vitro models were employed to simulate human skin lacking keratinized barriers. The first model was an ex vivo human skin culture with the keratinized dermal layer (stratum corneum) removed. The second model was a 3D keratinocyte/dermal fibroblast cell co-culture model with stratified keratinocytes on the top and a monolayer of skin fibroblast cells co-cultured at the bottom. The penetration and accumulation of the NPs in different cell types were observed using fluorescent microscopy, confocal microscopy, and cryogenic electron microscopy (cryo-EM). The cellular responses of keratinocytes and dermal fibroblast cells to stress induced by NPs stress were measured. The genetic regulatory pathway of keratinocytes to the intracellular NPs was identified using transcript analyses and KEGG pathway analysis. The cellular uptake of NPs by skin cells was confirmed by imaging analyses. Transepidermal transport and penetration of NPs through the skin epidermis were observed. According to the gene expression and pathway analyses, an IL-17 signaling pathway was identified as the trigger for cellular responses to internal NP accumulation in the keratinocytes. The transepidermal NPs were also found in co-cultured dermal fibroblast cells and resulted in a large-scale transition from fibroblast cells to myofibroblast cells with enhanced production of α-smooth muscle actin and pro-Collagen Ia. The upregulation of inflammatory factors and cell activation may result in skin inflammation and ultimately trigger immune responses.

ИССЛЕДОВАНИЕ ВЛИЯНИЯ ЭТАНОЛА НА ТРАНСЭПИДЕРМАЛЬНЫЙ ТРАНСПОРТ ИНДОЦИАНИНОВОГО ЗЕЛЁНОГО С ПОМОЩЬЮ СПЕКТРОСКОПИИ ОБРАТНОГО РАССЕЯНИЯ

ИССЛЕДОВАНИЕ ВЛИЯНИЯ ЭТАНОЛА НА ТРАНСЭПИДЕРМАЛЬНЫЙ ТРАНСПОРТ ИНДОЦИАНИНОВОГО ЗЕЛЁНОГО С ПОМОЩЬЮ СПЕКТРОСКОПИИ ОБРАТНОГО РАССЕЯНИЯ

Artificial membranes as models in penetration investigations

In vitro methods used in the research of transepidermal transport of active substances generally rely on the penetration rate of test compounds through standard membranes. Models typically used in penetration experiments are specially prepared human or animal skin samples or synthetic membranes. The objective of this study was to establish if the test results for an artificial liposome membrane can be extrapolated to determine the actual bioavailability of active substances. Tests were conducted in a side-by-side diffusion cell. As model membranes, a liquid-crystal lipid membrane (LM), phospholipid membrane (PM) and pig skin sample were used. The test compounds were eight synthetic dyes used in hair colouring products. Research findings reveal that membranes composed of lipids, identical to those present in the epidermis and forming analogical liquid-crystal structures provide a close approximation of the actual bioavailability of active substances (correlation between the results obtained for pig skin and LM was significant: R = 0.95 and R = 0.93 in the presence of a 1% Sodium dodecyl sulphate in donor system). Unlike biological membranes, intercellular cement does not contain phospholipids. The observed correlation between penetration coefficients through the PM and pig skin was not significant (R = 0.82). The experiments confirm that the PM constitutes a less credible model for the studies of transepidermal transport in real life conditions.

Influence of DMPS on the water retention capacity of electroporated stratum corneum: ATR-FTIR study

Anionic lipids like phosphatidylserine are known to significantly enhance electroporation mediated transepidermal transport of polar solutes of molecular weights up to 10 kDa. The underlying mechanism of the effect of anionic lipids on transdermal transport is not fully understood. The main barrier to transdermal transport lies within the intercellular lipid matrix (ILM) of the stratum corneum (SC) and our previous studies indicate that dimyristoyl phosphatidylserine (DMPS) can perturb the packing of this lipid matrix. Here we report on our investigation on water retention in the SC following electroporation in the presence and the absence of DMPS. The water content in the outer most layers of the SC of full thickness porcine skin was determined using ATR-FTIR-spectroscopy. The results show that in the presence of DMPS, the SC remains in a state of enhanced hydration for longer periods after electroporation. This increase in water retention in the SC by DMPS is likely to play an important role in trans-epidermal transport, since improved hydration of the skin barrier can be expected to increase the partitioning of polar solutes and possibly the permeability.

Ultrasound enhanced skin‐lightening effect of vitamin C and niacinamide

Cutaneous hyperpigmentation occurs in multiple conditions. There is a strong need for the improvement of hyperpigmentation especially among Asian women. However, the effect of existing skin-lightening agents is not sufficient. One reason attributes to the limited capability of active agents to be delivered transepidermally. Ultrasound is one promising approach to enhance transepidermal transport. In this work, we investigate the effect of the use of high-frequency ultrasound together with coupling gel containing skin-lightening agents (ascorbyl glucoside and niacinamide) on facial hyperpigmentation in vivo in Japanese women. The effect of ultrasound on the absorption of skin-lightening agents into the stratum corneum was evaluated in a tape-stripping method on human forearms in vivo. The skin efficacy was assessed in a facial clinical trial involving 60 subjects with hyperpigmentation in a paired design. Subjects were assigned to two groups, each group using two treatments (one on each facial cheek): (1) skin-lightening gel with ultrasound vs. no treatment or (2) skin-lightening gel with ultrasound vs. skin-lightening gel treatment. Changes in facial hyperpigmentation were objectively quantified by computer analysis and visual grading of high-resolution digital images of the face in addition to the subjective assessment via questionnaire. Ultrasound radiation enhanced the absorption of skin-lightening agents in the stratum corneum in a radiation-time-dependent manner. In the facial clinical trial, use of ultrasound radiation together with the skin-lightening gel significantly reduced facial hyperpigmented spots compared with both no treatment and skin-lightening gel alone after 4 weeks. The data suggest that use of high-frequency ultrasound radiation together with skin-lightening gel is effective to reduce hyperpigmentation via enhancing transepidermal transport of skin-lightening agents.

Modulation of accelerated repopulation in mouse skin during daily irradiation

Background and purpose: The timing of acceleration of repopulation in the epidermis during daily irradiation is related to the development of skin erythema and epidermal hypoplasia. Therefore, the relationship between impairment of the epidermal barrier function, the dermal inflammatory response and epidermal hypoplasia with the acceleration of repopulation was investigated. Materials and purpose: Skin fields of approximately 1 cm 2 on the thighs of TUC mice were given five daily fractions of 3 Gy in each week followed by top-up doses at the end of the first, the second, or the third week to determine residual epidermal tolerance and to calculate repopulation rates in weeks 1, 2, or 3. Systemic modulation of repopulation was attempted by daily indomethacine during fractionated irradiation whereas tape stripping or UV-B exposure before the start of fractionated irradiation attempted local modulation. In parallel experiments, the water permeability coefficient of the epidermis was determined ex vivo by studying transepidermal transport of tritiated water. Results: Without modulation, no repopulation was found in the first week of daily fractionation but repopulation compensated 30% of the dose given in week two and 70% of the dose given in week three. Only tape stripping before the start of fractionated irradiation accelerated repopulation in week one. UV-B had no effect on repopulation although it stimulated proliferation as much as tape stripping. Indomethacin did not suppress acceleration of repopulation. A significant increase in transepidermal water loss was found but only after repopulation had already accelerated. Conclusions: Acceleration of repopulation in mouse epidermis during daily-fractionated irradiation is not related to the simultaneous development of an inflammatory response. Also, the loss of the epidermal barrier function is not involved in the development of the acceleration response, which rather seems to be triggered directly by the decreased cellularity of the epidermis.

Removal of amino acid during a single passage of water across the gill of marine mussels

AbstractThe clearance of amino acid during a single passage of inhalent water across the gills of marine mussels was studied. Samples of excurrent water were collected and analyzed for their content of 14C‐labeled amino acids and total primary amines. Comparison with concentrations measured in the ambient medium revealed that mussels are capable of removing large fractions of the amino acid present in inhalent water in a single passage across the gill surfaces. At low rates of pumping, Modiolus demissus was capable of removing 95% of the 14C‐glycine from a 1μ M solution. Efficiency of clearance of amino acid was inversely related to pumping rate; absolute rate of uptake was directly related to pumping rate. Concentration related changes in uptake rate and efficiency of clearance indicated that the effective Kt of glycine uptake by intact Mytilus californianus and Modiolus demissus is between 2 and 5μ M. These values are an order of magnitude lower than published figures for Kt of glycine uptake by preparations of isolated gill tissue from these animals. This discrepancy is probably the result of an increase in the effective thickness of unstirred layers on the surfaces of isolated gills. Net clearance of glycine by M. californianus was observed to occur from concentrations as low as 0.28μ M and, from concentrations of 1μ M and less, an average of 20% of the total primary amine offered initially as glycine was removed during a circuit of inhalent water through the animal. Hence, transepidermal transport processes for amino acids in marine mussels are adapted to function efficiently at substrate concentrations found in their normal environment, and can provide a significant supplemental input of reduced carbon and amino nitrogen for these animals.

Characteristics of influex and net flux of amino acids in Mytilus californianus.

1. Intact mussels and an in vitro preparation of isolated gill tissue were employed to study characteristics of the influx and net flux of amino acids in Mytilus californianus. 2. The kinetics of influx of 14C-labelled amino acids were complicated by the presence of a rapidly labelled extracellular compartment. 3. Correction of influx data for the extracellular compartment revealed influx of 14C-cycloleucine and 14C-glycine to be mediated by a transport mechanism adequately described by Michaelis-Menten kinetics. Passive diffusion plays no significant role in influx. 4. Influx and net flux of glycine into intact Mytilus were examined. From high concentrations (greater than 20 micronM) the influx of 14C-glycine was equivalent to the net influx of primary amine as determined by fluorescamine. At low ambient concentrations (greater than 20 micronM), influx of 14C-glycine occurred more rapidly than net influx as determined by fluorescamine. The data suggest that influx of labelled substrate is accompanied by efflux of unknown primary amine(s). In the absence of labelled substrate, efflux continues until a steady-state concentration of 2-5 micronM in the medium is achieved.5. The rate of influx of 14C-cycloleucine into isolated gill tissue, and the concentration gradient which can be developed by gill tissue are reduced when sodium is replaced in the medium. 6. The efflux of primary amines from isolated gill tissue is stimulated by the replacement of sodium in the medium with choline. 7. The data are consistent with a sodium-coupled mechanism for the transport of amino acid into gill tissue. However, energetic considerations cast doubt on the adequacy of such a mechanism to account fully for the observed trans-epidermal transport.