A New Ex Vivo Human Skin Burn Model
Ania LabouchèreDaniel HaselbachMurielle MichettiCatherine PythoudWassim RaffoulLee Ann ApplegateNathalie Hirt‐BurriAnthony de Buys Roessingh
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Abstract:
Abstract Currently, most burn models for preclinical testing are on animals. For obvious ethical, anatomical, and physiological reasons, these models could be replaced with optimized ex vivo systems. The creation of a burn model on human skin using a pulsed dye laser could represent a relevant model for preclinical research. Six samples of excess human abdominal skin were obtained within one hour after surgery. Burn injuries were induced on small samples of cleaned skin using a pulsed dye laser on skin samples, at varying fluences, pulse numbers and illumination duration. In total, 70 burn injuries were performed on skin ex vivo before being histologically and dermato-pathologically analyzed. Irradiated burned skin samples were classified with a specified code representing burn degrees. Then, a selection of samples was inspected after 14 and 21 days to assess their capacity to heal spontaneously and re-epithelize. We determined the parameters of a pulsed dye laser inducing first, second, and third degree burns on human skin and with fixed parameters, especially superficial and deep second degree burns. After 21 days with the ex vivo model, neo-epidermis was formed. Our results showed that this simple, rapid, user-independent process creates reproducible and uniform burns of different, predictable degrees that are close to clinical reality. Human skin ex vivo models can be an alternative to and complete animal experimentation, particularly for preclinical large screening. This model could be used to foster the testing of new treatments on standardized degrees of burn injuries and thus improve therapeutic strategies.Keywords:
Ex vivo
Human skin
Epidermis (zoology)
Artificial skin
Pig skin
Severe burn
A simulant which precisely mimics the linear and non-linear mechanical properties of “the human skin” would be indispensable for assessment of load responses (and discomfort) produced by wearable technologies in the aerospace industry, be it an electronic sensor or a full body suit. In the current work, for the first time, a methodology has been developed to fabricate customized human skin simulants for any person and part of the body. The material comprises of four parts of silicone, which when mixed in different ratios, produces skin simulants with different stiffness properties. Extensive mechanical tests have been performed on different variants of the human skin simulants, and their stress versus strain responses have been matched with actual human skin test data from the literature. Also, the fracture properties of the simulants have been found to be in close range of the actual human skin. Additionally, non-linear hyperelastic constitutive models were used to fully characterize the mechanical behavior of the skin simulant variants. Mechanical tests on freshly excised porcine skin were conducted to validate our test results. To date, such accurate skin simulants has not been developed anywhere to the best of our knowledge. The material is cheap ($15/lb), has no biosafety or handling issues (unlike cadavers, cowhides or porcine skin), and castable to any shape or size. Besides testing, the skin simulants could also be used to develop liners for the wearable technologies, which are just like the skin of the person wearing it, thus reducing discomfort due to material mismatch and friction. Additionally, the skin simulants find applications in the area of manufacturing of prosthetics (liners) and orthotics, military grade armors and personal protection equipment (PPE), and testing of non-lethal and less-lethal ballistics.
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The technique of obtaining human skin with dermis and epidermis reconstructed from cells isolated from patients can enable autologous skin grafting on patients with few donor sites. It also enables in vitro trials on chemicals and drugs. The objective of this work was to demonstrate a method for obtaining human skin composed of associated dermis and epidermis, reconstructed in vitro.Experimental laboratory study, in the Skin Cell Culture Laboratory of Faculdade de Ciências Médicas, Universidade Estadual de Campinas.Cells from human fibroblast cultures are injected into bovine collagen type I matrix and kept immersed in specific culturing medium for fibroblasts. This enables human dermis reconstruction in vitro. On this, by culturing human keratinocytes and melanocytes, differentiated epidermis is formed, leading to the creation of human skin composed of associated dermis and epidermis, reconstructed in vitro.We showed that human skin composed of associated dermis and epidermis can be successfully reconstructed in vitro. It is histologically formed in the same way as human skin in vivo. Collagen tissue can be identified in the dermis, with cells and extracellular matrix organized in parallel to multilayer epidermis.It is possible to obtain completely differentiated human skin composed of associated dermis and epidermis, reconstructed in vitro, from injection of human fibroblasts into bovine collagen type I matrix and culturing of human keratinocytes and melanocytes on this matrix.
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We have developed an artificial skin that mimics the morphological and mechanical properties of human skin. The artificial skin comprises a polyurethane block possessing a microscopically rough surface. We evaluated the tactile sensations when skin-care cream was applied to the artificial skin. Many subjects perceived smooth, moist, and soft feels during the application process. Cluster analysis showed that these characteristic tactile feels are similar to those when skin-care cream is applied to real human skin. Contact angle analysis showed that an oil droplet spread smoothly on the artificial skin surface, which occurred because there were many grooves several hundred micrometers in width on the skin surface. In addition, when the skin-care cream was applied, the change in frictional force during the dynamic friction process increased. These wetting and frictional properties are important factors controlling the similarity of artificial skin to real human skin.
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The expression of gene K51 in the cells of human normal epidermis and epithelial skin tumors was investigated using in situ hybridization method with radioactive probe. The K51 gene transcripts were detected in the epidermis, sebaceous and sweat glands of human embryo and adult skin. The level of gene expression was higher in the stratum granulosum than in the basal layer of the skin. K51 gene expression was also found in the basal cell and metatypical carcinomas, with the level of expression lower than in the neighbouring epidermis and higher than in the surrounding skin stromal cells. Thus, K51 gene is expressed in the skin epidermis of human embryo and adults but the level of its activity is dramatically decreased in the cells of skin epithelial tumors. This potentially may be important as a diagnostic test.
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Exposure to the sun affects the skin and may eventually result in UV-induced skin damage. It is generally known that hyaluronan (HA) is one of the main structural and functional components of the skin. However, UV-related changes in the HA metabolism in the skin have not yet been elucidated. Using qRT-PCR, confocal microscopy and LC-MS/MS we compared the naturally sun-exposed (SE), sun-protected, experimentally repeatedly UVA + UVB-exposed and acutely (once) UVA + UVB irradiated skin of Caucasian women. The epidermis was harvested by means of suction blistering 24 h after the acute irradiation. In addition, the epidermis was compared with a UV-irradiated in vitro reconstituted 3D epidermis (EpiDerm) and an in vitro 2D culture of normal human keratinocytes (NHEK). The amount of HA was found to be statistically significantly enhanced in the acutely irradiated epidermis. The acute UV evinced the upregulation of HA synthases (HAS2 and HAS3), hyaluronidases (HYAL2 and HYAL3), Cluster of differentiation 44 (CD44), and Cell Migration Inducing Proteins (CEMIP and CEMIP2), while only certain changes were recapitulated in the 3D epidermis. For the first time, we demonstrated the enhanced gene and protein expression of CEMIP and CEMIP2 following UV irradiation in the human epidermis. The data suggest that the HA metabolism is affected by UV in the irradiated epidermis and that the response can be modulated by the underlying dermis.
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Artificial human skin is available commercially or can be grown in the laboratory from established cell lines. Standard microscopy techniques show that artificial human skin has a fully developed basement membrane that separates an epidermis with the corneal, granular, spinosal, and basal layers from a dermis consisting of fibroblasts in an extracellular matrix. In this chapter, we show how modeling can integrate microscopy data to obtain a better understanding of the development and aging of artificial human skin. We use the time-dependent structural information predicted by our model to show how irradiation with an electron beam at different times in the life of artificial human skin affects the amount of energy deposited in different layers of the tissue. Experimental studies of this type will enable a better understanding of how different cell types in human skin contribute to overall tissue response to ionizing radiation.
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There is an interest within forensic science to understand the physical and mechanical properties of human skin and the natural and synthetic simulants used to represent it, particularly with reference to reconstruction studies that consider injury to humans, for example during sharp-weapon and ballistic impact assaults. This paper discusses literature in the broad area of (i) human skin, (ii) animal skin and products such as leather and (iii) synthetic polymeric skin simulants. The physical and mechanical properties of human skin appear to be reasonably well documented in the literature. Animal models discussed appear to be restricted primarily to pig and to a lesser extent goat, plus some data on different types of leather. All skin (human and animal) and derivatives such as leather (from various animal sources) are natural materials and therefore variable in their physical and mechanical properties. The variability of commonly used simulants for human skin such as various types of leather could impact on the confidence of any reconstruction study data obtained by using such simulants. While it is recognized that synthetic simulants (polymers such as silicone and polyurethane) do not have the structure of human skin, their physical and mechanical properties can be manipulated relatively easily to match those of skin and are typically of low variability, providing confidence in the repeatability and reproducibility of reconstruction studies.
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Reconstructed human epidermis models are used as epidermis alternatives in skin research studies.
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Barrier function
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Skin electronics provides remarkable opportunities for non-invasive and long-term monitoring of a wide variety of biophysical and physiological signals that are closely related to health, medicine, and human-machine interactions. Nevertheless, conventional skin electronics fabricated on elastic thin films are difficult to adapt to the wet microenvironments of the skin: Elastic thin films are non-permeable, which block the skin perspiration; Elastic thin films are difficult to adhere to wet skin; Most skin electronics are difficult to work underwater. Here, a Wet-Adaptive Electronic Skin (WADE-skin) is reported, which consists of a next-to-skin wet-adhesive fibrous layer, a next-to-air waterproof fibrous layer, and a stretchable and permeable liquid metal electrode layer. While the electronic functionality is determined by the electrode design, this WADE-skin simultaneously offers superb stretchability, wet adhesion, permeability, biocompatibility, and waterproof property. The WADE-skin can rapidly adhere to human skin after contact for a few seconds and stably maintain the adhesion over weeks even under wet conditions, without showing any negative effect to the skin health. The use of WADE-skin is demonstrated for the stable recording of electrocardiogram during intensive sweating as well as underwater activities, and as the strain sensor for the underwater operation of virtual reality-mediated human-machine interactions.
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