Immunoarchitectural characterization of a human skin model reconstructed in vitro

Souto, Luís Ricardo Martinhão; Vassallo, José; Rehder, Jussara; Pinto, Glauce Aparecida; Puzzi, Maria Beatriz

doi:10.1590/s1516-31802009000100007

Cited by 11 publications

(9 citation statements)

References 27 publications

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“…In turn, leftovers of cosmetic surgery and circumcision are used as a main source of keratinocytes. Depending on the objectives of a study; three-dimensional skin models may also include macrophages [15], melanocytes [16], and dendritic cells [17]. Thus, the TMME that we are proposing in this study is made of mouse keratinocytes; thus, this model does not rely on any human tissue material or cells.…”

Section: Discussionmentioning

confidence: 99%

Three-Dimensional Model of Mouse Epidermis for Experimental Studies of Psoriasis

et al. 2013

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Three-dimensional models of skin and epidermis imitate the structure of real tissues and provide accurate information about certain skin conditions, such as psoriasis. A three-dimensional model of mouse epidermis was generated from the epidermal keratinocytes of newborn mice and treated with cytokines. The aim of this study was to evaluate this model as an experimental model of psoriasis and to assess the changes occurring in its structure and gene expression after the exposure to proinflammatory cytokines. Treatment of the three-dimensional model with either interleukin 17 or a combination of tumor necrosis factor and interferon γ was shown to produce morphological changes, which were similar to acanthosis in psoriatic skin. The observed changes in gene expression of metalloproteinases and certain psoriasis biomarkers, such as mki67, krt16 and fosl1, were similar to the changes in patients’ skin. Notably, changes caused by interleukin 17 were less evident than those caused by the combination of interferon γ and tumor necrosis factor. On the contrary, HaCaT cells exhibited no significant changes in the expression of fosl1 and had decreased levels of mki67 after being treated with a combination of TNF and IFNG. Moreover, treatment with IL17 had no significant effect on krt16 and mki67 expression and even reduced the fosl1 levels. The findings suggest that artificially generated three-dimensional models of murine skin can be used to study psoriasis.

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Section: Discussionmentioning

confidence: 99%

Three-Dimensional Model of Mouse Epidermis for Experimental Studies of Psoriasis

et al. 2013

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“…A current solution to this problem consists in the transplantation of human skin grafts, whether or not autologous, which accommodate the connective tissue and stimulate the development of blood vessels 5, 6. However, this solution is limited by the scarcity of donors and always involves a considerable risk of rejection.…”

Section: Introductionmentioning

confidence: 99%

“…However, this solution is limited by the scarcity of donors and always involves a considerable risk of rejection. Because skin grafts are, in general, degraded prematurely,6 they normally work only as temporary replacements, but while covering the lesion, these devices are capable of reducing fluid loss and the occurrence of infections, allowing for improved patient outcomes and reduced length of stay in hospitals.…”

Section: Introductionmentioning

confidence: 99%

Comparison of the properties of compacted and porous lamellar chitosan–xanthan membranes as dressings and scaffolds for the treatment of skin lesions

Bellini

Pires

Vasconcelos

et al. 2012

J of Applied Polymer Sci

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Compacted and porous lamellar membranes of xanthan (Xn) and chitosan (Ch) at mass ratios of 1 : 1 and 1.2 : 0.8 were prepared and tested to verify possible applications in the treatment of skin lesions. All membranes were prepared by complexation of the polysaccharides in solution and subsequent casting. The porous membranes were obtained by adding either Tween 80 or Pluronic F68 to the polysaccharide complexes before casting. Membranes prepared in the absence of surfactants at a mass ratio of Xn to Ch of 1 : 1 proved ideal for use as wound dressings, as they were thin (around 0.10 mm in thickness) and transparent and had low in vitro cytotoxicity to L929 cells, a tensile strength at break of 25 MPa, water absorption after 24 h of around 86 g/g and simulated body fluid absorption of 16% and adequate stability in the presence of the same solutions. Membranes prepared at the mass ratio of Xn to Ch of 1 : 1 in the presence of Pluronic F68 showed the most favorable characteristics for application as scaffolds for tissue engineering. These membranes consisted of a matrix with interconnected pores which were distributed homogeneously throughout the structure and had a thickness of 1.84 mm, high capacity for FBS uptake (around 18 g/g) and cell culture medium uptake (8.6 g/g), a loss of mass in the culture medium of 33% after 144 h, and low in vitro toxicity to L929 cells. In conclusion, membranes of Ch and Xn produced in the presence or absence of the surfactant Pluronic F68 have a high potential for use as scaffolds in tissue engineering or as dermal dressings, respectively, whereas in contrast, membranes prepared in the presence of Tween 80, regardless of the mass ratio of Xn to Ch, were very cytotoxic to L929 cells and therefore were not appropriate for any of the proposed applications. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

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“…When a lesion occurs, the skin repairs itself through the proliferation and growth of dermal and epidermal cells [3]. In deep skin lesions, destruction of the dermal and epidermal elements may occur.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Culture Medium for Human Keratinocytes

Batista

Rehder²,

Puzzi³

2011

J Stem Cell Res Ther

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Human keratinocytes are needed for tissue engineering of skin applied in tissue repair and regeneration aimed at clinical application. The need to have high cell concentrations within a short time when performing cell proliferation is vital. The growth and maintenance of these cells commonly involve the use of MCDB 153 medium, which is supplemented with animal-derived components, such as growth factors and fetal bovine serum (FBS). The aim of this work was to evaluate different formulations based on MCDB 153 medium for keratinocyte proliferation. For that, several experiments were realized to define which supplements are important for skin cell culture using 2 4 factorial design. Skin samples were obtained from four consenting patients submitted to dermolipectomy, the plastic surgery operation. These fragments were treated with 0.25% trypsin-EDTA for four hours, at 37ºC. Furthermore, the epidermis was separated from the dermis, providing skin cells. Batch cultures were performed in 6 and 96-well plates, in addition to 25 cm 2 flasks. Results concerning cell growth and metabolism showed that keratinocytes were full grown up in MCDB 153 medium containing insulin (7.5 µg.mL-1), bovine pituitary extract-BPE (80 µg.mL-1), epidermal growth factor-EGF (0.08 µg.mL-1), hydrocortisone (0.63 µg.mL-1) and glutamine (1 g.L-1). On the other hand, culture media proposed in the experimental design did not resulted in satisfactory cell growth. In addition, although, the initial glucose concentration was low in the most of cases, the lactate was strongly produced by cells reaching 1 g.L-1 .

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Immunoarchitectural characterization of a human skin model reconstructed in vitro

Cited by 11 publications

References 27 publications

Three-Dimensional Model of Mouse Epidermis for Experimental Studies of Psoriasis

Three-Dimensional Model of Mouse Epidermis for Experimental Studies of Psoriasis

Comparison of the properties of compacted and porous lamellar chitosan–xanthan membranes as dressings and scaffolds for the treatment of skin lesions

Evaluation of Culture Medium for Human Keratinocytes

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