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Mechanical Stretch on Human Skin Equivalents Increases the Epidermal Thickness and Develops the Basement Membrane.

Tokuyama E, Nagai Y, Takahashi K, Kimata Y, Naruse K - PLoS ONE (2015)

Bottom Line: HSEs were prepared in a gutter-like structure created with a porous silicone sheet in a silicone chamber.Transmission electron microscopy revealed that the structure of the basement membrane was more developed in HSEs subjected to stretching.This experimental system may be useful for analysis of the effects of stretch stimuli on skin properties and wound healing and is also expected to be applicable to an in vitro model of a hypertrophic scar in the future.

View Article: PubMed Central - PubMed

Affiliation: The Department of Plastic and Reconstructive Surgery, Okayama University Graduate School of Medicine, Okayama, Japan.

ABSTRACT
All previous reports concerning the effect of stretch on cultured skin cells dealt with experiments on epidermal keratinocytes or dermal fibroblasts alone. The aim of the present study was to develop a system that allows application of stretch stimuli to human skin equivalents (HSEs), prepared by coculturing of these two types of cells. In addition, this study aimed to analyze the effect of a stretch on keratinization of the epidermis and on the basement membrane. HSEs were prepared in a gutter-like structure created with a porous silicone sheet in a silicone chamber. After 5-day stimulation with stretching, HSEs were analyzed histologically and immunohistologically. Stretch-stimulated HSEs had a thicker epidermal layer and expressed significantly greater levels of laminin 5 and collagen IV/VII in the basal layer compared with HSEs not subjected to stretch stimulation. Transmission electron microscopy revealed that the structure of the basement membrane was more developed in HSEs subjected to stretching. Our model may be relevant for extrapolating the effect of a stretch on the skin in a state similar to an in vivo system. This experimental system may be useful for analysis of the effects of stretch stimuli on skin properties and wound healing and is also expected to be applicable to an in vitro model of a hypertrophic scar in the future.

No MeSH data available.


Related in: MedlinePlus

Stretching chamber and stretch device.(A) Over and side view of the chamber. Scale bar = 10 mm. (B) Overhead view of the stretch device. Yellow arrow shows the stretching direction. (C) Schema of the chamber.
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pone.0141989.g001: Stretching chamber and stretch device.(A) Over and side view of the chamber. Scale bar = 10 mm. (B) Overhead view of the stretch device. Yellow arrow shows the stretching direction. (C) Schema of the chamber.

Mentions: In this experiment, we devised a stretch chamber for applying a mechanical stretch to HSEs while they form. Three porous silicone sheets (pore diameter, 1 mm) were attached to the inside of a conventional silicone chamber (Menicon, Aichi, Japan) to form a gutter-like shape. A silicone resin (TSE3032; GE Toshiba Silicones, Tokyo, Japan) was used as an adhesive. The “gutter” was placed such that its bottom was located 4 mm above the bottom of the chamber (Fig 1A and 1C). Prior to use, the chamber was subjected to a 90-seconds plasma treatment with a vacuum plasma apparatus (YHS-R; SAKIGAKE-Semiconductor, Kyoto, Japan) to impart hydrophilicity to the silicone sheet surface.


Mechanical Stretch on Human Skin Equivalents Increases the Epidermal Thickness and Develops the Basement Membrane.

Tokuyama E, Nagai Y, Takahashi K, Kimata Y, Naruse K - PLoS ONE (2015)

Stretching chamber and stretch device.(A) Over and side view of the chamber. Scale bar = 10 mm. (B) Overhead view of the stretch device. Yellow arrow shows the stretching direction. (C) Schema of the chamber.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4631345&req=5

pone.0141989.g001: Stretching chamber and stretch device.(A) Over and side view of the chamber. Scale bar = 10 mm. (B) Overhead view of the stretch device. Yellow arrow shows the stretching direction. (C) Schema of the chamber.
Mentions: In this experiment, we devised a stretch chamber for applying a mechanical stretch to HSEs while they form. Three porous silicone sheets (pore diameter, 1 mm) were attached to the inside of a conventional silicone chamber (Menicon, Aichi, Japan) to form a gutter-like shape. A silicone resin (TSE3032; GE Toshiba Silicones, Tokyo, Japan) was used as an adhesive. The “gutter” was placed such that its bottom was located 4 mm above the bottom of the chamber (Fig 1A and 1C). Prior to use, the chamber was subjected to a 90-seconds plasma treatment with a vacuum plasma apparatus (YHS-R; SAKIGAKE-Semiconductor, Kyoto, Japan) to impart hydrophilicity to the silicone sheet surface.

Bottom Line: HSEs were prepared in a gutter-like structure created with a porous silicone sheet in a silicone chamber.Transmission electron microscopy revealed that the structure of the basement membrane was more developed in HSEs subjected to stretching.This experimental system may be useful for analysis of the effects of stretch stimuli on skin properties and wound healing and is also expected to be applicable to an in vitro model of a hypertrophic scar in the future.

View Article: PubMed Central - PubMed

Affiliation: The Department of Plastic and Reconstructive Surgery, Okayama University Graduate School of Medicine, Okayama, Japan.

ABSTRACT
All previous reports concerning the effect of stretch on cultured skin cells dealt with experiments on epidermal keratinocytes or dermal fibroblasts alone. The aim of the present study was to develop a system that allows application of stretch stimuli to human skin equivalents (HSEs), prepared by coculturing of these two types of cells. In addition, this study aimed to analyze the effect of a stretch on keratinization of the epidermis and on the basement membrane. HSEs were prepared in a gutter-like structure created with a porous silicone sheet in a silicone chamber. After 5-day stimulation with stretching, HSEs were analyzed histologically and immunohistologically. Stretch-stimulated HSEs had a thicker epidermal layer and expressed significantly greater levels of laminin 5 and collagen IV/VII in the basal layer compared with HSEs not subjected to stretch stimulation. Transmission electron microscopy revealed that the structure of the basement membrane was more developed in HSEs subjected to stretching. Our model may be relevant for extrapolating the effect of a stretch on the skin in a state similar to an in vivo system. This experimental system may be useful for analysis of the effects of stretch stimuli on skin properties and wound healing and is also expected to be applicable to an in vitro model of a hypertrophic scar in the future.

No MeSH data available.


Related in: MedlinePlus