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Generation of a Fibrin Based Three-Layered Skin Substitute.

Kober J, Gugerell A, Schmid M, Kamolz LP, Keck M - Biomed Res Int (2015)

Bottom Line: Our results show that ASCs and fibroblasts were viable, proliferated normally, and showed physiological morphology in the skin substitute.ASCs were able to differentiate into mature adipocytes during the course of four weeks and showed morphological resemblance to native adipose tissue.On the surface keratinocytes formed an epithelial-like layer.

View Article: PubMed Central - PubMed

Affiliation: Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.

ABSTRACT
A variety of skin substitutes that restore epidermal and dermal structures are currently available on the market. However, the main focus in research and clinical application lies on dermal and epidermal substitutes whereas the development of a subcutaneous replacement (hypodermis) is often disregarded. In this study we used fibrin sealant as hydrogel scaffold to generate a three-layered skin substitute. For the hypodermal layer adipose-derived stem cells (ASCs) and mature adipocytes were embedded in the fibrin hydrogel and were combined with another fibrin clot with fibroblasts for the construction of the dermal layer. Keratinocytes were added on top of the two-layered construct to form the epidermal layer. The three-layered construct was cultivated for up to 3 weeks. Our results show that ASCs and fibroblasts were viable, proliferated normally, and showed physiological morphology in the skin substitute. ASCs were able to differentiate into mature adipocytes during the course of four weeks and showed morphological resemblance to native adipose tissue. On the surface keratinocytes formed an epithelial-like layer. For the first time we were able to generate a three-layered skin substitute based on a fibrin hydrogel not only serving as a dermal and epidermal substitute but also including the hypodermis.

No MeSH data available.


Related in: MedlinePlus

(a) Three-layered skin substitute (5 mm3) constructed of fibrin hydrogel for in vitro analysis. (b) Lower layer (yellow) consists of fibrin clot with ASCs and mature adipocyte as hypodermal layer. Upper layer (white) consists of hydrogel mixed with fibroblasts mimicking the dermal layer. On top keratinocytes were seeded as epidermal layer.
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Related In: Results  -  Collection


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fig1: (a) Three-layered skin substitute (5 mm3) constructed of fibrin hydrogel for in vitro analysis. (b) Lower layer (yellow) consists of fibrin clot with ASCs and mature adipocyte as hypodermal layer. Upper layer (white) consists of hydrogel mixed with fibroblasts mimicking the dermal layer. On top keratinocytes were seeded as epidermal layer.

Mentions: For the hypodermal layer ASCs and mature adipocytes were embedded in the fibrin hydrogel. On top, another fibrin clot with fibroblasts was placed for the construction of the dermal layer. Keratinocytes were added on top of the two-layered construct to form the epidermal layer. The three-layered construct was cultivated for up to 3 weeks in cell culture inserts with keratinocytes being exposed to air according to the air-liquid interface cultivation model [20]. Constructs measured 4-5 mm3 for the following in vitro experiments (Figure 1).


Generation of a Fibrin Based Three-Layered Skin Substitute.

Kober J, Gugerell A, Schmid M, Kamolz LP, Keck M - Biomed Res Int (2015)

(a) Three-layered skin substitute (5 mm3) constructed of fibrin hydrogel for in vitro analysis. (b) Lower layer (yellow) consists of fibrin clot with ASCs and mature adipocyte as hypodermal layer. Upper layer (white) consists of hydrogel mixed with fibroblasts mimicking the dermal layer. On top keratinocytes were seeded as epidermal layer.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4508374&req=5

fig1: (a) Three-layered skin substitute (5 mm3) constructed of fibrin hydrogel for in vitro analysis. (b) Lower layer (yellow) consists of fibrin clot with ASCs and mature adipocyte as hypodermal layer. Upper layer (white) consists of hydrogel mixed with fibroblasts mimicking the dermal layer. On top keratinocytes were seeded as epidermal layer.
Mentions: For the hypodermal layer ASCs and mature adipocytes were embedded in the fibrin hydrogel. On top, another fibrin clot with fibroblasts was placed for the construction of the dermal layer. Keratinocytes were added on top of the two-layered construct to form the epidermal layer. The three-layered construct was cultivated for up to 3 weeks in cell culture inserts with keratinocytes being exposed to air according to the air-liquid interface cultivation model [20]. Constructs measured 4-5 mm3 for the following in vitro experiments (Figure 1).

Bottom Line: Our results show that ASCs and fibroblasts were viable, proliferated normally, and showed physiological morphology in the skin substitute.ASCs were able to differentiate into mature adipocytes during the course of four weeks and showed morphological resemblance to native adipose tissue.On the surface keratinocytes formed an epithelial-like layer.

View Article: PubMed Central - PubMed

Affiliation: Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.

ABSTRACT
A variety of skin substitutes that restore epidermal and dermal structures are currently available on the market. However, the main focus in research and clinical application lies on dermal and epidermal substitutes whereas the development of a subcutaneous replacement (hypodermis) is often disregarded. In this study we used fibrin sealant as hydrogel scaffold to generate a three-layered skin substitute. For the hypodermal layer adipose-derived stem cells (ASCs) and mature adipocytes were embedded in the fibrin hydrogel and were combined with another fibrin clot with fibroblasts for the construction of the dermal layer. Keratinocytes were added on top of the two-layered construct to form the epidermal layer. The three-layered construct was cultivated for up to 3 weeks. Our results show that ASCs and fibroblasts were viable, proliferated normally, and showed physiological morphology in the skin substitute. ASCs were able to differentiate into mature adipocytes during the course of four weeks and showed morphological resemblance to native adipose tissue. On the surface keratinocytes formed an epithelial-like layer. For the first time we were able to generate a three-layered skin substitute based on a fibrin hydrogel not only serving as a dermal and epidermal substitute but also including the hypodermis.

No MeSH data available.


Related in: MedlinePlus