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Ex Vivo Prefabricated Rat Skin Flap Using Cell Sheets and an Arteriovenous Vascular Bundle.

Fujisawa D, Sekine H, Okano T, Sakurai H, Shimizu T - Plast Reconstr Surg Glob Open (2015)

Bottom Line: Recently, research on tissue-engineered skin substitutes have been active in plastic surgery, and significant development has been made in this area over the past several decades.These results show that we were able to produce native-like skin.We have succeeded in creating regenerative skin flap ex vivo that is similar to native skin, and this technique could be applied to create various tissues in the future.

View Article: PubMed Central - PubMed

Affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan; and Department of Plastic and Reconstructive Surgery, Tokyo Women's Medical University, Tokyo, Japan.

ABSTRACT

Background: Recently, research on tissue-engineered skin substitutes have been active in plastic surgery, and significant development has been made in this area over the past several decades. However, a regenerative skin flap has not been developed that could provide immediate blood flow after transplantation. Here, we make a regenerative skin flap ex vivo that is potentially suitable for microsurgical transplantation in future clinical applications.

Methods: In rats, for preparing a stable vascular carrier, a femoral vascular pedicle was sandwiched between collagen sponges and inserted into a porous chamber in the abdomen. The vascular bed was harvested 3 weeks later, and extracorporeal perfusion was performed. A green fluorescent protein positive epidermal cell sheet was placed on the vascular bed. After perfusion culture, the whole construct was harvested and fixed for morphological analyses.

Results: After approximately 10 days perfusion, the epidermal cell sheet cornified sufficiently. The desquamated corneum was positive for filaggrin. The basement membrane protein laminin 332 and type 4 collagen were deposited on the interface area between the vascular bed and the epidermal cell sheet. Moreover, an electron microscopic image showed anchoring junctions and keratohyalin granules. These results show that we were able to produce native-like skin.

Conclusions: We have succeeded in creating regenerative skin flap ex vivo that is similar to native skin, and this technique could be applied to create various tissues in the future.

No MeSH data available.


Related in: MedlinePlus

Cross-sections of the epidermal cell sheets and SIEA adiposal flaps after 3 days of perfusion culture. A, Reverse-transplanted group (hematoxylin and eosin stain). B, Normally transplanted group (hematoxylin and eosin stain). C, Reverse-transplanted group. The epidermal cell sheet (green color) and laminin 332 (red color); arrowheads. Nuclei were counterstained with DAPI (blue color). D, Normally transplanted group. The epidermal cell sheet (green color) and laminin 332 (red color) arrowheads. Nuclei were counterstained with DAPI (blue color).
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Figure 3: Cross-sections of the epidermal cell sheets and SIEA adiposal flaps after 3 days of perfusion culture. A, Reverse-transplanted group (hematoxylin and eosin stain). B, Normally transplanted group (hematoxylin and eosin stain). C, Reverse-transplanted group. The epidermal cell sheet (green color) and laminin 332 (red color); arrowheads. Nuclei were counterstained with DAPI (blue color). D, Normally transplanted group. The epidermal cell sheet (green color) and laminin 332 (red color) arrowheads. Nuclei were counterstained with DAPI (blue color).

Mentions: In the reverse-oriented transplant group, the epidermal cell sheets were rounded and detached from the bed (Fig. 3A). In contrast, engraftment of the epidermal cell sheets oriented normally was successful (Fig. 3B). In the former group, the cell sheet was thin and fragmented, and basal membrane protein laminin 332 was observed on both sides of the cell sheet (Fig. 3C). In the latter group, the cell sheet was uniformly thick, and laminin 332 was observed only on the basal side of the cell sheet (Fig. 3D).


Ex Vivo Prefabricated Rat Skin Flap Using Cell Sheets and an Arteriovenous Vascular Bundle.

Fujisawa D, Sekine H, Okano T, Sakurai H, Shimizu T - Plast Reconstr Surg Glob Open (2015)

Cross-sections of the epidermal cell sheets and SIEA adiposal flaps after 3 days of perfusion culture. A, Reverse-transplanted group (hematoxylin and eosin stain). B, Normally transplanted group (hematoxylin and eosin stain). C, Reverse-transplanted group. The epidermal cell sheet (green color) and laminin 332 (red color); arrowheads. Nuclei were counterstained with DAPI (blue color). D, Normally transplanted group. The epidermal cell sheet (green color) and laminin 332 (red color) arrowheads. Nuclei were counterstained with DAPI (blue color).
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Related In: Results  -  Collection

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Figure 3: Cross-sections of the epidermal cell sheets and SIEA adiposal flaps after 3 days of perfusion culture. A, Reverse-transplanted group (hematoxylin and eosin stain). B, Normally transplanted group (hematoxylin and eosin stain). C, Reverse-transplanted group. The epidermal cell sheet (green color) and laminin 332 (red color); arrowheads. Nuclei were counterstained with DAPI (blue color). D, Normally transplanted group. The epidermal cell sheet (green color) and laminin 332 (red color) arrowheads. Nuclei were counterstained with DAPI (blue color).
Mentions: In the reverse-oriented transplant group, the epidermal cell sheets were rounded and detached from the bed (Fig. 3A). In contrast, engraftment of the epidermal cell sheets oriented normally was successful (Fig. 3B). In the former group, the cell sheet was thin and fragmented, and basal membrane protein laminin 332 was observed on both sides of the cell sheet (Fig. 3C). In the latter group, the cell sheet was uniformly thick, and laminin 332 was observed only on the basal side of the cell sheet (Fig. 3D).

Bottom Line: Recently, research on tissue-engineered skin substitutes have been active in plastic surgery, and significant development has been made in this area over the past several decades.These results show that we were able to produce native-like skin.We have succeeded in creating regenerative skin flap ex vivo that is similar to native skin, and this technique could be applied to create various tissues in the future.

View Article: PubMed Central - PubMed

Affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan; and Department of Plastic and Reconstructive Surgery, Tokyo Women's Medical University, Tokyo, Japan.

ABSTRACT

Background: Recently, research on tissue-engineered skin substitutes have been active in plastic surgery, and significant development has been made in this area over the past several decades. However, a regenerative skin flap has not been developed that could provide immediate blood flow after transplantation. Here, we make a regenerative skin flap ex vivo that is potentially suitable for microsurgical transplantation in future clinical applications.

Methods: In rats, for preparing a stable vascular carrier, a femoral vascular pedicle was sandwiched between collagen sponges and inserted into a porous chamber in the abdomen. The vascular bed was harvested 3 weeks later, and extracorporeal perfusion was performed. A green fluorescent protein positive epidermal cell sheet was placed on the vascular bed. After perfusion culture, the whole construct was harvested and fixed for morphological analyses.

Results: After approximately 10 days perfusion, the epidermal cell sheet cornified sufficiently. The desquamated corneum was positive for filaggrin. The basement membrane protein laminin 332 and type 4 collagen were deposited on the interface area between the vascular bed and the epidermal cell sheet. Moreover, an electron microscopic image showed anchoring junctions and keratohyalin granules. These results show that we were able to produce native-like skin.

Conclusions: We have succeeded in creating regenerative skin flap ex vivo that is similar to native skin, and this technique could be applied to create various tissues in the future.

No MeSH data available.


Related in: MedlinePlus