Limits...
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

Polarity analysis of epidermal cell sheets. A, Macroscopic appearance of the SIEA adiposal flap and 2 cell sheets after 3 days of perfusion culture. B, Fluorescent observation after 3 days of perfusion culture. C, GFP-positive pixels of reversed and normal transplanted groups. Asterisk indicates that the P value was less than 0.05. The error bars on histograms indicate standard errors of the means.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4494494&req=5

Figure 2: Polarity analysis of epidermal cell sheets. A, Macroscopic appearance of the SIEA adiposal flap and 2 cell sheets after 3 days of perfusion culture. B, Fluorescent observation after 3 days of perfusion culture. C, GFP-positive pixels of reversed and normal transplanted groups. Asterisk indicates that the P value was less than 0.05. The error bars on histograms indicate standard errors of the means.

Mentions: The epidermal cell sheets transplanted in the normal orientation were observed to be partially keratinized after 3 days culture. On the other hand, the cell sheets transplanted upside down appeared wet (Fig. 2A). Fluorescent observation under a surgical microscope showed that the normally orientated sheets were strongly luminescent and uniform over most of the cell sheet, but the reverse sheets had weak mist-like emission intensity (Fig. 2B).


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)

Polarity analysis of epidermal cell sheets. A, Macroscopic appearance of the SIEA adiposal flap and 2 cell sheets after 3 days of perfusion culture. B, Fluorescent observation after 3 days of perfusion culture. C, GFP-positive pixels of reversed and normal transplanted groups. Asterisk indicates that the P value was less than 0.05. The error bars on histograms indicate standard errors of the means.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4494494&req=5

Figure 2: Polarity analysis of epidermal cell sheets. A, Macroscopic appearance of the SIEA adiposal flap and 2 cell sheets after 3 days of perfusion culture. B, Fluorescent observation after 3 days of perfusion culture. C, GFP-positive pixels of reversed and normal transplanted groups. Asterisk indicates that the P value was less than 0.05. The error bars on histograms indicate standard errors of the means.
Mentions: The epidermal cell sheets transplanted in the normal orientation were observed to be partially keratinized after 3 days culture. On the other hand, the cell sheets transplanted upside down appeared wet (Fig. 2A). Fluorescent observation under a surgical microscope showed that the normally orientated sheets were strongly luminescent and uniform over most of the cell sheet, but the reverse sheets had weak mist-like emission intensity (Fig. 2B).

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