Our objective is to determine the quality of tissue engineered human skin via immunostaining, RT-PCR and electron microscopy (SEM and TEM). Culture-expanded human keratinocytes and fibroblasts were used to construct bilayer tissue-engineered skin. The in vitro skin construct was cultured for 5 days and implanted on the dorsum of athymic mice for 30 days. Immunostaining of the in vivo skin construct appeared positive for monoclonal mouse anti-human cytokeratin, anti-human involucrin and anti-human collagen type I. RT-PCR analysis revealed loss of the expression for keratin type 1, 10 and 5 and re-expression of keratin type 14, the marker for basal keratinocytes cells in normal skin. SEM showed fibroblasts proliferating in the 5 days in vitro skin. TEM of the in vivo skin construct showed an active fibrocyte cell secreting dense collagen fibrils. We have successfully constructed bilayer tissue engineered human skin that has similar features to normal human skin.
Fibroblasts/*cytology ; Keratinocytes/*cytology ; Mice, Nude ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Quality Control ; Regeneration/physiology ; Skin/pathology ; Skin Transplantation/pathology ; Skin Transplantation/*standards ; Tissue Engineering/*standards

OBJECTIVETo improve the paramedian forehead flap for nasal reconstruction.

METHODSBased on the findings of the Cutaneous branch of supratrochlear artery, the forehead musculo-cutaneous flap was divided into the musculo-flap and the skin-flap in 3 patients. The musculo-flap were used to reconstruct the septi-bone structure, and envelop silicon nasal-frame, while the skin-flap were used to reconstruct the nose. In 6 patients, the forehead flap was used as a skin-flap just with the muscle pedicle to reconstruct a nose with good appearance of nasal subunits, without the secondary operation.

RESULTSIn all of 9 cases, the cutaneous-branch of supratrochlear artery was found 1.7-2.0 cm above the orbital-rim. Eight patients got good results, only 1 patient had to do the secondary operation to coverage the exposed nasal frame.

CONCLUSIONSThe cutaneous-branch of supratrochlear artery could be an anatomical finding to form the forehead skin-flap and it is a good choice to be used to reconstruct a nose, even in complicated cases.

Adolescent ; Adult ; Female ; Forehead ; Humans ; Male ; Muscles ; transplantation ; Nose ; surgery ; Patient Satisfaction ; Reconstructive Surgical Procedures ; methods ; standards ; Skin Transplantation ; methods ; Surgical Flaps ; Treatment Outcome