[Histologic characteristics of in vitro constructed skin basement membrane].
OBJECTIVE
To observe the histological characteristics of constructed basement membrane in tissue-engineered skin.


METHODS
Forskins from circumcision in normal children were obtained with informed consent of the parents, and then the epidermal keratinocytes (KC) and dermal fibroblasts (Fb) were isolated with trypsin and collagenase D digestion in sequence. Tissue engineered skin with composite chitosan was maintained in a submerged state for 3 days, and then at the air-liquid interface. The tissue-engineered skins were fixed in neutral formalin and then embedded in paraffin after culture for 7, 10 and 15 days, respectively for immunohistological examination of the basement membrane component,including the condition of collagen type IV (COL-IV), collagen type VII (COL-VII), and laminin (LN).


RESULTS
HE staining showed that the keratinocytes formed a fine stratified squamous epithelium with the presence of basal, spinous, granular and corneous cell layers, and there was various amount of cells in flat and fusiform shape in each layer. It was found that a regular red staining strip situated at the dermal epidermal junction. Positive staining of collagen IV, collagen VII as well as LN was observed by immunohistological examination.


CONCLUSIONS
The results suggest that the composite chitosan tissue engineered skin has a good prospect for clinical use because it presents a perfect reconstruction of basement membrane.
Lu
Yuan-gang
YG
Department of Dermatology, Daping Hospital, Field Surgery Institute, the Third Military Medical University, Chongqing, PR China.
Wu
Jin-jin
JJ
Zhang
Jiao-yang
JY
Yang
Gui-hong
GH
Yang
Tao
T
D001485:Basement Membrane / Q000166:cytology*; D002478:Cells, Cultured; D002648:Child; D048271:Chitosan / Q000378:metabolism; D024141:Collagen Type IV / Q000378:metabolism; D024143:Collagen Type VII / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000378:metabolism; D009924:Organ Culture Techniques; D017768:Skin, Artificial; D023822:Tissue Engineering / Q000379:methods*
D024141:Collagen Type IV; D024143:Collagen Type VII; D007797:Laminin; D048271:Chitosan