Primary Study On Reconstruction Of Composite Chitosan Artificial Skin After Grafted Onto Experimental Rabbit

Background and Objectives:Ideal skin equivalent should consist of epidermis and dermis which can reconstruct slowly to resemble the structure of normal skin afer grafted on experimental animals.We aim to study the transformation of epidermal differentiation marker and the reformation of blood vessel and basement membrane after composite chitosan artificial skin grafted onto experimental rabbit.So we can understand the reconstruction of composite chitosan artificial skin after grafted and provide experimental evidences for its clinical use.Methods:We respectively make paraffin sections of composite chitosan artificial skin before grafted, one month or two month after grafted,natural human skin and natural rabbit skin,then divide them into experimental group and control. By immunohistochemical staining, the expression of epidermal keratin was identified with broad-spectrum polyclonal antibody to keratin,basal cells and suprabasal cells with monoclonal antibodies to cytokeratin K5 and cytokertin K10 respectively to show the differentiation of the epidermis,vascular endothelial cells and basement membrane with polyclonal antibodies to factor â…§ related antigen and laminin respectively to show the reformation of blood vessles and basement membrane.Results:1. The staining of broad-spectrum polyclonal antibody to keratin of keratinocytes of composite chitosan artificial skin before grafted ,one month ortwo month after grafted were positive,resembling natural epidermal keratinocytes. 2. The suprabasal cells of composite chitosan artificial skin before grafted, one month or two month after grafted were stained positively with monoclonal antiby to cytokeratin K10,closer to the surface of epidermis,the staining was deeper,resembling natural human epidermis.The basal cells were negative. With monoclonal antiby to cytokeratin K5 ,the basal cells of composite chitosan artificial skin before grafted,one month or two month after grafted were stained weakly,obviously weaker than natural human skin.The suprabasal cells were stained negative. 3. The staining of polyclonal antibody to factor â…§ related antigen of composite chitosan artificial skin before grafted was negative, one month or two month after grafted in the dermis were positive.We can find neonate blood vessles in the dermis,which were specialy obvious in one month after grafted.The quantities of blood vessles in the superficial dermis were more than natural dermis.4. With polyclonal antibodies to laminin,composite chitosan artificial skin before grafted was negative, one month or two month after grafted at the junction of epidermis and dermis were partly positive.we can find discontinous basement membrane. The basal cells were positive. The basement membrane of natural skin was continous, and its basal cells were negative. Furthermore, the basement membrane of blood vessles were positive resembling to the staining with polyclonal antibody to factor â…§ related antigen.5. The stainings of controls were all negative. Conclusions:The epidermis of composite chitosan artificial skin after grafted can express keratin,with a good differentiation of the epidermis.There are obvious neonate blood vessles in the dermis,indicating composite chitosan artificial skin can achieve reconstruction of blood vessles.Although thereformation of basement membrane is partial and discontinous,the reconstruction of composite chitosan artificial skin after grafted was well in general,resebling natural human skin.Composite chitosan artificial skin is an useful substitute for human skin,it has a good prospect for clinical use.