The Construction Of Skin Model And Its Testing Application In Cosmeceutical

The methods of constrcting skin model have been developed for more than30years.The technologies have also tended to be mature from a single epidermal model to complexlayer of skin substitute with the development of tissue engineering. While the research ofskin model is in the initial stage in domestic and establishing Chinese skin alternativemodel is on focus currently. According to the complex degree, the skin model is dividedinto the epidermis model, dermis model and epidermis-dermis model as the complex layerskin model. These in vitro alternative models paly an important role for the evaluatingtoxicology in vivo and in vitro. The purpose of this paper is to constrct skin model usingthe primary skin keratinocytes and fibroblasts cells. First of all, selecting appropriatebiological material as scaffold, then formulating optimize chemical composition definedmedium and taking normal tissue with no lesions, finally isolating and culturing primarykeratinocytes and fibroblasts cells from tissue. When the two kinds of primary cellsachieved optimal growth conditions, then transfer the transwell dish and continue inoculateunder gas-liquid cultivation, in oder to form a multilayer and highly differentiatedorganization model. Then start to verify the model: first, HE sections,immunohistochemical observation were done for the growth of state of skin model, thenused the constrcted skin model for testing skin toxicity and11kinds of chemical wereselected for testing substances appointed from European Centre for the Validation ofAlternative Methods; then choosing the MTT methods and the ELISA release assaymeasured the IL-1α factor from skin model in oder to determine the stability, sensitivityand accuracy of the skin model, and gradually establish alternatives methods and platformof skin toxicity instead of animal testing. Finally, the application of toxicity evaluation ofskin model: in order to evaluate the system as the basis, choosing8kinds of cosmeceuticalraw materials from our laborary to detect the toxicity.The main results are as follows:(1)The successful construction of the2strains of different age of fibroblast cell line;(2)the use of rat tail collagen constructs a collagenscaffold;construction of the skin model;(1) HE staining, immunohistochemical observation model and normal skin similar;(2) Model validation process, compared with the animal experiment,70%accuracy rate;(3) Application of detecting drug cosmetics;...