Study On Chitosan And Its Composite Membranes Used For Melanocyte Culture And Transplantation

Vitiligo is a common skin disease with local or generalized depigmentation attributed by disappearance or loss of epidermal melanocytes, and hard to cure. Its incidence worldwide is 0.1-2%. Although vitiligo is not a fatal disease, it might result in social and psychological impact on patients who may be more vulnerable to sunburn with increased risk of skin cancer. Utilization of biomaterial membranes for melanocyte culture and transfer offers a new approach for transplantation treatment of vitiligo to minimize cell damage and improve engraftment efficiency. The membranes should possess good mechanical properties for cell culture and transfer applications and function as an efficient wound dressing in addition to proper biological properties. It is hence a great challenging to design and produce appropriate membranes.In this work, chitosan, gelatin, and PEG were used as substrate materials to fabricate crosslinked membranes via physical crosslinking with sodium sulfate. Physical and mechanical properties of the membranes were studied. Morphology, proliferation, and activity of human skin melanocytes cultured on the membrances were also investigated. The crosslinked chitosan membranes were found to have good physical and mechanical properties and support the adhesion, growth and proliferation of the melanocytes. Animal experiments showed that the melanocytes cultured on the crosslinked chitosan membranes maintained their activities after transplantation, and the membranes promoted wound healing. Crosslinked chitosan/gelatin composite membranes possessed higher water absorption and water vapor transmission rate with the increase of gelatin ratio. The melanocytes cultured on the crosslinked chitosan/gelatin composite membranes had better cell morphology and activity. When the melanocytes cultured on the crosslinked chitosan/gelatin composite membranes were transplanted on mouse skin, a continuous layer of melanocytes with good activity was formed on the mouse skin. The crosslinked chitosan/PEG composite membranes had better hydrophilic properties and increase the water absorption and water vapor transmission rate. A membrane with good hydrophilic property is in favor of cell culture and control of cell morphology.The innovation in this work is following:the membranes are crosslinked via physical crosslinking to enhance their mechanical properties. The membranes are applicable to melanocyte adhesion, culture, transfer, and transplantation. The membranes can also protect and support the melanocytes during melanocytes culture, transfer, and transplantation, and supply nutrition for the cells. Moreover, the membranes can function as an efficient wound dressing, and have great potential to be applied for vitiligo treatment.