Abstract: Background: Preparation of the keratinocytes in vitro is an important problem for reconstructing of the skin. Therefore, in this study was tried by cultivating of mesenchymal stem cells derived from bone marrow of Wistar rat on the scaffold of human gingiva, the ability of these cells for differentiating towards keratinocytes was investigated. Materials and methods: To reach this goal, the tissues obtained from gingival surgeries in periodontal clinic were decellularized using two detergents including sodium dodecyl sulfate and Triton X-100 and, were used as a scaffold for cultivating with rat’s mesenchymal stem cells after washing and sterilization procedures. The scaffolds were studied by light and electron microscopy before and after 1, 2, and 4 weeks culture. Results: Scanning electron microscopy showed epithelium matrix and collagen fibers in connective tissue remained intact, cells penetrated into scaffold, and pseudo pods were formed. Observation of prepared sections from scaffolds by light microscope identified the cells could form a layer on the previous epithelium matrix a week after culture. In second week, the cells proliferated and created irregular multi-layer structure, but the cells were spindle and layers were more regular in 4 weeks after culture. Transmission electron microscopy of the scaffolds indicated desmosomal adhesions formed which illustrated a degree of differentiation towards keratinocytes. Conclusion: To regard the results is suggested bone marrow mesenchymal stem cells have the ability to differentiate towards keratinocytes. However, immunohistochemistry and molecular studies would help our knowledge about this differentiation. Keywords: bone marrow mesenchymal stem cells, keratinocyte, human gingival scaffold

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