Human-like Skin Tissue Graft Generated Using Human UmbilicalCordBlood Mesenchymal Stem Cells and Acellular Amniotic Membrane: A New Approach in Reconstructive Medicine

Background: Due to its outstanding biological properties, the Human Amniotic Membrane (HAM) has been a topic of medical research for many years to treat, repair, and regenerate damages related to the epithelial surface, corneal epithelial, orthopedic, and infected body parts. The HAM shows crucial anti-inflammatory, anti-bacterial, and anti-viral properties and immunomodulating qualities. Therefore, the HAM is one of the most used natural biomaterials in tissue engineering to generate bio-scaffolds. Objective: This study used HAM as a raw material to produce a collagen membrane that fully meets the criteria for creating a bioscaffold with suitable mechanical strength and limited risk of graft rejection. This collagen membrane would be a bioscaffold to carry Human Umbilical Cord Blood Mesenchymal Stem Cells (hUCB-MSCs) to generate human epithelial-like cell sheets. Shortly, these cell sheets can repair and regenerate the epithelial layer in trauma cases, severe damage from accidents, burns, and other post-operational conditions. Methods: Among the varieties of stem cells currently being researched, MSCs are the most studied and applied due to their outstanding bio-qualities and regenerative features. We used International Society for Cells and Gene Therapy (ISCT) standards to identify and characterize the hUCB-MSCs obtained from a donor's umbilical cord blood who provided consent. We used an inverted phase contrast microscope to evaluate morphological changes and epithelial stratification of cultured growing cells on the collagen scaffold. Further, staining techniques, such as H&E and Trichrome, were used to assess the morphology and structure of the new epithelial construct; histochemical staining with p63 and ck 1/10 markers was used to evaluate the cellular characteristics and specific traits. Transmission electron microscopy was used to validate cell junctions. Results: This study successfully obtained human epithelial-like cell sheets with multiple cell layers, morphology, and characteristics comparable to human epithelial tissue. Conclusion: The results of this study will constitute a potential alternative to autologous skin grafting in heavy skin loss and deep lesions.