The effects of peripheral mononuclear stem cell and differentiated langerhans cells application on wound models in experimental diabetics created rats

Chronic injuries that are difficult to heal, such as diabetes, are a problem in medical treatment. Seeking new treatment protocols is an important issue that has begun to be used in stem cell clinics. Peripheral Blood Mononuclear Cell (PBMC) is a good source of stem cells that can easily be obtained from peripheral circulation and can be easily applied to patients under good production conditions. Our study aimed to compare PBMC and Langerhans cells alone and in combination to determine whether these two stimuli were stimulating each other and to apply them to skin grafts in an experimental animal model to demonstrate their benefit in the treatment of chronic wounds. Diabetes Mellitus is now becoming an increasingly international health problem. Diabetic complications include numerous accompanying problems, including severe wound healing, chronic ulceration, and severe complications of the resulting limb amputation. Cellular products have begun to be used in an increasingly broad spectrum of disease treatments in recent years. Among these cellular products, PBMC is used in the treatment of various diseases, especially hematological malignancies. It is also known that langerhans cells, which play an active role in wound healing, especially in the inflammatory phase, have important immunological functions. In addition, in vivo experiments have shown that Langerhans cells play an important role in epidermal homeostasis and wound healing is particularly beneficial in early stages. The specificity of this study is to investigate the effects of the langerhans cells differentiated from the root cell in difficult-to-heal wounds and the mechanisms of their action in the treatment unknown in the literature. In this study, PBMC isolation, culture propagation and differentiation of langerhans cells (dLC) will be provided. These cells will be applied on the grafts in the culture medium and applied to the wound area. The back region of Wistar albino rats will be cut and four (four) full fold skin grafts of 1 x 1 cm size will be reciprocally replaced. Then the rats will be divided into 10 groups; only the graft wound model will be applied to the control group, only the feeder will be applied to the sham group and the other groups will be applied to the wound area on the PBMC and dLC grafts in the feeder. Cellular therapy will be repeated intradermally in a single dose for 14 days. On the 3rd and 7th days from the grafts on the back, samples will be taken with biopsy. In cellular therapy, 1x106 cells will be administered to the side of each wound of PBMC and dLC. During the wound healing process, cell application will be repeated to the biopsied areas. Also histochemically, hematoxylin-eosin, PAS and masson trichrome staining,; immunohistochemically, e-NOS, i-NOS for oxidative stress, PCNA as proliferative marker,VEGF marker for vascularization, IL-1 as inflammation markers and TUNEL staining for apoptosis will be performed. The images taken under the microscope will be transferred to the digital platform and evaluated by morphometric and H-score. The difference between the groups will be statistically analyzed. Success will be achieved when our study shows that PBMC and dLC contributes to supportive care in the treatment of chronic wounds such as diabetic foot. This success is thought to be an application that reduces costs and increases patient quality of life. It is expected that it will contribute to the literature by publishing the quality journal and be an effective product in clinical use.