In vivo application of tarantula cubensis venom theranekron, chitosan, and hyaluronic acid coated wound covering material by nanofabrication method

This thesis aims to produce wound dressing materials coated with the multilayer coating method (LbL) containing different doses of theranekron (T), which is the tarantula cubensis venom, a tarantula species whose proliferative effect has been proven in cell culture studies, and to examine the effect of these materials on healing using a wound model. to demonstrate with in vivo experiments. In the in vitro wound dressing material production of the thesis, fabrics cut in 2X2 cm dimensions were coated with LbL solutions created by adding different concentrations of T to the hyaluronic acid (HA) and chitosan (K) content required for the multilayer coating process and left to dry. Scanning electron microscopy (SEM), energy dispersive spectrum (EDS), and Fourier transform infrared spectroscopy (FTIR) analyses were applied to some of the prepared materials for characterization. The other parts were used in four circular wounds with a diameter of 1 cm opened on the back of each of 16 animals in vivo experiments in a total of 8 groups: control, HA+K, T10, T20, T50, T100, T200, T500. Histopathological hematoxylin-eosin, Masson trichrome, and Picro Sirius staining were performed on half of the skin tissues taken from the rats sacrificed on the 8th day after dressing and photography were taken two days apart. In the other halves, the expressions of the growth factors fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), epidermal growth factor (EGF), and transforming growth factor-β (TGF-β) were examined. As a result of the pre-treatment with polyethyleneimine (PEI) before the coating process, cationic charges were created on the surface of the cotton fibers. In the SEM-EDS analysis, the elemental analysis content of the untreated cotton fabric was found to be 54% C and 46% O, while the elemental analysis content of the untreated cotton fabric was found to be 44.79% C in the elemental analysis as a result of the pre-treatment. , 48.56% O, 6.19% N, and 0.46% Ca elements were found, and these results prove that the pre-treatment was successful. Especially in HA-K-coated fibers, chitosan particles in crystal form are seen homogeneously distributed on the fiber surface. In the fabrics impregnated with 6 different doses of T, as the amount of T dose increased, the crystalline structures of chitosan could not be seen in the SEM images. In wound area measurements, it is seen that the healing rate continues to increase significantly the wound surface area decreases rapidly in the T20 and T50 groups, and the change in healing between days 1 and 7 is approximately 41.32% and 44.59%, respectively. The decreased wound surface areas in the T100, T200, and T500 groups are remarkable, and the change in healing between days 1 and 7 was determined to be approximately 16.12%, 15.63%, and 13.43%, respectively. In histopathological staining, it was observed that in the T10, T20, and T50 groups, the improvement became evident depending on the dose and neovascularization, collagen production and granulation tissue formation increased. In contrast, in the T100, T200, and T500 groups, the healing was negatively affected. The amount of defect closure and neovascularization, collagen production, and connective tissue development decreased, resulting in hyperemic and necrotic lesions. field formations were detected. EGF gene expression levels evaluated by PCR were found to be particularly high in the T10, T20, and T50 groups, with the highest levels belonging to the T20 group. Growth factor expressions were detected very low in the T100, T200, and T500 groups. As a result, in vivo and in vitro phases of the wound dressing material containing Theranekron+HA+K were performed and it was concluded that the most ideal wound healing dose was 20 mcg. These results need to be supported by more detailed research.