Sinir Kondüiti Olarak Metakrilatlanmış Jelatin Modifiye Hücresizleştirilmiş Koyun İnce Bağırsak Submukozasının Geliştirilmesi

Abstract

Peripheral nerve damage is a common injury because of trauma. The slow regeneration of the complicated structure of the peripheral nervous system highly affects the regainment of function. Hence nerve tissue engineering studies regarding peripheral nerve injury have importance. GelMA-modified decellularized SIS hybrid scaffolds have been designed for peripheral nerve injury. The first part of the thesis is about the decellularization of sheep SIS by two methods. A conventional method involving the combinations of physical, chemical, and enzymatic applications, which is also used as a control group to develop an original scCO2 technology for SIS decellularization. The genomic DNA decrement was %91,13 after conventional method decellularization. The optimum method for scCO2 decellularization was decided with the genomic DNA decrease of %92,77. Both protocols were characterized and compared by H&E and DAPI histological stainings to investigate the qualitative effect of the used protocols and to assess the ECM after decellularization. GAG amount assay showed a %75,47 decrease in the conventional method decellularization group whereas no decrement was observed in the scCO2 decellularization group. In this regard, it was shown that the scCO2 technology was an efficient and high-yielding method for the decellularization of SIS. SEM and tensile testing were used to assess the morphologic and mechanical effects of the decellularization methods. The synthesis of GelMA prepolymer is conducted in the second part of the study which is followed by the optimization of hydrogel synthesis parameters. Gelatin is modified with varying ratios of MA and the ideal MD was determined by 1H-NMR. The pre-polymer group with %75,76 MD was used for hydrogel synthesis. Then the in situ crosslinking of GelMA pre-gels with varying GelMA concentrations was carried out by photo-initiator I2959 under UV irradiation. The obtained hydrogels are characterized by FT-IR, swelling, and mechanical analyses. The direct proportion between the increasing gelMA concentration and mechanical strength was shown. In the third part of the study SIS was incubated with GelMA solutions for UV crosslinking using I2959 photo-initiator. FT-IR and SEM were used as a means to prove the success of hybrid tissue scaffold synthesis as well as to investigate structural changes. The water retention capacity, degradation properties, surface wettability and mechanical strength of the hybrid scaffolds were investigated and compared with the GelMA hydrogels and acelluar SIS. Lastly the ability of hybrid membranes to induce neural cell adhesion and proliferation is proved via in vitro cell cultre of PC12 cell line in terms of cell viability, proliferation and morphological changes.