The synthesis and development of nanobioparticles in vitro applications with human cancer cell for boron neutron capture therapy

In this thesis project, it is aimed to develop a boron neutron capture therapy (BNCT) agent, boron nitride nanotube basis nanobioconjugate which has high sensitivity and specificity to glioblastome multiforme cells for monitoring them. The affinity of the synthesized agent to glioblastoma cells was examined in vitro. The obtained results were compared with the 4-borono-L-phenylalanine, which is used as BNCT agent in human. BNNTs were synthesized via chemical vapor deposition (CVD) method with the catalysis of lithium in NH3 atmosphere. The synthesized BNNTs were modified with PAMAM for increasing the solubility. L-Phe was conjugated by EDC-NHS chemistry. The conjugation yield was determined with HPLC. The optimization and characterization of obtained BNNT/PAMAM/L-Phe were carried out with XRD, SEM, EDX, TEM and FTIR. The nanoconjugates were radioiodinated with 125/131I via iodogen method. The quality control of radioiodination was carried out with TLRC and HPLRC. Furthermore the lipophilicity and paper electrophoresis of radioiodinated samples were performed. The stability tests of radioiodinated samples were carried out at room temperature and the results showed that they were stable in 24 hours. In order to evaluate the cytotoxic effects of the nanoconjugates different concentrations and time were investigated. Whether the cytotoxicity of iodine, which participated in the structure, or not. The incorporation of radioiodinated BNNT/PAMAM[G-2]/L-Phe, BNNT/PAMAM[G-2], L-BPA and L-Phe with 125/131I were examined by the time dependent on U87-MG and C8-D1A cells. The fluorescent cell imaging studies were performed with nanoconjugates after FITC conjugation. In the imaging study the cell nucleus was stained blue with DAPI and fluorescent labeled samples were demonstrated green imagine. While the images were evaluated the fluorescent nanoconjugates were migrated to the cytoplasm. The apoptotic effects of nanoconjugates were examined in U87-MG and C8-D1A cells. It was detected that there was no apoptotic effect. The obtained results were showed that the BNNT was biocompatible. PAMAM increased the solubility and bioaffinity of BNNTs. The modification of PAMAM was supplied synergistic effect to the BNNTs. While PAMAM was reducing the cytotoxic effects on cells, at the same time increase the solubility and cell affinity of BNNTs. In addition, the conjugation of L-Phe also have a positive effect, it has become more selective in U87-MG cells. Radioiodination has provided good carrier motifs.