Synthesis of fL and radionuclide labeling HPG (Hyperbranched Polyglycerol) polymers and using blood cells and labeling in cell culture

In this study, it was aimed to develop SPECT / Fluorescent(FL) multifunctional diagnostic agents and to investigate their usage for monitoring blood cells. For this reason, hyperbranched polyglycerols have been synthesized by anionic ring opening polymerization due to their global nature, functionality in terms of versatility and excellent biocompatibility profiles. Succinic anhydride was used for the functionalization of the polymer. Indocyanine green (ICG) is bound to HPG for fluorescence imaging. As a result, the binding efficiency was found as 73.59%. Deferoxamine was selected as the chelating agent and linked to succinate functionalized (SS-HPG) HPG. The structural characterizations of HPG, SS-HPG, HPG-ICG-DFO samples were determined by fourier transform infrared spectroscopy (FT-IR). The molecular weight determination of HPG was determined by high pressure liquid chromatography (HPLC) and found to be 6800 Da. The structural properties were determined by dynamic light scattering (DLS), scanning probe microscopy (SPM). Structural properties of dynamic light scattering (DLS) were determined by scanning end microscopy (SPM). The quality control of samples with radiolabeling [99mTcO4]- was verified by obtaining 100% efficiency with HPLRC and TLRC analyzes. Stability test results were stable for 6 hours. In vitro examination of the biological behavior of HPG-ICG-DFO samples were performed using K562 chronic myeloid leukemia cells. Cytotoxicity study showed that cytotoxicity increased with increasing substance concentration. The highest cell viability was observed at a concentration of 5 µg / mL. Time variation of the binding efficiency of 99mTc (HPG-ICG-DFO) samples to K562 cells was investigated. As a result, the percentage of binding was increased over time and found as 30% at 6 hours. The results showed that fluorescent stained HPG samples showed a promising study of the visualization of blood cells and the presence of bleeding focus.