INVESTIGATION OF LIPID NANOPARTICLE:PDNA COMPLEX INTERACTIONS WITH BIOLOGICAL MATERIAL
Bildiriler > Özet Bildiri · 2024
Yazarlar: Sahar Tarkavannezhad, Mustafa KOTMAKÇI
14th International Symposium on Pharmaceutical Sciences (ISOPS), Türkiye, Ankara, Uluslararası, Sözlü Sunum, Özet Bildiri, 2024-2024, ss 201-201
Özet
<p>Introduction: Low transfection efficiency and high toxicity are two of the</p><p>major challenges in non-viral gene delivery. Serum nucleases and biological</p><p>polyanions lower the efficiency of pDNA therapeutics. In this study, novel</p><p>lipid nanoparticle (LNP):pDNA complexes were developed as non-viral gene</p><p>delivery systems with the aim to investigate their protective effect against protease</p><p>degradation and polyanionic stress.</p><p>Materials and Methods: LNP’s were prepared by freez drying- rehydrationsonication</p><p>method using DOPC, DOTAP and Cholesterol at different molar</p><p>ratios (Oner et al., 2021). Samples were frozen at -80ºC, dried under deep</p><p>vacuum for 24 h, rehydrated in HEPES buffer and probe-sonicated. LNP:</p><p>pDNA complexes were formed at RT by incubating LNPs with pDNA at</p><p>N/P ratios between 0.5 and 12. Complex integrity and pDNA stability in</p><p>the presence of serum were assessed for different incubation periods (Zhang</p><p>et al., 2017). Complexes were incubated with heparin to test the resistance</p><p>to anionic stress. FBS and heparin treatments were analyzed by agarose gel</p><p>electrophoresis (Yadav et al., 2021).</p><p>Cytotoxicity studies were performed on PC-3 cells. Cells were treated with</p><p>complexes and cell viability was assessed using resazurin.</p><p>Results: Obtained LNPs effectively complexed pDNA above N/P ratio of 3.</p><p>No pDNA decomplexation was observed in the presence of 10% and 50%</p><p>FBS. Intentional release of pDNA revealed supercoiled pDNA conformation</p><p>for all complexes. Complexes at higher N/P ratios were more stable against</p><p>polyanionic stress. The complexes were non-toxic on PC-3 cell line at the</p><p>tested concentrations.</p><p>Conclusions: The prepared LNP’s successfully formed complexes with</p><p>pDNA and protected it against serum protease degradation. pDNA was not</p><p>released from the omplexes at N/P 12 ratio. All complexes were biocompatible</p><p>and suitable for transfection.</p><p>Acknowledgment: This study was supported by Ege University Scientific Research</p><p>Projects Department, project no. 30842.</p>