Preparation and characterization of PDR (IMAGE)
Caption
(A and B) Preparation process of the self-assembly of peptide, DNR prodrug, and siRNA, along with the corresponding formulation ratios. (C) DLS detection of the particle size of PDR at different ratios. (D) Flow cytometry detection of the uptake efficiency of THP-1 cells for PDR at different molar ratios. Cells were transfected with 100 nM siRNA for 4 h. (E) RT-qPCR detection of gene inhibition efficiency in THP-1 cells 24 h after transfection with PDR at various ratios (n = 2). (F) TEM image showing the morphology of PDR nanoassembly (peptide:DNR-progrug:siRNA at 10:50:1, the final formulation). Scale bar, 50 nm. (G) Hydrodynamic size, polydispersity index (PDI), zeta potential, and encapsulation efficiency of PDR nanoassembly. (H) Release efficiency of PDR in PBS buffer at pH 5.5 and pH 7.4. (I) Evaluation of PDR stability by detecting changes in particle size at different times (n = 3). (J and K) The presence of π–π stacking interactions in PDR was confirmed by comparing UV absorption wavelengths (J) and fluorescence intensities (K) before and after self-assembly. The concentration of siRNA was 100 nM. All characterization data in (F) to (K) correspond to the final formulation of the PDR nanoassembly at a peptide:DNR-prodrug:siRNA ratio of 10:50:1. *P < 0.05, **P < 0.01, ****P < 0.0001. P value determined by one-way ANOVA followed by Tukey’s multiple comparisons test.
Credit
Haiyin Yang, Beijing Institute of Technology.
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