Development of a simple, biocompatible and cost-effective Inulin-Diethylenetriamine based siRNA delivery system

Area of Science:

• Biomaterials Science
• Nanotechnology
• Molecular Biology

Background:

• Small interfering RNAs (siRNAs) offer therapeutic potential but require effective delivery systems.
• Current delivery methods face challenges in protecting siRNAs from degradation and ensuring cellular uptake.
• Natural polysaccharides represent a promising platform for developing novel siRNA delivery vehicles.

Purpose of the Study:

• To develop and characterize a novel siRNA delivery system based on functionalized inulin.
• To evaluate the efficacy, safety, and cellular uptake mechanisms of the inulin-based system.
• To compare the performance of the delivery system in different cell lines.

Main Methods:

• Inulin was functionalized with diethylenetriamine (DETA) to create Inu-DETA copolymers.
• Inu-DETA/siRNA complexes were formed and characterized for size, charge, and shape.
• Cytotoxicity, gene silencing efficacy, and cellular uptake mechanisms were assessed in 16HBE and JHH6 cells.

Main Results:

• Inu-DETA effectively bound siRNAs, forming polyplexes with optimal delivery at a 4:1 weight ratio.
• The complexes demonstrated high cytocompatibility and efficient siRNA delivery in JHH6 cells.
• Uptake mechanisms in JHH6 (micropinocytosis, clathrin-mediated endocytosis) facilitated cytoplasmic siRNA diffusion and activity.

Conclusions:

• A novel inulin-based siRNA delivery system (Inu-DETA) was successfully developed.
• The system exhibits efficient siRNA delivery and gene silencing in JHH6 cells with negligible cytotoxicity.
• Further in vivo studies are warranted to explore the therapeutic potential of this inulin-based delivery system.