Hyper-Branched Cationic Cyclodextrin Polymers for Improving Plasmid Transfection in 2D and 3D Spheroid Cells

Area of Science:

• Biomaterials Science
• Gene Therapy
• Nanotechnology

Background:

• Gene delivery systems are crucial for genetic therapies.
• Cationic polymers offer potential as non-viral vectors.
• Understanding delivery in 3D cell models is vital for in vivo applications.

Purpose of the Study:

• To evaluate a novel modified cationic hyper-branched cyclodextrin-based polymer (Ppoly) for plasmid DNA (pDNA) gene delivery.
• To compare the gene delivery efficiency and cytotoxicity of Ppoly-pDNA nanocomplexes in 2D monolayer and 3D multicellular spheroid models.
• To assess the biocompatibility and transfection capabilities of Ppoly as a non-viral gene delivery vector.

Main Methods:

• Utilized 2D monolayer and 3D multicellular spheroid cell culture models.
• Developed Ppoly-pDNA nanocomplexes loaded with Enhanced Green Fluorescent Protein (EGFP) encoding plasmid DNA.
• Assessed cytotoxicity and transfection efficiency using comparative analysis.
• Performed histopathology to visualize cellular uptake in 3D spheroids.

Main Results:

• Ppoly-pDNA nanocomplexes exhibited lower cytotoxicity and higher transfection efficiency in 2D cultures compared to 3D spheroids.
• Histopathology confirmed Ppoly-pDNA complex penetration into the inner layers of 3D spheroids.
• Ppoly demonstrated hemocompatibility, non-toxicity, high encapsulation efficiency, and good transfection efficiency in both 2D and 3D models.

Conclusions:

• The Ppoly-pDNA system is a promising non-viral gene delivery vector with good performance in both 2D and 3D cell models.
• The reduced toxicity in 3D spheroids suggests potential for improved in vivo gene therapy applications.
• Ppoly offers a safe and effective alternative to traditional gene delivery methods like lipofectamine.