Contact-Triggered Lipofection from Multilayer Films Designed as Surfaces for in Situ Transfection Strategies in...

Area of Science:

• Biomaterials Science
• Gene Delivery Systems
• Regenerative Medicine

Background:

• Biomaterials releasing active compounds are crucial for targeted regenerative medicine.
• In situ transfection using surface coatings offers potential for tissue engineering.

Purpose of the Study:

• To design and characterize thin multilayer films loaded with lipid/DNA complexes (lipoplexes) for in situ transfection.
• To evaluate the efficiency, biocompatibility, and gene transfer capabilities of these films.

Main Methods:

• Layer-by-layer (LbL) deposition using hyaluronic acid (HA) and chitosan (CHI).
• Complexation of plasmid DNA with a novel cationic lipid formulation (OH4/DOPE 1/1).
• Surface characterization using fluorescence microscopy, SPR, ellipsometry, zeta potential, AFM, and SEM.
• In vitro cell culture studies with C2C12 cells and in vivo studies using the chicken embryo chorion allantois membrane model.

Main Results:

• LbL films successfully embedded lipoplexes with efficient lipid and DNA deposition.
• Optimized films demonstrated excellent C2C12 cell attachment and biocompatibility.
• Efficient transfection of a reporter gene (green fluorescent protein) in hard-to-transfect C2C12 cells.
• Successful in vivo gene transfer in complex tissues using stored DNA-loaded films.

Conclusions:

• Lipoplex-loaded polyelectrolyte multilayers (PEMs) composed of HA and CHI are effective for in situ transfection.
• These biomaterials show significant potential as tools in regenerative medicine and tissue engineering.
• The novel cationic lipid formulation contributes to the efficiency of gene delivery.