Layered Double Hydroxide as a Potent Non-viral Vector for Nucleic Acid Delivery Using Gene-Activated Scaffolds for...

Area of Science:

• Biomaterials Science
• Gene Therapy
• Regenerative Medicine

Background:

• Nonviral vectors are sought for gene therapy due to safety concerns with viral vectors, but often have lower transfection efficiencies.
• A need exists for nonviral delivery systems with low cytotoxicity and high transfection efficacy for safe, transient gene delivery.

Purpose of the Study:

• To evaluate MgAl-NO3 layered double hydroxide (LDH) as a nonviral vector for delivering nucleic acids (pDNA, miRNA, siRNA) to mesenchymal stromal cells (MSCs).
• To assess LDH's efficacy in both 2D cell culture and 3D tissue engineering scaffolds for gene therapy applications.

Main Methods:

• Nanoparticles were formulated by complexing LDH with pDNA, miRNA mimics/inhibitors, and siRNA at various LDH:nucleic acid mass ratios.
• Delivery and cytotoxicity were assessed in MSCs cultured in 2D monolayers and within collagen-nanohydroxyapatite scaffolds (3D approach).

Main Results:

• LDH-pDNA nanoparticles exhibited significant cytotoxicity and low transfection in 2D MSC cultures, attributed to poor physicochemical properties.
• Lower mass ratios enabled efficient complexation and delivery of miRNA and siRNA to MSCs, with minimal cytotoxicity.
• Incorporation of LDH-miRNA nanoparticles into 3D scaffolds successfully resulted in miRNA overexpression in MSCs.

Conclusions:

• MgAl-NO3 LDH is a promising nonviral vector for delivering miRNA and siRNA to MSCs, particularly in 3D tissue engineering scaffolds.
• This study demonstrates an effective miRNA delivery platform for regenerative medicine applications, overcoming limitations of pDNA delivery.