3-D Scaffold Platform for Optimized Non-viral Transfection of Multipotent Stem Cells

Area of Science:

• Biomaterials science
• Gene therapy
• Tissue engineering

Background:

• Non-viral gene delivery optimization is crucial for biomaterial applications.
• Existing 2D screening methods lack relevance for 3D tissue engineering.
• There is a need for 3D screening systems to assess gene delivery in complex environments.

Purpose of the Study:

• To develop an enhanced throughput, 3D scaffold platform for screening gene delivery parameters.
• To investigate the influence of mineral coating properties on stem cell transfection.
• To optimize gene delivery for enhanced protein production in tissue engineering scaffolds.

Main Methods:

• A 96-well plate compatible 3D scaffold platform was created with interconnected macroporous structures.
• Mineral coatings with varied physicochemical properties were formed on the scaffolds.
• Screening of gene delivery parameters (plasmid amount, N/P ratio, cell density) and mineral coatings was performed using luciferase and BMP-2 encoding plasmids in human mesenchymal stem cells (hMSCs).

Main Results:

• The 3D scaffold platform enabled efficient screening of gene delivery parameters.
• Optimization of mineral coatings led to over 5-fold increase in BMP-2 production in hMSCs.
• Secreted BMP-2 was largely incorporated into the mineral coating, indicating successful gene delivery and protein expression.

Conclusions:

• The developed 3D mineral-coated scaffold platform accelerates gene delivery optimization.
• This platform improves the predictability of screening systems for gene delivery applications.
• The findings facilitate the translation of gene delivery strategies to clinical applications in tissue engineering.