Two and three-dimensional gene transfer from enzymatically degradable hydrogel scaffolds

Area of Science:

• Biomaterials Science
• Stem Cell Biology
• Gene Therapy

Background:

• Genetically modifying mesenchymal stem cells (MSCs) within scaffolds can guide differentiation.
• Synthetic hydrogels offer a platform for controlled gene delivery to cells.

Purpose of the Study:

• To explore gene transfer efficiency to MSCs within matrix metalloproteinase (MMP)-degradable hydrogels.
• To compare gene transfer in 2D versus 3D cell cultures within hydrogels.

Main Methods:

• MSCs were seeded on or within MMP-degradable poly(ethylene glycol) (PEG) hydrogels.
• DNA/poly(ethylene imine) (PEI) polyplexes were encapsulated within the hydrogels.
• Gene transfer was assessed using vectors encoding for green fluorescent protein and luciferase.

Main Results:

• Successful gene transfer was achieved for MSCs in both 2D and 3D hydrogel environments.
• Luciferase expression per cell was higher in 2D cultures compared to 3D, despite higher cell density in 3D.
• This indicates greater gene transfer efficiency in 2D cultures.

Conclusions:

• Gene transfer to MSCs within synthetic hydrogels is achievable in both 2D and 3D configurations.
• Hydrogel scaffolds facilitating cellular infiltration can potentially provide sustained gene delivery for tissue regeneration applications.
• Optimizing gene transfer methods for 3D cultures is crucial for maximizing therapeutic potential.