Efficient In Vitro Electropermeabilization of Reconstructed Human Dermal Tissue

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Tissue Engineering

Background:

• DNA electrotransfer is a promising gene delivery method with clinical limitations due to poorly understood tissue mechanisms.
• The extracellular matrix (ECM) is known to influence in vivo gene electrotransfer efficiency, particularly in tumors.

Purpose of the Study:

• To investigate the role of ECM composition and organization in DNA electrotransfer within normal tissues.
• To develop and validate an innovative 3D reconstructed human connective tissue model for studying electrotransfer mechanisms.

Main Methods:

• Developed a 3D in vitro model of human dermal tissue using primary dermal fibroblasts embedded in a collagen-rich ECM.
• Assessed cell-cell contacts, intercellular junctions, communication, and ECM organization within the 3D model.
• Demonstrated electropermeabilization of cells within the 3D tissue model using millisecond electric pulses.

Main Results:

• The 3D reconstructed human dermal tissue model accurately represents in vivo cell organization and complex ECM.
• Cells within the standardized 3D tissue model were efficiently electropermeabilized.
• The model provides a tool to study skin DNA electrotransfer mechanisms and the influence of ECM.

Conclusions:

• A better understanding of gene electrotransfer in complex tissue environments is crucial for improving electrogene therapy.
• This 3D human tissue model serves as a valuable platform for elucidating DNA electrotransfer mechanisms.
• Improved comprehension can enhance electrogene therapy approaches like systemic protein delivery and DNA vaccination.