Cell membrane damage and cargo delivery in nano-electroporation

Area of Science:

• Biotechnology
• Cell Biology
• Nanotechnology

Background:

• Nanochannel electroporation (NEP) is an advanced cell transfection technique.
• NEP utilizes an asymmetric electric field for targeted membrane poration, enhancing gene delivery and cell viability over conventional bulk electroporation (BEP).

Purpose of the Study:

• To systematically investigate the transfection window of NEP for delivering molecules of varying sizes.
• To analyze the relationship between applied voltage, molecule size, cell membrane damage, and transfection efficiency.
• To develop a predictive model for cell membrane damage during NEP using numerical simulations.

Main Methods:

• Experimental investigation of NEP with different molecule sizes (0.6 kDa and 6 kDa).
• Systematic variation of working voltage to determine optimal transfection parameters.
• Numerical analysis of transient transmembrane potential (t-TMP) and dynamic transmembrane potential (d-TMP) to model membrane poration and damage.

Main Results:

• Small molecules (∼0.6 kDa) can be delivered efficiently at lower voltages with minimal impact on non-transfection membrane integrity.
• Delivery of larger molecules (∼6 kDa) requires higher voltages, leading to increased non-transfection membrane damage and reduced cell viability.
• Numerical simulations of t-TMP and d-TMP accurately predict membrane damage on both transfection and non-transfection sides of the cell.

Conclusions:

• The study establishes a clear transfection window for NEP, crucial for optimizing cargo delivery based on molecule size.
• Predictive modeling of transmembrane potential provides a comprehensive understanding of cell membrane responses during NEP.
• NEP offers a tunable platform for efficient and safe genetic material delivery, with potential for broad biotechnological applications.