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Gene electrotransfer enhanced by nanosecond pulsed electric fields.

Guo S, Jackson DL, Burcus NI, Chen YJ, Xiao S, Heller R - Mol Ther Methods Clin Dev (2014)

Bottom Line: We demonstrated that the effect of nsPEFs on gene transfection was time restricted.The effect of nsPEFs on cell viability is also dependent on the specific pulse parameters.We also determined that both calcium independent and dependent mechanisms are involved in nsPEF effects on gene electrotransfer.

View Article: PubMed Central - PubMed

Affiliation: Frank Reidy Research Center for Bioelectrics, Old Dominion University , Norfolk, Virginia, USA.

ABSTRACT
The impact of nanosecond pulsed electric fields (nsPEFs) on gene electrotransfer has not been clearly demonstrated in previous studies. This study was conducted to evaluate the influence of nsPEFs on the delivery of plasmids encoding luciferase or green fluorescent protein and subsequent expression in HACAT keratinocyte cells. Delivery was performed using millisecond electric pulses (msEPs) with or without nsPEFs. In contrast to reports in the literature, we discovered that gene expression was significantly increased up to 40-fold by applying nsPEFs to cells first followed by one msEP but not in the opposite order. We demonstrated that the effect of nsPEFs on gene transfection was time restricted. The enhancement of gene expression occurred by applying one msEP immediately after nsPEFs and reached the maximum at posttreatment 5 minutes, slightly decreased at 15 minutes and had a residual effect at 1 hour. It appears that nsPEFs play a role as an amplifier without changing the trend of gene expression kinetics due to msEPs. The effect of nsPEFs on cell viability is also dependent on the specific pulse parameters. We also determined that both calcium independent and dependent mechanisms are involved in nsPEF effects on gene electrotransfer.

No MeSH data available.


The Profiles of nanosecond electric pulses. (a) A representative 60 ns pulse with applied electric field 3 KV or 24 KV/cm. (b) A representative 60 ns pulse with applied electric field 4 KV or 32 KV/cm.
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fig8: The Profiles of nanosecond electric pulses. (a) A representative 60 ns pulse with applied electric field 3 KV or 24 KV/cm. (b) A representative 60 ns pulse with applied electric field 4 KV or 32 KV/cm.

Mentions: NsPEFs were produced by a custom-designed nanosecond pulse generator that can generate fixed 60 ns PEFs with adjustable pulse frequency and applied electric field. The pulse profile was monitored and recorded by a digital phosphor oscilloscope (TDS3052B, Tektronix, OR) (Figure 8), and power was provided by a high voltage power supplier (EH60R1.5, Glassman High Voltage, NJ).


Gene electrotransfer enhanced by nanosecond pulsed electric fields.

Guo S, Jackson DL, Burcus NI, Chen YJ, Xiao S, Heller R - Mol Ther Methods Clin Dev (2014)

The Profiles of nanosecond electric pulses. (a) A representative 60 ns pulse with applied electric field 3 KV or 24 KV/cm. (b) A representative 60 ns pulse with applied electric field 4 KV or 32 KV/cm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362372&req=5

fig8: The Profiles of nanosecond electric pulses. (a) A representative 60 ns pulse with applied electric field 3 KV or 24 KV/cm. (b) A representative 60 ns pulse with applied electric field 4 KV or 32 KV/cm.
Mentions: NsPEFs were produced by a custom-designed nanosecond pulse generator that can generate fixed 60 ns PEFs with adjustable pulse frequency and applied electric field. The pulse profile was monitored and recorded by a digital phosphor oscilloscope (TDS3052B, Tektronix, OR) (Figure 8), and power was provided by a high voltage power supplier (EH60R1.5, Glassman High Voltage, NJ).

Bottom Line: We demonstrated that the effect of nsPEFs on gene transfection was time restricted.The effect of nsPEFs on cell viability is also dependent on the specific pulse parameters.We also determined that both calcium independent and dependent mechanisms are involved in nsPEF effects on gene electrotransfer.

View Article: PubMed Central - PubMed

Affiliation: Frank Reidy Research Center for Bioelectrics, Old Dominion University , Norfolk, Virginia, USA.

ABSTRACT
The impact of nanosecond pulsed electric fields (nsPEFs) on gene electrotransfer has not been clearly demonstrated in previous studies. This study was conducted to evaluate the influence of nsPEFs on the delivery of plasmids encoding luciferase or green fluorescent protein and subsequent expression in HACAT keratinocyte cells. Delivery was performed using millisecond electric pulses (msEPs) with or without nsPEFs. In contrast to reports in the literature, we discovered that gene expression was significantly increased up to 40-fold by applying nsPEFs to cells first followed by one msEP but not in the opposite order. We demonstrated that the effect of nsPEFs on gene transfection was time restricted. The enhancement of gene expression occurred by applying one msEP immediately after nsPEFs and reached the maximum at posttreatment 5 minutes, slightly decreased at 15 minutes and had a residual effect at 1 hour. It appears that nsPEFs play a role as an amplifier without changing the trend of gene expression kinetics due to msEPs. The effect of nsPEFs on cell viability is also dependent on the specific pulse parameters. We also determined that both calcium independent and dependent mechanisms are involved in nsPEF effects on gene electrotransfer.

No MeSH data available.