The role of pH fronts in tissue electroporation based treatments

PLoS One. 2013 Nov 21;8(11):e80167. doi: 10.1371/journal.pone.0080167. eCollection 2013.

Abstract

Treatments based on electroporation (EP) induce the formation of pores in cell membranes due to the application of pulsed electric fields. We present experimental evidence of the existence of pH fronts emerging from both electrodes during treatments based on tissue EP, for conditions found in many studies, and that these fronts are immediate and substantial. pH fronts are indirectly measured through the evanescence time (ET), defined as the time required for the tissue buffer to neutralize them. The ET was measured through a pH indicator imaged at a series of time intervals using a four-cluster hard fuzzy-c-means algorithm to segment pixels corresponding to the pH indicator at every frame. The ET was calculated as the time during which the number of pixels was 10% of those in the initial frame. While in EP-based treatments such as reversible (ECT) and irreversible electroporation (IRE) the ET is very short (though enough to cause minor injuries) due to electric pulse characteristics and biological buffers present in the tissue, in gene electrotransfer (GET), ET is much longer, enough to denaturate plasmids and produce cell damage. When any of the electric pulse parameters is doubled or tripled the ET grows and, remarkably, when any of the pulse parameters in GET is halved, the ET drops significantly. Reducing pH fronts has relevant implications for GET treatment efficiency, due to a substantial reduction of plasmid damage and cell loss.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Dogs
  • Electrochemotherapy / methods*
  • Electroporation*
  • Fuzzy Logic
  • Hydrogen-Ion Concentration*

Grants and funding

This work was supported by grants from: CONICET (PIP 1087/09), Universidad de Buenos Aires (UBACyT X132/08), ITBACyT and Argentina-Slovenia Scientific Collaboration Project (SLO0802/2008). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.