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Histopathology of normal skin and melanomas after nanosecond pulsed electric field treatment

Chen, Xinhuaa b; James Swanson, R.b c; Kolb, Juergen F.b; Nuccitelli, Richardb; Schoenbach, Karl H.b

doi: 10.1097/CMR.0b013e32832f1558
ORIGINAL ARTICLES: Translational research

Nanosecond pulsed electric fields (nsPEFs) can affect the intracellular structures of cells in vitro. This study shows the direct effects of nsPEFs on tumor growth, tumor volume, and histological characteristics of normal skin and B16-F10 melanoma in SKH-1 mice. A melanoma model was set up by injecting B16-F10 into female SKH-1 mice. After a 100-pulse treatment with an nsPEF (40-kV/cm field strength; 300-ns duration; 30-ns rise time; 2-Hz repetition rate), tumor growth and histology were studied using transillumination, light microscopy with hematoxylin and eosin stain and transmission electron microscopy. Melanin and iron within the melanoma tumor were also detected with specific stains. After nsPEF treatment, tumor development was inhibited with decreased volumes post-nsPEF treatment compared with control tumors (P<0.05). The nsPEF-treated tumor volume was reduced significantly compared with the control group (P<0.01). Hematoxylin and eosin stain and transmission electron microscopy showed morphological changes and nuclear shrinkage in the tumor. Fontana–Masson stain indicates that nsPEF can externalize the melanin. Iron stain suggested nsPEF caused slight hemorrhage in the treated tissue. Histology confirmed that repeated applications of nsPEF disrupted the vascular network. nsPEF treatment can significantly disrupt the vasculature, reduce subcutaneous murine melanoma development, and produce tumor cell contraction and nuclear shrinkage while concurrently, but not permanently, damaging peripheral healthy skin tissue in the treated area, which we attribute to the highly localized electric fields surrounding the needle electrodes.

aDepartment of Hepatobiliary Surgery, the First Affiliated Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, China

bFrank Reidy Research Center for Bioelectrics

cDepartment of Biological Sciences, Old Dominion University, Norfolk, Virginia, USA

Correspondence to Dr Xinhua Chen, MD, PhD, Department of Hepatobiliary Surgery, the First Affiliated Hospital Zhejiang University School of Medicine, 79 Qingchun Road, Hangzhou, Zhejiang 310000, China

Tel: +1 757 683 2439; e-mail:

Received 26 February 2009 Accepted 29 May 2009

© 2009 Lippincott Williams & Wilkins, Inc.