American Society for Laser Medicine and Surgery Abstracts

A DOSE-RESPONSE STUDY OF A NOVEL NON-THERMAL METHOD OF SELECTIVELY MODIFYING CELLULAR STRUCTURES IN SKIN WITH LOW ENERGY NANOSECOND ELECTRICAL STIMULATION David Kaufman, Michelle Martinez, Brian D. Zelickson, Edward A. Ebbers, Richard Nuccitelli, Lauren Jauregui, Darius Mehregan Kaufman Plastic Surgery, Inc., Folsom, CA; Zel Skin and Laser Specialists, Edina, MN; Pulse Biosciences, Inc., Hayward, CA; Wayne State University School of Medicine, Detroit, MI Background: A novel modality for modifying cellular structures of the epidermis and dermis through the application of a nano-pulse stimulation (NPS) electrical energy device to normal skin was evaluated in a first-in-human clinical study of dose-response effects. Study Design/Materials and Method: Five patients with healthy tissue planned for abdominoplasty excision were enrolled in a 60-day study of effects of a novel method for modifying tissue using low energy, high voltage NPS. A total of 30 squares of 25mm 2 or less within the planned excision area were treated at 60 days, 30 days, 15 days, 5 days and 1 day prior to surgery, using 6 progressively higher NPS energy levels. Photographs were taken of each treated area. Five different staining methods were used to assess tissue changes. Results: The majority of test sites exhibited delayed epidermal loss followed by re-epithelialization by Day 15 and a normal course of healing. Histologic analysis identified a nucleolysis effect on epidermal cells. Minimal effects were seen on melanocytes, elastic fibers and collagen when compared with controls at day 60 in the majority of evaluated tissue samples, with notable exceptions of 2 tested areas treated at the higher energy levels that showed signs of dermal damage. Conclusion: The selective effect of NPS on cellular structures in the epidermal and dermal layers suggests a non-thermal mechanism for targeting cellular structures that does not affect the viability of dermal tissue within a range of energy levels. These histology results indicate that the lowest effective energy level was not established in this study. The specificity of effects and a favorable healing response lends itself to cellular targets in the epidermal or dermal layers of the skin, including treatment of benign and non-benign lesions. NPS skin treatments provide a promising, non-thermal method for treating skin conditions and removing epidermal lesions. EXTRACELLULAR VESICLE EXOCYTOSIS ARE UPREGULATED IN PORT WINE STAIN BLOOD VESSELS Rong Yin, Wenbin Tan, Shawn Rice, Xin Liu, J. Stuart Nelson Beckman Laser Institute and Medical Clinic, University of California, Irvine, CA; Penn State Cancer Institute, Hershey, PA Background: Port wine stain (PWS) is characterized by a progressive dilatation of immature venule-like vasculatures which result from differentiation-impaired endothelial cells. In this study, we aimed to identify the major biological pathways accounting for the pathogenesis of PWS. Study Design/Materials and Method: Sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS) was used to identify differentially expressed proteins in PWS lesions, followed-up by confirmative studies with immunohistochemistry, immunoblot and Transmission Electron Microscopy (TEM). Results: One hundred seven out of 299 identified proteins showed differential expressions in PWS lesions as compared to normal skin, mainly involving the functions of biosynthesis, membrane trafficking, cytoskeleton and cell adhesion/migration. The confirmative studies showed that expressions of cell adhesion/ migration/exocytosis related proteins such as VAT1, IQGAP1, HSC70, clathrin, perlecan, spectrin a1 and GDIR1 were significantly increased in PWS blood vessels; while collagen subtypes 6A1 and 6A3 were decreased in PWS skin. Furthermore, TEM studies showed there is a significant upregulation of extracellular vesicle exocytosis from PWS blood vessels as compared to control. Conclusion: Our results showed that (1) PWS blood vessels have increased expressions of cell adhesion/migration/exocytosis related molecules; (2) extracellular vesicles show increased exocytosis from PWS blood vessels, which carry and mediate critical intercellular signaling in the pathogenesis of PWS; and (3) dysregulation of collagen 6 contributes to collagenous alterations in PWS skin.