RESUMO
PURPOSE: The application of nanosecond pulsed electric fields (nsPEFs) could be an effective therapeutic strategy for peritoneal metastasis (PM) from colorectal cancer (CRC). The aim of this study was to evaluate in vitro the sensitivity of CT-26 CRC cells to nsPEFs in combination with chemotherapeutic agents, and to observe the subsequent in vivo histologic response. METHODS: In vitro cellular assays were performed to assess the effects of exposure to 1, 10, 100, 500 and 1000 10 ns pulses in a cuvette or bi-electrode system at 10 and 200 Hz. nsPEF treatment was applied alone or in combination with oxaliplatin and mitomycin. Cell death was detected by flow cytometry, and permeabilization and intracellular calcium levels by fluorescent confocal microscopy after treatment. A mouse model of PM was used to investigate the effects of in vivo exposure to pulses delivered using a bi-electrode system; morphological changes in mitochondria were assessed by electron microscopy. Fibrosis was measured by multiphoton microscopy, while the histological response (HR; hematoxylin-eosin-safran stain), proliferation (KI67, DAPI), and expression of immunological factors (CD3, CD4, CD8) were evaluated by classic histology. RESULTS: 10 ns PEFs exerted a dose-dependent effect on CT-26 cells in vitro and in vivo, by inducing cell death and altering mitochondrial morphology after plasma membrane permeabilization. In vivo results indicated a specific CD8+ T cell immune response, together with a strong HR according to the Peritoneal Regression Grading Score (PRGS). CONCLUSIONS: The effects of nsPEFs on CT-26 were confirmed in a mouse model of CRC with PM.
Assuntos
Antibióticos Antineoplásicos/uso terapêutico , Morte Celular , Terapia por Estimulação Elétrica/métodos , Mitomicina/uso terapêutico , Oxaliplatina/uso terapêutico , Neoplasias Peritoneais/patologia , Neoplasias Peritoneais/terapia , Linfócitos T Citotóxicos , Animais , Neoplasias Colorretais/patologia , Terapia Combinada , Modelos Animais de Doenças , Imunocompetência , Camundongos , Neoplasias Peritoneais/secundário , Fatores de Tempo , Resultado do TratamentoRESUMO
Dual-energy mammographic imaging experimental tests have been performed using a compact dichromatic imaging system based on a conventional x-ray tube, a mosaic crystal, and a 384-strip silicon detector equipped with full-custom electronics with single photon counting capability. For simulating mammal tissue, a three-component phantom, made of Plexiglass, polyethylene, and water, has been used. Images have been collected with three different pairs of x-ray energies: 16-32 keV, 18-36 keV, and 20-40 keV. A Monte Carlo simulation of the experiment has also been carried out using the MCNP-4C transport code. The Alvarez-Macovski algorithm has been applied both to experimental and simulated data to remove the contrast between two of the phantom materials so as to enhance the visibility of the third one.