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    Micronutrient-Enhanced Hyperthermic Intraperitoneal Chemotherapy for Treatment of Peritoneal Metastasis: A Novel Experimental Design

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    Author
    Cucher, Daniel Jeremy
    Issue Date
    2014
    Keywords
    HIPEC
    Micronutrient
    Mouse model
    Medical Sciences
    3D printing
    Advisor
    Heimark, Ronald L.
    
    Metadata
    Show full item record
    Publisher
    The University of Arizona.
    Rights
    Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
    Abstract
    Introduction: Peritoneal carcinomatosis is an end stage sequela occurring in 10% of patients with colorectal cancer. Palliative approaches have evolved over the past several decades and the role for surgical cytoreduction with hyperthermic intraperitoineal chemotherapy (HIPEC) has proven efficacy in several studies. Optimization of HIPEC therapy includes the addition of adjuncts to the carrier solution of intraperitoneal chemotherapy to improve tumor cell killing. In this study the addition of vitamin C, selenium, and quercetin ("micronutrient combination") to mitomycin C is evaluated in-vitro, and a novel murine model of HIPEC is developed using a hyperthermic chemotherapy infuser device designed de novo and printed on a 3D resin printer. Methods: HCT-116 cells were grown in culture and divided into treatment groups including: control, micronutrient combination, mitomycin C, and mitomycin C + micronutrient combination. Groups were cultured up to 72 hours after treatment and then subjected to MTT assay, crystal violet assay, trypan blue synergy assay, clonogenicity assay, cell cycle assessment by flow cytometry with propidium iodide, and western blotting for cleaved caspase-3. The infuser device was designed in a CAD environment, printed on a 3D resin printer, and underwent fluid temperature stability analysis and flow experiments by infusing methylene blue into live mice followed by necropsy and analysis of dyeing patterns. Results: MCC treated cells proliferated at 32.7%, and tumor cells treated with MCC + MNC carrier solution proliferated at 27.3%. Normothermic MCC and the MNC alone caused a 26.8% and 33.3% reduction in cell survival, and MCC delivered to cells in the micronutrient combination solution decreased cell survival by 53.2%. 95.3% and 99% of cells treated with MCC or MNC alone demonstrated viability, and 85% of cells treated with MCC + MNC demonstrated short term viability, suggesting synergy. HCT-116 clonogenicity is disrupted by MCC and MNC individually, and nonexistent in the MCC + MNC treatment group. Cleaved caspase-3 mediated apoptosis is upregulated by MCC, and by MNC to a lesser extent. Flow cytometry apoptosis demonstrates increased S-phase cell cycle arrest in the MCC + MNC sample. The mouse infuser HIPEC apparatus demonstrated an thorough distribution of blue dye in predictable regions of the abdomen with an acceptable range of hyperthermic regulation.
    Type
    text
    Electronic Thesis
    Degree Name
    M.S.
    Degree Level
    masters
    Degree Program
    Graduate College
    Medical Sciences
    Degree Grantor
    University of Arizona
    Collections
    Master's Theses

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