Echogenic liposomes for ultrasound-triggered drug delivery
- Authors: Izuchukwu, Ezekiel Charles
- Date: 2021-10
- Subjects: Liposomes , Drug delivery systems , Colon (Anatomy) Cancer Treatment , Transmission electron microscopy , Fourier transform infrared spectroscopy , Liquid chromatography , Echogenic liposomes , Ultrasound-triggered drug delivery
- Language: English
- Type: Masters theses , text
- Identifier: http://hdl.handle.net/10962/188997 , vital:44805
- Description: Colorectal cancer is one of common cancers worldwide. It is the third most diagnosed cancer and the second leading cause of death. The use of 5-fluorouracil (5-FU) alone or in a chemotherapy regime has been the effective treatment of colorectal cancer patients. The efficacy of 5-FU in colorectal cancer treatment is significantly limited by drug resistance, gastrointestinal, and bone marrow toxicity through high-level expression of thymidylate synthase, justifying a need to improve its therapeutic index. Liposomes are colloidal membranes comprising of one or more lipid bilayers enclosing an aqueous core. They have been used to improve the therapeutic index of many anti-cancer drugs by changing drug absorption, elongating biological half-life, reducing metabolism, and reducing toxicity to healthy tissues. Echogenic liposomes are specifically designed to respond to external triggering like ultrasound stimulation by entrapping a gas or an emulsion that can vaporize. A liposome's unique property is that it can entrap both hydrophobic and hydrophilic substances simultaneously in the lipid bilayer and the aqueous core, respectively. These stimuli-responsive liposomes can be triggered externally with ultrasound, to release the chemotherapeutic cargo only at the required site. This research aims to formulate echogenic liposomes encapsulating 5-FU for potential ultrasound triggered release (echogenic). Liposome formulations wereprepared with lipid composition of crude soybean lecithin and cholesterol by thin-filmhydration method and the drug was passively loaded in the formulation. The 5-FU loadedliposomes were evaluated by dynamic light scattering (DLS) for particle size, polydispersityindex, and zeta potential and transmission electron microscopy (TEM) for morphology.Encapsulated liposomal formulations were also evaluated using physicochemical techniquesincluding thermogravimetric analysis (TGA), differential scanning calorimetry (DSC),Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). Theencapsulation efficiency and release kinetics were studied using a validated high-performanceliquid chromatography (HPLC) method. Echogenic properties were explored by entrapping abiocompatible gas (argon) at the same time as the drug (5-FU) using a pressure/freezemethodology. The liposomal formulations were typically spherical with a size of about 150 nmand encapsulation efficiency of 62%. Low-frequency ultrasound (20 kHz) was used to triggerthe drug release from the complete formulation at 10%, 15%, and 20% amplitude and exposuretime of 5 min and 10 min. The rate of drug release from the nano-carrier was a function of theultrasound amplitude and exposure time and reached a maximum of 65% release under theconditions investigated. The cumulative release was investigated, with and without theapplication of ultrasound. It was demonstrated that the application of ultrasound resulted in complete release (99%) after 12 h while this dropped to 70% without ultrasound. These results are encouraging for optimizing ultrasound parameters for triggered and controlled release of the 5-FU, for conditions such as the management of cancer where low-power ultrasound can be applied. , Thesis (MSc) -- Faculty of Science, Chemistry, 2021
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- Date Issued: 2021-10
Analysis of the role of Hsp90 in colon cancer and cancer stem-like cell biology in vitro using a genetically paired cell line model
- Authors: Slater, Cindy
- Date: 2015-04-09
- Subjects: Heat shock proteins , Heat shock protein Hsp90 family , Colon (Anatomy) Cancer Treatment , Cell lines , Cancer stem-like cell , SW480 , SW620
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/480345 , vital:78433
- Description: Colon cancers are commonly associated with mutations or changes in signaling in the Wnt/β-catenin pathway. Cancer treatments, such as chemotherapy and radiation, are challenged by the limited methods of disease detection and the insufficient elimination of contributing factors, such as cancer stem-like cells (CSC), to metastatic disease. CSC are characterised by their ability to survive anchorage-independently and attribute to therapeutic resistance. To examine the biological changes associated with the progression of human colon cancers and the role of Hsp90 in cancer and CSC biology, SW480 and SW620 genetically paired (isogenic) colon cancer cell lines from the same patient were characterised for populations of putative CSC, tumoursphere (TS) forming ability, cell growth, behaviour and response to anti-cancer therapeutics. The SW480 cell line was established from a primary colon adenocarcinoma and the SW620 was established from a lymph node metastasis of the primary cancer one year later. To address the role of Hsp90 in colon cancer, the sensitivity of cells and TS were analysed in response to geldanamycin and novobiocin, and an isoform-specific approach to the targeting of Hsp90α was developed. Flow cytometric analysis of putative CSC by phenotype revealed variable proportions of cells bearing the CD44+/CD133+ surface protein marker, widely used in the identification of colon CSC, in SW480 and SW620 cells. The paired cell lines maintained similar proportions of putative CSC, identified by the expression of the ABCG2 protein (side population; 1 %) and through high aldehyde dehydrogenase activity (ALDEFLUOR; 6 %). SW480 cells demonstrated greater TS forming efficiency than SW620 cells (49.9 and 35.5 %, respectively) and observations of wound-healing showed SW480 cells to be more migratory than SW620 cells. No difference in response to Hsp90 inhibition was observed between paired cell lines, however SW480 TS resisted treatment with geldanamycin. This the first study to report a dose-dependent increase in TS growth in response to novobiocin inhibition of Hsp90, and to demonstrate that that the sensitivity of SW480 and SW620 TS to oxaliplatin, a common drug for the treatment of metastatic colon cancers, was enhanced by novobiocin, providing promise for the elimination of CSC with combined chemotherapeutics. We analysed the Wnt/β-catenin pathway in response to expression of short hairpin RNA (shRNA) against Hsp90α or a control non-targeting shRNA, under the control of a tetracycline-responsive promoter. Hsp90α knockdown contributed to a deregulated stress response, presenting with reduced Hsp27 and β-catenin protein, but corresponded to an increase in the association between Hsp90, β-catenin and Hsp27 in vitro. The reduction of Hsp90α did not influence sensitivity of the colon cancer cells to activators or inhibitors of the Wnt pathway, but rather correlated to reduced TS formation, cell adhesion and spreading, identifying potential therapeutic benefit to the controlled reduction of Hsp90α for the deregulation of colon cancer characteristics. Given that Hsp27 and β-catenin are both involved in cell adhesion, cytoskeletal dynamics and interact directly with each other, we propose a role for targeting Hsp90α in the regulation cell adherence indirectly via reductions in levels of β-catenin and Hsp27, rather than by modifying the transcriptional activity of β-catenin. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2015
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- Date Issued: 2015-04-09