Targeted chemotherapy: Trastuzumab tailored Docetaxel loaded PLGA nanoaparticles for HER2 positive breast cancer
| dc.contributor.committeeMember | Haddadi, Azita | |
| dc.contributor.committeeMember | Krol, Ed. | |
| dc.contributor.committeeMember | Nazarali, Adil J | |
| dc.creator | Sadat, Sams 1984- | |
| dc.date.accessioned | 2020-02-11T21:12:36Z | |
| dc.date.available | 2020-02-11T21:12:36Z | |
| dc.date.created | 2015-06 | |
| dc.date.issued | 2015-08-17 | |
| dc.date.submitted | June 2015 | |
| dc.date.updated | 2020-02-11T21:12:36Z | |
| dc.description.abstract | This study was designed to explore the rationale of using trastuzumab (TmAb) tailored docetaxel (Doc) loaded poly-D, L-lactide-co-glycolide (PLGA) nanoparticles (NPs) to target the human epidermal receptor-2 (HER2) receptors of breast cancer cells. Utilizing emulsification solvent evaporation technique, pre-activated drug loaded NPs were prepared by embedding a homo bi-functional spacer, bis(sulfosuccinimidyl) suberate (BS3) non-covalently onto the NP surface. Freeze-dried pre-activated NPs were then decorated with TmAb followed by one step covalent attachment method. Prepared NPs were characterized for size, surface charge, polydispersity index (PDI), drug loading, drug entrapment efficiency, and antibody (Ab) quantification. All the results after physicochemical characterization were found in a desired range. The covalent attachment between TmAb and BS3 embedded NPs was confirmed by both Fourier transform infrared (FTIR) spectroscopy and Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Quantitative assay was performed to measure the amount of TmAb attached per mg of NPs. In vitro cell uptake studies were evaluated for fluorescent coumarin-6 (Coum-6) dye instead of drug loaded into the PLGA NPs with flow cytometry and confocal microscopy. Higher cellular uptake was resulted for Ab decorated NPs in both cell types which support the potential use of this formulation strategy as a targeted chemotherapeutic drug delivery system against HER2 positive breast cancer. Cell uptake of Coum-6 loaded NPs of different formulations was also observed after preincubation of both cell lines to investigate the targeting efficiency of the TmAb. Preincubation of HER2 overexpressed SKBR-3 cells with TmAb demonstrated a significant reduction in cellular uptake of Coum-6 loaded TmAb modified NPs. The level of HER2 expression was assessed for the targeted NPs comparing other NP formulations, free Doc, filtered TmAb, and Herceptin® by flow cytometry, Western blot, and fluorescence microscopy. Relative HER2 expression levels were observed for all treatment groups, however HER2 expression was lower after treating the cells with TmAb modified Doc loaded NPs. To explore the possible reasons of reduced HER2 expression and chemotherapeutic responses, cell viability and apoptosis study were considered after treating both cell lines with the formulations. Apoptosis studies revealed the chemotherapeutic responses for different formulations to the HER2 overexpressed and moderate expressed cell lines. | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | http://hdl.handle.net/10388/12632 | |
| dc.subject | HER2, Breast cancer, Nanoparticle, PLGA, Docetaxel, Chemotherapy, Trastuzumab | |
| dc.title | Targeted chemotherapy: Trastuzumab tailored Docetaxel loaded PLGA nanoaparticles for HER2 positive breast cancer | |
| dc.type | Thesis | |
| dc.type.material | text | |
| thesis.degree.department | Pharmacy and Nutrition | |
| thesis.degree.discipline | Pharmacy | |
| thesis.degree.grantor | University of Saskatchewan | |
| thesis.degree.level | Masters | |
| thesis.degree.name | Master of Science (M.Sc.) |