以乙二醇幾丁聚醣、氧化鐵與聚吡咯製備而成之奈米團 簇應用於協同性化學動力及光熱療法

Abstract

Arising novel nanotechnologies have been shown to be exceptional in the treatment of difficult to tackle diseases such as cancer. One of the most promising technics for treatment is the use of photothermal therapy (PTT) to target cancer tissue while at the same time using the intrinsic mechanism of cancer cells to weaken the cells ahead of the treatment reducing the need of excessively high temperatures being applied for a long time. The current study presents Glycol Chitosan Polypyrrole Iron Oxide nanoclusters (GCPI NC´s) as an innovative material for the synergetic treatment of reactive oxygen species (ROS)-driven chemodynamic therapy (CDT) to inactivated HSPs with photothermal-hyperthermia achieved through the development of pH-targeting glycol chitosan/iron oxide enclosed core polypyrrole nanoclusters (GCPI NCs). In the current study Pyrrole was polymerized into Polypyrrole (Ppy) in the presence of an acidic solution of Glycol Chitosan (GC) using iron (III) chloride Hexahydrate (〖FeCl〗_3∙6H_2 O) and Iron (II) chloride tetrahydrate (〖FeCl〗_2∙4H_2 O) as oxidaxing agents. To form Glycol Chitosan Polypyrrole nanoclusters (GCP NC´s). Followed by the in situ synthesis of Iron oxide (Fe3O4) using the residual iron ions from the polymerization of Pyrrole. Taking advantage of the anionic nature of Iron Oxide and cationic nature of Glycol Chitosan to self-assemble the nanoclusters. The nanoclusters take advantage of the pH responsiveness nature of the Glycol Chitosan to enhance the targeting ability of the nanoclusters in the acidic microenvironment of the cancer cells. Similarly, the NIR heat absorbance capabilities of Polypyrrole ensure that the nanoparticle reaches temperatures were hyperthermia affects the cells. While the Iron Oxide generates ROS via Fenton reaction in the cells bringing about oxidative damage and initiates the chemodynamic therapy which in turn inactivate the heat shock proteins. The production of ROS in cancer cells was measured by dyeing with 2ʹ,7ʹ-Dichlorofluorescin Diacetate (DCFH-DA) and the fluorescence intensity was measured. While HSP´s were measured using a fluorescence microscope after applying HSP antibody dye. The effectiveness of the treatment was evaluated in vitro by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to measure the cytotoxicity in both Henrietta Lacks cell line (HeLa) and Normal Mouse fibroblast cell line (L929). Significant reduction in cell viability was exhibited after the treatment in HeLa cells and minor effect on non-cancer L929 cells. Cell experiments also displayed the targeting ability of the nanoclusters to cancer cells by fluorescence microscopy in addition of DAPI dyeing of the cells and Cyanine5 (Cy5) dyeing of the nanoclusters.