THE DEVELOPMENT OF ALPHA MANGOSTIN ENCAPSULATED IN CHITOSAN NANOPARTICLES CONJUGATED WITH FOLIC ACID AND ITS EFFECT ON PROLIFERATION BREAST CANCER CELL LINE MCF -7

• Main Author: Yudhasasmita, Swara
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/36089
• Institution: Institut Teknologi Bandung

Breast cancer is a disease caused by uncontroled cell growth from the mammary gland. The uncontroled cell growth is caused by mutations in the protooncogene and tumor suppressor gene. Breast cancer causes the highest death after lung, intestine, stomach and liver cancer. The treatments such as chemotherapy, surgery and radiation can cause side effect for patients. Therefore, the search for natural compound still needs to be done to overcome the adverse effects on patients. ? mangostin is a natural compound found in the skin of the mangosteen fruit. This compound has a high anticancer activity. However, this compound has a low solubility and bioavailability. To increase the bioavailability of this compound into the target cell, alpha mangostin can be encapsulated in chitosan nanoparticles conjugated with folic acid as an activated drug delivery system. The addition of folic acid ligand on the surface of chitosan nanoparticles can increase the efficiency of nanoparticles internalization by the target cells. The aim of this study is to synthesize chitosan nanoparticles conjugated with folic acid containing ? mangostin and to study the physicochemical properties, drug release and their effects on cytotoxicity, and internalization by MCF 7 breast cancer line cells. Conjugation of chitosan with folic acid is conducted using covalent linker (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide) EDC.Chitosan nanoparticles synthesis was carried out by ionic gelation method using TPP. The parameters observed were nanoparticles size & charge using zetasizer, and morphology of nanoparticles using SEM. Drug release of chitosan nanoparticles conjugated with folic acid encapsulating alpha mangostin was measured by dialysis method. Cytotoxicity of chitosan nanoparticles conjugated with folic acid containing alpha mangostin was conducted using the MTT assay method. Internalization of chitosan nanoparticles conjugated folic acid encapsulating alpha mangostin was carried out by labeling nanoparticles with Rhodamin B. Internalization was observed using a confocal microscope. The presence of nanoparticles inside the MCF-7 cells was observed by using TEM. Analysis of Ki 67 gene expression was conducted by using RT-qPCR at the mRNA level. Based on the results of the study it was known that chitosan nanoparticles conjugated folic acid and chitosan nanoparticles conjugated folic acid containing alpha mangostin were successfully synthesized and had a size of ± 120 nm, with a nanoparticle charge above ± 25 mV and spherical shape. Alpha mangostin from chitosan nanoparticles conjugated with folic acid can be released rapidly in acidic pH rather than in neutral pH. The MTT assay results showed that the value of IC50 alpha mangostin and chitosan nanoparticles conjugated with folic acid containing alpha mangostin were 13±0,46 ?g/ ml and 17,03±1,6 ?g / ml respectively. The use of nanoparticles as a drug delivery system did not reduce the IC50 value. However, the use of these nanoparticles can deliver ? mangostin directly to the target cell. Based on observations using confocal microscope, chitosan nanoparticles conjugated with folic acid which contain alpha mangostin could probably enter the MCF 7 breast cancer cell line through receptor mediated endocytosis especially using folic acid which was conjugated to chitosan. Furthermore, chitosan nanoparticles can be found in the cytoplasm instead of the endosome. Administration of chitosan nanoparticles conjugated with folic acid containing alpha mangostin can inhibit Ki-67 gene expression. This result was in line with MTT result which showed the inhibiton of proliferation in MCF-7 cells. In summary, Chitosan nanoparticles conjugated with folic acid containing alpha mangostin has been successfully synthesized and characterized. This nanoparticles can be internalized by MCF-7 cells and inhibit cell proliferation.