Synthesis and characterization of a targeted nanoniosome based on anti-CD44 antibody and investigation of its binding to breast cancer cells (MCF-7) and induction of apoptosis

Abstract: Targeted nanoniosomes are novel structures for cancer treatment and many researchers are working on it .The aim of this study was to construct and characterize a vaccine containing anti-CD44 antibody based on nanoniosomes and to investigate their binding to breast cancer cells (MCF-7) and induction of apoptosis .After preparing anti-CD44 antibody, one ml of a 5 mg/ml solution was prepared from it, then one ml of a special nanoniosome synthesis kit with a concentration of 100 mg/ml was mixed with one ml of a 1% solution of FITC fluorescent dye and mixed vigorously for 5 minutes. Then the obtained mixture was centrifuged at high speed at 10,000 rpm and the supernatant was discarded and added to the precipitate of the prepared antibody solution and mixed vigorously again for 5 minutes. After washing the nanoniosomes, they were evaluated using a DLS device to determine the size range, a TEM microscope to examine its shape, and a spectrophotometer at a wavelength of 280 nm to determine the loading rate. Then, the breast cancer cell line MCF-7 was prepared and cultured in RPMI1640 culture medium enriched with 10% FBS, and half a milliliter of nanoniosomes at concentrations of 100, 50, and 25 mg/ml were added to half a milliliter of the prepared cell suspension and incubated for 24 hours. After treating breast cancer cells with synthesized nanoniosomes, 50 microliters of the treated cells were examined under a fluorescent microscope and photographs were taken of the cells attached to the nanoniosomes. Also, apoptosis testing was performed using annexin and PI dyes by flow cytometry . This study showed that the synthesized nanoniosomes were spherical in shape and ranged in size from 85 to 180 nm with an average size of 132 nm, a DPI of 0.2, and a positive zeta potential of 30 mV. In group one, i.e., cancer cells exposed to CD44-conjugated nanoniosomes at a concentration of 100 mg/mL, the highest number of apoptotic and peri-apoptotic cells was observed, and decreasing the nanoniosome concentration had a decreasing effect on the number of apoptotic and peri-apoptotic cells. Nanoniosomes without CD44 did not have much effect on the number of apoptotic and peri-apoptotic cells. Antibody alone (group 5) was also almost similar to nanoniosomes at a concentration of 25 mg/mL (group 3). The results of the binding and entry test in different groups also showed that in group one, i.e. cancer cells exposed to CD44-conjugated nanoniosomes at a concentration of 100 mg/ml, the highest amount of stained and luminous cells (+++) was observed, and to a lesser extent, luminous cells were also seen in groups 2 and 3 .It seems that nanoniosomes conjugated with anti-CD44 antibody have the ability to induce apoptosis, but it is suggested that the efficacy of this nanoparticle be proven in vitro and in vivo with other molecular tests.