Prezentare

1) With more than 14 million new cases and 8.8 million deaths per year worldwide, cancer
remains one of the most dangerous threats to human health. Currently, surgical
intervention is a main method for cancer therapy, and chemotherapy has been increasingly
refined, resulting in great progress in treating cancer. However, chemotherapy alone usually
causes severe side effects, as chemotherapeutic drugs can kill not only cancer cells but also
normal cells. An improvement strategy in the use of chemotherapeutic drugs is the use of a
targeted drug delivery system, which can reduce therapeutic doses and transport
therapeutic drugs directly into the tumor tissue under various conditions such as external
magnetic field or ultraviolet or infrared illumination, thus reducing side effects.
Breast cancer is the most frequent cancer among women, with 2.1 million new cases each
year
At the moment, certain oxide nanoparticles are used both for diagnostic and therapeutic
purposes.

2) Thus, we choose to test effectiveness of a nanoparticles based drug delivery system for a
chemotherapeutic anthracyclinic (violamycin B1) on a tumorigenic standardized cell lines of
breast adenocarcinoma MCF7, but also in comparison with the normalized breast cell line
(MCF12A). We delivered the drug on a system of iron oxide nanoparticles coated by L-DOPA
and compared in preliminary tests cellular parameters such as: cytotoxicity, viability, cell
proliferation rate and the phenomena of cellular up taking nanoparticles and chemotherapy
by videomicroscopy.

3) The drug delivery system was obtained by laser pyrolysis, having an optimum size so
being to small and can be filtered renal or above 200 nm is phagocytosed by macrophages.
Our system formes clusters having maximum dimensions of 100 nm. Also, by coating the
systems in L-DOPA, the positive charges are neutralized, but at the same time the rate of
uptake of drugs for MCF7 increases - breast adenocarcinoma overexpresses the LAT1
protein responsible for the cellular uptake of L amino acids such as L-DOPA, wich was
observed in other cancer also

4) Viability was measured by the MTS method, wich measures the oxidative-reducing
capacity of enzymatic systems such as NADH, being an end poit method. Cytotoxicity was
measured by the LDH method of care by measuring the amount of lactate dehydrogenase
enzyme excreted by the cells in the environment. This two methods evolves differently, one
going up and one going down, being . Thus, it is observed that the cytotoxicity of
the nanoparticles coated with L-DOPA is lower than of the uncoated ones, whereas the
viability of the cells was higher in the case of NPs coated with L-DOPA. These results can be
seen on both MCF7 and MCF12A.

5) Also, preliminary tests have shown there may be a significant difference in cytotoxicity
and viability when nanoparticles were coated by violamycin. Tumor cells are found to be
more affected than non-tumor cells, with lower viability and higher cytotoxicity. It is
important because adenocarcinoma cells in vivo will be surrounded by normal breast cells.

6) To see the anti-proliferative effects of the NP system with violamycin for the use of the
xCELLIGENCE platform, wich measures cell impedance. It can be seen that the drugs that we
delivere are clearly antiproliferative, reusing the tumor cells detaching form the plate after
72 hours. When the cell index is zero it means that the cells have detached and no longer
make contact with electrodes, so we need to objectify cell death, but also to be sure about
the cellular up-take of the system.

7) Thus, these things are objectified by videomicroscopy for 72 hours Biostation system,
using a specific lysosome dye, which is the desdination of our delivery sistem. We can see
the uptake by MCF7 cells and survival rate in the presence of nanoparticles coated or
uncoated with L-DOPA, but when the system was loaded with violamycin it is seen that
more than half of the cells die after 24 min. , confirming the delay of approximately 10
mines between free and loaded violamycin on delivery system.

8) In conclusion, L-DOPA coating NPs reduce cytotoxicity on both cell lines; increasing the
specificity, affecting more adenocarcinoma cells, compared to normal cells, affecting also
proliferation rate more on MCF7 than on MCF12A. Therefore, in the future this system of
nanoparticles with L-DOPA and VB1 which can be used to treat the treatment of breast
cancer, but also other solid tumors.