Introduction: Polymeric nano-particles having several properties represent a path towards promising future to improve the health quality using the novel nano-medical methods. The bio-degradable polymers include the synthetic polymers such as Poly (lactide) (PLA), poly (Lactic cogly colloid) Copolymers (PLGA), polycarbonate (PLC), poly (Amino acids) and the natural polymers such as chitosan, gelatin and albumin. The transfer in cells and the particles absorption are influenced by the geometry, structure, the process of creating nano-carrier and the selected materials as carriers. The characteristics including bio-compatibility, bio-degradability, the prescription method of final formulation and the drug release are required to be considered in selecting the primary materials to prepare the nano-particles.In this study, synthesis and assessment of the morphological and physical characteristics and transferring capability of nano-particle FeCO-Chitosan-PLA-PEG-FA within the cancerous cells have been carried out. The nano-particle based on FeCO-Chitosan can be used as an effective nano-carrier to transfer curing agents and drugs to the cancerous cells due to the effective transferring capability of the curing agents, the ability to move toward a magnet, and the high efficiency in the cellular absorption. Furthermore, the nano-particles of FeCO-Chitosan have been blended with PLA-PEG-FA polymers to improve the properties such as permeability, bio-compatibility, capability of long circulation in blood, and the targeted transfer of nano-particles into cancerous cells.
Methods: In order to synthesize the nano-particle FeCO-Chitosan-PLA-PEG-FA, firstly the copolymer PLA-PEG-FA was synthesized so that after synthesizing the copolymer acrylate-PLA-PEG-alkyn to connect FA to the mentioned copolymer, 0.1 mmol of the copolymer acrylate-PLA-PEG-alkyne and 0.12 mmol of folic acid adjusted by Azide group were dissolved in 30 mm of 10 mmol NH4HCO3. Then, 50 mol of ascorbic sodium and 20 mol of cuSO4 were added to the copolymer acrylate-PLA-PEG-alkyne undergoing the shake, the product was filtered using the 0.2 µm (micrometer) mesh, and after adding the appropriate quantity of NaCl, it was deposited several times by dichloro methane. Then, the nano-particle of FeCO-Chitosan-PLA-PEG-FA was synthesized by connecting the amino group in the nano-particle of FeCO-Chitosan and the alkyne group in the copolymer of acrylate-PLA-PEG-alkyne. The nano-particles were charged by the certain quantity of siRNA-FAM and their transfer into cells were traced back by Florence microscope to assess the transferring capability of the nano-particle of FeCO-Chitosan-PLA-PEG-FA into the cancerous cells.
Results: The results obtained from the NMR spectroscopy of copolymer of PLA-PEG-FA indicates validity of the synthesis of this copolymer. The observed peak at 3.6 was related to (-CH2CH2O-) of PLA-PEG. Furthermore, the observed peaks at range of 5.2 and 1.5 were related to the protons of CH and CH3 in PLA, respectively. The peaks observed at the range of 4.5 and 8.6 were related to the protons of folic acid, also the peak at 8.1 was related to the triazole ring existed in the copolymer of PLA-PEG-FA. The results of FTIR spectroscopy for the polymers of PEG, PLA-PEG, PLA-PEG-FA, and nano-particles of FeCO, the chitosan of FeCO, as well as FeCO-Chitosan-PLA-PEG-FA signified the observed peak at 1712 in PEG was related to the carboxyl group of PEG, also the peak observed at the 1100, 1647, 1762 in the copolymer of PLA-PEG were related to the groups of C=O, C-O-C and COOH in the copolymer of PLA-PEG, respectively.The emergence of the weak peaks in the range of 1580 and 1650 indicates that the connection of folic acid to the copolymer of PLA-PEG was successful. The image of the nano-particle of FeCO-Chitosan-PLA-PEG-FA from the electron microscope showed that this nano-particle has the spherical structure with the size between 20 and 40 nm. These results agrees well with the results from the DLS instrument showing that the nano-particles have the size around 28 nm and the cationic surface charge of 5.2 mV. Moreover, the photo from Florence microscope represents that the release of green color was observed in some cells treated with the nano-particles of FeCO-Chitosan-PLA-PEG-FA/siRNA-FAM signifying the transferring capability of these nano-particles in cells.
Conclusion: The assessment results showed that the synthesized nano-particles have the proper capabilities regarding the size, surface charge, charging capacity of Folic acid as well as transferring into the cancerous cells.