Preparation Of Iron-Containing Polymer Nano-Materials And Radiation Therapy For Cell Model

ObjectiveTo get the iron-containing polymer nanomaterial-- polyethylene glycol- ferrocene nanoparticles(mPEG-Fc NPS) from The ferrocene(C11H10FeO2) acid surface-modified polyethylene glycol(mPEG),and to study intake capacity of the nanoparticles to breast cancer tumor cells; also to study the effect of mPEG-Fc NPS as a radiosensitizer on the breast cancer cell(4T1 cell) under the irradiation of X-ray.Method1.The experimental materials were represented by Transmission Electron Microscopy(TEM),Fourier Transform Infrared(FT-IR) Spectroscopy,Dynamic Light Scattering(DLS),Nuclear Magnetic Resonance(NMR) Spectroscopy,Matrix-assisted laser desorption/ionization time-of-flight(MALDI-TOF) mass Spectroscopy and so on.2.Based on the principle of recombination of nanoscale with “enhanced permeability and retention effect(EPR) [1-2]”,the 4T1 cells were incubated for 24 hours with polyethylene glycol-ferrocenen nano particles(mPEG-Fe NPS) of different concentration,the capability of cell to ingest mPEG-Fe NPS of different concentration had been tested by the method of inductively coupled plasma emission spectrum(ICP-OES).3.The working concentration of mPEG-Fc NPS having a good cytocompatibility had been selected by CCk-8 Assay,it was regarded as a safe concentration;by the use of cell clonal formation test and CCk-8 Assay,the ability of mPEG-Fc NPS to radiosensitize the breast cancer cell(4T1 cell) had been studied;its sensibilization effect had been verified by live-dead experiment and testing cellular free radical.Result1.The findings of TEM:mPEG-Fc NPS has a good dispersity,about 100 nm in diameter,mainly distributed within 80-120nm;after exposure to X-ray,it also remained a good dispersity and size.2.Tthe findings of CCk-8 Assay:when the concentration of mPEG-Fc NPS was 0.80mmol/L,its cellular relative growth rate was about 65.88%,which had an obvious cytotoxicity,with the decline of concentration of mPEG-Fc NPS,the relative growth rate of 4T1 cell gradually went up,the cellular relative growth rate reached to 84.62% at the concentration of 0.60mmol/L,mPEG-Fc NPS had little influence on the growth of 4T1 cell and a good cytocompatibility.3.Findings of Matrix-assisted laser desorption/ionization time-of-flight(MALDI-TOF) mass Spectroscopy:as the time marched on,the quantity of mPEG-Fc NPS ingested by cells gradually increased,in the case of same concentration,as compared with the control group(pure ferrocenecarboxylic acid),the quantity of iron from mPEG-Fc NPS entering the cells clearly increased.4.It was shown by CCk-8 Assay that,under the irradiation of 4Gy of X-ray,mPEG-Fc NPS had a clear effect of reducing the cellular relative growth rate and showed an effect of radiosensitization. Meanwhile,the 4gy X- ray exposure, the concentrations of mPEG-Fc NPS were 0.1,0.2,0.6 NPS also a drop in the cell proliferation rate trend.5.Findings of clonal experiment:the difference in cell clonal formation rate between the experiment group and control group was significant when the mPEG-Fc NPS with a concentration of 0.60mmol/L was irradiated by 4Gy X-ray.6.It was shown by the result of DCFH-DA assay that,with the exposure to 4Gy X-ray radiation,Fc-COOH and mPEG-Fc NPS can induce the elevated intracellular level of ROS,but mPEG-Fc NPS can induce a clearly elevated intracellular level of ROS.7.It was shown by Annexin V-PE/7-AAD apoptosis Detection kit test tha,mPEG-Fc NPS hardly had any cytotoxicity tests before exposure to X-ray radiation,meanwhile,after exposure to X-ray,the radiosensitivity of 4T1 cells had been enhanced.Conclusion1.mPEG-Fc NPS had a good dispersity and homogeneity.2.mPEG-Fc NPS enhanced the ability of cell to ingest Fe.3.mPEG-Fc NPS showed an excellent radiosensitization effect,and it was shown by the experimental result that,the sensibilization of mPEG-Fc was clearly positively correlated with the dose of X-ray irradiation and concentration of poly-mPEG-Fc NPS.