Biological Effects Of Nano-water Suspension Of Fullerene Phosphonic Acid Derivatives

So far, the research of fullerene has come down to many subjects and applied fields, such as physics, chemistry, materials science and life science. Recently, there are several papers reporting the biological activities including cytotoxicity, genotoxicity and anti-bacteria of fullerene nanoparticle suspension, which draws a wide attention.In this dissertation, preparations and biological activities of aqueous suspension of three fulleren phosphonate derivatives, MMPF,BMPF and C₆₀>C(PO₃Et₂)₅ have been described. The results are presented as follows:1. Nanoparticle suspensions, including n-MMPF,n-BMPF,n-C₆₀>C(PO₃Et₂)₅ were made successfully and the evaporation-dialysis method was developed to remove the organic solvent resident which couldn't be detected by UV-Vis spectra . The characterization of those fullerene nanoparticle suspensions were characterized by UV-Vis, zeta-potential, dynamic light scattering (DLS) and transmission electron Microscopy (TEM). The results showed that the properties of those nanoparticle suspensions were related to the materials.In addition, TEM study showed a good shape and well dispersion.2. An aqueous nanoparticle suspension of a bis-methanophosphonate fullerene(n-BMPF) was tested to clarify its effects on DNA restrictive endonucleases BamHâ… ,EcoRâ… å'ŒCfr9â… and Xmaâ… and the template of plasmid pEGFP-N1 with single restrictive site for all the enzymes. It was found that the products is different among the different restricive reactions. The inhibition by the n-BMPF was dose-dependent and IC₅₀ values for reactions of EcoRâ… were 4.3μmol/L, but the IC₅₀ values for reactions of BamHâ… was higher than 30μmol/L. It could be concluded from tne inhibition of Cfr9â… and Xmaâ… that the correlation between the inhibition and the content of G/C of the DNA identified sequence was not the linerity. When added more pEGFP-N1, the linetype plasmid still could not be seen, the results suggested that the it was not a competed inhibition, it may be a non-competed and absorptive inhibition. Addition of two scavengers of reactive oxygen species (ROS), mannitol and sodium azide at the concentrations of 2―10 mmol/L did not antagonize the activities of the n-BMPF against reaction. However, increase of EcoRâ… amounts in the reaction antagonzied the activities of the n-BMPF .These data implied that this inhibition probably did not correlate to ROS. Meanwhile, Activites of the aqueous nanoparticle suspension of a mono-methanophosphonate fullerene(n-MMPF) and the other new methanophosphonate fullerene on DNA restrictive endonucleases EcoRâ… and the template of pEGFP-N1.It was suggested that the inhibition lied on the structure of the fullerene derivatives. Finally, the n-BMPF was tested clarify its effects on polymerase chain reaction (PCR) with the catalyst of Taq DNA polymerase and DNA exonuclease Exoâ…¢. The inhibition by the n-BMPF was dose-dependent and IC₅₀ values for reactions of PCR were 2.7μmol/L. Increase of Taq DNA polymerase amounts in PCR system antagonized the activities of the n-BMPF. However, Addition of two scavengers of reactive oxygen species (ROS), mannitol and sodium azide at the concentrations of 2―10 mmol/L did not antagonize the activities of the n-BMPF against PCR. These data implied that this inhibition probably did not correlate to ROS. Meanwhile, the inhibition for the DNA exonuclease Exoâ…¢by the n-BMPF was evident and dose-dependent.3. The interaction of three kinds of phosphonate fullerene derivatives with two kinds of blood proteins BSA,BHb was studied by UV difference spectra respectively. Then, the combination of BSA to three kinds of phosphonate fullerene derivatives was investigated by using TCA deposition method. It showed that there was no function between the blood proteins and three kinds of phosphonate fullerene derivatives.