Studies On The Preparation,Cytotoxicity, And Gene Transfection Ability Of Carbon Nanomaterial-Derivative Chitosan Hybrids

In this paper, combining the advantages of carbon nanomaterials, chitosan (CS) and RGD peptide, we synthesized the CS NPs, CS-RGD NPs, CS-oleate NPs, Graphene-CS NPs, and Graphene-CS-RGD NP hybrids. The morphologies/microstructures, surface groups, and surface potential/particle size distribution of the materials were characterized by transimition electron microscopy (TEM), atomic force microsocopy (AFM), Fourier transform infrared spectroscopy (FTIR), and Zeta potential instruments. Additionally, cytotoxicity and transfection ability for enhanced green fluorescence protein plasmid (pEGFP-N1) of the materials on A375cells, MCF-7cells, and HeLa cells were investigated in detail. The main works are summarized as follows:1. The MWCNTs and Graphene were treated with mixed acids (concentrate HNO3:H2SO4, v/v,1:3) for48h,60h and2.5h respectively. There kinds of materials were characterized by different characterization methods. The cytotoxicity to the different cell lines and the pEGFP-N1gene transfection property of the prepared three kinds of materials were investigated in detail. The results demonstrate that the mixed acid treatment can change the micro-morphologies and surface functional groups of MWCNTs and Graphene. All the three kinds of materials can transport the pEGFP-N1into cells, and the gene molecules can be expressed successfully. The cytotoxicity of MWCNTs-COOH to HeLa cells is related to its length. The Graphene after mixed acid treatment show lower cytotoxicity than pristine Grapheme at high concentrations. The cytotoxicity of Graphene treated by mixed acid oxidation is little dependent on the cell lines.2. The CS NPs, CS-oleate NP hybrids and CS-RGD NP hybrids were prepared by ionotropic gelation process. CS NPs and CS NP hybrids were characterized by different characterization methods. The cytotoxicity to the different cells and the pEGFP-Nl gene transfection property to A375cells of CS NPs and CS NP hybrids were investigated in detail. The results of AFM and TEM suggest that the CS NPs and CS NP hybrids were successfully prepared. CS-RGD NP hybrids are positive charged in ultrapure water. All the three kinds of CS NP hybrids can transport the pEGFP into A375cells, and the gene molecules can be expressed successfully. The cytotoxicity of CS NPs and CS-RGD NP hybrids has no significant relationship with cell lines, but is dependent on the concentration of samples.3. GO-CS NP hybrids and GO-CS-RGD NP hybrids were prepared by ionotropic gelation process. Four kinds of GO-CS-RGD NP hybrids were prepared by changing TPP concentration. GO-CS NP and GO-CS-RGD NP hybrids were characterized by different characterization methods. The pEGFP gene transfection property of the GO-CS NP and GO-CS-RGD NP hybrids to A375cells were investigated in detail. The cytotoxicity to the different cells of GO-CS NP and GO-CS-RGD NP hybrids were investigated by MTT and WST-8assays. The results suggest that the best transfection efficiency of GO-CS NP and GO-CS-RGD NP hybrids was3.2%and2.8%for A375cells. When the TPP concentration is1.0mg mL-1, GO-CS-RGD NP hybrids possess largest inhibition on A375cell viability. The results of MTT and CCK-8assays suggest that the cytotoxicity of GO-CS-RGD NP hybrids has significant relationship with cell lines.