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Studies on Preparation and Characterization of Multi-walled Carbon Nanotube-Chitosan Nanoparticle Hybrids for Protein Immobilization

Author: YangKeQin
Tutor: TangHao
School: Hunan Normal University
Course: Analytical Chemistry
Keywords: Multi-walled carbon nanotubes Chitosan nanoparticles Polyamidoamine Nanocomposite materials Ionotropic gelation HeLa cell cytotoxicity BSA immobilization efficiency
CLC: TB383.1
Type: Master's thesis
Year: 2011
Downloads: 84
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Abstract


In this paper, combined the special chemical/physical properties of carbon nanotubes (CNTs) with the good biological performance of chitosan (CS) and polyamidoamine (PAMAM), the CS nanoparticles (CS NPs), multi-walled CNTs-CS NPs (MWCNT-CS NPs), CS NPs-PAMAM, and MWCNT-CS NPs-PAMAM hybrids were prepared by ionotropic gelation process. The micromorphologies of prepared hybrids were characterized by atomic force microscopy (AFM), transmission electron microscopy (TEM). The content ratio of CS in the MWCNT-CS NP hybrids was analyzed by thermogravimetric analysis (TGA). The dispersive property, cytotoxicity to HeLa cells, and bovine serum albumin (BSA) immobilization efficiency were investigated by ultraviolet-visible (UV-vis) spectroscopy, fluorescence microscopy, and MTT assay and so on. The effects of the ion concentration, adsorption time, and BSA concentration on the BSA immobilization efficiency of the CS NPs and MWCNT-CS NP hybrids were evaluated. Additionally, the micromorphologies and BSA immobilization efficiency of CS NPs-PAMAM and MWCNT-CS NPs-PAMAM hybrids were also investigated. The main work is summarized as follows:1. The CS NPs and MWCNT-CS NP hybrids were prepared by ionotropic gelation process. The morphologies of these hybrids were characterized by AFM and TEM. The effects of the concentrations of sodium tripolyphosphate (TPP) and MWCNTs on the diameter of CS NPs and the distribution of CS NPs on the surface of MWCNTs were investigated in detail. On the other hand, the content ratio of CS in the hybrids was analyzed by TGA and the dispersive property of the MWCNT-CS NP hybrids in water was evaluated. The results suggest that the CS NP particle size increase with the increase of TPP concentration. Under optimal conditions, the CS NPs were tethered onto MWCNT surface in relatively high density and with uniform coverage. The TG A results further confirm the successful coating of CS NPs on the surface of MWCNTs. The MWCNT-CS NP hybrids possess good dispersibility and stability in an aqueous suspension and no obvious accumulation of the hybrids were observed after stored 30 days at room temperature.2. The cytotoxicity to HeLa cells and protein (BSA as a model protein herein) immobilization efficiency of the MWCNT-CS NP hybrids were investigated in detail. The results demonstrate that the MWCNT-CS NP hybrids possess higher BSA immobilization efficiency (58.7%), being 1.3,4.5 times as large as those of CS NPs and carboxylated MWCNTs (MWCNT-COOH). The MTT assay and optical microscopic observation results suggest the MWCNT-CS NP hybrids exhibit better biocompatibility than MWCNT-COOH and CS NPs. In the studied concentration range (5-100μg mL-1), the hybrids show a little effect on the cellular toxicity; at the same concentration, the cytotoxicity of the MWCNT-CS NP hybrids decreases about 15,26%as compared with those of CS NPs and MWCNT-COOH.3. The CS NPs-PAMAM and MWCNT-CS NPs-PAMAM hybrids were also prepared by ionotropic gelation process and their morphologies were characterized by TEM. The BSA immobilization efficiency of the two hybrids were investigated in detail and the results suggest that under the same preparation conditions, the addition of PAMAM increase the particle size of CS NPs and MWCNT-CS NPs-PAMAM hybrids. As compared with CS NPs and MWCNT-CS NP hybrids, the CS NPs-PAMAM and MWCNT-CS NPs-PAMAM hybrids possess higher BSA immobilization efficiency.

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