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Research on Surface Modification and Transfection of Nano-hydroxyatite as Gene Carrier

Author: ChenZheDong
Tutor: YuanTieChui；HuangSuPing
School: Central South University
Course: Materials Science and Engineering
Keywords: nano-hydroxyapatite dolyethylenimine dexametaphosphate dovineserum album dumanserumalbum dispersionstability transfection
CLC: TB383.1
Type: Master's thesis
Year: 2011
Downloads: 82
Quote: 0
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Abstract


As a gene carrier, Nano-hydroxyapatite (HAP) has many excellent features, but there are still many obstacles on the application of hydroxyapatite as gene carrier, the key issue now is to improve the dispersionstability of nano-hydroxyapatite and its ability of both carrying genes and transfection. polyethylenimine (PEI), sodium hexametaphosphate, bovine serum albumin, human serum albumin were respectively taken as dispersant in this study, HAP colloid is prepared via ultrasonic dispersion and Mechanical Stirring. The colloidal properties are detected by UV spectrophotometer, infrared spectroscopy, nano-particle size and zeta potential analyzer, transmission electron microscopy. And the modified HAP nanoparticles were experimented in biocompatibility, cell toxicity and transfection efficiency. Primary conclusions are as follows:From the analysis of HAP/PEI dispersion we found that when the PEI concentration is 4‰, the system has favorable stability; moderate acidic environment is conducive to system stability; PEI was adsorbed on the surface of HAP particles, PEI’s positive charge and the polymer chain makes the dispersion property of HAP particles improved due to the electrostatic repulsion and Spatial steric hindrance. TEM analysis shows that the dispersion property of HAP has been improved. hexametaphosphate is 4%, pH is 9.5,we acquire the stable suspension.The hexametaphosphate adsorptions at the surface of HAP particles provide a high potential which reduce congregation. TEM analysis reveals that the dispersion property of HAP has been improved.From HAP/bovine serum album suspension we lean that, under neutral environment, when the concentration of bovine serum albumin is 2.0mg/ml, the suspension obtain good dispersion stability, zeta potential and particle size analysis illuminates that the defect of bovine serum album on HAP is the effect of electrostatic repulsion and Spatial steric hindrance. The TEM analysis indicated that the dispersion property of bovine serum album adsorbed HAP was improved.The result of human serum album was similar to bovine serum album, when the concentration of human serum album is 2.0mg/ml and pH is 7.4 the suspension achieve the best dispersion stability. Zeta potential and particle size analysis points out the influence of the concentration and pH on HAP. infrared spectroscopy confirms HAP was modified because the adsorb of human serum album. TEM displays the good dispersion property.HAP modificated by PEI can load DNA effectively, and also adapt to the environment protect gene from decomposing enzyme effectively; Lower concentration of PEI result in little influence on cytotoxicity; HAP perpared in this experiment would transfect target genes into cells effectively.

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