| Nanomaterial was firstly used in biomedical field as the carriers of genes and drug molecules.Since then, it has been paid extensive attention and provided versatile vector media for the drug or genetherapy, diagnose, molecular detection of human disease. Its nano effects including small size, largespecific surface area, controlled release and ease of modification, make it not only possess considerablecapacity of loading functional molecules, but also can be endowed special targeting ability to certaintissue or organ even at subcellular level. As one kind of nanomaterials, besides some propertiesdescribed above, magnetic nanomaterial has been researched and applied in many areas for itssuperparamagnetism, good biocompatibility (wildly used for contrast medium in medical imaging), andhigh efficiency of delivery. Liposome is a traditional and mature vector for gene or pharmaceutical, ithas high delivery efficiency and easy operability, however, its constraints of toxicity and transientexpression have become the bottleneck for application. This work integrated the liposome transfectagent with magnetic nanomaterial, exploring the advantages and operability of the liposomal nanomagnetic complexes used for Eukaryotic animal cell transfection. Such process using liposomal nanomagnetic complexes to transport genes is called liposomal magenetofection. This article hassystematically explored in different aspects of the liposomal magnetofection, which would layfoundation of the mammal cell multi-gene co-transfection. Some results are below:(1) Characterizations of the liposomal nano magnetic gene vectors complexes have involved themicro morphology, diameters distribution, zeta potentials and the capacity of loading DNA molecules.From atomic force microscopy, liposomal nano magnetic gene vectors complexes displayed thenanoscale assembly and formed compact fishing-net structure after combining with plasmid DNA,which may largely enhance the loading capacity of DNA molecules. Nano-zeta analysis was used todetermine the average hydrophobic diameter and ζ potentials. Average hydrophobic diameters ofcombiMAG, combiMAG/DNA, combiMAG/liposome/DNA were161.7nm,170.5nm、189.9nmrespectively; ζ potentials of liposome、liposome/DNA、combiMAG/lipo/DNA were49.4mV、24.3mV、33.9mV respectively. These data can demonstrate that liposomal nano magnetic vector complexeshave good properties of small size and positive potential, which does good to condense negative DNAsadhereing onto the cell surface and then and deliver it into the cytoplasm.(2) Optimizing of the liposomal magenetofection and fulfillment the double-gene co-transfection.The experiment on the time under a magnetic field revealed that the optimal time would be4h while theliposomal magenetofection had the highest efficiency. The cell co-transfection results showed that,co-transfected percentages in total and positive cells reached to about29.6%and71.4%respectively.(3) Through fluorescent location trafficking, the representative zones were zoomed out andmagnified to illustrate the location moving of the fluorescent MNPs at one certain areas of cells. We canclearly notice that most particles have moved from intercellular space or membrane into the cytoplasm,and some of them appeared in the nuclease at12h post magnetofection. This may lead to further research on the mechanism of cellular uptake process.Above all, those good characters of liposomal magenetofection indicate it could be a suitable wayto gene delivery, especially for cell transfection. Moreover, the facile availability, well maneuverability,less time-intensity, good biocompatibility and larger gene capacity of LMF thus could arouse moreapplication for nucleic acid delivery or multi-gene co-transfection. |
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