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A Study of Targeting Distribution in Injured Spimal Cord Region with TAT-mediated Magnetic Liposomes

Author: WeiZhiJian
Tutor: FengShiQing
School: Tianjin Medical University
Course: Surgery
Keywords: Spinal cord injury Magnetic nano- liposomes TAT Schwann cells Targeting
CLC: R651.2
Type: Master's thesis
Year: 2011
Downloads: 38
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Abstract


[Objective] Preparation and Characterization of a FITC-labeled TAT-mediated magnetic nano-liposomes in vitro and in Schwann cells co-cultured with AR coating capacity; while observing their spinal cord tissue after spinal cord injury in the local gathering to study the distribution of rat spinal cord damage targeting, determined to provide experimental evidence for the liposome-mediated drug treatment of spinal cord injury. [Materials and Methods] In this study, inverting evaporation method both transmembrane a long circulatory function and superparamagnetic Fe304 particles coated with hydrophobic the TAT magnetic nano-liposomes, transmission electron microscopy, particle size analyzer and zeta potential was measured liposome particle size and characterization. Saphenous nerve ligation Wistar adult rats with unilateral activated Schwann cells, the dual enzymatic digestion block method is a combination of mechanical separation isolated and cultured Schwann cells; application of low concentrations of collagenase, trypsin fast digestion and purified by differential adhesion method Schwann cells. S-100 antibody and DAPI staining to identify the purity of the Schwann cells. Schwann cells and the external the TAT magnetic nano liposome and missed TAT magnetic nano-liposomes were incubated in the control group to join magnetic nano liposome (59μg/ml) 0.5ml missed TAT: experimental group on external TAT magnetic nano-liposomes (59μg/ml) 0.5ml, 5% CO2, 37 ℃ were incubated for 1 hour, the Schwann cell uptake of liposomes was observed under a fluorescence microscope. In this study, 16 female Wistar rats, Impactor Model-II blow T10 spinal cord injury (spinal cord injury, SCI)) model (10g × 25mm). After the modeling were randomly divided into 2 groups, respectively, within 24 hours via the tail vein injection of FITC-labeled magnetic nano-liposomes (1mg/kg), the control group was injected missed TAT magnetic nano-liposomes, the experimental group external TAT the magnetic nanoliposomes 1 hour after injection the rats were sacrificed, remove the spinal cord tissue for 5 Frozen section liposomes gathered in the spinal cord tissue was observed under a fluorescence microscope, while taking in rat liver, spleen, kidney and frozen Slice and liposome uptake by other organs was observed under a fluorescence microscope. Adult female Wistar rats were 16, were randomly divided into two groups, the control group did not for the treatment of spinal cord injury, only to take the same dose of anesthesia, the experimental group to establish a model of spinal cord injury, the two groups of animals were injected in the tail vein route external TAT magnetic nano liposomes (1mg/kg), the point in time of the experimental group is selected as 24 hours after injury, the time required to increase production model of the control group 24 hours after anesthesia. 1 hour after injection the rats were sacrificed, remove the spinal cord tissue for 5 Frozen section liposomes gathered in the spinal cord tissue was observed under a fluorescence microscope. [Results] 1. Magnetic lipid generally spherical, well dispersed, smaller particle size, Zeta potential, no significant difference in external TAT group and the the missed TAT group of particle size and zeta potential. 2 activated Schwann cells were separated and cultured after purification, cell growth is strong. Transmitted to the 4th generation, low magnification almost no fibroblast-like cells. Schwann cell purity of 95%. Cellular uptake experiments external TAT liposomes gathering inside the Schwann cells, and most of them gathered around the nucleus, without access TAT ??liposomes less into the Schwann cells, the comparison of both microscopic FITC fluorescence average optical density (average optical density, AOD), both statistically significant. In the blue fluorescence excitation of fluorescence microscopy, FITC magnetic nano-liposomes showed green fluorescence. In the injured spinal cord tissue, external TAT liposomes more gathered around the spinal cord injury, neuronal cell cytoplasm there are a large number of fluorescent particles gathered into a sheet showing the tendency of nuclear, without connected TAT liposomes is less distributed in the damaged spinal cord. Calculated both AOD values, the results were statistically significant. In the blue fluorescence excitation of fluorescence microscopy, missed the TAT group's liver, spleen, kidneys are visible bright green fluorescence, external TAT green fluorescent relatively weak. Blue fluorescence excitation of fluorescence microscopy, normal rat spinal cord tissue less green fluorescence, and gathered a lot of green fluorescence in the spinal cord, spinal cord injury in rats, calculated both AOD values, the results were statistically significant. [Conclusion] This study demonstrates that TAT-mediated liposome can be poured into the damaged spinal cord, the neurons, and has targeted gathered damaged spinal cord tissue characteristics, its loading the treatment of spinal cord injury drug carrier Feasibility basis.

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CLC: > Medicine, health > Surgery > Of surgery > Head and Neurosurgery > Spinal cord
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