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Study on the Nanoparticles of Norcantharidin-hydroxypropyl Chitosan and Its in Vitro Anti-cancer Activity

Author: FengZuoZuo
Tutor: ChenZhiWei
School: Shandong University of Technology
Course: Biochemistry and Molecular Biology
Keywords: norcantharidin hydroxypropyl-chitosan nanoparticles controlled-release anti-cancer
CLC: R94
Type: Master's thesis
Year: 2012
Downloads: 82
Quote: 1
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Purpose: In this paper, the Norcantharidin-Hydroxypropyl-Chitosan-Nanoparticles (NCTD-HPCS-NPs) were prepared from Hydroxypropyl-Chitosan (HPCS), which was made of Chitosan. In addition, the in vitro release characteristics and antineoplastic activities of NPs were evaluated respectively, which was compared with NCTD under the same treatment conditions.Methods:HPCS was synthesized by the method of alkalization and crosslinking. The NCTD-HPCS-NPs were prepared by ionic crosslinking method with this glycan, and each method was optimized by the orthogonal test. Character of NPs was detected by scanning electron microscope, and the in vitro drug-release of was measured by dialysis method, which was compared with NCTD. Finally, the in vitro anti-tumor activity of NCTD and NCTD-HPCS-NPs were tested by MTT assay, and the IC50of each dosage form was calculated with Origin8.0software.Results:1. The results of preparation for HPCS showed that the best reaction conditions were as follows:the ratio of Mass (chitosan):Volume (propylene oxide):Volume (tetramethylammonium-hydroxide) were1:10:0.5, the temperature was60℃, and reaction time was6h. Under these conditions, the degree of substitution(DS) of HPCS was0.42on average with a better solubility of7.5g/L.2. The results of preparation for NPs showed that when the concentration of NCTD was2.0mg/mL, HPCS was1.Omg/mL and tripolyphosphate(TPP) was1.Omg/mL. The product of NCTD-HPCS-NPs was in the best conditions, and the size was95.15±3.81nm. Entrapment efficiency(EE) was23.68±1.79%and drug-loading(DL) was54.53±2.61%on average.3. The in vitro drug release assay showd that as the control group, release of NCTD was last for3hours, and no more than3.5hours in the three release medium contained0.1MHCI, PBS(pH=6.8), PBS (pH=7.4). Compared with NCTD, release of NPs was6hours for a longer time in the same conditions. One hour later, it became controlled-release obviously, and the totally capacity was80%at most.4. The results of MTT tests showed that which dealed with NCTD&NPs for24h compared with the normal group. Under this condition, the inhabitation ratio(IR) of NCTD was just little higher than NPs in the low drug-concentration group(CNCTD=10μg/mL). The IR of NPs was better, especially the IR was between160and640μg/mL, and the IR of NPs could be66.46%in totally in the middle concentration group(20μg/mL≤CNCTD≤80μg/mL). According to the results of Origin8.0software, IC50of NCTD and NPS were283.72±4.55μg/mL and194.26±3.69μg/mL, respectively. Compared with NCTD, IC50of NPs was reduced about30%.Conclusion:1. HPCS had a better solubility.2. The size of NCTD-HPCS-NPs was small, which had a higher level DL and better stability.3. NPs has a better drug-release than NCTD in vitro. The release time was three times than that of NCTD, and the totally drug-capacity was better.4. Compared with NCTD, NCTD-HPCS-NPs have a better in vitro anti-tumor activity than NCTD to the human hepatoblastoma BEL-7402cell line. In the case of one time dose, IC50of NPs was much less than NCTD. Thus the potency of the drug was enhanced in a certain extent by NPs, and the toxicity of NCTD was decreased at the same time.

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