一种光聚合PEGDA-HA水凝胶递送纳米粒药物用于非小细胞肺癌的治疗

发布时间：2018-05-22 13:13

本文选题：水凝胶 + 局部凝胶药物递送　；参考：《华东师范大学》2017年硕士论文

【摘要】：局部药物递送系统应用于肿瘤的化疗是一种将药物直接递送并作用于肿瘤细胞的化疗手段。相比较于传统的化疗手段,其主要特点为局部药物浓度高,相对药物剂量低,生物毒性低,机体适应性好。近年来,有越来越多的学者研究将水凝胶作为一种局部药物递送载体递送抗肿瘤药用于肿瘤的化疗。水凝胶作为一种新型的生物友好型材料越来越被普遍研究用于组织功能领域和药物递送载体领域。水凝胶是一种以水为介质的亲水性聚合物,在水中能够溶胀并保持大量水分而又不能溶解的具有三维网络结构的交联体,并且在水中能有足够的稳定性,柔软而富有弹性的功能高分子。高分子链上的亲水基团以氢键与水分子相连接,使其具有强大的锁水功能,能保住90%以上的水分,因此水凝胶兼具固体和流体的性质。水凝胶拥有良好的生物亲和性和生物粘附性,独特的物理化学结构使得它可以作为一种局部药物递送系统用于肿瘤的化疗。本课题设计的是一种光聚合水凝胶PEGDA-HA。PEGDA-HA水凝胶由聚乙二醇二丙烯酸酯(PEGDA),低分子量的HA,和光引发剂Irgacure 2959通过紫外光激发聚合而成。在制备过程中,我们探究了聚乙二醇二丙烯酸酯和光引发剂Irgacure2959不同质量比例聚合成凝胶的凝胶时间和化学降解速率的关系,并确定了比较合适的引发剂和单体质量比。为了提高水凝胶的生物相容性,在凝胶体系中加入了一种聚合物填充材料低分子量的HA。实验过程中我们探究了不同低分子量HA含量的水凝胶的溶胀速率及其降解速率与HA含量的关系。由此确定了 PEGDA、HA、和Irgacure2959三者的比例关系。从而制备了 PEGDA-HA光聚合水凝胶。我们将PEGDA-HA凝胶递送紫杉醇纳米粒PLGA-PTX,共同组成了PEGDA-HA/PLGA-PTX局部凝胶药物递送系统。实验过程中,我们通过乳化-溶剂挥发法制备了 PLGA包裹紫杉醇的纳米粒PLGA-PTXNPs。通过TEM,DLS对该NPs进行了表征。实验结果表明PLGA-PTX NPs形态呈现非常明显的核壳结构,包封率为74.04%,载药量为7.4%,纳米粒粒径为170±10.2nm,n=3,PDI=0.309±0.089,n=3,粒径分布比较均一。zeta 电位为-4.64±0.05mV,n=3。通过CCK8实验,我们测得了该PLGA-PTXNPs的IC50值为10-5 527mg/mL。通过包载Dio荧光素,我们测得PLGA-PTXNPs能够被NCI-H460细胞摄取,通过流式细胞仪的定量数据与激光共聚焦数据,我们可以看到NCI-H460细胞对PLGA-PTXNPs的摄取比率达到了 96%。由PEGDA-HA/PLGA-PTX凝胶的紫杉醇药物释放曲线可知,药物释放速度呈现先快后慢的趋势,在前20小时释放了近30%的药量,在给药后40小时内释放了近50%的药量,此后药物释放逐渐减缓。至给药后100小时内释放了 65%药量。虽然此后释药速度有所减缓,但是一直呈现了上升趋势,可以预计,该局部凝胶药物递送系统可以维持肿瘤部位20天的持续药物释放。通过药效学实验数据我们可以发现,实验组PEGDA-HA/PLGA-PTX组在没有对老鼠造成减重的毒性反应情况下,实现了抑制肿瘤细胞生长和杀灭肿瘤细胞的效果。观察对比实验组PEGDA-HA/PLGA-PTX组和对照组PBS组的HE染色荧光图片我们可以发现,实验组除了肿瘤组织,均没有观察到有组织坏死现象。对比实验组PEGDA-HA/PLGA-PTX组和对照组PBS组的TUNEL染色荧光图片,我们可以发现,只有在实验组的肿瘤组织观察到组织细胞凋亡现象。以上实验结果说明,我们课题设计的PEGDA-HA/PLGA-PTX局部凝胶药物递送系统是一种生物相容性非常好的局部药物递送系统,通过局部直接给药的方式直接作用于肿瘤部位,在给药后20天内均表现了良好的抑制肿瘤生长和杀灭肿瘤细胞的药效学结果。
[Abstract]:Local drug delivery system used in tumor chemotherapy is a means of direct delivery and action to tumor cells. Compared with traditional chemotherapy, the main characteristics are high local drug concentration, low drug dose, low biological toxicity and good adaptability. In recent years, more and more scholars have studied water coagulation. Gelatin is used as a local drug delivery carrier to deliver chemotherapeutic agents for cancer. Hydrogels are widely used as a new biofriendly material to be widely used in the field of tissue function and drug delivery carriers. Hydrogel is a hydrophilic polymer with water as medium, which can expand and maintain a large amount of water in water. Water can not be dissolved in a three-dimensional network structure of the cross body, and in the water can have enough stability, soft and elastic functional polymer. The hydrophilic group on the polymer chain connects with the water molecule by hydrogen bonds, making it strong water locking function and can hold more than 90% water, so the hydrogel has both solid and solid. The properties of fluid. Hydrogels have good biocompatibility and bioadhesion. The unique physical and chemical structure makes it a local drug delivery system for cancer chemotherapy. This topic is designed for a polyhydrogel PEGDA-HA.PEGDA-HA hydrogel consisting of polyglycol two acrylate (PEGDA), low molecular weight H A, and photopolymerization of photoinitiator Irgacure 2959 by UV excitation. In the process of preparation, we explored the relationship between the gel time and the chemical degradation rate of the gels with different mass ratios of polyethylene glycol two acrylate and photoinitiator Irgacure2959, and determined the suitable initiator and monomer mass ratio. The biocompatibility of hydrogels by adding a low molecular weight HA. experiment with a polymer filling material in the gel system, we explored the relationship between the swelling rate of the hydrogels with different low molecular weight HA content and the relationship between the degradation rate and the HA content. Thus, the proportion relationship between the PEGDA, HA, and Irgacure2959 three was determined. PEGDA-HA photopolymerization hydrogel. We sent the PEGDA-HA gel to paclitaxel nanoparticles PLGA-PTX, together to form a PEGDA-HA/PLGA-PTX local gel delivery system. In the experiment, we prepared the PLGA encapsulated paclitaxel nanoparticles PLGA-PTXNPs. via the emulsification solvent evaporation method, and DLS was used to characterize the NPs. The results show that the morphology of PLGA-PTX NPs has a very obvious nuclear shell structure, the encapsulation efficiency is 74.04%, the drug loading is 7.4%, the nano particle size is 170 + 10.2nm, n=3, PDI=0.309 0.089, n=3, the uniform.Zeta potential is -4.64 + 0.05mV, and n=3. through CCK8 experiment, we have measured the PLGA-PTXNPs. Fluorescein, we measured that PLGA-PTXNPs was able to be absorbed by NCI-H460 cells. Through quantitative data of flow cytometry and confocal laser confocal data, we can see that the ratio of NCI-H460 cells to PLGA-PTXNPs uptake reached 96%. by the paclitaxel release curve of PEGDA-HA/PLGA-PTX gel, and the release rate of the drug was fast and then slow. The trend, releasing nearly 30% of the dose in the first 20 hours and releasing nearly 50% of the dose within 40 hours after the administration, the release of the drug slowed down gradually. 65% doses were released within 100 hours after the administration. Although the release rate has slowed down since then, the drug delivery system has been on the rise, and it is expected that the local gel delivery system can be maintained. After 20 days of sustained drug release on the tumor site, we can find that in the experimental group PEGDA-HA/PLGA-PTX, the effect of inhibiting the growth of tumor cells and killing the tumor cells was achieved without the toxic reaction to the mice caused by weight reduction in the experimental group. The H of group PEGDA-HA/PLGA-PTX and the control group PBS group were observed and compared. We can find that there is no tissue necrosis in the experimental group except the tumor tissue. Compared with the PEGDA-HA/PLGA-PTX group in the experimental group and the PBS group in the control group, we can find that the apoptosis of the tissue cells is observed only in the tumor tissue of the experimental group, and the results of the experiment are explained by the experimental results. The PEGDA-HA/PLGA-PTX local gel delivery system, designed by us, is a highly biocompatible local drug delivery system, which directly acts on the tumor site by local direct drug delivery, and shows good pharmacodynamic results in the 20 days after administration to inhibit tumor growth and kill tumor cells.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943;R96

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