發(fā)布時(shí)間：2018-12-15 09:08

【摘要】：目的1、制備一種新型空心磁性聚合物微球(Hollow magnetic polymer microspere, HMPMS),并將相變材料全氟戊烷(Perfluoropentane, PFP)包裹于其內(nèi),檢測(cè)其基本特性,并通過體外實(shí)驗(yàn)驗(yàn)證其相變及對(duì)微波消融(Microwave ablation, MWA)效果的影響。2、研究自制包裹PFP的空心磁性聚合物微球(PFP-HMPMS)被HepG2肝癌細(xì)胞吞噬的能力,并對(duì)其細(xì)胞毒性進(jìn)行評(píng)價(jià)。研究PFP-HMPMS增強(qiáng)HepG2肝癌細(xì)胞MWA效果的可行性。3、通過在體實(shí)驗(yàn)研究自制PFP-HMPMS增效裸鼠皮下HepG2移植瘤模型MWA效果的可行性。材料和方法1、采用模板沉積法制備HMPMS,對(duì)其外觀、粒徑進(jìn)行分析。研究不同溫度促進(jìn)其相變的情況及PFP-HMPMS對(duì)MWA溫升速率的影響。2、 用普魯士藍(lán)染色觀察HepG2細(xì)胞對(duì)PFP-HMPMS的吞噬能力；用四唑鹽(Methyl thiazolyl tetrazolium, MTT)比色法檢測(cè)PFP-HMPMS的細(xì)胞毒性；用流式細(xì)胞儀檢測(cè)HepG2細(xì)胞吞噬PFP-HMPMS后MWA在不同溫控條件下的細(xì)胞存活狀態(tài)的比例,并與對(duì)照組相比較。3、建立裸鼠皮下HepG2移植瘤模型,對(duì)其行MWA治療,比較局部注射PFP-HMPMS的實(shí)驗(yàn)組與對(duì)照組溫升速率的差異,并通過組織病理學(xué)檢查考察PFP-HMPMS增強(qiáng)MWA治療腫瘤的效果。結(jié)果1、本研究制備出的HMPMS球形度較好、粒徑均一,且包裹PFP后,可在溫度超過40℃時(shí)發(fā)生相變,并能在微波(Microwave, MW)場(chǎng)中增強(qiáng)MW能量的吸收而快速升溫。2、自制的HMPMS可被HepG2細(xì)胞所吞噬,標(biāo)記陽性率為71.7士3.5%；當(dāng)PFP-HMPMS濃度為100μg/ml時(shí)對(duì)HepG2細(xì)胞的生存具有輕微的生長抑制作用(P0.05);MWA2h后流式結(jié)果顯示HepG2細(xì)胞吞噬PFP-HMPMS后,當(dāng)加熱溫度為45℃、50℃及60℃時(shí),正常存活細(xì)胞及早期凋亡細(xì)胞的百分比明顯降低；而50℃及60℃時(shí),HepG2細(xì)胞早期凋亡、晚期凋亡或死亡的比例明顯提高(P0.05)。3、實(shí)驗(yàn)組荷瘤裸鼠局部注射PFP-HMPMS后,溫度升高到50℃所需要的時(shí)間與對(duì)照組相比差別具有統(tǒng)計(jì)學(xué)意義(P0.05)；組織病理學(xué)結(jié)果顯示PFP-HMPMS沉積區(qū)域腫瘤細(xì)胞結(jié)構(gòu)變化顯著。結(jié)論1、自制的PFP-HMPMS能夠通過溫度的升高發(fā)生相變,并且在MW場(chǎng)中可增強(qiáng)MW能量的吸收而快速升溫。2、自制的HMPMS可以被HepG2細(xì)胞所吞噬；PFP-HMPMS對(duì)HepG2細(xì)胞增殖影響較小；在相同溫控條件下,PFP-HMPMS具有增強(qiáng)腫瘤細(xì)胞凋亡和死亡的比例的作用。3、自制的PFP-HMPMS能夠提高M(jìn)WA溫升效率,增強(qiáng)MWA對(duì)在體HepG2皮下移植瘤的治療效果。
[Abstract]:Objective 1. To prepare a new hollow magnetic polymer microsphere (Hollow magnetic polymer microspere, HMPMS),) and encapsulate the phase change material perfluoropentane (Perfluoropentane, PFP) in it. The phase change and the effect of microwave ablation on (Microwave ablation, MWA) were tested in vitro. 2. The ability of self-made hollow magnetic polymer microspheres (PFP-HMPMS) coated with PFP to be swallowed by HepG2 hepatoma cells was studied. Its cytotoxicity was evaluated. To study the feasibility of PFP-HMPMS to enhance the MWA effect of HepG2 hepatoma cells. 3. To study the effect of self-made PFP-HMPMS on the effect of subcutaneous HepG2 transplantation in nude mice with MWA in vivo. Materials and methods 1. HMPMS, was prepared by template deposition method to analyze its appearance and particle size. The effect of PFP-HMPMS on the temperature rise rate of MWA was studied. 2 the phagocytosis of HepG2 cells to PFP-HMPMS was observed by Prussian blue staining. Cytotoxicity of PFP-HMPMS was detected by tetrazolium (Methyl thiazolyl tetrazolium, MTT) colorimetry. Flow cytometry was used to detect the survival rate of MWA after PFP-HMPMS phagocytosis by HepG2 cells under different temperature control conditions. Compared with the control group, a subcutaneous HepG2 transplanted tumor model was established in nude mice and treated with MWA. The difference of temperature rise rate between the experimental group and the control group was compared, and the effect of PFP-HMPMS enhanced MWA on tumor was investigated by histopathological examination. Results 1. The HMPMS prepared in this study has good sphericity and uniform particle size, and after encapsulating PFP, the phase transition can take place when the temperature exceeds 40 鈩，

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