DHA藻油脂质体制备及性质的研究

以大豆卵磷脂、胆固醇、吐温-80、DHA藻油为原料,探讨了薄膜超声法制备DHA藻油脂质体的工艺条件;分析了原料配比、温度、pH、超声时间、超声功率等因素对藻油脂质体包封率、粒径、电位的影响,对其氧化稳定性进行了初探,并研究其体外模拟消化行为。结果表明,藻油脂质体的最佳制备条件为:大豆卵磷脂与胆固醇比为3∶1(g/g)、大豆卵磷脂与藻油比为5∶1(g/g)、水化温度为40℃、pH为7.4、超声时间为120 s、超声功率350 W,在此条件下,所制备的脂质体呈椭圆球形,包封率为(91.55±0.4)%,粒径为(224.5±0.21)nm,PDI为0.224±0.003,电位(-32.4±0.03) mV;体外模拟小肠消化性能良好;室温放置一周后脂质体氧化稳定性良好。     

Practical cell labeling with magnetite cationic liposomes for cell manipulation

Personalization of the cell culture process for cell therapy is an ideal strategy to obtain maximum treatment effects. In a previous report, we proposed a strategy using a magnetic manipulation device that combined a palm-top size device and a cell-labeling method using magnetite cationic liposomes (MCLs) to enable feasible personalized cell processing. In the present study, we focused on optimizing the MCL-labeling technique with respect to cell manipulation in small devices. From detailed analysis with different cell types, 4 pg/cell of MCL-label was found to be obtained immediately after mixing with MCLs, which was sufficient for magnetic cell manipulation. The amount of label increased within 24 h depending on cell type, although in all cases it decreased along with cell doubling, indicating that the labeling potential of MCLs was limited. The role of free MCLs not involved in labeling was also investigated; MCLs' role was found to be a supportive one that maximized the manipulation performance up to 100%. We also determined optimum conditions to manipulate adherent cells by MCL labeling using the MCL dispersed in trypsin solution. Considering labeling feasibility and practical performance with 10³–10⁵ cells for personalized cell processing, we determined that 10 μg/ml of label without incubation time (0 h incubation) was the universal MCL-labeling condition. We propose the optimum specifications for a device to be combined with this method.     

柠檬烯脂质体的乙醇注入法制备及其体外释放

为提高柠檬烯稳定性,增加其水溶性,采用乙醇注入法制备了柠檬烯脂质体。以包封率为指标,考察了影响脂质体制备的一些重要的工艺参数,优化了制备柠檬烯脂质体的工艺条件,得到了最佳的工艺条件为:芯材:胆固醇:吐温80:大豆卵磷脂=0.2:0.1:1:10(质量比),醇水体积比1:10,最大包封率为86.6%。体外释放结果显示,经过25h,脂质体在模拟胃液和模拟肠液中的释放率分别为12%和18%;因而,脂质体改善了柠檬烯的水溶性,增加了其稳定性,对柠檬烯有较好的保护及缓释效果,这有利于提高其生物利用率。     

维生素E脂质体的制备工艺筛选、优化及其性质研究

目的优化维生素E（VE）脂质体的制备工艺并考察其性质。方法采用乙醇注入法、乙醚注入法、逆相蒸发法、薄膜水化法和复乳法分别制备v。脂质体,以包封率和保留率为考察指标,选择最优制备方法;经L9（3^4）正交试验设计优化选择,确定脂质体的最佳配方。结果薄膜水化法制备所得的VE脂质体包封率最高,VE保留率较高;用薄膜水化法制备脂质体的最佳配方为磷脂：VE：胆固醇=20：0．8：1．5;用透射电镜观察最佳实验组VE脂质体发现其具有指纹状结构,Zeta电位为．30．9±0．9mV,平均粒径为33．7nm。结论薄膜水化法制得的V。脂质体具有包封率高,VE保留率高,粒径均匀等特点。     

Application of Liposomes in Some Dairy Products

The application of liposomes as potential carriers to deliver food components is considerably an innovative technology. While the application of liposome technology has been very limited to date, researches indicating the potential of liposomes for improving the flavor of ripened cheese using accelerated methods, the targeted delivery of functional food ingredients, the synergistic delivery of ascorbic acid and tocopherols for promoting antioxidant activity in foods, and the stabilization of minerals (such as iron) in milk have been performed. In the food industry, liposomes and nanoliposomes have been employed to encapsulate flavoring and nutritive agents, and also, they have been suitable candidates to deliver antimicrobials. In this paper, application of lipase, proteinase, nisin, and flavor-containing liposomes in products during the processing (such as cheese maturity) as well as the application of liposomes-encapsulated micronutrients (such as iron) in milk are reviewed.     

钼蓝分光光度法分析测定空白脂质体中的磷含量

建立了空白脂质体中磷含量的测定方法,并进行了方法学验证。该方法采用钼蓝分光光度法,将样品用硫酸、过氧化氢消化,冷却后,加入显色试剂,用紫外可见分光光度计在波长822nm处测定吸收值。该方法的线性方程为y=0．0684x＋0．0086,线性关系良好（r=0．9993）,精密度为1．08％;显色后1h内比色结果稳定,回收率为95．1％～103．2％。     

丁二酸酐改性磷脂及其对脂质体稳定性的影响

研究了酰化改性磷脂对包封鱼腥草挥发油的脂质体稳定性的影响。采用单因素试验优选酰化改性磷脂的最佳工艺条件,以逆相蒸发搅拌超声法制备脂质体,过柱分离检测包封率,在-10℃、40℃和常温下放置24h后,通过离心沉淀测定检测稳定性,显微镜法观察形态。丁二酸酐酰化改性磷脂制备的脂质体比未改性的稳定,其中酰化率为77%的改性磷脂包封鱼腥草挥发油脂质体的综合性能较好,其平均粒径为97nm。     

Differential Inhibitory Effect of alpha-, beta-, gamma-, and delta-Tocopherols on the Metal-Induced Oxidation of Cholesterol in Unilamellar Phospholipid-Cholesterol Liposomes

ABSTRACT: Cholesterol oxidation products (COPs) are present in biological tissues and in foods. The inhibitory effect of antioxidants, such as tocopherols, on COPs formation has been only partially investigated. The antioxidant effect of dl alpha-, dl beta-, dl gamma-, and dl-delta tocopherol on the metal-induced oxidation of phosphatidylcholine (PC): cholesterol liposomes was assayed. Formation during liposome oxidation of six different COPs was monitored by gas chromatography. dl alpha-, and dl gamma-tocopherol show good inhibitory effect against PC-fatty acid oxidation and also on COPs formation. dl delta-Tocopherol is less effective than the alpha-and gamma-homologous, beta-tocopherol being unable to prevent PC and cholesterol oxidation. dl alpha-, and dl gamma-Tocopherol are more effective to prevent the oxidation of the lateral chain of cholesterol molecule. At the highest tocopherol concentration assayed, dl alpha-tocopherol shows prooxidant effect, enhancing liposomal oxidation and COPs formation. It is concluded that the tocopherols assayed can inhibit cholesterol oxidation but to a different degree.     

化妆品用果酸脂质体的制备研究

用反相蒸发法制得平均粒径为 164nm的果酸脂质体。研究了果酸的质量浓度对包封率的影响 ,脂质体溶液的质量浓度和温度对泄漏率的影响 ,稳定剂丙二醇的较佳质量浓度。结果表明 ,当果酸质量浓度是0 3mg/mL时可以得到 2 8%的包封率 ,丙二醇的较佳质量浓度是 5g·L-1～ 15g·L-1,温度升高使脂质体的泄漏率增加。     

Preparation of liposome-coated oligonucleotide labeled with ^99mTC and its uptake in vascular smooth muscle cells

To explore the preparation method of liposome-coated 99mTc-labeled antisense oligonucleotide (ASON),targeteing the proliferating cell nuclear antigen (PCNA), and to explore the biological characteristics and the uptake kinetics of a radiolabeled probe in vascular smooth muscle cells, an 18-base single-stranded antisense oligonucleotide targeting PCNA mRNA and the complementary strand (sense oligonucleotide, SON) were synthesized. The ASON (SON) was labeled with 99mTc, by conjugating the bifunctional chelator (hydrazino nicotinamide, HYNIC), and purified through a gel filtration column of Sephadex G-25. The product was then encapsulated in cationic liposome (oligofectamineTM). The radiolabeling efficiency, radiochemical purity, stability of the liposome-coated 99mTc-HYNIC-ASON in a phosphate buffered solution (PBS), and fresh human serum and its uptake rate were studied. There was no significant difference between the 99mTc radiolabeling efficiencies of HYNIC-ASON and HYNIC-SON, which were 60.04% ± 1.92% and 59.60% ± 2.53%, respectively (P ＞ 0.05, n = 5). The radiochemical purity of the liposome-coated 99mTc-HYNIC-ASON was 94.70% ± 1.90% (n = 5). And after incubation with PBS and fresh human seAt 90 min after transfection, the uptake rate of the liposome-coated 99mTc-HYNIC-ASON reached its peak of 83.8% ±5.92% in vascular smooth muscle cells (VSMCs) and was much higher than that of the nonliposome-coated 99mTc-HYNIC-ASON, which was 11.16% ± 0.54% (P ＜ 0.01, n = 4). The labeling method of PCNA ASON (SON) conjugated by HYNIC has been proved successful. The liposome was able to enhance the ASON (SON) uptake in VSMCs,and could be widely used as a safe, convenient, effective gene transfer carrier.