载阿霉素介孔二氧化硅纳米粒的细胞毒性及细胞摄取

目的制备载阿霉素的介孔二氧化硅纳米粒(mesoporous silica nanoparticles,MSN),对其理化性质及细胞摄取行为进行初步研究。方法通过聚合法制备MSN,透射电镜表征纳米粒的形态,动态光散射粒径测定仪测定粒子的平均粒径及分布。紫外分光光度计测定阿霉素的负载行为,MTT比色分析法研究粒子的细胞毒性,激光共聚焦显微镜观察其人乳腺癌MCF-7细胞对载药粒子的摄取。结果纳米粒分布均一,平均粒径约70 nm(PDI<0.1),载药量质量分数达到20%。MCF-7细胞对载药粒子的摄取较快,空白纳米粒具有较低的细胞毒性。结论介孔二氧化硅纳米粒具有较高的药物载药量和良好的生物相容性,能较快地被对人乳腺癌MCF-7细胞摄取,有望成为一种新型的药物化疗载体。     

Box-Behnken设计-效应面法优化吴茱萸碱-羟基乙酸共聚物纳米粒处方及体外释药研究

目的：Box-Behnken设计-效应面法优化吴茱萸碱聚乳酸-羟基乙酸共聚物[poly （lactic-co-glycolic acid）,PLGA]纳米粒处方（吴茱萸碱-PLGA纳米粒）,考察体外释药行为。方法：单因素考察PLGA用量,油水体积比,泊洛沙姆188浓度,超声功率和时间等因素的影响,采用Box-Behnken响应面法优化吴茱萸碱-PLGA纳米粒处方。采用甘露醇为冻干保护剂,制备吴茱萸碱-PLGA纳米粒冻干粉末,并考察体外释药情况及释药模型。结果：吴茱萸碱-PLGA纳米粒最佳处方为：PLGA用量为445.1 mg、油水体积比1：5.2、泊洛沙姆188质量分数为1.2%。包封率和粒径分别为（75.73±1.33）%和（173.27±6.86） nm,与模型预测值接近。体外释药符合Higuchi模型：M_t/M_∞=0.109 9t^1/2+0.081 6,缓释特征明显。结论：Box-Behnken实验设计可用于吴茱萸碱-PLGA纳米粒处方研究,为进一步研究奠定了基础。     

替尼泊苷多层包衣白蛋白纳米粒的制备工艺及体外抗肿瘤活性

目的以人血清白蛋白为载体包载替尼泊苷,经过包衣修饰后制备包载替尼泊苷的多层包衣纳米粒(teniposide-encapsulated multilayer nanoparticles,P-CS-NP),以期降低药物的不良反应并改善其体外抗肿瘤活性。方法以粒径、多分散指数和载药率为评价指标,采用单一因素法筛选出替尼泊苷白蛋白纳米粒的最优处方工艺,通过加入壳聚糖和聚谷氨酸聚乙二醇共聚物进一步制备多层包衣白蛋白纳米粒,筛选得到最优包衣量。以游离的替尼泊苷作为参比,用MTT法测定纳米粒对人肺癌A549细胞的体外细胞毒性,并用流式细胞仪和共聚焦显微镜测定和观察多层包衣纳米粒的细胞摄取率和细胞摄取行为。结果确定了多层包衣纳米粒的处方及制备工艺。多层包衣纳米粒的体外细胞毒性比游离的替尼泊苷小,摄取具有时间依赖性,与壳聚糖共孵育的纳米粒的细胞摄取量增加,入胞后纳米粒主要分布在细胞质。结论白蛋白纳米粒能被壳聚糖和聚谷氨酸聚乙二醇共聚物包衣修饰,多层包衣纳米粒可以作为替尼泊苷的药物递送载体,其体外细胞毒性降低。     

阿霉素壳寡糖纳米粒的制备及体外抗肿瘤研究

目的制备负载阿霉素的壳寡糖纳米粒,并研究其理化性质和体外抗肿瘤细胞毒性。方法采用离子凝胶法制备负载阿霉素的壳寡糖纳米粒;透射电镜观察纳米粒形态,激光粒度仪测定粒径和表面电位,紫外分光光度法测量包封率、载药量,考察载药纳米粒的体外释药特性;采用MTT法对载药壳寡糖纳米粒在体外乳腺癌细胞株MCF-7的细胞毒作用进行评价。结果制得的阿霉素壳寡糖纳米粒呈球形或类球形,形态较为完整,平均粒径为(136.77±1.21)nm,表面电位为(20.53±0.31)m V,包封率为(56.99±1.40)%,载药量为(15.49±0.38)%,168 h的累积释放率为72.15%;阿霉素和载药纳米粒对MCF-7细胞增殖的抑制作用存在明显的浓度和时间依赖性,且载药纳米粒对MCF-7细胞增殖的抑制作用随时间增加而逐渐强于游离阿霉素。结论此方法制备的阿霉素壳寡糖纳米粒粒径较小,药物释放具有明显的缓释作用,并具有较好的抗肿瘤作用。     

卡莫氟固体脂质纳米粒的制备及工艺研究

目的:制备卡莫氟固体脂质纳米粒并考察其药剂性质。方法:采用高压均质法制备卡莫氟固体脂质纳米粒混悬液,以单因素考察和正交设计法筛选处方和工艺,并考察其形态、粒径、载药量及包封率。结果:优选出的较佳处方大豆卵磷脂、泊洛沙姆188、吐温-80和硬脂酸用量分别为8.0、12.0、1.0、7.5mg·mL-1;所制得的固体脂质纳米粒为圆整的实体粒子,表面光滑,平均粒径为78.7nm,载药量为23.47%,包封率为82.33%。结论:高压均质法可用于卡莫氟类脂溶性药物固体脂质纳米粒的制备。     

壳寡糖-水杨酸接枝物纳米粒用于释放阿霉素的研究

目的 考察壳寡糖/水杨酸纳米粒负载碱化阿霉素的可能性,评价制备而得的微粒给药系统理化性质及其体外释放行为。方法 以碳二亚胺为交联偶合剂合成壳寡糖/水杨酸接枝共聚物,三硝基苯磺酸法测定水杨酸接枝率。运用超声分散法制备壳寡糖/水杨酸空白纳米粒,芘荧光法测定纳米粒临界聚集浓度,动态光散射法测定微粒粒径和表面电位,MTT法考察空白纳米粒的细胞毒性。以碱化阿霉素为模型药物,透析法制备壳寡糖/水杨酸载药纳米粒,经透射电镜考察载药纳米粒的形态,对其体外释放行为进行了研究。结果 合成得到的壳寡糖分子量=9000/水杨酸理论投料量=50%的实际接枝率为16.92%,空白纳米粒的临界聚集浓度为867.0 μg/mL,空白纳米粒的粒径和表面Zeta电位分别为434.0 nm和48.6 mV,对人肝癌细胞Hep-G2的半数抑制浓度为1745μg/mL。在碱化阿霉素理论投药量为10%时壳寡糖/水杨酸载药纳米粒的实际载药量为8.52%,包封率为93.15%。;载药纳米粒的粒径和表面电位分别为214.2 nm和33.6 mV。体外释放结果表明药物的释放呈现pH敏感性;并主要以溶蚀的方式从载体内部释放出来。结论 壳寡糖/水杨酸接枝物可以有效包裹碱化阿霉素并成为粒径均一的纳米粒给药系统。载药纳米粒具有pH敏感和缓释作用。壳寡糖/水杨酸接枝物有望成为潜在的难溶性药物的载体材料。     

长春新碱固体脂质纳米粒的制备工艺优化

目的制备长春新碱固体脂质纳米粒(VCR-SLN)并优化其处方组成和制备工艺。方法单因素考察筛选载体、稳定剂及制备工艺,用正交试验进行优化,以包封率、载药量和粒径为指标,筛选最佳处方和制备工艺,并对在优化条件下制备的VCR-SLN进行质量评价。结果以单硬酸酯甘油酯为载体,大豆卵磷脂、泊洛沙姆188为乳化剂,采用复乳-溶剂扩散法制备得VCR-SLN,其平均粒径为156.3 nm,包封率为55.12%,载药量为3.09%。结论复乳-溶剂扩散法适用于制备VCR-SLN。     

咖啡酸苯乙酯纳米混悬剂的制备及其体外抑制乳腺癌细胞作用研究

目的 优化咖啡酸苯乙酯纳米混悬剂的处方,并考察其体外抑制乳腺癌细胞作用.方法 以泊洛沙姆188、蜂胶作为载体,采用反溶剂沉淀法制备,通过星点设计–效应面法优化最佳制备工艺参数,并考察咖啡酸苯乙酯纳米混悬剂的稳定性、载药量、包封率以及冻干保护剂的筛选,同时考察对4T1乳腺癌细胞的生长抑制作用和细胞摄取情况.结果 咖啡酸苯...     

盐酸表柔比星-聚乳酸-羟基乙酸共聚物纳米粒的制备

摘要目的制备盐酸表柔比星 聚乳酸 羟基乙酸（PLGA）共聚物纳米粒,对其进行质量评价。方法采用乳化 溶剂挥发法制备盐酸表柔比星纳米粒;对主要处方因素如PLGA用量、外水相中聚山梨酯 80用量、泊洛沙姆188和聚山梨酯 80比例进行正交设计,以药物的包封率、载药量和药物利用率等为考察指标。结果采用优化后处方制得的纳米粒药物包封率为（32.6±1.2）%,载药量为（7.2±0.5）%,药物利用率为（51.6±3.4）%,纳米粒平均粒径166.6 nm,药物可持续160 h释放。结论该方法制备盐酸表柔比星纳米粒工艺简单,无需使用聚乙烯醇,药物释放缓慢。     

载阿霉素PLGA-PLL-PEG纳米粒的制备及抑瘤作用研究

目的:制备载阿霉素聚乳酸羟基乙酸-聚赖氨酸-聚乙二醇(PLGA-PLL-PEG)纳米粒,并研究其抑瘤作用。方法:应用PLGA-PLL和活化PEG聚合而成的PLGA-PLL-PEG为载体包载阿霉素,制得载阿霉素PLGA-PLL-PEG纳米粒。检测纳米粒的形态大小、粒径分布、阿霉素的含量,计算载药量和包封率,比较纳米粒和阿霉素在144 h内的累积释放率(Q)和对乳腺癌HeLa细胞的增殖抑制率,计算半数抑制率(IC_(50))。结果:所制载阿霉素PLGA-PLL-PEG纳米粒为规则圆形,分散性良好,无黏连,平均粒径为(136.7±9.3)nm(n=5),平均包封率为(76.67±8.63)%,平均载药量为(3.86±0.55)%(n=3);阿霉素的Q_(12h)达100%,载阿霉素PLGA-PLL-PEG纳米粒的Q_(24h)为52.9%、Q_(144h)为81.2%。载阿霉素PLGA-PLL-PEG纳米粒对HeLa细胞的增殖抑制率较阿霉素增长缓慢,二者的IC_(50)分别为1.844、0.345μg/mL。结论:成功制得载阿霉素PLGA-PLL-PEG纳米粒,其具有良好的缓释效果,其抑瘤作用强于阿霉素。     

TPGS修饰脂质体对阿霉素抗肿瘤活性的增敏作用

目的制备聚乙二醇1000维生素E琥珀酸酯(TPGS)修饰的阿霉素脂质体并考察其对阿霉素抗肿瘤活性的增敏作用。方法用阳离子树脂吸附法测定阿霉素脂质体的包封率;MTT法测定对MCF-7和MCF-7/ADR的毒性;用荧光显微镜观察阿霉素的细胞摄取,并用HPLC测定细胞内的阿霉素含量。结果 TPGS修饰的阿霉素脂质体增加了MCF-7/ADR对阿霉素的摄取,并增强了对MCF-7和MCF-7/ADR细胞的毒性。结论 TPGS修饰脂质体能显著增强MCF-7和MCF-7/ADR对阿霉素的敏感性。     

The effect of poloxamer 188 on nanoparticle morphology,size, cancer cell uptake,and cytotoxicity

The aim of this work was to investigate the effect of triblock copolymer poloxamer 188 on nanoparticle morphology, size, cancer cell uptake, and cytotoxicity. Docetaxel-loaded nanoparticles were prepared by oil-in-water emulsion/solvent evaporation technique using biodegradable poly(lactic-co-glycolic acid) (PLGA) with or without addition of poloxamer 188, respectively. The resulting nanoparticles were found to be spherical with a rough and porous surface. The nanoparticles had an average size of around 200 nm with a narrow size distribution. The in vitro drug-release profile of both nanoparticle formulations showed a biphasic release pattern. An increased level of uptake of PLGA/poloxamer 188 nanoparticles in the docetaxel-resistant MCF-7 TAX30 human breast cancer cell line could be found in comparison with that of PLGA nanoparticles. In addition, the docetaxel-loaded PLGA/poloxamer 188 nanoparticles achieved a significantly higher level of cytotoxicity than that of docetaxel-loaded PLGA nanoparticles and Taxotere (P < .05). In conclusion, the results showed advantages of docetaxel-loaded PLGA nanoparticles incorporated with poloxamer 188 compared with the nanoparticles without incorporation of poloxamer 188 in terms of sustainable release and efficacy in breast cancer chemotherapy.From the Clinical EditorThe effects of poloxamer 188, a triblock copolymer were studied on nanoparticle morphology, size, cancer cell uptake and cytotoxicity. An increased level of uptake of PLGA/poloxamer 188 nanoparticles in resistant human breast cancer cell line was demonstrated, resulting in a significantly higher level of cytotoxicity.     

Doxorubicin-loaded solid lipid nanoparticles to overcome multidrug resistance in cancer therapy

In the present study we developed doxorubicin-loaded solid lipid nanoparticles (SLN-Dox) using biocompatible compounds, assessed the in vitro hemolytic effect, and examined their in vivo effects on drug retention and apoptosis intensity in P-glycoprotein-overexpressing MCF-7/ADR cells, a representative Dox-resistant breast cancer cell line. Our SLNs did not show hemolytic activity in human erythrocytes. In comparison with Dox, SLN-Dox efficiently enhanced apoptotic cell death through the higher accumulation of Dox in MCF-7/ADR cells. Therefore, SLN-Dox have potential to serve as a useful therapeutic approach to overcome the chemoresistance of adriamycin-resistant breast cancer.From the Clinical EditorDoxorubicin loaded solid lipid nanoparticles (SLN-Dox) were studied in a cell line representative of doxorubicin resistant breast cancer. The nanoparticles did not show hemolytic activity; furthermore, they efficiently enhanced apoptotic cell death through higher accumulation of doxorubicin in cancer cells. This approach may be viable in overcoming the chemoresistance of adriamycin resistant breast cancer.     

Doxorubicin-loaded galactose-conjugated poly(d,l-lactide-co-glycolide) nanoparticles as hepatocyte-targeting drug carrier

The objective of this work is to produce doxorubicin-loaded galactose-conjugated poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles (NPs) to be specifically recognised by human hepatoma cellular carcinoma (Hep G2) cells and assess NPs cytotoxicity. Doxorubicin-unloaded and doxorubicin-loaded galactose-conjugated PLGA NPs were prepared using an emulsion method and characterised for morphology, size, drug release behaviour, Hep G2 recognition and cell cytotoxicity. The produced doxorubicin-loaded PLGA-galactose-conjugate nanoparticles (PLGA-GAL NPs) are spherical in shape with a size of 365?±?74?nm, a drug encapsulation efficiency of 69% and released in a biphasic pattern with higher release rates at pH 5. In vitro cell studies confirmed the specific interaction between the receptors of Hep G2 and the PLGA-GAL NPs. Cell cytotoxicity tests showed that unloaded NPs are non-toxic and that doxorubicin-loaded NPs caused a cellular viability decrease of around 80%, therefore representing a promising approach to improve liver-specific drug delivery.     

Folate-modified poly(2-ethyl-2-oxazoline) as hydrophilic corona in polymeric micelles for enhanced intracellular doxorubicin delivery

The transmembrane transport of drug loaded micelles to intracellular compartment is quite crucial for efficient drug delivery. In the current study, we investigated the cellular internalization and anticancer activity of doxorubicin loaded micelles with folate modified stealthy PEOz corona. Folate-decorated micelles incorporating doxorubicin were characterized for particle size, degree of folate decoration, drug loading content and encapsulation efficiency, morphology, and surface charge. The targeting capability and cell viability were assessed using HeLa, KB, A549 and MCF-7/ADR cell lines. In vitro study clearly illustrated the folate receptor (FR) mediated targeting of FA modified micelles to FR-positive human HeLa, KB and MCF-7/ADR cells, while specific delivery to FR-negative A549 cells was not apparently increased at the same experimental conditions. Cytotoxicity assay showed 60% and 58% decrease in IC₅₀ values for HeLa and KB cells, while only a slight decrease for A549 cells, following treatment with folate modified formulations. The enhanced intracellular delivery of FA modified micelles in MCF-7/ADR cells was also observed. In vivo antitumor tests revealed DOX entrapped FA-PEOz-PCL micelles effectively inhibited the tumor growth and reduced the toxicity to mice compared with free DOX. The current study showed that the targeted nano-vector improved cytotoxicity of DOX and suggested that this novel PEOz endowed stealthy micelle system held great promise in tumor targeted therapy.     

Emodin loaded solid lipid nanoparticles: preparation, characterization and antitumor activity studies

The objective of the present study was to prepare and characterize emodin (EMO)-loaded solid lipid nanoparticles (E-SLNs) and evaluate their antitumor activity in vitro. EMO and pharmaceutical lipid material were used to prepare E-SLNs by high pressure homogenization (HPH). Poloxamer 188 and Tween 80 were used as surfactants. The physicochemical properties of the E-SLNs were investigated by particle size analysis, zeta potential measurement, drug entrapment efficiency (EE), stability and in vitro drug release behavior. The E-SLNs showed stable particle size at 28.6 ± 3.1 nm, ideal drug EE and relative long-term physical stability after being stored for 4 months. The drug release of E-SLNs could last 72 h and exhibited a sustained profile, which made it a promising vehicle for oral drug delivery. MTT assay showed that E-SLNs could significantly enhance the in vitro cytotoxicity against human breast cancer cell line MCF-7 and MDA-MB-231 cells compared to the EMO solution, while free EMO, blank SLNs (B-SLNs) and E-SLNs all showed no significant toxicity to human mammary epithelial line MCF-10A cells. Flow cytometric analysis demonstrated that E-SLNs also showed more significant cell cycle arrest effect in MCF-7 cells compared to bulk EMO solution. Hoechst 33342 staining and Annexin V-FITC/PI double staining further confirmed that E-SLNs induced higher apoptotic rates in MCF-7 cells, indicating that cell cycle arrest and apoptosis maybe the underlying mechanism of the enhanced cytotoxicity. Taken together, it seems that HPH was a simple, available and effective method for preparing high quality E-SLNs to enhance its aqueous solubility. Moreover, these results suggest that the delivery of EMO as lipid nanoparticles maybe a promising approach for cancer therapy.     

Preparation of psoralen polymer–lipid hybrid nanoparticles and their reversal of multidrug resistance in MCF-7/ADR cells

Multidrug resistance (MDR) is the leading cause of failure for breast cancer in the clinic. Thus far, polymer–lipid hybrid nanoparticles (PLN) loaded chemotherapeutic agents has been used to overcome MDR in breast cancer. In this study, we prepared psoralen polymer–lipid hybrid nanoparticles (PSO-PLN) to reverse drug resistant MCF-7/ADR cells in vitro and in vivo. PSO-PLN was prepared by the emulsification evaporation-low temperature solidification method. The formulation, water solubility and bioavailability, particle size, zeta potential and entrapment efficiency, and in vitro release experiments were optimized in order to improve the activity of PSO to reverse MDR. Optimal formulation: soybean phospholipids 50?mg, poly(lactic-co-glycolic) acid (PLGA) 15?mg, PSO 3?mg, and Tween-80 1%. The PSO-PLN possessed a round appearance, uniform size, exhibited no adhesion. The average particle size was 93.59?±?2.87?nm, the dispersion co-efficient was 0.249?±?0.06, the zeta potential was 25.47?±?2.84?mV. In vitro analyses revealed that PSO resistance index was 3.2, and PSO-PLN resistance index was 5.6, indicating that PSO-PLN versus MCF-7/ADR reversal effect was significant. Moreover, PSO-PLN is somewhat targeted to the liver, and has an antitumor effect in the xenograft model of drug-resistant MCF-7/ADR cells. In conclusion, PSO-PLN not only reverses MDR but also improves therapeutic efficiency by enhancing sustained release of PSO.     

Reversal activity of nanostructured lipid carriers loading cytotoxic drug in multi-drug resistant cancer cells

To overcome multi-drug resistance (MDR) of cancer cells, paclitaxel (PTX) and doxorubicin (DOX)-loaded nanostructured lipid carriers (NLC) were prepared by solvent diffusion method using monostearin as solid lipid and oleic acid as liquid lipid matrix. The cytotoxicities and reversal activity of drug-loaded NLC were tested against human breast cancer (MCF-7) cells, human ovarian cancer (SKOV3) cells and their multi-drug resistant (MCF-7/ADR and SKOV3-TR30) cells. The chemical conjugant of folic acid and stearic acid (FA-SA) was further synthesized to prepare folated NLC. Comparing with taxol and doxorubicin solution, the NLC loading PTX exhibited high cytotoxicities in MCF-7 and MCF-7/ADR cells, while the NLC loading DOX only indicated high cytotoxicity in MCF-7/ADR cells. The reversal powers of the NLC loading PTX and DOX were 34.3 and 6.4 folds, respectively. The NLC loading PTX and DOX showed the same trends of enhanced cytotoxicity against SKOV3 and SKOV3-TR30 cells. The reversal powers were 31.3 and 2.2 folds for the NLC loading PTX and DOX, respectively. The modification of NLC with FA-SA could further enhance the cytotoxicities of drug in drug sensitive and drug resistant cells.     

Using doxorubicin and siRNA-loaded heptapeptide-conjugated nanoparticles to enhance chemosensitization in epidermal growth factor receptor high-expressed breast cancer cells

Abstract

The aim of this study was to develop the heptapeptide-conjugated active targeting nanoparticles for delivery of doxorubicin and siRNA to epidermal growth factor receptor (EGFR) high-expressed breast cancer cells. The active targeting nanoparticles were prepared by using a synthesized poly(D,L-lactide-co-glycolide)-poly(ethylene glycol) (PLGA-PEG) copolymer conjugated with a heptapeptide. The particle size of peptide-conjugated nanoparticles was less than 200?nm with narrow size distribution and the surface charge was negative. The uptake of peptide-conjugated nanoparticles was more efficient in EGFR high-expressed MDA-MB-468 cells than in EGFR low-expressed HepG2 cells by 3.9 folds due to peptide specific binding to EGF receptor followed by EGF receptor-mediated endocytosis. The nanoparticles were used to deliver doxorubicin and siRNA, and their in vitro release was faster in pH 4.0 (500?U lipase) than in pH 7.4. The IC₅₀ of doxorubicin-loaded peptide-conjugated nanoparticles was lower than that of peptide-free nanoparticles by 2.3 folds in MDA-MB-468 cells. Similarly, the cellular growth inhibition of siRNA/DOTAP-loaded peptide-conjugated nanoparticles was 2.1 folds higher than that of peptide-free nanoparticles. In conclusion, the heptapeptide-conjugated PLGA-PEG nanoparticles provided active targeting potential to EGFR high-expressed MDA-MB-468 breast cancer cells, and a synergistic cytotoxicity effect was achieved by co-delivery of doxorubicin and siRNA/DOTAP-loaded peptide-conjugated nanoparticles.