茶多酚锰合成、表征及络合和诱导肿瘤细胞凋亡的研究

合成了茶多酚锰(TP-Mn)和茶多酚锗(TP-Ge), 并采用红外光谱法及液相色谱质谱联用技术表征TP-Mn的理化特性. 选用反向液相色谱(RP-HPLC)和电感藕合等离子体质谱(ICP-MS)技术研究牛血清白蛋白(BSA)络合TP-Mn的能力, 指出BSA可直接络合于TP-Mn. 采用MTT法研究TP和TP-Mn诱导Hela卵巢癌细胞的凋亡率. 结果表明, TP和TP-Mn均能诱导Hela卵巢癌细胞凋亡, 但TP-Mn诱导凋亡率约为TP的2倍. 比较TP-Ge和TP-Mn诱导Raji人B淋巴瘤细胞凋亡率发现, 两者诱导肿瘤细胞的凋亡率几乎相同, 均高达86%左右. 人血清白蛋白(HSB)可作为输送TP-Ge和TP-Mn的药物载体.     

微乳液成核-离子交联制备阿司匹林/壳聚糖纳米微球及其体外释放行为

以自制阿司匹林为药物模型,壳聚糖(CS)为载体源,采用微乳液成核-离子交联法制备了阿司匹林/壳聚糖纳米缓释微球.分别用傅里叶变换红外(FTIR)光谱、场发射扫描电子显微镜(FESEM)、透射电子显微镜(TEM)、动态激光光散射(DLLS)、X射线粉末衍射(XRD)等表征了纳米微粒的化学组成、外观形貌、平均粒径和粒径分布、微球中壳聚糖的晶体结构以及阿司匹林的分布形态.结果表明,利用微乳液成核-离子交联法制备的阿司匹林/壳聚糖微球平均粒径约为88nm且粒径分布均匀,成核后壳聚糖结晶形态基本未变,阿司匹林以分子形态分布于微粒中,分子间未形成堆砌,为无定形态.采用UV-Vis分光光度计考察了微球的药物包封率、载药量,并对微球在生理盐水和葡萄糖溶液中的释药行为进行跟踪.结果表明,微球的载药量可达55%,药物包封率可达42%,实验条件下具有较好的药物缓释作用.     

壳聚糖接枝β-环糊精聚合物空心微球的制备及其载药性能

以壳聚糖(CS)和β-环糊精(β-CD)为原料,制备了壳聚糖接枝β-环糊精聚合物(CS-g-CD),然后以三聚磷酸钠(TPP)为交联剂,通过静电自组装的方法得到了聚合物空心微球,并以5-氟尿嘧啶(5-FU)为模型药物,研究了聚合物空心微球对药物的负载性能。利用红外光谱(FT-IR)、元素分析、X-射线衍射(XRD)表征了聚合物的结构和组成,激光粒度仪和透射电子显微镜(TEM)表征了聚合物微球的粒径及形貌。通过紫外可见分光光度计研究了微球对药物的载药量和包封率。结果表明,得到的微球呈现较为规则的空心结构,粒径在300~350nm左右,分散性较好,对药物载药量和包封率较高。     

微乳液法制备可共载水溶和脂溶药物的壳聚糖季铵盐乙醇脂质体的载药性能表征

采用微乳液法制备了可包载脂溶性和水溶性药物的羧甲基壳聚糖十八烷基季铵盐(OQCMC)乙醇脂质体,研究了OQCMC乙醇高分子脂质体的相图、粒径和电位、对药物的包封及释放能力及共载水溶性和脂溶性荧光染料后的细胞内递送能力.结果表明:OQCMC上长链季铵盐分子的取代度和共乳化剂乙醇的加入量对相图中微乳区域的面积影响不大;微乳液法可制备包载水溶性长春新碱(VCR)、脂溶性消炎痛(IMC)或二者共载的OQCMC载药微球,微球粒径为(52.40±0.55)nm,分布均匀;微乳液体系对VCR的最大载药率为22.7%,对IMC的最大载药率为20.1%,二者共载时,VCR的最大载药率为12.2%,IMC的最大载药率为10.0%;载药微球对药物具有缓控释功能.OQCMC乙醇高聚物脂质体可有效地包载荧光染料异硫氰酸荧光素FITC(水溶性)和尼罗红(脂溶性),并将二者递送到卵巢癌HO8901细胞内.     

双重载药多层海藻酸盐-壳聚糖缓释微球的制备及体外释放

采用滴注法将海藻酸钠与钙离子交联,制成负载血管内皮生长因子(VEGF)的藻酸钙核心球,利用层层自组装技术在核心球的表面依次包覆壳聚糖、海藻酸和壳聚糖,壳聚糖中负载万古霉素(VAN),形成多药载药缓控体系.采用正交实验考察海藻酸钠浓度、钙离子浓度及壳聚糖浓度对VEGF和VAN的药物包封率和载药量的影响,优化了制备工艺.采用扫描电子显微镜观察多层微球的表面、截面形貌及粒径,采用傅里叶变换红外光谱检测海藻酸盐与壳聚糖的自组装情况,分别采用酶联免疫吸附(ELISA)双抗体夹心法和紫外分光光度法检测VEGF和VAN的包封率、载药量及体外释放情况.结果表明,海藻酸钠最优浓度为0.04g/mL,氯化钙最优浓度为0.15g/mL,壳聚糖最优浓度为0.01g/mL.微球光滑圆整,均质实心,直径900~1100μm,VEGF的包封率达61.31%,VAN的包封率为3.48%.体外释放实验结果表明,VEGF缓释时间为15.5d,并出现2个释放高峰;VAN缓释时间为4.5d,释药情况平稳持续,无明显突释.双重载药多层包覆微球兼具控制感染和促进血管生成两种潜能,有望应用于组织工程骨的基础研究和临床实践.     

香草醛交联壳聚糖载药微球的性能及其成球机理分析

以壳聚糖溶液为水相、液体石蜡为油相形成油包水型乳液, 以香草醛为交联剂, 采用乳化交联法制得壳聚糖微球. 结合IR光谱和XRD测试, 分析了壳聚糖交联固化成球的机理: 壳聚糖和香草醛之间所发生的Schiff碱反应和氢键的形成以及缩醛化反应, 以此为基础共同形成交联结构从而使壳聚糖交联固化成球. 探讨了交联后壳聚糖微球结晶度降低的原因: 壳聚糖固化时分子链未充分进行有序的结晶排列, 交联后的壳聚糖结构较复杂, 从而破坏了原壳聚糖分子的规整性. 选用盐酸小檗碱为模型药物, 制备了香草醛交联的壳聚糖载药微球, SEM结果显示, 载药微球表面致密且球形度好, 微球粒径在5-15 μm之间. 此外, 采用分光光度计对载药微球的载药率、药物包封率和药物体外释放性质进行了测试和分析, 结果表明载药微球缓释效果明显.     

羧甲基壳聚糖/有机累托石纳米复合材料及其药物缓释性能研究

利用简单的溶液插层法制备了羧甲基壳聚糖/有机累托石纳米复合材料,其中累托石(REC)用十六烷基三甲基溴化铵进行改性.用X-射线衍射(XRD)、红外光谱(FTIR)和扫描电镜(SEM)表征了该纳米复合材料的微观结构和形态,实验表明羧甲基壳聚糖插层进入了累托石层间,增大了累托石的层间距,并且累托石均匀地分布在羧甲基壳聚糖基体中.以牛血清蛋白(BSA)为药物模型,研究了纳米复合材料与海藻酸钠形成的微球的药物缓释性能.结果显示,该微球对药物的包封率及缓释性能与纯羧甲基壳聚糖微球相比都有较大改善,包封率从56%提高到86%,药物缓释时间从24 h上升到72 h.并且纳米复合材料/海藻酸钠微球的释药具有pH响应性,在pH为1.2的条件下释药慢,而在pH为7.4时释药快,可用于小肠或结肠定位缓释系统.因此,羧甲基壳聚糖/有机累托石纳米复合材料很有潜力作为药物载体.     

淀粉基荧光微球的制备及其布洛芬包载和释药性能

以淀粉为原料,用乙酸酐酰化后,将异硫氰酸荧光素（FITC）接枝到淀粉醋酸酯大分子链上,以自组装的方法制备出包载布洛芬的淀粉基荧光微球,同时考察了淀粉基荧光纳米微球对布洛芬的控制释放性能。 荧光淀粉酯的用量、布洛芬的加入量及丙酮与水的体积比可影响载药微球的包封率和药物释放速度。 研究结果表明,当荧光淀粉酯用量为200 mg、布洛芬的量为60 mg、水和丙酮体积分别为50和20 mL时所合成的载药微球包封率最高,为69.5%,其药物体外释放也最快,48 h可释放62.7%。 用扫描电子显微镜和激光共聚焦显微镜(CLSM)对包药微球的体外释放过程进行了表征。     

阿司匹林-蒙脱土-壳聚糖缓释微球的制备及其体外释放性能

利用溶液法预先制备壳聚糖(Cs)-蒙脱土(MMT)复合材料(Cs-MMT),以Cs-MMT、Cs为原料,采用反相悬浮聚合法制得一种新型药物缓释体系阿司匹林-蒙脱土-壳聚糖载药微球(Asp-MMT-Cs)。采用FT-IR、SEM表征了Cs-MMT和Asp-MMT-Cs载药微球的结构及形态;设计正交实验优化了Asp-MMT-Cs载药微球的制备工艺;通过体外释放实验探讨了载药微球在不同模拟释放液中的释药规律。结果表明:所得微球球形度好,粒径分布较均匀;最优工艺制得的载药微球平均粒径为81.20μm,载药量为9.61%,包封率为76.78%。该缓释体系具有pH敏感性,更倾向于在pH较高的磷酸盐缓冲溶液中释放。     

含胶原蛋白/壳聚糖微球保湿面膜的制备及其性能研究

采用喷雾干燥法制备胶原蛋白/壳聚糖复合微球,用纳米喷枪将复合微球的乙醇溶液均匀喷洒于粘胶纤维面膜上,制备了一种新型的含胶原蛋白/壳聚糖微球保湿面膜,并研究复合微球和面膜的形貌,面膜的保湿、透湿以及生物相容性。结果表明:喷雾干燥法制备的复合微球大小较为均一,形貌完整,含胶原蛋白/壳聚糖微球的面膜保湿率为85.3%,透湿率为4 300g·M~(-2)·h~(-1),细胞活性测试第七天仍有0.98,细胞活性大于0.8,表明该胶原蛋白;壳聚糖微球保湿面膜具有良好的生物相容性。     

羟基磷灰石／胶原／植物多酚复合材料的研究

以原花色素、茶多酚等植物多酚为交联剂,采用低温原位合成法制备羟基磷灰石/胶原/植物多酚(HA/COL/PP)复合材料。对材料的形貌、热稳定性、溶胀性质进行了表征。结果表明,植物多酚的加入使复合材料中各成分结合更紧密,增加了复合材料的热稳定性,降低了复合材料的溶胀度。比较研究表明,添加原花色素对上述性能的改善更有效。为了考察加入植物多酚后复合材料的生物活性,分别对羟基磷灰石/胶原/原花色素(HA/COL/PA)、羟基磷灰石/胶原/茶多酚(HA/COL/TP)复合材料进行了体外矿化能力研究,观察到两种材料的表面都形成了新的矿化沉积层,说明加入了植物多酚不影响复合材料的体外矿化能力。因而,羟基磷灰石/胶原/植物多酚复合材料是一种有潜力的骨替代材料。     

Synthesis, characterization and cytotoxicity studies of chitosan-coated tea polyphenols nanoparticles

Chitosan nanoparticles (CS-NPs) were prepared by ionic gelation method using carboxymethyl chitosan and chitosan hydrochloride as carriers of tea polyphenols. The characteristics of chitosan-coated tea polyphenols nanoparticles (CS-TP NPs) were determined by using transmission electron microscopy (TEM) and FT-IR spectroscopy. It was found that the synthesized CS-TP NPs were non-spherical in shape with an average size of 407±50nm. Meanwhile, the drug content and encapsulation rate of the nanoparticles was 8-16% and 44-83%, respectively. These CS-TP NPs also demonstrated sustained release of tea polyphenols in PBS. The antitumor of CS-TP NPs towards HepG2 cancer cells was investigated. The result showed that CS-TP NPs retained significant antitumor activities.     

Efficient Separation and Purification of Epigallocatechin Gallate (EGCG) Based on EGCG-Imprinted Polymer Prepared with Chitosan as Matrix

A molecularly imprinted polymer (MIP), which was suitable for recognizing epigallocatechin gallate (EGCG), was prepared by using EGCG as template molecule and biocompatible chitosan as a functional matrix in aqueous medium. Molecular recognition ability of the EGCG-imprinted polymer (EIP) was evaluated by high performance liquid chromatography (HPLC). The results show that the EIP has a high imprinting factor (1.32) for EGCG and was used to purify EGCG from crude tea polyphenol efficiently. The percentage of EGCG can be improved from 78.6% in crude tea polyphenol (TP) to 90.1% in product and the adsorption quantity per unit can reach 4.02 mg · g^?1. EIP shows potential excellent prospect in the application of separating and purifying EGCG from TP.     

Characterization and biological evaluation of paclitaxel-loaded poly(L-lactic acid) microparticles prepared by supercritical CO2

In this study, paclitaxel loaded poly( L-lactic acid) (PTX-PLLA) microparticles were prepared using solution enhanced dispersion by supercritical CO2(SEDS) technique. This supercritical antisolvent technique offers the advantage of negligible organic solvent residua in the drug loaded microparticles. Scanning electron microscopy (SEM) showed that microparticles exhibited rather spherical shape and small particle size with narrow particle size distribution. X-ray diffraction (XRD) and differential scanning calorimeter (DSC) indicated that PTX was amorphously dispersed in the PLLA matrix. The drug loading and encapsulation efficiency of PTX-PLLA microparticles were 14.33% and 62.68%, respectively. In vitro cytotoxicity evaluation of PTX-PLLA microparticles against nonsmall-cell lung cancer A549 and ovarian cancer SKOV3 cell lines indicated that PTX-PLLA had superior antiproliferation activity against the A549 and SKOV3 cell lines, compared with free PTX formulations. The cellular internalization of fluorescent microparticles was evidenced by fluorescence microscope and further confirmed by transmission electron microscopy (TEM). This was attributed to the efficient intracellular accumulation of PTX via cell phagocytosis and sustained release of PTX from PLLA matrix. The anticancer activity of PTX-PLLA was associated with PTX-induced cell apoptosis such as nuclear aberrations, condensation of chromatin and swelling damage in mitochondria. The cell apoptosis index detected by flow cytometry was higher in PTX-PLLA group than in free PTX. The PTX-PLLA formulation, which was obtained through micronization of PTX and encapsulation of micronized PTX into PLLA simultaneously in the SEDS process, significantly potentiated the anticancer activity of PTX.     

胺基化壳聚糖树脂吸附分离茶多酚的研究

对珠状壳聚糖树脂进行胺基化改性,制备了胺基化珠状壳聚糖树脂,并用FTIR对胺基化珠状壳聚糖树脂进行了结构表征。利用该树脂对绿茶中茶多酚进行吸附分离,探讨了其吸附条件,考察了其吸附性能。结果表明,胺基化壳聚糖树脂对茶多酚的吸附既符合Langmuir等温式,也符合Freundlich等温式;本实验最佳吸附条件为:温度25℃,茶汤溶液pH值5,吸附时间2h;最大吸附量达到486.0mg/g。     

UF-两段厌氧处理茶多酚废水的研究

采用前置超滤膜(UF)的两相厌氧工艺对原水COD为18362.6mg/L,荼多酚为3608.3mg/L、色度为2624.2倍的茶多酚生产废水进行为期90d的实验研究.结果表明,当实验压力为0.2Mpa时,膜组件对COD去除率为63.4%,茶多酚去除率为95.1%,色度去除率为93.4%.然后,对两相厌氧工艺的投配率、P含量和酸化段水力停留时间(HRT1)对废水COD、色度与茶多酚去除率和产气率的影响进行了研究.当投配率为15.0%、P含量为38.1mg/L、HRT1=24h,该工艺达到最佳处理效果,出水COD为1288.1mg/L,COD去除率为80.8%,色度为95.6倍,色度去除率为44.6%,残余茶多酚为119.8mg/L,茶多酚去除率为32.3%,产气率为0.85m3/kg COD,与未采用UF预处理的两相厌氧水解工艺相比,COD、色度和荼多酚去除率分别提高23.40%,10.2%和1613%,产气率增加0.15m3/kg COD.     

Preparation of PLLA/PLGA microparticles using solution enhanced dispersion by supercritical fluids (SEDS).

In this work, poly(L-lactic acid)/poly(lactide-co-glycolide) (PLLA/PLGA) microparticles were prepared using the technique of solution-enhanced dispersion by supercritical fluids (SEDS). For comparison, separate PLLA and PLGA microparticles were also produced by the same SEDS process. The produced microparticles were characterized by scanning electron microscopy, laser particle size analyzer, X-ray diffraction, differential scanning calorimetry, Fourier transform infrared spectroscopy, and gas chromatography. Results indicate that PLLA/PLGA microparticles possess sphere-like shapes with smooth surfaces. The mean particle size of PLLA/PLGA microparticles ranges from 1.76 to 2.15 microm, depending on the feeding ratio of PLLA to PLGA used in the SEDS process. The crystallinity of PLLA/PLGA microparticles decreases after the SEDS processing, so that the produced microparticles are in an amorphous state. Pure PLGA was hard to precipitate in small, fine microparticle form without the presence of PLLA. A model drug, paclitaxel, was encapsulated into PLLA/PLGA microparticles by the same SEDS process, and the in vitro release rate of paclitaxel from these PLLA/PLGA composites could be modulated by variation of the mixing ratio PLLA:PLGA. The prepared microparticles have negligible residual organic solvent. Drug-loaded PLLA/PLGA microparticles produced by SEDS have potential as an advanced colloidal suspension for pharmaceutical applications.     

Protein delivery based on uncoated and chitosan-coated mesoporous silicon microparticles

Mesoporous silicon is a biocompatible, biodegradable material that is receiving increased attention for pharmaceutical applications due to its extensive specific surface. This feature enables to load a variety of drugs in mesoporous silicon devices by simple adsorption-based procedures. In this work, we have addressed the fabrication and characterization of two new mesoporous silicon devices prepared by electrochemistry and intended for protein delivery, namely: (i) mesoporous silicon microparticles and (ii) chitosan-coated mesoporous silicon microparticles. Both carriers were investigated for their capacity to load a therapeutic protein (insulin) and a model antigen (bovine serum albumin) by adsorption. Our results show that mesoporous silicon microparticles prepared by electrochemical methods present moderate affinity for insulin and high affinity for albumin. However, mesoporous silicon presents an extensive capacity to load both proteins, leading to systems were protein could represent the major mass fraction of the formulation. The possibility to form a chitosan coating on the microparticles surface was confirmed both qualitatively by atomic force microscopy and quantitatively by a colorimetric method. Mesoporous silicon microparticles with mean pore size of 35 nm released the loaded insulin quickly, but not instantaneously. This profile could be slowed to a certain extent by the chitosan coating modification. With their high protein loading, their capacity to provide a controlled release of insulin over a period of 60-90 min, and the potential mucoadhesive effect of the chitosan coating, these composite devices comprise several features that render them interesting candidates as transmucosal protein delivery systems.     

新型微球载体材料——PACA-co-PEG的性能

聚氰基丙烯酸烷基酯;药物载体;生物降解性     

In Vitro Biomineralization and Bulk Characterization of Chitosan/Hydroxyapatite Composite Microparticles Prepared by Emulsification Cross-Linking Method: Orthopedic Use

Chitosan/hydroxyapatite composite microparticles were prepared by a solid-in-water-in-oil emulsification cross-linking method. The characteristics and activity in presence of simulated body fluid for 14 and 21?days were investigated. The size distribution, surface morphology, and microstructure of these biomaterials were evaluated. The scanning electron microscopy revealed an aggregate of microparticles with a particle size, ranged from 4 to 10???m. The deposited calcium phosphate was studied using X-ray diffraction analysis, Fourier transform infrared spectroscopy, and inductively coupled plasma/atomic emission?spectroscopy analysis of phosphorus. These results show that the mineral, formed on microparticles, was a mixture of carbonated hydroxyapatite and calcite. Scanning electron microscopy revealed that calcium phosphate crystals growth was in form of rods organized as concentric triangular packets interconnected to each other by junctions. Interaction between chitosan and growing carbonated hydroxyapatite and calcite crystals are responsible for a composite growth into triangular and spherical shapes. The results demonstrated that these microparticles were potential materials for bone repair.