温度敏感葡聚糖凝胶的合成及其细胞相容性研究

合成了N-异丙基丙烯酰胺（NIPAAm）与葡聚糖的共聚水凝胶，利用红外光谱表征了单体和凝胶结构，研究了水凝胶在去离子水和细胞培养基中的温度响应性，并对凝胶进行了细胞培养和脱附研究．结果表明，共聚凝胶具有很好的温度敏感性和细胞相容性，可以通过控制温度变化，实现对细胞的无损伤脱附．     

Partial or total silylation of dextran with hexamethyldisilazane

The silylation reaction of dextran with 1,1,1,3,3,3-hexamethyldisilazane (HMDS) in DMSO was studied as the first step of the synthesis of new amphiphilic polyester-grafted dextrans. According to the experimental conditions, i.e. dextran molar weight, medium temperature and reaction time, HMDS/OH ratio, addition of a catalyst and co-solvent, partially or totally silylated dextrans were recovered. The highest silylation yields were obtained with the lowest molecular weight dextrans. The increase in temperature medium and/or reaction time, the presence of catalyst or co-solvent favored the protection yield. Whatever the dextran used, complete silylation of the polysaccharide chain could be achieved by adequate selection of the experimental conditions. The thermal properties of resulting silylated polysaccharides were investigated by temperature modulated DSC. It was observed that T_g values of partially silylated dextran were maintained between 120 and 140 °C, independently of the dextran molecular weight. Interestingly, DMSO proved to behave as an efficient plasticizer of (partially) silylated dextrans. The partially silylated dextrans were efficiently used as multifunctional macroinitiators for the controlled ring-opening polymerization (ROP) of lactone. The ROP was then promoted from the remaining hydroxyl groups in the presence of tin or aluminium activator. After polymerization and ultimate deprotection of the silylated dextran backbone, amphiphilic polyester-grafted dextrans were readily recovered.     

葡聚糖修饰漆酶的制备及其催化特性和稳定性

将经NaIO₄氧化处理的葡聚糖（500×10³）连接在漆酶分子上以提高漆酶在酸性溶液环境中的稳定性。HPLC和SDS-PAGE分析表明葡聚糖修饰漆酶的分子量在200×10³以上。荧光光谱和圆二色性光谱分析表明修饰漆酶保持了天然漆酶的三级和二级结构。酶催化反应显示修饰漆酶与天然漆酶具有类似的底物亲和性和催化活性，而其在酸性条件下的稳定性显著提高。通过葡聚糖修饰使得漆酶在pH 3,50℃下的酶活半衰期从0. 07 h提高到17. 1 h，在含30%（体积）乙腈, pH 3下的酶活半衰期从0. 11 h提高到10. 3 h。在不同pH下测定修饰漆酶在216 nm处的圆二色性光谱，结果显示偶联葡聚糖增强了漆酶分子的结构稳定性，提高了其在高温及有机溶剂存在下的催化活性。     

生长抑素-葡聚糖的99Tcm标记及体外结合分析

采用还原氨化方法，将天然生长抑素（SMS）与葡聚糖(Dx20)偶联，并以125I－奥曲肽（125I-Tyr3-Octreotide)为放射配基，进行受体竞争结合实验，测定了SMS-Dx50的IC50；以SnCl2为还原剂对SMS-Dx50进行99Tcm标记，并进行99Tcm- SMS-Dx50受体结合分析，考察其对生长抑素受体的亲合能力。结果表明：SMS-Dx50保持了对SMS 2型受体的高亲和力；其IC50与奥曲肽的IC50（0.79 nmol/L）相近,为5.95 nmol/L；99Tcm- SMS-Dx50标记率>85％, 经PD-10柱分离纯化，其放化纯度>98%；99Tcm- SMS-Dx50对生长抑素2型受体特异性结合为25%～40％，保持了较高的亲和力；SMS-Dx50适合于99Tcm标记，可用于生长抑素受体阳性肿瘤诊断的进一步研究     

凝胶渗透色谱法检测淀粉酶解产物相对分子质量及其分布

运用普适校正原理,不需要Mark常数K、α值,使用示差折光检测仪及凝胶渗透色谱系统,直接测定了葡聚糖的相对分子质量及相对分子质量分布,优化了色谱操作条件,证明凝胶渗透色谱是监控淀粉酶解反应程度的一种有效手段.     

Creep-resistant dextran-based polyurethane foam as a candidate scaffold for bone tissue engineering: Synthesis,chemico-physical characterization,and in vitro and in vivo biocompatibility

A highly crosslinked composite dextran-based scaffold (named DexFoam) was tailored to overcome specific deficiencies of polymeric and ceramic bone scaffolds and to guarantee a bone-mimicking microenvironment for the proliferation of human mesenchymal stem cells in vitro. The creep resistance for up to 90% compressive stain, the capability to regain the original shape after deformation, and the good thermal stability in both physiological and “body limit” conditions make DexFoam a valid alternative to the currently available bone scaffolds. Histopathological evaluation for host reaction and tissue colonization of DexFoam scaffold, implanted subcutaneously in mice, demonstrated its in vivo biocompatibility and biodegradability.     

pH敏感型交联微球的制备

采用铈盐-羟基氧化还原引发体系,以对苯乙烯磺酸钠(SSS)和葡聚糖(Dex)为原料,通过自由基接枝聚合将聚对苯乙烯磺酸钠(PSSS)接枝到Dex上,得到接枝共聚物Dex-g-PSSS,再通过交联反应合成C(Dex-g-PSSS)微球。利用FTIR、光学显微镜、Zeta电位仪表征了试样的结构和形态,研究了反应条件对PSSS接枝度的影响,考察了C(Dex-g-PSSS)微球在模拟胃液和结肠液中的溶胀率。表征结果显示,C(Dex-g-PSSS)微球为交联微球,其Zeta电位为绝对值较大的负值。实验结果表明,在适宜的反应条件(硫酸铈铵浓度12 mmol/L、H+浓度0.08 mol/L、Dex 5 g、SSS 5 g、50℃,6 h)下,可得到接枝度为11.86 g(基于100 g Dex-g-PSSS)的PSSS。C(Dex-g-PSSS)微球有较明显的p H敏感性,它在模拟胃液中的溶胀率仅有134%,在模拟结肠液中的溶胀率可达到678%,是潜在的口服结肠定位给药系统基质材料。     

微结构葡聚糖凝胶浓缩分离蛋白质的性能

采用UV,HPLC等测试方法研究了含微结构的葡聚糖凝胶（BHMs）对蛋白的浓缩分离性能。结果表明：含微结构的葡聚糖凝胶在室温下对牛血清蛋白的浓缩分离效率达95.3%以上,随温度的升高,浓缩分离效率降低,且当温度超过BHMs凝胶的低临界溶液温度(40℃)后,浓缩分离效率发生突变;和普通交联葡聚糖凝胶相比,通过控制流动相的温度,葡聚糖BHMs凝胶可实现对多种不同分子量蛋白的高效分离,提高了分离效率,分离时间由24h缩短至9h。吸水溶胀动力学以及SEM分析表明,BHMs凝胶的溶胀满足指数动力学方程,疏松多孔性结构及由其决定的温度敏感性是BHMs凝胶具备良好浓缩分离性能的原因。     

热疗用葡聚糖磁流体的制备及表征

用化学共沉淀法,以不同浓度的葡聚糖40000和20000水溶液作为分散剂,制备纳米铁氧体磁流体。铁氧体粒子的平均直径为15 nm左右,其分散主要依靠空间位阻作用维持。葡聚糖磁流体中铁氧体粒子与葡聚糖以氢键、范德华力结合,葡聚糖包覆量约为8%。随葡聚糖相对分子质量和浓度的升高,磁流体(2.45%)的稳定性上升,用50%葡聚糖40000水溶液制备的磁流体,在室温静置72 h以上,没有分层现象。     

葡聚糖接枝型高载量金属螯合介质的制备与性能

对琼脂糖凝胶微球进行烯丙基活化,再接枝葡聚糖分子,考察葡聚糖分子量等因素对葡聚糖接枝过程的影响;以葡聚糖接枝琼脂糖凝胶微球为基质,制备亚氨基二乙酸型金属螯合介质,考察葡聚糖接枝过程对金属螯合介质的孔道结构、流通性能和载量等的影响. 结果表明,分子量20~500 kDa的葡聚糖都能均匀分布于琼脂糖凝胶微球内,葡聚糖接枝量随分子量增加而增大,所制的金属螯合介质形貌、粒径及其分布基本不受影响,且具有更好的流通性能,孔道结构比商品介质Ni Sepharose 6FF更丰富. 葡聚糖接枝的金属螯合介质对带组氨酸标签的乳酸脱氢酶和睫状神经营养因子的载量分别达到19和27 mg/mL,较Ni Sepharose 6FF的载量分别提高26.6%和42.0%.