Biocompatibility studies of silk fibroin-based artificial nerve grafts in vitro and in vivo

Silk fibroin (SF) has been used extensively in the biomedical field including tissue engineering for the generation of artificial bones, skins or ligaments. We have previously reported on good in vitro biocompatibility of SF fibers with peripheral nerve tissues and cells. In the present study, we developed a novel design of the SF-based artificial nerve graft (SF graft) which was composed of a SF-nerve guidance conduit (NGC) inserted with SF fibers. MTT assay was performed to determine the cytotoxicity of the SF-NGC extract fluid on the cultured L929 cells derived from an immortalized mouse fibroblast cell line. In addition, this SF graft was implanted into adult rats for bridging a 10-mm long sciatic nerve defect. The following-up experiments at initial stage (1—4 week) of nerve regeneration including routine blood tests and histochemical investigation were conducted to evaluate the in vivo biocompatibility of the SF graft with peripheral nerves. The results demonstrated that the SF-NGC graft was biocompatible with the surrounding tissues and cells due to its low inflammatory potential with a grade 0 under the U.S. Pharmacopeia guidelines and it was generally suitable to a certain degree for bridging peripheral nerve defects in virtue of supporting Schwann cell adherence, expansion and migration. Therefore the SF graft is a promising alternative to classical autografts for peripheral nerve repair.     

羟基磷灰石/丝蛋白复合骨材料的生物相容性

以共沉淀法制备了羟基磷灰石/丝蛋白(HA/SF)复合骨修复材料,并对材料进行了XRD、TEM、SEM、孔径分布和孔隙率等相关的测试和表征。结果表明:该复合材料的无机组分为20~30 nm长、5 nm宽的棒状羟基磷灰石(HA)结晶,这些晶粒沿c轴自组装团聚成簇分散在丝蛋白基质中形成三维多孔结构,其孔径分布在0.3~115μm之间,开口孔隙率达66%。该材料植入动物体内,未出现明显的排异反应,在植入部位有连续的骨性连接和新生骨形成,说明HA/SF复合材料具有良好的生物相容性和骨诱导活性。     

Cu（Ⅱ） effect on the conformation of regenerated silk fibroin in dilute aqueous solution

Much attention has been paid to the natural mechanism of silkworm spinning due to the impressive mechanical properties of the natural fibers. In this work, we studied the effect of Cu（Ⅱ） ions on the secondary structure of Bombyx mori regenerated silk fibroin （SF） in dilute solution by circular dichroism （CD）. The results indicate that a given amount of Cu（Ⅱ） induces the SF conformational transition from random coil to β-sheet, however, further addition of Cu（Ⅱ） is unfavorable for this conversion. Meanwhile, the conformational changes induced by Cu（Ⅱ） follow a nucleation-dependent aggregation mechanism, which is similar to that found in Prion protein （PrP） denaturation and Aβ-peptide aggregations, leading to the neurodegenerative disease. This work would help one understand further the natural spinning process of silkworm. Additionally, it would be significant for the study of the nervous system diseases, because silk fibroin, extracted in large amounts from Bombyx mori silkworm gland, could be a proper model to study PrP denaturation and Aβ-peptide aggregations.     

Cu(II) effect on the conformation of regenerated silk fibroin in dilute aqueous solution

In nature, metal ions play a vital role in the function of protein and nucleic acid. It has been demonstrated that there are many kinds of metal ions in the gland of Bombyx mori (B. mori) silkworm, and their contents are different in different parts of silk gland[1]. These ions may promote the transition of gel to sol in the gland of silkworm when the silk fibroin (SF) goes forward to the spinneret. Many investigations have shown that K(I) and Ca(II) can induce the conformational transitio…     

丝素在NMMO·H2O中的溶解及溶液流变性能的研究

探讨了丝素在N-甲基吗啉-N-氧化物一水合物(NMMO.H2O)中的溶解行为,结果发现:天然蚕丝纤维脱胶后得到的丝纤维可以直接溶解在NMMO.H2O中,但是很难得到高于10%浓度的溶液,因而无法用于纺丝.而由再生丝素水溶液制得的再生丝素膜则易于溶解在NMMO.H2O中,并可以得到浓度高达25%的再生丝素/NMMO.H2O溶液,在上述研究的基础上,本文还对再生丝素/NMMO.H2O的流变行为进行了初步研究.     

基于遗传算法和神经网络优化的故障诊断专家系统

传统故障诊断专家系统中存在知识获取困难和推理能力弱的问题,采用模糊神经推理控制器作为推理机可改善此类问题,为进一步提高系统的性能,用遗传算法对推理机进行离线训练．在系统中,使用了传统故障诊断专家系统给予规则的推理方式,使系统在提高灵活性的基础上保持了透明性。     

Role of humidity on the structures and properties of regenerated silk fibers

Silk fiber was processed from highly concentrated spinning dope to solid fibers along with water removal. To understand the mechanism of water removal during silk fiber spinning process, a microfluidic chip was designed and applied to investigate the structures and mechanical properties of two kinds of regenerated silk fibroin fibers dry-spun at different relative humidity. The experimental results showed that the diameters of the fibers spun at 40% RH are always larger than the fibers spun at 50% RH due to different removal rates of water. The fibers spun at low humidity contain more β-sheet structure and lower degree of chain orientation and crystalline orientation. These results indicate that the fast phase transition of silk fibroin from sol–gel to silk fiber undergoes with rapid water removal and higher fiber orientation relates to more residue water and drawing force.     

蚕丝蛋白仿生多肽RAGA16的纳米结构与性能研究

利用天然蛋白质氨基酸序列设计生物仿生材料是近年来兴起的一个热门研究领域.本文中将蚕丝蛋白特征氨基酸序列(Gly-Ala-Gly-Ala-Gly-Ser)部分引入离子互补型多肽RA-DA16-Ⅰ中,设计了新型多肽RAGA16.采用原子力显微镜(AFM)、旋转流变仪(Rheometer)、傅立叶变换红外光谱仪(FTIR)、倒置荧光显微镜等技术对多肽RAGA16的自组装结构、肽链的二级结构、流变学性能以及细胞相容性等进行研究.结果表明,蚕丝蛋白多肽序列引入后,多肽RAGA16具有自组装特性,能够自组装形成致密的纳米级三维网络结构,所形成的水凝胶力学性能明显提高,通过分析得知这可能是由于多肽二级结构中Silk Ⅰ结构成分比例增加所致.荧光染色显示,绝大多数接种于多肽水凝胶中的小鼠前成骨细胞MC3T3-E1能存活,且在不同的三维平面上生长增殖.综上,利用蚕丝蛋白特征序列对RADA16-Ⅰ进行仿生改性后,对拓展多肽材料在生物医学领域的应用具有重要意义.     

不同聚合物修饰的金纳米棒及其生物相容性研究

十六烷基三甲基溴化铵(CTAB)在制备金纳米棒时常常作为一种阳离子表面活性剂,但CTAB本身对细胞有毒害作用,降低其偶联的金纳米棒(GNR)的毒性是很必要的.通过层层组装的方法合成了不同聚合物修饰的金纳米棒,并采用TEM、UV-vis吸收光谱、Zeta电位和MTT比色法进行了表征.结果表明:通过聚合物修饰,金纳米棒细胞毒性明显降低,具有良好的生物相容性.     

人工肛门括约肌性能与生物相容性的动物实验

设计动物实验,验证优化设计后人工肛门括约肌系统功能性和生物相容性.实验分别在术前和术后第1、4d,第1、2、4、5周,从2条Beagle犬颈静脉抽取血样进行血清化学分析.分析参数包括:总蛋白,白蛋白,总胆红素,天冬氨酸转氨酶,尿素氮,肌酐酸和高敏C反应蛋白.实验结果表明:系统各模块功能正常,肝功能指标总胆红素含量正常,天冬氨酸转氨酶术后略高于正常范围,术后2周恢复正常;肾功能指标尿素氮正常,肌酐酸含量在正常范围内波动;总蛋白和白蛋白含量术后虽有波动,但均恢复至正常范围;炎性因子高敏C反应蛋白含量未发生变化,表明体内无感染炎症现象;各血样指标显示人工肛门括约肌对Beagle犬生理功能并未造成不良影响.     

胶原修饰快速成形PLGA/TCP人工骨支架体外生物相容性研究

探讨经Ⅰ型胶原修饰的快速成型(RP)聚乳酸-羟基乙酸/磷酸三钙(PLGA/TCP)支架的生物相容性,为进一步体内实验提供研究基础.以经Ⅰ型胶原修饰改性的PLGA/TCP支架作为实验组,未经胶原修饰的原始支架作为对照组,检测两组支架材料的亲水性.BMSCs接种两组支架,分别测定细胞黏附率、细胞增殖率.扫描电镜(SEM)观察细胞黏附形态,以检验支架材料的细胞相容性.结果表明胶原改性支架具有较高的亲水性,实验组细胞黏附率在接种4、8和12 h者,明显优于对照组(P<0.05).实验组在培养2、4、6和8 d的细胞增殖率也明显高于对照组(P<0.05).扫描电镜可见骨髓基质细胞在实验组支架上的增殖数量多于对照组.说明Ⅰ型胶原改性的PLGA/TCP支架具有良好的细胞生物相容性,可作为骨组织工程支架应用于骨缺损的修复研究.     

NKA-9树脂对血浆中胆红素的吸附及生物相容性

将NKA-9树脂装柱处理后,应用重度黄疸性疾病新生儿患者的换血血浆分别在静态和灌流状态下进行吸附,分别测定不同时间、不同胆红素浓度下的吸附率.并以吸附柱直径、灌流速度、温度三因素设计三水平正交实验,探索较佳的吸附条件.再测定灌流过程中血浆有效成分的变化量,并做生物相容性实验.结果显示NKA-9树脂对胆红素在静态和灌流状态下的吸附率分别在36%～45%和70%～90%之间.较佳吸附时间为40 min、吸附柱直径与体积比为0.20 cm-2、灌流速度为100.56 cm/h、温度25℃.在较佳吸附条件下NKA-9树脂对血浆中胆红素的最大吸附率接近95%.实验证明NKA-9树脂对血浆中胆红素的吸附率较高,且血液相容性和生物相容性良好,可作为临床血液灌流法治疗新生儿黄疸性疾病的吸附剂.     

Structure and Properties of Porous Silk Fibroin Materials

In this article the structure and properties of freeze-dried porous silk fibroin materials were tested and analyzed.The results indicated that for porous silk fibroin materials prepared by means of freeze-drying,if freez-ing temperature was below —20℃,the structure of silk fibroin was mainly amorphous with a little silk Ⅱ crystal structure,and if freezing temperature was above —20℃,quite a lot of silk Ⅰ crystal structure formed.Porous silk fibroin materials,with average pore diameter be-     

几种玉米须黄酮分子结构性质及抗氧化活性的量子化学研究

采用理论计算化学手段结合分子动力学与量子化学方法,对玉米须相关的几种黄酮类化合物槲皮素、木犀草素、芹菜素分子结构和活性进行了理论计算.从几何结构、自由基稳定性、前线轨道能等角度分析了影响黄酮类化合物抗氧化活性的因素.在分子层面上探讨了玉米须黄酮的抗氧化作用机理,预测的黄酮类化合物的抗氧化活性与实验结果一致.     

羧甲基壳聚糖接枝聚丙烯酸水凝胶的制备及体内外评价

以丙烯酸(AA)和羧甲基壳聚糖(CMC)为单体,通过热引发自由基接枝共聚反应,制备了一种新型pH敏感的羧甲基壳聚糖接枝聚丙烯酸(CMC-g-PAA)水凝胶。FT-IR结果表明成功实现了聚合反应,溶胀性实验表明CMC-g-PAA水凝胶具有明显的pH敏感性。以胰岛素(INS)为模型药物,将其负载到CMC-g-PAA水凝胶中,得到了载药量为216.5mg/g的载药凝胶。体外释放曲线表明:在pH值为1.2的条件下,2h后INS的累计释放量为(16.3±2.6)%;在pH值为7.4的环境中,2h后INS的累计释放量达到(57.2±3.5)%。说明载药凝胶可以在肠道环境中靶向释放INS,避免INS被胃酸和胃蛋白酶破坏。动物实验结果表明,负载INS水凝胶具有良好的降血糖效果。CMC-g-PAA水凝胶与Caco-2细胞共同培养,细胞存活率接近100%,表明水凝胶对Caco-2细胞没有细胞毒性。CMC-g-PAA水凝胶在蛋白或多肽类药物定位递送方面具有优良的应用前景。     

基于人工鱼群的交通诱导系统最优查询研究

与齐齐哈尔市公安交通警察支队交通控制中心合作开发,并以齐齐哈尔市路况为实际进行凋研,将改进的人工鱼群算法应用于交通路径诱导系统数据库优化查淘中,算法提高了最优路径查询的效率。对人工鱼群算法进行了改进,引入贝叶斯变异算子和十字交叉变异算子,避免了算法局部寻优能力差和收敛速度慢等弊端。仿真实验表明算法是正确有效性的。     

In vitro and in vivo studies on biodegradable magnesium alloy

The microstructure, mechanical property, electrochemical behavior and biocompatibility of magnesium alloy(Bio De MSM^TM) were studied in the present work. The experimental results demonstrated that grain refining induced by extrusion improves the alloy strength significantly from162 MPa for the as-cast alloy to 241 MPa for the as-extruded one. The anticorrosion properties of the as-extruded alloy also increased.Furthermore, the hemolysis ratio was decreased from 4.7% for the as-cast alloy to 2.9% for the as-extruded one, both below 5%. Bio De MSM^TMalloy shows good biocompatibility after being implanted into the dorsal muscle and the femoral shaft of the New Zealand rabbit, respectively, and there are no abnormalities after short-term implantation. In vivo observation indicated that the corrosion rate of this alloy varies with different implantation positions, with higher degradation rate in the femur than in the muscle.     

多孔柞蚕丝素支架的制备及其在药物缓释中的应用

采用冷冻干燥法制备了多孔柞蚕丝素支架,分析了丝素溶液浓度与支架孔径、孔隙率之间的关系,并研究了多孔柞蚕丝素支架负载5-氟尿嘧啶的缓释性能.研究表明,当丝素浓度较低时,支架中的孔洞分布较为均匀,孔径和孔隙率也较大,而随着丝素溶液浓度增大,支架的孔径和孔隙率逐渐减小;支架的药物释放过程可分为快速释放和缓慢释放两个阶段.     

春秋战国时期丝绸纹样的构成

为全面了解春秋战国时期丝绸纹样风格的形成及影响因素,以考古资料为依据,从纹样的总体文化内涵上,分析其人文背景、造型特征、组织形式及寓意.归纳出这一时期丝绸纹样的构成特点,为继承和创新现代丝绸纹样的设计提供借鉴.     

异叶败酱化学成分的研究及体外细胞毒性检测

采用硅胶柱色谱法对异叶败酱（Patrinia heterophylla Bunge.）化学成分进行了研究．通过理化性质及波谱学方法对分离得到的化合物进行结构鉴定，并运用SRB法对分离的部分化合物进行体外细胞毒性测试．从该植物中分离鉴定了10个化合物，分别为：软木三萜酮（I），齐墩果酸（Ⅱ），熊果酸（Ⅲ），常春藤皂甙元（Ⅳ），海棠果醇（V），齐墩果酸-3—O-α—L-吡喃阿拉伯糖苷（Ⅵ），β-香树精（Ⅶ），α-香树精（Ⅷ），β-谷甾醇（Ⅸ），胡萝卜苷（X），除化合物Ⅱ之外，其余化合物都是首次从该植物中分离得到．SRB法结果表明：化合物Ⅱ、Ⅲ、Ⅳ和Ⅵ有较强的细胞毒性．