Green Electrospun Silk Fibroin/Galactose Chitosan Composite Nanofibrous Scaffolds for Hepatic Tissue Engineering

The electrospun nanofibrous scaffolds made of proteins and polysaccharides were thought to be able to simulate the structure of natural extracellular matrix well.Silk fibroin(SF)and chitosan(CS)are probably the most widely used natural materials in biomedical fields including liver tissue engineering for their good properties and wide variety of sources.The asialoglycoprotein receptors of hepatocyte were reported to specifically recognize and interact with galactose.In this work,a green electrospun SF/galactosylated chitosan(GC)composite nanofibrous scaffold was fabricated and characterized.The data indicated that the addition of GC greatly influenced the spinning effect of SF aqueous solution,and the average diameter of the composite nanofibers was about 520nm.Moreover,the green electrospun SF/GC nanofibrous scaffolds were demonstrated significantly enhancing the adhesion and proliferation of hepatocyte(RH35)according to our data.The present study did a useful exploration on constructing scaffolds for liver regeneration by green electrospinning,and also laid a good foundation for the further applicative research of this green electrospun scaffolds in liver tissue engineering.     

Disc-Electrospun PCL Nanofiber Formed Micro-patterned Structure Scaffold

Disc-electrospinning using a disc as spinneret and a rotary drum as collector is a novel technology to prepare nanofiber which has been applied in tissue engineering scaffolds. In this study, nanofibrous mats with mlcro-patterned structure were fabricated via disc-electrospinning. Poly （ε-eaprolactone） （PCL） was dissolved in trifluoroethanol （TFE） at various concentrations （ 2 %-7 % ） （w/v） for electrospinning and the applied voltage ranged from 40 to 70 kV. Scanning electron microscopy （SEM） was employed to observe the morphology of the nanofibrous scaffolds. SEM images illustrated that the nanofibers with beads formed micro-patterned structure such as triangles and other polygons. The average diameter of nanofibers presented various size with the concentration increased from 2% to 7%. The beads on the nanofibers constructed the vertexes of the polygons, while nanofibers bridged between adjacent vertexes. The concentration of solution and applied voltage may be two dominant factors to influence the topological structure of the nanofibrons scaffolds. Cells cultured on the micro-patterned scaffold spread along the edges of the polygons. The scaffold with patterned structure may have a promising application in tissue engineering.     

壳聚糖/聚乙烯醇/氧氟沙星共混无纺布的制备及体外释放研究

以静电纺丝法制备了具有抗菌性能的壳聚糖/聚乙烯醇/氧氟沙星共混无纺布,测定了共混无纺布的力学性能,通过红外光谱(FT-IR) 、X-射线衍射(XRD)和扫描电子显微镜(SEM)对其结构性能进行了表征,选出了载药量较大、力学性能较好(壳聚糖/聚乙烯醇质量比为7,氧氟沙星质量分数为4%)的共混无纺布,在不同pH值的缓冲溶液中对其进行了体外药物释放实验。结果表明,所得的无纺布纤维直径均匀,平均为200nm。聚乙烯醇可以改善氧氟沙星/壳聚糖/聚乙烯醇共混无纺布的力学性能;氧氟沙星在壳聚糖/聚乙烯醇共混膜中有良好的相容性;氧氟沙星在pH为7.40的缓冲溶液中达到释放平衡时需要6h。     

新型前交叉韧带支架材料的构建及其生物相容性的实验研究

探讨以聚羟基丁酸己酯/聚左旋乳酸(PHB/PLLA1∶1)胶原杂化支架作为前交叉韧带组织工程载体材料的可行性。制备"三明治"样结构PHB/PLLA共聚物并测量其孔隙率等指标。以I型胶原对制备的PHB/PLLA支架进行杂化,获得PHB/PLLA胶原杂化支架。扫描电镜观察其表面结构。将兔皮肤成纤维细胞(SF)接种于PHB/PLLA胶原杂化支架,共培养5d后,扫描电镜下观察其在材料上生长情况。PHB/PLLA支架杂化后胶原填充于纤维空隙,分布比较均匀。体外培养的皮肤成纤维细胞成功种植在支架材料上,在材料上粘附、生长良好。说明构建的支架材料具有良好的三维构型和生物相容性,有望为前交叉韧带损伤的修复提供了一种新型的支架材料。     

Hippocampal Neuron-Derived Extracellular Matrix Coated Nanofibrous Scaffold for Neural Tissue Engineering

The aim of this study is to prepare poly-L-lactide( PLLA) electrospun nanofibrous scaffolds coated with hippocampal neuron-derived extracellular matrix( N-ECM)and construct a novel neural tissue engineering scaffold.Neonatal rat hippocampal neurons were seeded on PLLA nanofibers,and then decellularized to derive a cell-free extracellular matrix loaded N-ECM/PLLA modified scaffolds. The morphology and ingredients of N-ECM/PLLA were observed by scanning electron microscopy( SEM) and immunofluorescence staining respectively, and the cytocompatibility of the composite scaffolds was characterized by cell count kit-8( CCK-8) assay. The N-ECM was clearly identified loading on scaffolds when being imaged via SEM and immunofluorescence staining results showed that the N-ECM was made up of fibronectin and laminin. Most importantly, compared with tissue culture polystyrene and pure scaffolds, N-ECM/PLLA scaffolds could effectively facilitate the proliferation of rat adrenal neuroma cells( PC12 cells),indicating their better cell compatibilities. Based on the combination of N-ECM and PLLA biomaterials,the present study has fabricated a unique and versatile neural tissue engineering scaffold,offering a new thought for future neural tissue engineering.     

Dexamethasone-Loaded PLGA Microspheres Incorporated PLLA/PLGA/PCL Composite Scaffold for Bone Tissue Engineering

The combination of micro-carriers and polymer scaffolds as promising bone grafts have attracted considerable interest in recent decades.The poly(L-lactic acid)/poly(lactic-co-glycolic acid)/polycaprolactone(PLLA/PLGA/PCL)composite scaffold with porous structure was fabricated by thermally induced phase separation(TIPS).Dexamethasone(DEX)was incorporated into PLGA microspheres and then loaded on the PLLA/PLGA/PCL scaffoldtopreparethedesiredcompositescaffold.The physicochemical properties of the prepared composite scaffold were characterized.The morphology of rat bone marrow mesenchymal stem cells(BMSCs)grown on scaffolds was observed using scanning electron microscope(SEM)and fluorescence microscope.The resultsshowedthatthePLLA/PLGA/PCLscaffoldhad interconnected macropores and biomimetic nanofibrous structure.In addition,DEX can be released from scaffold in a sustained manner.More importantly,DEX loaded composite scaffold can effectively support the proliferation of BMSCs as indicated by fluorescence observation and cell proliferation assay.The results suggested that the prepared PLLA/PLGA/PCL composite scaffold incorporating drug-loaded PLGA microspheres could hold great potential for bone tissue engineering applications.     

Incorporation of Sphingosine 1-Phosphate Loaded Mesoporous Silica Nanoparticles into Poly( L-lactic acid ) Nanofibrous Scaffolds for Bone Tissue Engineering

Controlled release of the functional factors is the key to improve clinical therapeutic efficacy during the tissue repair and regeneration.The three-dimensional(3D)scaffold can provide not only physical properties such as high strength and porosity but also an optimal environment to enhance tissue regeneration.Sphingosine1-phosphate(S1P),an angiogenic factor,was loaded into mesoporous silica nanoparticles(MSNs)and then incorporated into poly(L-lactic acid)(PLLA)nanofibrous scaffold,which was fabricated by thermally induced phase separation(TIPS)method.The prepared scaffolds were examined by attenuated total reflection Fourier transform infrared spectroscopy(ATR-FTIR),scanning electron microscopy(SEM),and transmission electron microscopy(TEM)and compressive mechanical test.The ATR-FTIR result demonstrated the existence of MSNs in the PLLA nanofibrous scaffold.The SEM images showed that PLLA scaffold had regular pore channel,interconnected pores and nanofibrous structure.The addition of MSNs at appropriate content had no visible effect on the structure of scaffold.The compressive modulus of scaffold containing MSNs was higher than that of the scaffold without MSNs.Furthermore,fluorescein isothiocyanate(FITC)was used as model molecule to investigate the release behavior of S1P from MSNsincorporated PLLA(MSNs/PLLA)nanofibrous scaffold.The result showed that the composite scaffold largely reduced the initial burst release and exhibited prolonged release of FITC than MSNs.Thus,these results indicated that S1P-loaded composite nanofibrous scaffold has potential applications for bone tissue engineering.     

Electrospun Nanofibers of Hydroxyapatite/Collagen/Chitosan Promote Osteogenic Differentiation of BMSCs

Bone tissue engineering, aiming at developing bone substitutes for repair and regeneration of bone defects instead of using autologous bone grafts, has attracted wide attention in the field of tissue engineering and regenerative medicine. Developing biomimetic biomaterial scaffolds able to regulate osteogenic differentiation of stem cells could be a promising strategy to improve the therapeutic efficacy. In this study, clectrospun composite nanofibers of hydroxyapatite/collagen/chitosan （ HAp/Col/CTS ） resembling the fibrous nanostructure and constituents of the hierarchically organized natural bone, were prepared to investigate their capacity for promoting bone mesenchymal stem cells （BMSCs） to differentiate into the osteogenic lineage in the absence and presence of the osteogenlc supplementation, respectively. Call morphology, proliferation and quantified specific osteogenic protein expression on the electrospun HAp/Coi/CTS scaffolds were evaluated in comparison with different controls including dectrospun nanofibrous CTS, HAp/CTS and tissue culture plate. Our remits showed that the nanofibrous HAp/Col/CTS scaffolds supported better spreading and proliferation of the BMSCs than other substrates （ P 〈 0.01 ）. Expressions of osteogenesis protein markers, alkaline phosphatase （ALP） and Col, were significantly upregulated on the HAp/Col/CTS than those on the CTS （P 〈0.01） and HAp/ CTS （P 〈 0. 05 ） scaffolds in the absence of the osteogeulc supplementation. Moreover, presence of osteogeulc supplementation also proved to enhance osteogeule differentiation of BMSCs on HAp/ Col/CTS scaffolds, indicative of a synergistic effect. This study highlights the potential of BMSCs/HAp/Col/CTS cell-scaffold system for functional bone repair and regeneration applications.     

Effect of Silica Fume on Mechanical Properties of Concrete Incorporating Steel Slag Powder

This study investigated the effect of silica fume(SF) on mechanical properties of concrete incorporating steel slag powder(SSP). The compressive strength and splitting strength tests of concrete with different content of SF(0%, 4%, 8% and 12%) and of SSP(0%, 10%, 20%, 30% and 40%) were carried out, and the test results were analyzed and fitted. Obtained results showed that the brittleness, compressive strength and compressive strength discreteness of concrete increased due to the incorporation of SF. SSP weakened the compressive strength of concrete, which reduced within 10% when the content of SSP was less than 20%. SF and SSP showed synergistic hydration effect when they were mixed, and the optimal group was SF8 SSP30, whose compressive strength was close to that of plain concrete, and whose brittleness as well as discreteness of compressive strength were lower relatively. With the content of SSF and of SSP as variables, the tension-compression ratio and compressive strength of concrete can be well estimated by surface fitting.     

透明质酸改性PHBV组织工程纳米纤维支架的研究

组织工程支架材料表面性质对细胞的黏附起着重要作用,进而影响细胞的增殖、分化等一系列生长过程.本研究采用天然生物大分子透明质酸(hyaluronic acid,HA)对聚(3-羟基丁酸酯-co-3-羟基戊酸酯)(Poly(3-hydroxybutyrate-co-3-hydroxyvalerate),PHBV)进行表面修饰以提高材料的表面生物活性.首先通过静电纺丝法制备PHBV纳米纤维支架,利用1,6-己二胺胺解在PHBV表面引入自由胺基,并以此为反应活性位点在水溶性的碳化二亚胺/N-羟基琥珀酰亚胺缩合剂体系中将透明质酸固定接枝在PHBV表面.SEM结果表明,静电纺丝制备的PHBV纳米纤维支架表面平滑程度高,纤维直径分布较均匀,且没有串珠;FTIR证明1,6-己二胺改性及透明质酸接枝改性PHBV纳米纤维支架材料均成功实现;茚三酮法表明PHBV表面胺基密度随胺解时间增加而增大,在胺解约50min时达到最大值;水接触角法表明固定接枝透明质酸后,表面亲水性明显改善;细胞实验表明透明质酸改性的PHBV纳米纤维支架可显著促进软骨细胞的体外增殖.综上所述,透明质酸改性的PHBV纳米纤维支架可望应用于软骨组织工程领域.     

Fabrication of PEI-Liposome Loaded Fish Gelatin Nanofibers Mats by Green Electrospinning

Polyethyleneimine （PEI） and cationic liposomes were widely used for gent delivery and the combination of PEI and liposome was reported to result in a higher efficiency of cell transfection in vitro. In recent years, better transfection was observed for the drug-loaded iiposome fixed on the tissue engineering scaffolds via embedding, surface adsorption or covalent grafting thus protected and bound by the scaffolds. In the present study, a novel PEI-liposome loaded fish gdatin composite nanofiber was successfully fabricated by a green electrosplnning process. The existence of PEI- liposome in the composite nanofibers was determined by Fourier transform infrared （ FTIR ） spectra, transmission electron microscopy （TEM）, and confoeal laser scanning microscopy （CLSM）. As shown by scanning electron microscopy （SEM）, the dectrospun composite nanofibers with uniform diameter were smooth and round, and the morphology of the fish gelatin fibers did not change significantly after the incorporation of PEI-liposomes. The transfection results in vitro suggest PEI-liposome loaded fish gelatin material may have a promising application in non-viral gene delivery systems.     

Effects of Quenching Temperature and Time on Pore Diameter of Poly（3-hydroxybutyrate-co-3-hydroxyhexanoate） Porous Scaffolds and MC3T3-E1 Osteoblast Response to the Scaffolds

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) scaffolds were prepared by thermally inducing phase separation (TIPS) for bone reconstruction. Scanning electron microscopy and porosity measurements were used to analyze the structure and properties of the scaffolds. The pore diameter of the scaffolds could be easily controlled by changing the quenching temperature and time. The biocompatibility was assessed by examining the proliferation and morphology of MC 3T3-E1 osteoprogenitor cells seeded on the scaffolds. Cultures grown in the presence of a source of phosphate ions showed the formation of a mineralized extracellular matrix. The results indicate that PHBHHx scaffolds prepared using TIPS are a promising candidate for bone reconstruction.     

水溶性维生素E的合成工艺研究

采用纯度较高的VE琥珀酸单酯作为原料、对甲苯磺酸作为催化剂与聚乙二醇1 000进行酯化反应得到TPGS.通过考察初始原料比、催化剂用量、反应温度和反应时间对酯化反应的影响,发现初始原料比、催化剂用量和反应温度对酯化反应的影响比较显著,而当反应达到平衡时,反应时间对结果影响不大.因此又采用正交设计对优化TPGS的合成工艺,得到最佳工艺为反应时间为4 h、反应温度为110℃、催化剂用量为0.5 g和原料配比为2∶1.考察其重现性,得到平均转化率为95.261%、选择率为94.280%、收率为95.067%.     

多孔钛植入体表层孔隙内TGF-β1缓释明胶微球涂层的工艺优化

采用正交试验法优化载转化生长因子β1(transforming growth factor-β1,TGF-β1)缓释明胶微球多孔钛植入体制备工艺,探讨多孔钛植入体孔隙内微球涂层的载药、释药特性.采用粉末注射成形(Metal Injection Molding, MIM)技术制备多孔钛植入体,选用明胶为TGF-β1缓释载体材料,乳化冷凝聚合交联法制备明胶微球,检测微球粒径与形貌以及载TGF-β1微球的包封率、载药率,采用渗涂法制备多孔钛表层孔隙内载TGF-β1明胶微球涂层,释放试验检测涂层的释药特性.实验结果表明,MIM技术制备的多孔钛植入体的孔隙度为(62.02±1.82)%,孔径为50～300 μm,抗压缩强度为(63.23±12.81) MPa,弹性模量为(0.95±0.61) GPa.明胶微球粒径随明胶浓度的减小、搅拌速度和交联时间的增加而减小,交联剂用量对微球粒径影响无显著性差异.制备的TGF-β1明胶微球为球形,平均粒径为(21.42±3.67) μm,载药量为(0.91±0.02) μg/g,包封率为(91.41±1.82)%.TGF-β1微球涂层体外14 d,时的TGF-β1释放率为(94.2±3.4)%;粒径为(21.42±3.67) μm的明胶微球的最佳工艺参数如下:明胶浓度为10%,搅拌速度为800 r/min,交联剂用量为0.1 mL,交联时间为2 h.多孔钛植入经5%(质量分数)明胶溶液预处理后用20 g/L微球渗涂可在表层孔隙内形成均匀微球涂层,且不阻塞表层孔隙,微球涂层TGF-β1释放时间为14 d.     

Distribution of bone marrow stem cells in large porous polyester scaffolds

This work examined the optimal syringing depth during in vitro cell loading in order to even cell distribution after syringing a drop of cell suspension in cylinder poly（lactide-co-glycolide） （PLGA） porous scaffolds. The scaffolds of 10 mm height and 10 mm diameter were fabricated via room-temperature compression molding ＆ particulate leaching technique based on spherical porogens. In vitro tests were employed for such examinations： a global observation of a cell-loaded scaffold stained by the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） technique and a quantitative measurement of spatial distribution of cells after slicing the cell-loaded scaffolds into layers. It was found that an even distribution of cells was soon achieved only if the initial cell suspension was seeded on the layer that was below the top surface but above the middle of scaffolds. The availability of in vitro osteoblastic differentiation of rat bone marrow stem cells in such a kind of spherical-pore PLGA scaffolds was meanwhile confirmed.     

Preparation of Nano Silver/Silk Fibroin Composite Films

In this study, the outstanding biocompatibility of silk fibroin （SF） and the highly efficient anti-bacterial effect of nano silver （NS） were utilized to prepare SF/NS composite film with anti- bacterial property. The structure and property of the film were characterized. The results showed that the structure of SF in the film was mainly silk I. SF in the film was almost insoluble in water. The tensile strength of film with NS was significantly lower than that of films without NS. When the addition of NS was within the range of 0%-0.6%, the elongation at break had no significant difference. The antibacterial rate of the film on staphylococcus aurens and escherichia coil increased with the amount of NS. The minimum amount of NS in the fdm was O. 1% and the maximum amount was 0.5%.     

维生素E及其与维生素C,维生素B2联用对小鼠抗氧化体系机能的影响

研究了维生素 E(VE)及其与维生素 C(VC) ,维生素 B2 (VB2 )联用对小鼠机体抗氧化机能的影响。结果表明 ,对小鼠单独饲喂 VC或 VE,VC,VB2 联合饲喂 ,均能显著提高小鼠心、肝组织超氧化歧化酶 (SOD)的活性作用 ;单独饲喂 VE,VC或 VE,VC,VB2 联合饲喂 ,其肝脏丙二醛 (MDA)含量均低于对照组 ,差异非常显著 ,肝脏 SOD/ MDA和 GSH-Px/ MDA比值升高 ;单独饲喂 VC组 GSH-Px/ MDA比值升高尤为明显 ;饲喂VE或 VC,VE,VB2 联合饲喂时 ,小鼠肝组织中谷胱甘肽过氧化物酶 (GSH-Px)活力有增强趋势。这表明 3种维生素联合使用时 ,可产生协同抗氧化、调血脂作用 ,这种作用有利于预防动脉粥样硬化的发生与发展。     

Control of the morphology of micro/nanostructures of polycarbonate via electrospinning

Many of the applications proposed for bioassays, scaffolds for tissue engineering, filtrations, and supports for catalysts require polymeric membranes with large specific surface areas. Polycarbonate （PC） is a possible candidate for these applications because of its excellent mechanical performance and good biocompatibility. Electrospinning is a simple and effective method for large-scale fabrication of micro-/nanofibrous membranes with large specific surface areas. How to control the morphology of electrospun PC fibers, however, has not been systematically investigated. We describe the controllable fabrication of continuous and uniform PC fibers. We electrospin PC/chloroform solutions doped with different types of surfactants including anionic, zwitterionic, nonionic and cationic surfactants. Only cationic surfactants can lead to the successful fabrication of uniform PC fibers. After the analysis of the correlation between solution properties such as viscosity, surface tension, and conductivity and the morphology of electrospun fibers, we conclude that the addition of cationic surfactants such as cetane trimethyl ammonium bromide （CTAB） that leads to a decrease in viscosity is the main factor responsible for the formation of PC fibers. The demonstration of the fabrication of uniform PC fibers will lend experience to processing other polymers into fibers via electrospinning.     

BHC,DDT和B（a）P在内蒙古草地牧草中污染现状的初步调查

本文通过对内蒙古草地上优良牧草中，有机氯农药（６６６，ＤＤＴ）和苯并（ａ）芘（简写Ｂ（ａ）Ｐ），污染现状的调查，对四个地带性草地类型十个调查区域内１５４份牧草样品中，两项有害物质含量超标情况及分布进行了分析研究．结果表明：６６６、ＤＤＴ和Ｂ（ａ）Ｐ平均含量在草甸草原牧草中分别为０．０９２７ｍｇ／ｋｇ、０．００８６ｍｇ／ｋｇ、０．００５０ｍｇ／ｋｇ，在典型草原牧草中分别为０．０４５０ｍｇ／ｋｇ、０．００３０ｍｇ／ｋｇ、０．００５２ｍｇ／ｋｇ，在荒漠化草原牧草中分别为０．１００３ｍｇ／ｋｇ、０．００１５ｍｇ／ｋｇ、０．００５６ｍｇ／ｋｇ．在草原化荒漠牧草中分别为０．２５２９ｍｇ／ｋｇ、０．００００ｍｇ／ｋｇ、０．００５６ｍｇ／ｋｇ．全区草地上生长的１６种牧草中，两项有害物质均被检出，大部分含量都比较低，但个别样品含量很高，说明内蒙古草地环境已经受到一定程度的污染，应引起高度的重视．     

Tensile properties of tungsten-rhenium wires with nanofibrous structure

In this study, the mechanical properties of tungsten-rhenium wires with nanofibrous microstructure were investigated at both room temperature (RT) and 800℃. The strengthening mechanism associated to the nanofibrous microstructure was discussed. The results showed that the tungsten-rhenium wires with nanofibrous grains exhibited a very high tensile strength reaching values of 3.5 GPa and 4.4 GPa for the coarse (grains diameter of 240 nm) and fine (grains diameter of 80 nm) wires, respectively. With increasing the temperature from RT to 800℃, the tensile strength decreased slightly but still held high values (1.8 GPa and 3.8 GPa). All the fracture surfaces exhibited apparent necking and characteristics of spear-edge shaped fracture surface, indicating excellent ductility of the wires. A model of the strengthening mechanism of these tungsten-rhenium wires was proposed.