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.     

Formation of drug-loaded electrospun twin fibers

Electrospinning is a well-known process for producing submicrometer fibers, which have wide applications in many fields, especially in tissue engineering scaffolds and drug-delivery systems. This paper presents the formation of drug-loaded electrospun twin fibers. The correlations between the twin fiber formation and the polymer materials or the loaded drugs were studied by using poly(l-lactide) and poly(l-lactide-co-glycolide) as electrospinning materials, and rifampin and paclitaxel as loaded drugs. Scanning electron microscopy showed that the formation of twin fibers is significantly affected by the loaded drug but not the polymer material. A possible reason for twin fiber formation was analyzed.     

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.     

3D printed titanium scaffolds with ordered TiO2 nanotubular surface and mesoporous bioactive glass for bone repair

Titanium alloy scaffolds have recently gained substantial interest for the treatment of critical-size bone defect, particularly along with the maturity of the 3D printing technology that is capable of turning scaffold design ideas into real implants. As titanium alloys lack surface osteogenic activity, for improved biological performance of such scaffolds, surface modification is necessary. Various coating materials and coating methods have been explored. In this study, we developed a unique surface modification method to provide the surface of 3D printed titanium scaffolds with nano-sized structure and bioactive agent. Uniform, ordered TiO₂ nanotube arrays were formed by applying two-step anodization, and then mesoporous bioactive glass (MBG) was loaded into nanotubes. The results of in vitro immersion testing showed that bioactive ions, i.e., Si and Ca ions, could be steadily and continuously released from MBG into basal medium. The assessment of the responses of hBMSCs confirmed that the surface-modified scaffolds supported the adhesion and proliferation of hBMSCs, indicating good surface cytocompatibility. The developed method of combining surface nanostructure and bioactive agent could be used as a new strategy to improve the osteogenic activity of 3D printed titanium alloy scaffolds.     

Covalently immobilized gelatin gradients within three-dimensional porous scaffolds

A stable gelatin gradient providing continuous increment of signaling for cell adhesion and proliferation was fabricated within 3D poly（L-lactic acid） （PLLA） scaffolds. The porous PLLA scaffold fabricated by NaCI particle leaching was vertically fixed on a glass vial. 1,6-Hexanediamine/propanol solution was continuously injected into the vial by a micropump to aminolyze the PLLA scaffold. As a result of reaction time difference, the introduced -NH2 groups increased continuously along with the longitude of the PLLA scaffold in the z-direction. After covalent immobilization of gelatin by glutaraldehyde coupling, the gelatin gradient scaffold was thus obtained. In vitro chondrocyte culture showed that the cells had higher viability and more extending morphology in the gelatin gradient scaffold than that in the uniform gelatin control.     

聚乙烯醇静电纺丝法固定葡萄糖氧化酶

利用静电纺丝纳米纤维具有高比表面积和多孔疏松结构的优势固定葡萄糖氧化酶，以提高酶电极的性能．通过聚乙烯醇（PVA）和葡萄糖氧化酶（GOD）共同静电纺丝的方法在金电极表面获得了固定化酶膜，用于构筑安培型葡萄糖生物传感器，膜的红外光谱、紫外-可见光谱和扫描电镜的分析均表明酶成功固定在静电纺丝形成的纳米纤维膜中．循环伏安测试表明固定化酶在静电纺丝纳米纤维中保持了活性，采用PVA静电纺丝法固定COD比利用浇铸膜法所得到的酶修饰电极对葡萄糖有更好的电流响应特性，通过在静电纺丝溶液中加入纳米金进一步提高了酶电极的电化学响应特性．     

VE TPGS-Loaded Silk Fibroin/Hydroxybutyl Chitosan Nanofibrous Scaffolds for Skin Care Application

Vitamin E（ VE） is an ideal antioxidant and a stabilizing agent in biological membranes. In this study,silk fibroin（ SF） /hydroxybutyl chitosan（ HBC） nanofibrous scaffolds are loaded with VE tocopherol polyethylene glycol 1000 succinate（ VE TPGS） via electrospinning. SEM images show that the average nanofibrous diameter has no significant difference when the content of VE TPGS increases to 4. 0%（ SF / HBC）. However,the average nanofibrous diameter decreases largely to 200 nm when the VE TPGS content reaches 6. 0%. Furthermore,VE TPGS presents a sustained release behavior from the nanofibrous scaffolds. Cell viability studies of mouse skin fibroblasts（ L929） demonstrate that VE TPGS loaded SF / HBC nanofibrous scaffolds present good cellular compatibility.Moreover,the incorporation of VE TPGS could strengthen the ability of SF / HBC nanofibrous scaffolds on protecting the cells against oxidation stress using the Tertbutyl hydroperoxide（ t-BHP）-induced oxidative injury model. Therefore,VE TPGS-loaded SF /HBC nanofibrous scaffolds might be potential candidates for personal skin care,wound dressing and skin tissue engineering scaffolds.     

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.     

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.     

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.     

HPEI接枝改性PAN/GO-PAA静电纺纳米复合纤维膜对Au(Ⅲ)的吸附

以丙烯酸(AA)为单体,过氧化苯甲酰(BPO)为引发剂,通过原位聚合法,在氧化石墨烯(GO)表面包覆聚丙烯酸(PAA),得到GO-PAA纳米复合物.将GO-PAA纳米复合物分散在聚丙烯腈(PAN)基体中,利用静电纺丝技术得到PAN/GO-PAA静电纺纳米复合纤维膜.利用化学接枝反应在PAN/GO-PAA静电纺纳米复合纤维膜表面接枝超支化聚乙烯亚胺(HPEI),构筑HPEI-g-PAN/GO-PAA静电纺纳米复合纤维膜.研究了HPEI-g-PAN/GO-PAA静电纺纳米复合纤维膜对Au(Ⅲ)的吸附性能.结果表明:HPEI-g-PAN/GO-PAA静电纺纳米复合纤维膜对Au(Ⅲ)的最大吸附量为1 808.60 mg·g^－1,在吸附过程中,部分Au(Ⅲ)被还原为片状和不规则颗粒状的Au单质.在共存离子体系中,HPEI-g-PAN/GO-PAA对Au(Ⅲ)具有较好的吸附选择性.     

In Vitro Cytotoxicity and Protein Drug Release Properties of Chitosan/Heparin Microspheres

Chitosan/heparin microspheres were prepared using the water-in-oil emulsification solvent evapo- ration technique. The microsphere diameters were controlled by selecting the fabrication process parame- ters. Scanning electron micrographs showed that the chitosan/heparin microspheres were regular and the surface morphology was smooth. Fourier transform infrared showed that the chitosan amino groups reacted with heparin carboxylic groups to form acylamides in the microspheres. Analysis of the microsphere cytotox- icity showed that they had no cytotoxic effect and behaved very similar to the negative control (polystyrene). To analyze the protein drug release profiles of the microspheres, bovine serum albumin was loaded as a model drug into the microspheres and released in vitro. Marked retardation was observed in the BSA re- lease profiles. The results show that chitosan/heparin microspheres may provide a useful controlled release protein drug system for used in pharmaceutics.     

组织工程用聚乳酸梯度支架的制备及分形研究

采用改进的溶液浇铸/粒子沥滤工艺,将聚乳酸（PLLA）与致孔剂粒子粉碎后,制备了双层和多层孔径梯度变化的PLLA泡沫支架。扫描电镜观察表明,材料内部界面处存在一定的扩散现象,且界面处孔的连通性好,通过调整致孔剂粒子的粒径以及PLLA与致孔剂粒子的比例,可有效地调控支架的各项参数（材料大小、孔径、孔隙率）。对材料进行分形研究,表明梯度多孔支架内部无明显界面,孔径过渡平缓。     

一步法制备有序排列的珠丝状纤维复合物

基于电纺丝和电喷雾技术的原理相似性,该文联合使用这2种技术来一步获得珠丝状的纤维复合物。通过使聚乙烯醇溶液和聚乙烯吡咯烷酮溶液分别从对喷电纺丝装置的两个喷丝头以电纺丝和电喷雾的模式进行喷射,获得纳米纤维和微米颗粒,纤维与颗粒相互碰撞结合从而得到表面带有颗粒的纤维复合物。结果表明:电喷雾得到的颗粒较均匀地附着于电纺丝纤维表面,复合纤维束经过旋转收集装置的牵引可以呈现良好的单方向有序排列;通过改变电喷雾的喷射介质的质量分数可以改变复合纤维的形貌;当提高溶液的质量分数使得射流的喷射模式由电喷雾转变为电纺丝时,纤维复合物的珠丝状结构将被消除,并且使得纤维束的排列有序度变差。     

Functionalization of PCL fibrous membrane with RGD peptide by a naturally occurring condensation reaction

Poly(e-caprolactone)(PCL)is widely adopted as an ingredient for tissue engineering scaffolds.To improve its cell affinity,in this study,we developed a new method to introduce bioactive RGD peptides onto the surface of PCL via condensation reaction between 2-cyanobenzothiazole(CBT)and D-cysteine.The PCL fibrous membranes were prepared by electrospinning,and RGD functionalization was characterized by fluorescence microscopy,scanning electron microscopy(SEM),X-ray photoelectron spectroscopy(XPS)and water contact angle(WCA).As expected,our results demonstrated the successful RGD immobilization on the surface of PCL.RGD modification improved the hydrophilicity of PCL,changing their WCA from 112.20°to38.35°.Cell adhesion,spreading and proliferation of 3T3fibroblasts were also enhanced.We therefore believe that the methods reported in this study was facile and effective for functional modification of the hydrophobic PCL scaffolds.The moderate reaction conditions are also suitable for covalent immobilization of bioactive molecules onto PCL.     

静电纺丝法制备溶解型胶原蛋白肽/丝素蛋白复合纤维面膜

采用静电纺丝技术制备了一种溶解型贴式面膜。将胶原蛋白肽、丝素蛋白两种精华成分加入8%聚乙烯吡咯烷酮（PVP）的甲酸溶液中作为纺丝液,以旋转滚筒作为接收装置进行静电纺丝。探究了共混比例、纺丝液浓度及透明质酸添加量对静电纺丝纤维的影响。通过扫描电子显微镜、电子万能实验机对纤维形貌和纤维膜的力学强度进行测试表征,随后进一步对面膜的保湿补水性能进行测试,分析了该纤维膜的实际功效。结果表明,复合纤维具有较均一的形貌,膜布本身可以起到精华液的作用,具有良好的溶解性,能够起到较好的保湿补水作用。     

几种骨组织工程复合支架材料在模拟生理溶液中的生物矿化性能比较

采用生物活性玻璃(BG)、磷酸三钙(TCP)、羟基磷灰石(HA)等生物活性陶瓷与3-羟基丁酸酯-3-羟基戊酸酯的共聚物(PHBV)复合,制备了性能良好的骨组织工程支架材料,分析了PHBV/BG,PHBV/TCP,PHBV/HA三种复合多孔支架在模拟生理溶液中的一系列化学反应,以及多孔材料在模拟生理溶液中浸泡后的成分、结构和微观形貌的变化.研究结果表明,三种复合支架材料在模拟生理溶液中发生了降解反应而失重;PHBV/BG和PHBV/TCP在模拟生理溶液中还发生了生物矿化反应,在表面形成矿化沉积层,为具有骨生物活性的结晶态类骨碳酸羟基磷灰石;而PHBV/HA在模拟生理溶液中没有明显的生物活性反应.     

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.     

Nanofibrous Scaffold Containing Osteoblast-Derived Extracellular Matrix for the Proliferation of Bone Marrow Mesenchymal Stem Cells

Extracellular matrix( ECM) plays a prominent role in establishing and maintaining an appropriate microenvironment for tissue regeneration. The aims of this study were to construct a tissue engineered scaffold by reconstituting osteoblast cell-derived ECM( O-ECM) on the electrospun nanofibrous scaffold,and further to evaluate its subsequent application for promoting the proliferation of bone marrow mesenchymal stem cells( BMSCs). To engineer a biomimetic scaffold, calvarial osteoblasts and electrospun poly-llactic acid( PLLA) nanofibers were prepared and subjected to decellularize for O-ECM deposition. To evaluate and characterize the O-ECM/PLLA scaffold, the morphology was examined and several specific mark proteins of osteoblasts matrix were evaluated.Furthermore,the cell counting kit-8( CCK-8) assay was used to detect the proliferation of the BMSCs cultivated on the O-ECM/PLLA scaffold. The results indicated O-ECM/PLLA scaffold was loaded with Collagen I, Fibronectin, and Laminin, as the composition of the marrow ECM. After decellularization,O-ECM deposition was observed in O-ECM/PLLA scaffold. Moreover,the O-ECM/PLLA scaffold could significantly enhance the proliferation of BMSCs,suggesting better cytocompatibility compared to the other groups tested. Taken together,a biomimetic scaffold based on the joint use of O-ECM and PLLA biomaterials,which represents a promising approach to bone tissue engineering, facilitates the expansion of BMSCs in vitro.     

采用S/W/O/W法制备BSA-PLG微球

以生物可降解材料PLG607为载体, 采用S/W/O/W溶剂挥发法制备牛血清白蛋白(BSA)微球, 对其性质进行表征, 并考察了处方中不同添加剂对BSA-PLG微球粒径、 包封率和体外释放的影响. 结果表明, 采用S/W/O/W法所得微球粒径为40～70 μm, 表面光滑圆整. 加入添加剂（m(羟丙基 β 环糊精) ∶m(牛血清白蛋白)=1 ∶1）后包封率可以从27.9%提高到46.3%, 加入添加剂羟丙基-β-环糊精和羟基磷灰石制备蛋白微球, 能减小突释, 提高蛋白的包封率, 改善体外释放行为, 达到均匀释放.