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.     

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.     

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.     

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.     

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.     

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.     

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.     

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。     

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.     

Preparation of fiber-microsphere scaffolds for loading bioactive substances in gradient amounts

Gradient scaffolds are needed for interface tissue regeneration. In this study, a technique combining electrospinning and electrospraying was developed for preparing poly(L-lactide-co-glycolide) (PLGA) fiber-microsphere scaffolds for loading bioactive substances in gradient amounts. The gradient fiber-microsphere scaffolds contain two sheets of electrospun membranes and a sheet of microspheres loaded with bioactive substances in gradient amounts between the electrospun membranes. The morphologies of the gradient scaffolds were characterized and bovine serum albumin (BSA) was loaded as a model bioactive substance. The amount of BSA-loaded microspheres decreased gradually along the length of the gradient scaffold. The addition of poly (ethylene glycol) significantly improved the hydrophilicity of the gradient scaffold and the release behavior of BSA with respect to the gradient became apparent, with differences in the release amounts along the length of the gradient scaffold being observed. The biocompatibility of the gradient scaffold was verified using MC3T3-E1 pre-osteoblastic cells. The study demonstrated that the combination of electrospinning and electrospraying was a feasible method for the preparation of gradient scaffolds for potential applications in interface tissue engineering.     

细菌纤维素组织工程支架的仿生矿化研究

骨骼创伤已经成为当今影响人类健康的一大病症。因此,骨修复材料就成为研究的一大热点。骨组织工程支架作为重要的骨修复材料,可以诱导成骨细胞生长并为新骨生长提供条件。传统的骨组织工程支架包括合成高分子(如聚乳酸、聚乙醇酸等)和天然高分子(如胶原、壳聚糖等)。与传统支架材料相比,细菌纤维素(BC)具有良好的生物相容性、精细的纳米空间三维网络结构,有作为组织工程支架的潜能。通过仿生矿化处理,BC纳米纤维表面可以生长出羟基磷灰石(HA)的晶体颗粒,且HA颗粒均匀覆盖在纳米纤维表面。通过热分析得出,仿生矿化处理会使BC的热稳定性得到一定的提升。     

聚氨酯改性及复合支架应用于血管组织工程

通过查阅近年来将聚氨酯作为血管组织工程支架材料的相关文献,介绍了聚氯酯改性的常用方法在血管组织工程支架中的应用情况,综述了聚氨酯表面改性支架和复合支架在血管组织工程中的应用.     

小直径血管支架的仿生构建及其种子细胞

静电纺丝仿生天然细胞外基质(ECM)结构,所制备的高度多孔、高比表面积的纳米级(50～500nm)纤维赋予了丰富的分子架构和生化信号,为种子细胞提供理想的生长微环境.不同组分、纤维尺寸及取向和种子细胞类型,可以裁剪获得不同生化和力学性能的电纺支架细胞复合物.总结了血管组织工程仿生ECM的设计与构建,并强调与支架复合的种子细胞在血管组织工程的作用.     

Preparation of composite tubular grafts for vascular repair via electrospinning

A novel type of composite vascular graft was developed via electrospinning in the present investigation.Collagen and chitosan were blended to form the inner and outer layer.Poly(1-lactide-co-caprolacto...     

3D打印制备HA/PCL复合材料组织工程支架的研究

应用熔融沉积成型技术（FDM）制备羟基磷灰石（HA）/聚己内酯（PCL）组织工程支架,探讨其内部结构和力学性能。以羟基磷灰石和聚己内酯为原料,采用熔融共混技术分别制备HA质量分数为20%的nano-HA/PCL和micro-HA/PCL复合材料,使用自主研发的熔体微分FDM 3D打印机制备HA/PCL复合材料组织工程支架。通过显微镜观察发现,所制备的nano-HA/PCL和micro-HA/PCL组织工程支架具有均匀分布且相互连通的近似矩形的孔隙。nano-HA/PCL和micro-HA/PCL组织工程支架的断面图分析结果表明,nano-HA/PCL组织工程支架中HA粒子分布均匀,而micro-HA/PCL组织工程支架中HA粒子发生了团聚,导致nano-HA/PCL组织工程支架的拉伸强度和弯曲强度均高于micro-HA/PCL组织工程支架。因此,利用熔体微分FDM 3D打印机打印生物活性nano-HA/PCL复合材料组织工程支架在骨组织工程中具有潜在的应用前景。     

Compatibility of Chitosan-Gelatin Films with Adipose Tissue Derived Stromal Cells

Introduction Chitosan, the partially N-deacetylated form of chitin, is the second most abundant biomaterial after cellulose. Due to its biocompatibility, biodegradability, and bio- activity, chitosan and its derivant complexes have been studied in various…     

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。     

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.     

Strontium Substituted Nanohydroxyapatite Incorporated 3D Printing Scaffold for Bone Tissue Engineering

The customized implants which are composed of polycaprolactone( PCL) and strontium substituted nanohydroxyapatite( SrHA) were fabricated successfully by using fused deposition modeling( FDM),which is a simple 3 D printing technology for fabricating personalized products. The physical and chemical properties of composite scaffolds were characterized by transmission electron microscopy( TEM), Fourier transform infrared spectroscopy( FTIR), X-Ray diffraction( XRD) and inductively coupled plasma-atomic emission spectroscopy( ICPAES). The results suggested that strontium element was successfully doped into nanohydroxyapatite and all scaffolds showed the homogeneous network structure. Furthermore, the in vitro biocompatibility of the scaffolds was evaluated by cell counting kit-8( CCK-8) assay. The data indicated that the prepared scaffolds exhibited excellent biocompatibility to bone marrow mesenchymal stem cells( BMSCs). Besides,strontium element can be released from PCL-SrHA scaffolds in a sustained manner. Therefore,the 3 D printing PCL-SrHA scaffolds hold great potential for bone tissue engineering.