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.     

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.     

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.     

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.     

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。     

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.     

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.     

Effects of Polyethylene Oxide Concentration on the Size of Beads in Electrospun Beaded Nanofibers

Embedding particle drugs in beaded nanofibers by electrospinning has been shown a potential approach to control drug release in tissue engineering. The bead size is one of the critical parameters in controlling the drug release rate. In this study,the relationship between polymer concentration and beads size was investigated. Aqueous polyethylene oxide（ PEO） solutions with different concentrations were prepared to obtain various beaded nanofibers by electrospnning. Optical microscope and scanning electron microscope（ SEM） were used to observe the variation tendency of bead size. With an increase in the polymer concentration,the diameter of fibers between beads became bigger,while the fiber uniformity improved. In addition, the polymer concentration influenced the distribution of bead diameter. Higher polymer concentration would reduce the possibility of small-sized beads formation and improve the uniformity of bead diameter. The study provides a possible way to control the size of beads,which is helpful for further research on the control of particle drug release.     

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.     

高链玉米淀粉对聚己内酯组织工程支架材料结构与性质的影响

采用高链玉米淀粉改性聚己内酯（PCL）,通过溶剂浇铸/粒子沥滤法制备聚己内酯/淀粉（SPCL）组织工程支架;考察了改性后支架材料的化学结构、结晶性、亲水性和细胞相容性.结果表明,适量淀粉的引入能够降低支架材料的结晶性、改善材料的亲水性和细胞相容性,70％PCL/30％淀粉支架材料的细胞粘附率、增殖率和细胞活性为最佳,有望应用在骨组织工程中.     

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。     

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.     

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.     

聚乙烯醇-透明质酸-胶原组织工程支架研究

以聚乙烯醇(PVA)、透明质酸(HA)和胶原蛋白(Col)为组分,构建组织工程三维多孔支架.将聚乙烯醇分别与不同质量的胶原和透明质酸复合,测定复合材料的含水率和膨胀率.扫描电镜观察材料横截面的表面形态.结果是不同分子质量的PVA与不同配比的Col和HA复合,得到的复合支架材料含水率为57.97%～81.26%,膨胀率为150.00%～459.20%.不同配比的复合支架材料具有不同的内部孔洞结构.结果表明,PVA-HA-Col复合支架材料具有较高的含水率和膨胀率,内部孔洞网络丰富,可作为组织工程支架材料.     

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.     

高性能聚合物电纺纳米纤维最新进展

近20年来,静电纺丝技术得到了快速发展和应用; 不同材料的电纺纳米纤维(包括聚合物基、金属基、陶瓷基、碳基及其复合材料等)已在能源、环境、生物医学和国防军工等领域得到了泛应用.通常,静电纺丝技术需将聚合物或聚合物前驱体原料溶解于溶剂中或者加热熔融进行电纺加工.然而,芳杂环高性能聚合物(如聚酰亚胺、芳香聚酰胺、聚苯等)由于其主链上的刚性环状结构,既难溶解于普通有机溶剂,也难加热熔融,没有流动性,故难以通过静电纺技术制备其纳米纤维.为了解决这个难题,研究人员努力探索了许多间接方法和途径来制备电纺高性能聚合物纳米纤维,并取得了突破性进展.如通过电纺前驱体法大规模地制备聚酰亚胺纳米纤维、利用热致重排进行分子转化制备了电纺聚苯并二噁唑纳米纤维、利用模板电纺法制备了聚苯基和聚吡咙基纳米纤维等.该文详细介绍了通过静电纺丝技术制备高性能聚合物纳米纤维的最新进展,具体包括聚酰亚胺、聚苯并咪唑、聚苯撑苯并二噁唑、芳香聚酰胺、聚苯、聚吡咙等芳杂环聚合物纳米纤维.此外,由于聚丙烯腈是制备碳纤维的重要前驱体,也对电纺聚丙烯腈纳米纤维的制备做了简要介绍.     

纳米支架在组织工程中的研究进展

纳米纤维支架技术是组织工程中为了支持细胞形成受损组织生物替代物的新兴技术。纳米支架能模拟天然的细胞外基质,作为三维的模板供细胞吸附、增殖以及分化。主要介绍了纳米纤维支架的结构特性、构建技术以及生物学效应。     

聚乙烯醇及改性聚乙烯醇/明胶多孔支架的体外生物相容性研究

以吐温80为发泡剂,采用乳化发泡/冷冻—解冻法制备多孔的聚乙烯醇(PVA)和改性聚乙烯醇/明胶(PVA/Gel)支架材料,用扫描电子显微镜(SEM)对其形貌进行观察,并通过体外细胞培养对支架的生物相容性进行评价.实验结果表明,用该方法制备的多孔支架具有良好的贯通性,且PVA/Gel支架贯通性高于PVA支架;细胞结果显示,乳化发泡法制备的多孔支架对细胞的黏附、增殖和形貌无不良影响;荧光染色结果表明,PVA/Gel支架由于引入了大量氨基,使得细胞的黏附力优于PVA支架,具有更好的生物相容性.     

Preparation of PLGA-collagen hybrid scaffolds with controlled pore structures for cartilage tissue engineering

Scaffolds used for cartilage tissue engineering should have high mechanical strength and well-controlled pore structure to provide suitable microenvironments for functional tissue regeneration. In this study, hybrid scaffolds which had open and interconnected pore structure and high mechanical property were prepared by hybridization of PLGA mesh and collagen using ice particulates as porogen templates. Embossing ice particulates template was used to form open pore structures on the scaffold surfaces. Free ice particulates were used to generate interconnected bulk pores in the scaffolds. Hybridization with PLGA mesh provided the scaffolds with high mechanical property. Bovine articular chondrocytes were cultured in the hybrid scaffolds. The unique pore structures facilitated the homogeneous distribution of chondrocytes and cartilaginous matrices throughout the scaffolds. Subcutaneous implantation demonstrated cartilage-like tissue regeneration. The hybrid scaffolds should have a high potential for cartilage tissue engineering.     

组织工程三维多孔支架制备技术的最新进展

理想的组织工程支架是由具有一定降解性能的高分子材料制成的,制备的支架除了要有一定的孔隙率和适于细胞生长的孔径外,还要有较高的机械性能和生物活性.为此介绍了用于组织工程中的可降解高分子材料,并详细阐述了最新的制备组织工程支架的方法:超临界CO2技术,计算机辅助成型技术,以及静电纺丝技术.