多孔BCP支架及其仿ECM涂层对MG63细胞功能的影响

通过改良冰模板法制备一种由内向外呈放射状排列、全开孔板层状多孔结构的仿生多孔双相磷酸钙(BCP)支架,并在支架表面制备不同的仿细胞外基质(ECM)涂层。通过体外细胞实验,评价支架结构及表面4种仿ECM生物涂层对人成骨肉瘤MG63细胞(MG63)的细胞功能影响。研究结果表明:支架及其仿ECM涂层均无细胞毒性,仿生ECM涂层支架对MG63细胞功能的作用优于未涂层支架,且明胶-透明质酸复合涂层试样对MG63细胞功能的作用优于单一涂层,明胶-透明质酸-掺RGD涂层试样对MG63细胞增殖、黏附、分化功能的作用均优于其他各涂层组。     

静电纺丝法制备组织工程纳/微米纤维支架

静电纺丝是一种简便易行的新型组织工程多孔支架制备方法,电纺支架具有独特的微观结构和适当的力学性能.由于具有与天然细胞外基质相近的纳米级结构,电纺支架能够仿生细胞外基质的结构特点,使之有望成为理想的组织工程支架.文中介绍了静电纺丝和同轴静电纺丝的基本原理和发展过程、电纺支架的加工方法和结构特点以及电纺纤维的定向收集技术,阐述了各种天然和合成聚合物纳/微米电纺支架在软骨、骨、血管、心脏、神经等组织工程领域的应用,并展望其应用前景.     

组织工程血管研究现状

组织工程化血管的产生和发展使人类最终解决完善血管替代物的问题成为可能.随着干细胞功能研究的进步和材料及塑型科学的不断发展,血管组织工程进展迅速,备受瞩目.文中就血管组织工程中关键的3大要素:支架材料、种子细胞和共培养构建的研究进展作一综述.     

组织工程血管研究现状

组织工程化血管的产生和发展使人类最终解决完善血管替代物的问题成为可能.随着干细胞功能研究的进步和材料及塑型科学的不断发展，血管组织工程进展迅速，备受瞩目.文中就血管组织工程中关键的3大要素：支架材料、种子细胞和共培养构建的研究进展作一综述.     

纤维基支架在骨和软骨组织再生中的应用

组织工程对于骨和软骨组织缺损的修复具有广阔的发展前景。在组织工程三要素之一的支架中,纤维基支架由于具有较大的比表面积、相互连通的多孔结构以及与天然细胞外基质相似等优势,在骨和软骨修复中展现出巨大的应用潜力。纤维支架的成型通常基于静电纺丝法、热致相分离法、自组装肽法和溶剂热法,血管化和软骨再生是此类支架在临床应用上面临的主要挑战。通过仿生天然骨和软骨组织的复杂结构、综合多种技术手段在结构和功能上设计纤维基三维多孔支架将是改善骨和软骨组织工程支架修复效果的主要发展方向。     

可降解聚醚酯弹性体PTCG的合成及初步生物学评价

用熔融缩聚法合成聚对苯二甲酸丁二醇酯-co-聚对苯二甲酸环己烷二甲醇酯-b-聚乙二醇(PTCG)聚醚酯弹性体.按照ISO 10993标准,采用L929小鼠成纤细胞对其进行体外细胞毒性测试;以狗的血管平滑肌细胞 (SMCs)为模板细胞,测试细胞在材料表面的贴附性能.采用旋转成型/粒子洗出法制备三维血管支架,接种SMCs后在生物反应器上进行动态培养,探索PTCG作为组织工程血管支架材料的可行性.结果表明,合成的PTCG聚醚酯弹性体无细胞毒性;SMCs在PTCG薄膜表面贴附生长良好;所制备支架的孔径、孔隙率和力学性能等满足组织工程血管支架的要求;体外培养3 d后,大量SMCs长入血管支架并开始分化.     

短肽自组装技术构建纳米三维细胞培养基质

绝大多数用于细胞3D(3-dimension,3D)培养的生物材料支架具有多孔渗水性的无定形结构,能被生物降解,但没有生物活性,也没有组织特异性,其支架显微结构与体内ECM(extracellular matrix,ECM)相差很大.目前,国际上兴起用短肽自组装技术构建纳米三维细胞培养基质,它具有纳米级结构、具有生物活性、组织特异性、成分清楚可控、生物力学、显微结构与体内一致等特征.纳米三维细胞培养基质在细胞生物学,肿瘤学,高通量药物筛选,组织工程,再生医学,以及生物纳米材料的发展具有重要的意义.     

点击化学法构建多生长因子序贯控释型ECM及骨组织工程化研究

正我校生命科学学院聂磊博士2017年获批国家自然科学基金项目:点击化学法构建多生长因子序贯控释型ECM及骨组织工程化研究,项目编号:31700840.骨组织工程可以解决临床上自体骨和异体骨供源不足、交叉感染等问题,采用支架支撑细胞,通过生长因子对细胞的扩散、迁移和分化调控,构建骨细胞外基质(Extracellular Matrix,ECM),完成血管化和成骨化等修复.从组织工程学和转化医学     

肌腱组织工程的研究概况及完善策略

从种子细胞、支架材料和生长因子3个方面概述了当前肌腱组织工程的研究进展,同时对肌腱组织工程今后的研究方向和一些完善策略进行了归纳和总结.     

肝组织工程支架的仿生设计与有限元分析

为了解决细胞在细胞支架复合培养中难以植入的问题,提出了一种卷裹型支架.将平面多孔支架采用卷裹的方式成型,使细胞在整个支架内部形成螺旋状三维分布,支架表面的管道结构可使培养液在支架内部渗透,从而提高物质交换效率.提出了一种具有仿生参数的流道设计方案,从肝脏血管铸型中提取血管的形状数据,作为仿生学依据进行流道设计,具体为树状多层分支结构.用此流道结构仿生了动脉、静脉的血液循环功能,并利用多孔材料仿生了毛细血管的渗透功能.建立了不同形状参数的流场分析模型,进行流体与多孔材料的渗流分析.结果表明,仿生设计方案的流场分布最为均匀,且平均流速较高,间接表明营养物质的输送效率最高.该支架能够使细胞呈三维分布,并扩大培养液在支架内的有效分布范围,更好地维持细胞的存活、增殖和迁移,为向组织化方向发展奠定了一定的基础.     

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.     

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.     

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.     

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.     

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.     

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.     

静电纺丝及其在纳米复合材料制备中的应用研究

本文介绍了静电纺丝的发展史、原理、设备及其在复合材料制备中的应用.借助于静电力的纺丝过程被称为静电纺丝.静电纺丝可以制备直径在微米以下甚至纳米级的纤维.近年来,随着纳米技术的兴起,静电纺丝受到研究者们的广泛重视.静电纺丝制备的纳米纤维可直接用于制备纳米复合材料,也可经高温处理后成为纳米碳纤维,后者在复合材料中也有很大的潜在用途.     

基于区域填充的纤维图像提取算法

对天然纤维显微图像中所有纤维对象的正确分离和提取,是棉麻纤维特征分析的必要前提.针对纤维图像中图像背景、纤维边缘和纤维内腔的灰度分布特性,提出基于区域填充的纤维图像提取算法.该算法以各个"纤维内腔"为图像分离提取的标准,采用膨胀区域填充的方法,并通过距离变换自动确定填充起始点,对经过二值化处理的纤维图像中的背景和纤维进行了区分,从而得到各个纤维对象的正确提取.该算法能够有效处理纤维图像间的黏连问题.     

Preparation and mineralization of PLGA/Gt electrospun fiber mats

Poly(lactic-co-glycolic acid)(PLGA)/gelatin(Gt) ultrafine composite fibers were fabricated via electro-spinning.The effect of gelatin on the morphology and tensile property of the electrospun fiber mats was investigated.Mineralization was carried out in 10×simulated body fluid(10SBF).The deposited calcium phosphate(CaP) was identified by scanning electron microscopy(SEM),energy dispersive spectroscopy(EDS),X-ray diffraction(XRD) and Fourier transform infrared spectroscopy(FTIR).Results indicated that the av...     

适用于不同尺寸血管的脱细胞方法研究

将胰酶消化与反复冻融相结合,旨在建立一种适用于各种类型血管的通用脱细胞方法,如隐静脉、颈动脉和主动脉。隐静脉、颈动脉和主动脉经胰酶消化和反复冻融脱细胞处理后,采用苏木精—伊红染色、Masson三色染色及弹性纤维染色来定性评价脱细胞效果和细胞外基质的保存效果,采用Image-Pro-Plus 5.1图像处理软件作进一步定量评价;扫描电子显微镜观察胞外基质的完整性。结果显示,组织染色及定量分析表明此胰酶消化与反复冻融相结合的方法完全脱除了隐静脉、颈动脉和主动脉得细胞,细胞外介质结构保存良好且完整。扫描电子显微镜观察亦表明细胞外基质保存良好,且基质纤维致密规整。表明胰酶消化与反复冻融相结合的脱细胞方法是一种很有前景的制备各种不同类型血管支架的方法。