明胶在聚氨酯表面的固定化及其对内皮细胞生长的促进作用

理想的组织工程支架材料应具备有效促进细胞生长的能力和良好的组织相容性 .然而现有的聚合物生物材料大多呈现疏水性 ,不能有效支持细胞的生长 [1,2 ] .细胞外基质和血清中含有对细胞粘附、生长和繁殖有显著促进作用的多种活性因子 ,如纤维粘连蛋白 ( Fn)、层粘连蛋白 ( L aminin)、胶原( Collagen)、多聚赖氨酸和冷析蛋白 ( CIG)等 [3～ 5] ,把这些因子固定到材料表面 ,可为细胞的粘附生长提供理想的条件 .本文通过碳二酰亚胺脱水缩合技术 ,将明胶 ( Gelatin)共价键合到聚甲基丙烯酸接枝改性的聚氨酯 ( PU- g- PMAA)薄膜表面 ,并初…     

可释放一氧化氮的芯-壳结构静电纺丝支架的制备（摘要）

本文制备了可释放一氧化氮（NO）的小口径人工血管.首先合成了氮烯醇化的N,N′-二丁基-1,6-己二胺（DBHD/N2O2）作为NO供体,并将其混入生物可降解材料聚己内酯中（PCL）,利用静电纺丝技术构建管状血管支架.这种血管材料展现了良好的抗血小板粘附的性能,但是却存在着细胞毒性及NO的突释现象.为了解决这两方面问题,利用同轴电纺技术构建了以PCL或明胶为壳,混有DBHD/N2O2的PCL为芯的芯-壳结构纳米纤维多孔血管支架.这种芯-壳纤维结构能够缓慢释放NO,并且细胞相容性也得到了明显的改善.     

聚羟基脂肪酸酯的亲水性改性及其与人脐静脉内皮细胞相容性的研究

用丝素蛋白(SF)对微生物合成的高分子聚合物聚(3-羟基丁酸酯-co-3-羟基己酸酯)(PHBHHx)进行亲水改性,以提高材料的生物相容性.水接触角测定和表面自由能分析表明,丝素蛋白在支架表面吸附,使PHBHHx材料表面的水接触角从90°降至51°,表面自由能从37.9 mJ/m2增至57.4 mJ/m2,因而增加了材料的亲水性.进一步对亲水性改性前后PHBHHx多孔支架与人脐静脉内皮细胞(HUVECs)的相容性进行了比较.MTT法细胞活力分析表明,细胞在支架上培养3,5,7天后,其在SF改性PHBHHx多孔支架上的活力显著高于在未改性的PHBHHx支架上的活力;扫描电镜观察细胞生长形貌表明,细胞在改性后多孔支架上黏附及生长5天后,形成了连续细胞单层,其生长状态优于在未改性的PHBHHx支架上的生长状态;胶原含量测定表明细胞在改性后支架上比在未改性支架上有更好的胶原分泌能力,即改性后支架更利于诱导HUVECs分泌细胞外基质(ECM)从而构建类似体内的生长环境.     

丝素蛋白修饰改性的聚羟基脂肪酸酯支架上人体平滑肌细胞的生长

聚三羟基丁酸脂和聚三羟基己酸脂的共聚物(PHBHH_x)是一种具有良好强度和韧性的生物可降解高分子材料, 可作为组织工程心脏瓣膜支架的选择材料之一. 但其生物相容性尚不甚理想. 为此, 本工作利用丝素蛋白修饰改性高分子多孔支架, 以提高支架的生物相容性. 并将人体平滑肌细胞接种在该复合支架上进行体外培养, 以证实改性效果. 其中, 用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)方法测试细胞生长, 评估复合支架的细胞相容性. 并用扫描电子显微镜观察细胞在支架上的生长形态. 结果显示, 丝素蛋白修饰改性后的复合支架更有利于细胞的粘附与生长, 平滑肌细胞在支架上表现出良好的生长形态. 这表明, 丝素能够改善多孔支架的生物相容性, 使PHBHH_x/丝素蛋白复合物能更适宜作为组织工程心脏瓣膜的支架材料. 结果对于进一步研究细胞外间质在复合支架上的生长以及体外培养的组织重建有重要的参考意义.     

聚乳酸-聚己内酯复合纳米纤维支架的制备和表征

在二氧六环/乙醇溶剂体系中,采用凝胶抽提相分离法制备了聚乳酸-聚己内酯(PLLA-PCL)复合纳米纤维支架,研究了凝胶温度、聚合物比例、聚合物浓度、致孔剂及二氧六环/乙醇(溶剂/非溶剂)比例对复合纳米纤维支架结构与性能的影响.结果表明,当凝胶温度处于-20~-10℃,PCL含量为30%~50%,非溶剂含量不超过15%,致孔剂与溶质质量比不超过20∶1时,均能得到具有类似于天然细胞外基质的纳米纤维(50~500 nm)结构的PLLA-PCL复合纤维支架.随着PCL含量的增加,复合纤维支架的弹性模量减小;PCL含量为30%时,复合支架的相容性和结晶性最好.该复合纤维支架具有良好的生物活性和一定的降解性能.     

新型改性聚己内酯型聚氨酯弹性体的合成及性能

用不同种类异氰酸酯[脂肪族六亚甲基二异氰酸酯(HDI)和脂环族异佛尔酮二异氰酸酯(IPDI)]对聚己内酯(PCL)进行改性,得到两端为羟基的异氰酸酯改性的PCL预聚体.将未改性和改性的PCL端羟基进行磷酸化后[磷酸化组分PCL210磷酸酯(A)、PCL205/HDI磷酸酯(B)和PCL205/IPDI磷酸酯(C)]与双官能度的环氧(1,4-丁二醇二缩水甘油醚,E)进行开环交联反应,得到生物相容且可降解的聚己内酯型聚氨酯弹性体材料(AE,BE和CE).聚己内酯型聚氨酶弹性体的力学性能、静态水接触角、体外降解/溶胀和细胞毒性测试结果表明,PCL异氰酸酯的改性有助于提高材料的强度、弹性、耐疲劳性和降解速率,同时未明显提高材料的细胞毒性.     

具有NO原位催化能力和内皮细胞特异性粘附的小口径血管支架的制备和表征

人工合成的小口径血管支架亟需攻克抗凝血和快速内皮化两大难题。负载有机硒催化剂聚乙烯亚胺(SePEI)能够催化一氧化氮(NO)的原位生成,从而有效地抑制血栓形成。层粘连蛋白来源多肽Tyr-Ile-Gly-Ser-Arg(YIGSR)具有内皮细胞选择性粘附功能,负载到支架表面能够诱导内皮层的快速形成。本实验采用静电纺丝构建聚己内酯(PCL)支架,通过层层自组装构建Se PEI和透明质酸(HA)的双电层催化NO的生成;最外层的HA接枝有YIGSR多肽促进内皮化。考察和比较了材料修饰前后的物化性能。修饰过程对纤维形貌无明显影响,负载稳定性经过65d仍能保持在99%以上。材料经修饰后力学性能有所提高,能够满足作为血管支架的要求,为下一步细胞培养和体内实验打下基础。     

间充质干细胞扩增载体材料

间充质干细胞(MSCs)具有高度自我更新能力、多分化潜能、体外易分离和培养的特性,是细胞治疗和组织工程重要的种子细胞来源,但如何大规模地获得具有可再生活性的MSCs一直是限制其临床应用的关键因素,近几年发展起来的贴壁动物细胞动态培养技术为MSCs的大规模体外扩增提供了一条重要的途径。本综述结合动物细胞扩增载体的发展现状,主要介绍了用于间充质干细胞三维动态培养的明胶载体、海藻酸盐载体、壳聚糖载体和其他多糖载体等常规载体及其表面修饰和改性方法,并进一步介绍了以非酶解途径回收扩 增细胞的新型干细胞载体的研究进展。随着新型载体材料的涌现以及人们对干细胞生长和扩增特点的了解,采用三维动态培养技术安全而有效地大规模体外扩增MSCs的必要性将得到进一步的确认。     

贯通多孔聚酯基PolyHIPE支架在肝细胞培养中的应用

以生物相容性好且可降解的端基双键化的聚(ε-己内酯)(PCL)或聚(4-甲基-ε-己内酯)(PMCL)为基体,通过乳化剂Hypermer T96制备高内相乳液(HIPE)。该乳液在紫外光引发下,通过后加入的季戊四醇四-3-巯基丙酸酯(PETMP)与碳碳双键化的PCL(PCL-AC)或者PMCL(PMCL-AC)发生巯基-烯点击反应而交联成聚高内相乳液(PolyHIPE),经冷冻干燥得到PolyHIPE三维多孔支架。采用扫描电子显微镜、差示扫描量热仪、万能材料试验仪对支架的微观形貌、热力学以及力学性能进行了表征。通过体外细胞毒性实验以及肝细胞培养实验对支架的生物学性能进行了评估。力学性能测试结果表明:在乳液制备过程中,去离子水温度对支架的力学性能影响更为显著。体外细胞毒性以及肝细胞培养结果表明:支架无细胞毒性,且支架的刚性越小越有利于肝细胞的增殖和功能表达。     

角膜基质细胞在p(HEMA-MMA)改性前后的粘附率、形貌与力学性能变化

采用原子力显微镜(AFM)对改性前后材料表面粘附生长的角膜基质细胞的亲和力、三维形貌和力学性能进行了分析。结果显示,改性后材料的细胞亲和力有较大改善,且材料表面细胞三维形态更加正常,铺展更为舒展,与材料的接触面积较大。细胞力学性能分析也发现改性后材料表面细胞具有更高的粘附力和杨氏模量,以及更低的硬度,说明未改性p(HEMA-MMA)材料具有明显的细胞毒性,这种毒性作用导致在其表面生长的细胞的细胞骨架遭到破坏,细胞健康状态明显不如改性后材料表面细胞。因此,改性后的Col/bFGF-p(HEMA-MMA)更适合作为人工角膜材料使用。     

Preparation of Poly(ε-caprolactone)/Poly(ester amide) Electrospun Membranes for Vascular Repair

With adjustable amphiphilicity and anionic/cationic charge, biodegradability and biocompatibility, amino acid-based poly(ester amide)s(PEAs) have drawn attention in the research of tissue engineered vascular grafts. In this work, L-phenylalanine-based PEAs with or without L-lysine were synthesized through polycondensation, and ultrafine fibrous grafts consisted of PEAs and poly(ε-caprolactone)(PCL) in given mass ratios were further prepared via blend electrospinning. Surface characterizations by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy confirmed the chemical structure, and the wettability indicated that the prepared PCL/PEAs electrospun membranes exhibited less hydrophobic than PCL. Tensile results showed that the PCL/PEAs membranes possessed suitable mechanical properties, which could meet the requirements of artificial blood vessels. Cell culture and hemolytic tests exhibited that the PCL/PEAs electrospun membranes are biocompatible. In general, the electrospun grafts of PCL/PEAs could be applied for vascular repair.     

Immobilization of anti-CD31 antibody on electrospun poly(ɛ-caprolactone) scaffolds through hydrophobins for specific adhesion of endothelial cells

Hydrophilicity improvement and bioactive surface design of poly(?-caprolactone) (PCL) grafts are of key importance for their application in tissue engineering. Herein, we develop a convenient approach for achieving stable hydrophilic surfaces by modifying electrospun PCL grafts with a class II hydrophobin (HFBI) coating. Static water contact angles (WCA) demonstrated the conversion of the PCL grafts from hydrophobic to hydrophilic after the introduction of amphiphilic HFBI. ATR-FTIR and XPS confirmed the presence of self-assembled HFBI films on the surface of the PCL nanofibers. The biocompatibility of the HFBI-modified PCL grafts was evaluated by cell proliferation in vitro, and by arteriovenous shunt (AV shunt) experiments ex vivo. Anti-CD31 antibody, which is specific for endothelial cells (ECs), was subsequently immobilized on the HFBI-coated PCL scaffolds through protein-protein interactions. This bioactive PCL graft was found to promote the attachment and retention of endothelial cells. These results suggest that this stepwise strategy for introducing cell-specific binding molecules into PCL scaffolds may have potential for development of vascular grafts that can endothelialize rapidly in vivo.     

Preparation and characterization of composite nanofibers of polycaprolactone and nanohydroxyapatite for osteogenic differentiation of mesenchymal stem cells

Nanocomposites of nanohydroxyapatite (nHAP) dispersed in poly(?-caprolactone) (PCL) were prepared by electrospinning (ES) to obtain PCL/nHAP nanofibers. Nanofibers with similar diameters (340 ± 30 nm) but different nHAP concentrations (0-50%) were fabricated and studied for growth and osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs). The nanofibrous membranes were subjected to detailed analysis for its physicochemical properties by scanning electron microscopy (SEM), thermogravimetric analysis, X-ray diffraction, Fourier-transform infrared spectroscopy, and mechanical tensile testing. nHAP particles (~30 nm diameter) embedded in nanofibers increased the nanofibrous membrane's ultimate stress and the elastic modulus, while decreased the strain at failure. When cultured under an osteogenic stimulation condition on nanofibers, MSCs showed normal phenotypic cell morphology, and time-dependent mineralization and osteogenic differentiation from SEM observations and alkaline phosphatase activity assays. The nanofibers could support the growth of mesenchymal stem cells without compromising their osteogenic differentiation capability up to 21 days and the enhancement of cell differentiation by nHAP is positively correlated with its concentration in the nanofibers. Energy dispersive X-ray analysis of Ca and P elements indicated mineral deposits on the cell surface. The mineralization extent was significantly raised in nanofibers with 50% nHAP where a Ca/P ratio similar to that of bone was found. The present study indicated that electrospun composite PCL/nHAP nanofibrous membranes are suitable for mineralization of MSCs intended for bone tissue engineering.     

Preparation of Small‐Diameter Tissue‐Engineered Vascular Grafts Electrospun from Heparin End‐Capped PCL and Evaluation in a Rabbit Carotid Artery Replacement Model

Aiming to construct small diameter (ID <6 mm) off‐the‐shelf tissue‐engineered vascular grafts, the end‐group heparinizd poly(ε‐caprolactone) (PCL) is synthesized by a three‐step process and then electrospun into an inner layer of double‐layer vascular scaffolds (DLVSs) showing a hierarchical double distribution of nano‐ and microfibers. Afterward, PCL without the end‐group heparinization is electrospun into an outer layer. A steady release of grafted heparin and the existence of a glycocalyx structure give the grafts anticoagulation activity and the conjugation of heparin also improves hydrophilicity and accelerates degradation of the scaffolds. The DLVSs are evaluated in six rabbits via a carotid artery interpositional model for a period of three months. All the grafts are patent until explantation, and meanwhile smooth endothelialization and fine revascularization are observed in the grafts. The composition of the outer layer of scaffolds exhibits a significant effect on the aneurysm dilation after implantation. Only one aneurysm dilation is detected at two months and no calcification is formed in the follow‐up term. How to prevent aneurysms remains a challenging topic.     

Synthesis and Characterization of Biodegradable Poly(ϵ-caprolactone)-b-Poly(L-lactide) and Study on Their Electrospun Scaffolds

A series of multi-block copolymers, poly(L-lactide)-b-poly (?-caprolactone) (PLLA-b-PCL) were synthesized. The first step of the synthesis consisted of the transesterification between the PLLA and 1,4-Butanediol, followed by the copolymerization of PLLA-diols and PCL, using isophorone diisocyanate (IPDI) as a coupling agent. The synthesized polymers were characterized by Fourier transform infrared (FTIR) spectra, differential scanning calorimetry (DSC) and wide angle X-ray diffraction (WAXD). PLLA/PCL block copolymers were electrospun into ultrafine fibers. The morphology of the electrospun fibrous scaffolds were investigated by Scanning Electron Microscopy (SEM). Results showed that the morphology and diameter of the fibers were affected by the electrospinning solution concentrationan and different weight ratio of PLLA/PCL. These electrospun PLLA-b-PCL fibrous membranes exhibited good flexibility and deformability. In comparison with the electrospun PLLA membrane, the electrospun fibrous membranes of PLLA-b-PCL demonstrated an enhanced elongation with still high tensile strength and Young's modulus to be beneficial for tissue engineering scaffolds.     

CONTROL OF DIMENSIONAL STABILITY AND DEGRADATION RATE IN ELECTROSPUN COMPOSITE SCAFFOLDS COMPOSED OF POLY(D,L-LACTIDE-co-GLYCOLIDE) AND POLY(ε-CAPROLACTONE)

The purpose of this study is to investigate the effect of composition poly(D,L-lactide-co-glycolide)/poly(ε- caprolactone)(PLGA/PCL)blending on the morphology,shrinkage and degradation behaviors of the electrospun fibers. With the increase of PLGA content in the composite fibers,the average diameter of the electrospun fibers increased from 1.35μm to 1.95μm.The serious shrinking of the electrospun PLGA meshes could be circumvented by adding 20% PCL in the fibers,resulting from the semi-crystalline nature ...     

CONTROL OF DIMENSIONAL STABILITY AND DEGRADATION RATE IN ELECTROSPUN COMPOSITE SCAFFOLDS COMPOSED OF POLY(D,L-LACTIDE-CO-GLYCOLIDE)AND POLY(Ε-CAPROLACTONE)

The purpose of this study is to investigate the effect of composition poly(D,L-lactide-co-glycolide)/poly(ε-caprolactone)(PLGA/PCL)blending on the morphology,shrinkage and degradation behaviors of the electrospun fibers.With the increase of PLGA content in the composite fibers,the average diameter of the electrospun fibers increased from 1.35 μm to 1.95μm.The serious shrinking of the electrospun PLGA meshes could be circumvented by adding 20% PCL in the fibers,resulting from the semi-crystalline nature of PCL.The degradation rate of the electrospun meshes could be modulated by PLGA/PCL composition.In addition,the electrospun meshes containing 20% PCL displayed stable dimensional morphology with degradation.     

Preparation and Characterization of Aligned PLLA/PCL/HA Composite Fibrous Membranes

Aligned poly(L-lactide) (PLLA)/poly(?-caprolactone) (PCL)/hydroxyapaite (HA) composite fibrous membranes were fabricated by electrospinning. Their morphology, thermal stability, mechanical properties, hydrophilic properties and biocompatibility were investigated. The electrospun fibers are highly aligned and the HA are oriented along the fiber axis. When HA are incorporated, the PLLA/PCL/HA composite fibers become thinner due to the increased conductivity. In addition, the aligned HA reinforce the electrospun fibrous membranes. The larger porosity and higher hydrophilic properties induced by HA in the electrospun fibers have improved the degradation of the PLLA/PCL/HA fibrous membranes which have no toxic effect on proliferation of adipose-derived stem cells.     

Poly(ε-caprolactone) filled with electrospun nylon fibres: A model for a facile composite fabrication

Electrospun fibres are very rarely used as reinforcing agents in polymer-based composites. A fabrication approach is presented that allows to easily prepare composites based on polycaprolactone (PCL) filled with nylon 6 electrospun fibres by compression moulding. At very low filler contents (3%), the obtained composites exhibited improved stiffness with a simultaneous increase in ductility, differently from what is usually found in PCL nanocomposites with a variety of fillers, in which increases in modulus happen at the expense of elongation at break. The presence of fibres with a very small diameter, typical of the products of electrospinning, favoured a good interfacial adhesion between matrix and filler. Being of a similar order of magnitude than polymer lamellae, electrospun fibres can be used to shape the morphology of lamellar stacks, and therefore the final properties of the composites.