Patterning the neuronal cells via inkjet printing of self-assembled peptides on silk scaffolds

The patterning of neuronal cells and guiding neurite growth are important for neuron tissue engineering and cell-based biosensors. In this paper, inkjet printing has been employed to pattern self-assembled I₃QGK peptide nanofibers on silk substrates for guiding the growth of neuron-like PC12 cells. Atomic force microscopy (AFM) confirmed the dynamic self-assembly of I₃QGK into nanofiber structures. The printed self-assembled peptide strongly adheres to regenerated silk fibroin (RSF) substrates through charge-charge interactions. It was observed that in the absence of I₃QGK, PC12 cells exhibited poor attachment to RSF films, while for RSF surfaces coated or printed with peptide nanofibers, cellular attachment was significantly improved in terms of both cell density and morphology. AFM results revealed that peptide nanofibers can promote the generation of axons and terminal buttons of PC12 cells, indicating that I₃QGK nanofibers not only promote cellular attachment but also facilitate differentiation into neuronal phenotypes. Inkjet printing allows complex patterning of peptide nanofibers onto RSF substrates, which enabled us to engineer cell alignment and provide an opportunity to direct axonal development in vitro. The live/dead assay showed that printed I₃QGK patterns exhibit no cytotoxicity to PC12 cells demonstrating potential for future nerve tissue engineering applications.     

Preparation and Characterization of Silk Fibroin Aerogel

Silk protein fibroin,as a biomedical material,has good biocompatibility,biodegradability,regulation and excellent physical and chemical properties. In this work,a low density porous silk fibroin material is prepared from fibroin solution by high-speed shearing with impeller. By adjusting shear rate of the stirrer,silk fibroin aerogel with different sizes of the aperture is prepared. In general, this aerogel has small porosity, uniform pores, good mechanical properties and slow rate of degradation. It is observed that increasing the shear rate results in higher porosity of aerogel,while the diameter of the aerogel becomes smaller. This silk aerogel may offer a new option as biomaterial for the tissue engineering application based on the information on the structural behaviors.     

Nanoscale control of silks for regular hydroxyapatite formation

The control of nanoparticle shape is still a challenge for different hydroxyapatite (HA) preparation ways.In the present study,silk fibroin with regular nanostructures was firstly prepared and then emp...     

丝素改性聚氨酯膜的物理性质及其细胞相容性

用丝素蛋白或丝素肽表面改性的聚氨酯膜表现出良好的柔韧性，它能够克服丝素膜刚而脆的弱点；由水分散型聚氨酯制备的改性膜的透气性高于疏水性聚氨酯膜；细胞培养实验结果表明：经丝素蛋白或丝素肽改性后，聚氨酯膜的生物相容性明显提高且处理效果是稳定的。     

明胶/丝素共混多孔膜的制备

采用冷冻干燥法制备明胶与家蚕丝素的共混多孔蛋白膜，探讨了交联剂用量，共混比例及冷冻温度对膜结构和性能的影响，指出戊二醛对共混多膜具有明显的交联作用，膜内丝素与明胶的相容性较好，蛋白质的结构以无定形为主，通过与明胶共混，能够改善多孔丝素膜的拉伸性能。     

Silk Fibroin/Poly-L-lactide Lactone Bi-layered Membranes for Guided Bone Regeneration

After removal of the caries or diseased teeth,the alveolar ridge will undergo absorption and atrophy.When the amount of alveolar bone is insufficient,it will cause an inability to perform effective dental implant restoration.In order to control the absorption and promote the repair and regeneration of alveolar ridge,a method of implanting guided bone regeneration(GBR) membranes at the extraction site is often used.In this study,silk fibroin(SF)and poly-L-lactide lactone(PLCL) were used to prepare bilayered guided bone regeneration membranes,and its morphology,hydrophilicity,surface roughness and mechanical properties were studied.At the same time,the drug release behaviors and cell compatibility of the bilayered membranes were studied.The results showed that SF/PLCL bi-layered membranes had good mechanical properties and surface hydrophilicity,and the drug-loaded bi-layered membranes had good cell compatibility.The bilayered membranes fabricated in this study are of potential for applying in the oral health field to promote bone regeneration.     

Examination of Subunit Composition of Bombyx Mori Silk Fibroin

Main subunits of the silk fibroin were separated by GFC(gel filtration chromatography)technigue.The nativesilk fibroin and α,c subunits were measured by gel elec-trophoresis.The aminoacid compositions of the nativesilk fibroin and α,c subunits were analyzed by means ofaminoacid measurement.Some properties of silk was inter-preted in view of subunit composition of silk fibroin.     

电纺聚-L-乳酸/β-磷酸三钙复合物纳米纤维膜

利用静电纺丝法制备了生物可吸收聚-L-乳酸(PLLA)/β-磷酸三钙(β-TCP)复合物纳米纤维膜.采用扫描电子显微镜(SEM)、场发射扫描电子显微镜(FESEM)等手段研究了复合物纳米纤维膜的结构和形态,详细探讨电纺工艺条件对制备PLLA/β-TCP复合物纳米纤维的形态影响.通过拉伸力学测试、噻唑蓝比色法(MTT)对复合物纳米纤维膜的力学性能和体外细胞相容性作了进一步研究.结果表明,PLLA/β-TCP复合物纳米纤维的几何结构与电纺条件有关,随着聚合物溶液浓度增加、溶液流速增大,纤维直径有不同程度的增大;复合物纳米纤维膜的拉伸强度和杨氏模量随β-TCP的含量增加而下降;复合物纳米纤维膜对L-929细胞系无细胞毒性,显示良好的细胞相容性.     

Cu（Ⅱ） effect on the conformation of regenerated silk fibroin in dilute aqueous solution

Much attention has been paid to the natural mechanism of silkworm spinning due to the impressive mechanical properties of the natural fibers. In this work, we studied the effect of Cu（Ⅱ） ions on the secondary structure of Bombyx mori regenerated silk fibroin （SF） in dilute solution by circular dichroism （CD）. The results indicate that a given amount of Cu（Ⅱ） induces the SF conformational transition from random coil to β-sheet, however, further addition of Cu（Ⅱ） is unfavorable for this conversion. Meanwhile, the conformational changes induced by Cu（Ⅱ） follow a nucleation-dependent aggregation mechanism, which is similar to that found in Prion protein （PrP） denaturation and Aβ-peptide aggregations, leading to the neurodegenerative disease. This work would help one understand further the natural spinning process of silkworm. Additionally, it would be significant for the study of the nervous system diseases, because silk fibroin, extracted in large amounts from Bombyx mori silkworm gland, could be a proper model to study PrP denaturation and Aβ-peptide aggregations.     

高含量再生丝素蛋白水溶液的热性能研究

用差示扫描量热仪(DSC),对不同含量的再生丝素蛋白水溶液(质量分数为12%~42%),以及同一含量不同剪切速率(1 000 s-1和2 000 s-1)下溶液热性能的变化进行了对比研究。结果表明,随着含量的增大,再生丝素蛋白水溶液发生相转变的温度逐渐降低;同样,经剪切后的溶液发生相转变的温度均向低温漂移,且随着剪切作用的加强,向低温的漂移越明显。试验表明,丝素水溶液含量的增加和剪切作用可加速溶液从无规线团或α-螺旋向β-折叠构象的转变速度,且随着剪切速率的提高,构象转变的速度也加快。     

丝素在NMMO·H2O中的溶解及溶液流变性能的研究

探讨了丝素在N-甲基吗啉-N-氧化物一水合物(NMMO.H2O)中的溶解行为,结果发现:天然蚕丝纤维脱胶后得到的丝纤维可以直接溶解在NMMO.H2O中,但是很难得到高于10%浓度的溶液,因而无法用于纺丝.而由再生丝素水溶液制得的再生丝素膜则易于溶解在NMMO.H2O中,并可以得到浓度高达25%的再生丝素/NMMO.H2O溶液,在上述研究的基础上,本文还对再生丝素/NMMO.H2O的流变行为进行了初步研究.     

家蚕丝素溶液的粘弹性研究

采用Coucttc流变仪对家蚕丝素溶液的粘弹性进行研究。对于生丝丝素溶液,当丝素浓度较稀时(如4w/v％),溶液表现出牛顿流动行为;当丝素浓度较高时(如25W/V％),溶液表现出复杂的非牛顿流动行为。成烈的家蚕丝腺中的丝素,在剪切速率大于0.25s~1时,能引起纤维化。     

Silk Fibroin-modified Poly（butadiene） urethane Films and Their Effects on Fibroblast Viability

Surface-modified poly（butadiene）urethane （PBTU） films with silk fibroin （SF） were prepared by simple chemical method under the normal temperature. The physical properties and biological behaviour of the SF-modified PBTU film were evaluated. The results showed that the SF-modified PBTU films kept the tenacity and pliability very well, and could overcome rigid and brittle weaks of silk fibroin films. The morphology of SF in the PBTU film was dendritic aggregations, and the water-contact angle measurement indicated that the surface hydrophilicity of modified films was apparently enhanced. The biocompatibility of PBTU films was improved due to the change of surface components. The degree of platelet adhesion and the cell viability of rat embryo dermal fibroblasts seeded on PBTU films, SF films, and SF-modified PBTU films were measured by counting platelets before and after they contacted the films and MTT assay, respectively. The results indicated that platelet adhesion resistance and cell viability on the modified film were greatly superior to those on the PBTU film and the compound interface had good stability in the air.     

Mechanism of silk processing in insects and spiders

Silk spinning by insects and spiders leads to the formation of fibres that exhibit high strength and toughness. The lack of understanding of the protein processing in silk glands has prevented the recapitulation of these properties in vitro from reconstituted or genetically engineered silks. Here we report the identification of emulsion formation and micellar structures from aqueous solutions of reconstituted silkworm silk fibroin as a first step in the process to control water and protein-protein interactions. The sizes (100-200 nm diameter) of these structures could be predicted from hydrophobicity plots of silk protein primary sequence. These micelles subsequently aggregated into larger 'globules' and gel-like states as the concentration of silk fibroin increased, while maintaining solubility owing to the hydrophilic regions of the protein interspersed among the larger hydrophobic regions. Upon physical shearing or stretching structural transitions, increased birefringence and morphological alignment were demonstrated, indicating that this process mimics the behaviour of similar native silk proteins in vivo. Final morphological features of these silk materials are similar to those observed in native silkworm fibres.     

天然丝素制备三维多孔支架

研究盐沥滤法制备三维多孔丝素支架,在丝素多孔支架上培养人成骨肉瘤细胞、成纤维细胞及肝细胞.研究材料的细胞相容性,发现多种动物细胞在盐沥滤法所制备的丝素多孔支架材料上能够很好地黏附和增殖.通过调整丝素水溶液的浓度、溶解丝素溶剂体系、NaCl添加量和NaCl粒径,制备出结构和性质可以调控的三维支架.该结果为丝素多孔支架材料的制备提供了新的研究思路和方法.     

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.     

丝素在生物医学领域中的应用

讨论了丝素（silk fibroin,SF)与生物医学领域应用相关的基本性质，以及在生物分析及药物缓释载体，人工器官及组织工程支撑体材料中的应用现状，结论认为：丝素具有固化结晶方式的多样化。易于保持高度生物亲和性以及形成特殊的多孔性网状结构。丝素还具有优良的成膜特性，并且通过与其它一些天然高分子材料的复合与化学改性，可以进一步改善其物理性能，从而适宜于构建新型药物载体与人工器官，同时，针对丝素蛋白在体的生物相容性，在体降解特性方面存在的问题，认为其在体变化机制不仅取决于与之接触体液的化学构成，也与温度，力学环境等生物物理因素有关，因此其在体变化机制是生物力学与生物流变学的因素与生物化学因素协同控制的。     

切变速度和Ca~(2+)对再生丝素蛋白溶液构象转变的影响

采用偏光显微镜和拉曼光谱研究了切变速度和Ca2+对再生丝素蛋白水溶液构象转变的影响.结果表明:切变速度和Ca2+是影响再生丝素蛋白溶液性质的两个主要因素,对于同一再生溶液,切变速度增大,溶液中丝素蛋白分子沿切变方向的有序排列也随之增加,丝素溶液更容易从无规线团或α-螺旋向β-折叠转变,并且转变的程度随着剪切速度的增加而加强;透析溶液中Ca2+含量增加,则样品出现偏光现象的临界切变速度有所降低.拉曼光谱显示丝素蛋白分子构象由SilkⅠ向SilkⅡ转变.     

Fine Structure of Regenerated Silk Fibroin Solids

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