双喷头静电纺丝法制备载软骨脱细胞基质的复合纳米纤维支架

背景：软骨脱细胞基质是一种理想的用于构建软骨组织工程支架的生物材料,可以用于软骨缺损的修复。目的：以聚乙烯醇为脱细胞基质的负载材料,3-羟基丁酸酯与4-羟基丁酸酯共聚物为框架材料,通过双喷头静电纺丝构建复合纳米纤维支架,并初步探索其体外生物活性。方法：通过酶、化学与超声振荡清洗结合的方法去除软骨组织中的细胞成分,制成软骨脱细胞基质。用双喷头静电纺丝的方法分别制备3-羟基丁酸酯与4-羟基丁酸酯共聚物(记为P34HB)、3-羟基丁酸酯与4-羟基丁酸酯共聚物-聚乙烯醇(记为P34HB-PVA)、3-羟基丁酸酯与4-羟基丁酸酯共聚物-聚乙烯醇-脱细胞基质(记为P34HB-PVA-dECM)纳米纤维支架,表征支架的纤维形貌、组成成分、亲水性能和力学性能。将人骨髓间充质干细胞与3种支架共培养,通过Live/Dead染色来检测细胞在支架的活力,扫描电镜观察细胞在支架上的黏附形态,阿尔玛蓝试剂盒检测细胞在支架上的增殖性能,Ⅱ型胶原免疫荧光评估细胞在支架上的成软骨分化。结果与结论：(1)扫描电镜下可见,各组支架纤维呈随机分布,纤维之间互相连接呈现多孔结构,其中P34HB-PVA-dECM支架的纤维直径最...     

静电纺纳米纤维基组织工程大孔支架的研究进展

静电纺丝作为一种纳米纤维支架的仿生构建方法,已在组织工程和再生医学领域中得到越来越多的应用和关注。但是,静电纺支架的主要问题是密集排列的纳米纤维之间的空隙很小,阻碍细胞的长入和三维(3 D)组织的形成。为了解决这一问题,近年来已发展了许多用于扩大静电纺纳米纤维支架孔尺寸的制备方法。首先概述组织工程支架中大孔对细胞行为的影响,然后对静电纺纳米纤维3 D大孔支架的制备方法和技术研究进展进行综述,讨论这些3 D大孔支架促进细胞长入的效果,最后对静电纺3 D大孔支架在组织工程中应用的主要挑战和前景,提出了看法。     

生物降解聚乳酸纤维的制备方法研究进展

聚乳酸（PLA）是具有代表性的可生物降解聚酯，聚乳酸纤维的纺丝成形加工也一直是研究和开发的热点。本文综述了可生物降解聚乳酸纤维的制备方法：熔融纺丝、溶液纺丝、静电纺丝等纤维成型的特点及进展。     

海藻酸钠/壳聚糖复合凝胶的制备与细胞毒性评价

背景：海藻酸钠和壳聚糖分别是聚阳离子和聚阴离子的天然高分子材料,二者可以互为交联剂形成复合凝胶,避免使用普通交联剂产生的细胞毒性。 目的：制备海藻酸钠壳聚糖复合凝胶并评价其体外细胞毒性。 方法：以0.25 mol/L乙酸溶解壳聚糖,制成质量浓度30 g/L溶液,再以0.1 mol/L的NaOH中和酸性得到壳聚糖絮状沉淀,将壳聚糖絮状沉淀与质量浓度3%海藻酸钠等比例混合,高频振动混匀,使二者形成复合凝胶。使用傅里叶变换红外光谱、扫描电镜分析观察凝胶成分和交联纤维网络结构。分别以海藻酸钠壳聚糖复合凝胶24,72 h浸提液、聚乙烯24,72 h浸提液及苯酚溶液培养L-929细胞,体外检测其细胞毒性。 结果与结论：傅里叶变换红外光谱检测到海藻酸钠壳聚糖复合凝胶特征性峰值改变,扫描电镜显示其内部形成丰富间隙的空间网络结构。浸提液法检测海藻酸钠壳聚糖复合凝胶的细胞毒性为合格,表明海藻酸钠/壳聚糖复合凝胶具有成为组织工程支架材料的良好条件。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

静电纺丝聚膦腈/明胶复合纤维支架的生物矿化行为**◆

背景：虽然静电纺丝高分子纤维的生物矿化研究文章已不少见,但国内外尚无关于静电纺丝聚膦腈及其与明胶复合纤维的生物矿化研究报道。 目的：考察聚膦腈/明胶复合纤维支架作为骨组织工程支架的可行性。 方法：静电纺丝法构建生物可降解聚膦腈/明胶复合纤维支架,采用5倍模拟体液,并结合扫描电镜、X射线能谱、X射线光电子能谱、傅里叶变换红外光谱等手段,观察其生物矿化行为。 结果与结论：与纯明胶的纤维膜相比,聚(丙氨酸乙酯-甘氨酸乙酯)膦腈(PAGP)和明胶混合溶液静电纺丝得到的复合纤维膜,经交联处理后仍能够保持良好的纤维形貌和多孔结构。在采用CO2平衡的改进5倍模拟体液中,纯PAGP和PAGP/明胶纤维表面沉积的矿物质都经历了片状二水合磷酸一氢钙前驱体的生成及其向羟基磷灰石转化的过程,但后者由于明胶成分的存在,整个过程发生发展的速度要明显快于前者。而对于纯明胶纤维,其在改进5倍模拟体液中浸泡24 h后,所生成矿物质仍主要为羟基磷灰石的前驱体二水合磷酸一氢钙。说明复合纤维中,疏水性PAGP的引入不仅有利于纤维形貌的保持,还能抑制明胶的溶出,使PAGP /明胶复合纤维的矿化性能明显改善。       

新型组织工程血管材料：静电纺复合纳米纤维小口径管状支架

背景：小口径人工血管对生物相容性和抗凝血的要求远远高于普通大口径人工血管,因此血管移植体内原位诱导组织再生成为了新的研究方向。 目的：总结近几年静电纺复合纳米纤维小口径管状支架的主要研究进展,并讨论其在体内原位诱导血管再生方面的重要应用。 方法：由第一作者检索中国期刊网CNKI全文数据库、万方数据库及ISI Web of Knowledge外文数据库,有关复合纳米纤维小口径管状支架的制备方法、血管支架仿生天然细胞外基质微环境的表面修饰以及种植体植入后生物相容性和安全性评价等方面的文献。 结果与结论：静电纺复合纳米纤维制备小口径管状支架,即将天然材料和合成材料共纺在一起,这样既能克服天然生物高分子材料力学性能的不足,又能避免合成材料在生物相容性和安全性的缺陷,成为制备小口径血管组织工程支架的必然趋势。同时制备多层血管,进行功能化修饰,模拟天然细胞外基质的结构和功能,将成为用于心血管组织修复及再生小口径血管组织工程研究的新方向。在获得上述新进展的同时,经动物实验检验的静电纺血管支架以聚合物为主。尽管这类支架采用了各种手段避免血栓、炎症等不良反应,其生物相容性仍旧无法与天然材料相比。由此可见,在天然材料与合成材料之间找到一个最佳比例,使复合材料的力学性能和血管相容性达到一个平衡,将会显著提高静电纺复合纳米纤维支架在小口径血管组织再生中的应用。     

乳酸-乙醇酸共聚物载药复合纤维制备及其释药行为研究

乳酸-乙醇酸共聚物(PLGA)复合纤维具有良好的生物相容性和生物可降解性,且降解速率可由相对分子质量和共聚物组成来调控.采用同轴静电纺丝法制备以PLGA为壳层材料、聚乙烯基吡咯烷酮(PVP)为内芯材料的复合纤维,研究两种模型药物(5-氟尿嘧啶和硝苯地平)在同轴复合纤维载体中的药物释放行为,并用扫描、透射电镜观察复合纤维的形貌与结构,用紫外分光光度计测量载药量和累积释放率.实验结果表明,通过改变芯、壳纺丝液浓度、PLGA相对分子质量以及共聚物中LA和GA的组成比,可制得具有芯-壳结构且直径大小不同的复合纤维.采用相同电纺丝条件,可以将不同药物以相同载药量包覆于复合纤维中,但药物的释放行为不相同.     

脱细胞皮肤基质/聚氨酯混纺纤维支架促进大鼠皮肤缺损的修复

背景：研究证实,将脱细胞基质与聚合物进行混合纺丝不仅可改善纤维的结构性质,还可保存脱细胞基质的生物学活性,但是目前将聚氨酯与脱细胞皮肤基质进行静电纺丝制备皮肤组织工程支架还没有相关报道。目的：观察脱细胞皮肤基质/聚氨酯混纺纤维支架对大鼠皮肤缺损的修复作用。方法：利用静电纺丝技术分别制备聚氨酯静电纺丝纤维支架和脱细胞皮肤基质/聚氨酯混纺纤维支架。扫描电镜下观察纤维结构;将大鼠脂肪间充质干细胞分别接种于两组支架上,扫描电镜下观察支架上的细胞形态。取10只SD大鼠,在每只大鼠背部制作3个1 cm×1 cm的全层皮肤缺损,其中2处缺损分别植入聚氨酯静电纺丝纤维支架(对照组)、脱细胞皮肤基质/聚氨酯混纺纤维支架(实验组),剩余缺损处不植入任何材料(空白对照组)。术后第1,2,3周,观察皮肤创面愈合情况;植入后第3周,进行创面苏木精-伊红染色,计算创面瘢痕面积。结果与结论：(1)扫描电镜下可见两种静电纺丝纤维均为网状结构,并且大鼠脂肪间充质干细胞在两种支架上沿纤维附着,黏附良好。(2)随着术后时间的延长,各组皮肤创面逐渐愈合,至术后第3周,实验组、对照组皮肤创面基本愈合,空白对照组创面可见小溃疡形...     

电纺丝技术制备组织工程食管仿生支架

背景：前期实验中曾发现纤维的取向可以引导平滑肌细胞的取向生长,因此,设想通过制备取向排列的电纺丝纤维支架,以引导食管平滑肌细胞的有序生长,从而有利于维持肌细胞的形貌及生物功能。 目的：以可降解聚己内酯、明胶、丝素蛋白为基材,采用自制的电纺丝系统制备无规和有序的纳米级多孔纤维。 方法：将聚己内酯与丝素蛋白以4∶1质量比混合,通过调整溶液浓度、电压、喷射速度等参数,采用自制的电纺丝系统制备聚己内酯/丝素蛋白电纺丝纤维。将聚己内酯与明胶分别以2∶1、1∶1、1∶2质量比混合,在金属平板接收器下,通过调整溶液浓度、电压、喷射速度等参数,采用自制的电纺丝系统制备聚己内酯/明胶无规电纺丝纤维;同时改用滚轴接收装置,通过调整滚轴转数、电压、喷射速度等参数,制备聚己内酯/明胶有序电纺丝纤维。 结果与结论：在溶液质量浓度为0.08 g/mL、纺丝液流速1.6 mL/h和电压22.5 kV的条件下,制得了均匀、无串珠、纤维直径为(535.9±126.7) nm的聚己内酯/丝素蛋白多孔纳米纤维膜。在溶液质量浓度为0.10 g/mL、纺丝液流速0.8 mL/h和电压22.5 kV的条件下,制得了无明显串珠、纤维直径为(257.9±117.8) nm的聚己内酯/明胶多孔纳米纤维膜;并且在1∶2质量比时更易成纤维,纤维尺寸更均匀。在滚轴转速3 000 r/min。溶液流速0.8 mL/h。电压15 kV的条件下,制得的聚己内酯/明胶有序电纺丝纤维排序更理想,纤维也更均匀。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

静电纺丝复合缓释抗菌微球的制备及性能分析

背景：临床静脉注射抗生素治疗骨髓炎时,由于骨骼血供较差,药物难以在骨髓炎局部病灶处达到有效治疗浓度,治疗效果不佳一直是临床瓶颈。目的：研制一种可局部缓释盐酸万古霉素、同时具备成骨作用的可降解生物材料植入物,在治疗骨髓炎的同时尽可能起到促进局部骨组织修复的作用。方法：将6,12,18 g/L的盐酸万古霉素-海藻酸钠溶液以电喷的方式制成微球,进行真空冷冻干燥处理;制备聚3-羟基丁酸酯4-羟基丁酸酯电纺溶液,电纺同时,向接收装置上加入载盐酸万古霉素微球,使其包裹于电纺丝中,制备了含6,12,18 g/L盐酸万古霉素微球的静电纺丝支架,分别记为P@VancoMs6、P@VancoMs12、P@VancoMs18。(1)体外实验：将MC3T3-E1细胞分别接种于3种支架上,检测细胞黏附、增殖,筛选适宜额盐酸万古霉素质量浓度,用于后续实验。将MC3T3-E1细胞接种于P@VancoMs6、P@VancoMs12上,加入成骨诱导培养基,通过Ⅰ型胶原和骨钙蛋白免疫荧光染色评价支架体外成骨性能。将P@VancoMs6、P@VancoMs12分别与金黄色葡萄球菌共培养,采用抑菌圈实验分析复合物的抗菌性能。...     

Electrohydrodynamic jetting behaviour of polyhedral oligomeric silsesquioxane nanocomposite

In this paper, we investigate in detail the electrohydrodynamic spraying of a nonbiodegradable nanocomposite polyhedral oligomeric silsesquioxane polymer developed in our laboratories and currently being explored for coating metallic stent materials. Different concentrations of the polymer have been dissolved to prepare, characterise, and electrohydrodynamically deposit the polymer on stainless steel. From the experiments, the solution containing 15 wt% polymer was selected for further investigation. The variation of film/coating thickness as a function of spraying time was studied and the structural features of the film were assessed using microscopy. Films were also tensile tested. This study has identified a process and conditions which can be used in our stent coating research.     

The degradative resistance of polyhedral oligomeric silsesquioxane nanocore integrated polyurethanes: an in vitro study

Polymer biostability is one of the critical parameters by which these materials are selected for use as biomedical devices. This is the major rationale for the use of polymers which are highly crystalline and stiff namely expanded polytetrafluoroethylene (ePTFE) and Dacron in particular, as arterial bypass grafts. While this is immaterial in high-flow states, it becomes critically important at lower flows with a greater need for more compliant vessels. Polyurethanes being one of the most compliant polymers known are as such, the natural choice to build such constructs. However, concerns regarding their resistance to degradation have limited their use as vascular prostheses and in order to augment their strength, herein a novel polyhedral oligomeric silsesquioxane integrated poly(carbonate-urea)urethane (POSS-PCU) nanocomposite was synthesised by our group. In the following series of experiments, the POSS-PCU nanocomposite samples were exposed to accelerated degradative solutions, in an 'in-house' established model in vitro for up to 70 days before being subjected to infra-red spectroscopy, scanning electron microscopy, stress-strain studies and differential scanning calorimetry. Our results demonstrate that these silsesquioxane nanocores shield the soft segment(s) of the polyurethane, responsible for its compliance and elasticity from all forms of degradation, principally oxidation and hydrolysis. These nanocomposites hence provide an optimal method by which these polymers may be strengthened whilst maintaining their elasticity, making them ideal as vascular prostheses particularly at low flow states.     

Herceptin functionalized polyhedral oligomeric silsesquioxane - conjugated oligomers - silica/iron oxide nanoparticles for tumor cell sorting and detection

Sorting and detection of circulating tumor cells (CTC) in peripheral blood as an efficient and non-invasive method to diagnose cancer have recently attracted much attention. In this article, we developed a multiply-engineered nanoparticle system for CTC sorting and detection, which consists of (1) conjugated oligomer (CO) as fluorescence signal source, (2) polyhedral oligomeric silsesquioxanes (POSS) scaffold for CO localization for better fluorescent effects, (3) silica nanoparticles (SiNPs) as formulation matrix of the POSS containing CO, (4) iron oxide (IO) layer on the silica nanoparticles (IO-SiNPs) for magnetic collection, and (5) herceptin surface functionalization of the IO-SiNPs to target cancer cells of HER2 overexpression. Such a multiply-engineered structure can be used for either traditional immunomagnetic methods or microfluidic devices for CTC sorting and detection.     

Electrospinning of poly(lactic acid)/polyhedral oligomeric silsesquioxane nanocomposites and their potential in chondrogenic tissue regeneration

The study was conducted to evaluate the cytocompatibility and hydrolytic degradability of the new poly(lactic acid)/polyethylene glycol-polyhedral oligomeric silsesquioxane (peg-POSS/PLLA) nanocomposite as potential material for cartilage regeneration. PLLA scaffolds containing 0 to 5% of peg-POSS were fabricated by electrospinning. Human mesenchymal stem cells (hMSC’s) were cultured in vitro to evaluate the cytocompatibility of the new nanocomposite material. Hydrolytic degradation studies were also carried out to analyze the mass loss rate of the nanocomposites through time. The addition of the peg-POSS to the PLLA did not affect the processability of the nanocomposite by electrospinning. It was also observed that peg-POSS did not show any relevant change in fibers morphology, concluding that it was well dispersed. However, addition of peg-POSS caused noticeable decrease in mean fiber diameter, which made the specific surface area of the scaffold to rise. hMSC’s were able to attach, to proliferate, and to differentiate into chondrocytes in a similar way onto the different types of electrospun peg-POSS/PLLA and pure PLLA scaffolds, showing that the peg-POSS as nano-additive does not exhibit any cytotoxicity. The hydrolytic degradation rate of the material was lower when peg-POSS was added, showing a higher durability of the nanocomposites through time. Results demonstrate that the addition of peg-POSS to the PLLA scaffolds does not affect its cytocompatibility to obtain hyaline cartilage from hMSC’s.     

Biocompatibility of novel polymer-apatite nanocomposite fibers

On the basis of the bioactivity of hydroxyapatite (HA) and the excellent mechanical and biocompatible performance of polyethylene terephthalate (PET), composite microfibers made of nanograde HA with PET was designed and fabricated to mimic the structure of biological bone, which exhibits a composite of nanograde apatite crystals and natural polymer. The PET/HA nanocomposite was molded into fibers so that the bulk structures' mechanical properties can be custom tailored by changing the final 3D orientation of the fibbers. This study focused on the in vitro biocompatibility evaluation of the PET/HA composite fibers as potential bone fixation biomaterial for total hip replacement prosthesis surfaces. The MTT assay was performed with the extracts of the composite fibers in order to evaluate the short-term effects of the degradation products. The cell morphology of L929 mouse fibroblast cell line was analyzed after direct contact with the fiber scaffolds for different time periods, and the cell viability was also analyzed by the Alamar Blue assay. The release of the inflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha), from RAW 264.7 macrophages in the presence of fiber extracts and fibers was used as a measure of the inflammatory response. The ability of the fiber matrices to support L929 attachment, spreading, and growth in vitro, combined with the compatible degradation extracts and low inflammation potential of the fibers and extracts, suggests potential use of these fibers as load-baring bone fixation biomaterial structures.     

Polyhedral oligomeric silsequioxane-polyurethane nanocomposite microvessels for an artificial capillary bed

Fabricating artificial vascularised tissue would involve tissue-engineering techniques, but current technology limits this as cultured cells depend on growth media in vitro and on diffusion in vivo. Therefore, there is a need to construct a synthetic microvascular network, which would sustain these cultured cells in a similar manner to normal tissue. This is again hampered by the poor patency rates of current microvascular grafts. Based on our previous work on polyhedral oligomeric silsesquioxane-polyurethane nanocomposites, which have shown the unique ability to repel coagulant proteins whilst still allowing endothelialisation, we have now developed a new generation of microvascular prosthesis using this polymer. Using these dip-coated nanocomposite microvessels, we have shown that it is possible to mimic the hydraulic conductivity and pressure-responsive radial compliance characteristics of biological microvessels. This would allow nutrient exchange across its walls as well as minimise compliance mismatch throughout the physiological pressure range thus reducing intimal hyperplasia in the long term. This microvessel would have the following implications: (1) as a microvascular substitute to vein grafts and (2) in the future as a component of a microvascular network.     

Preparation of hydroxyapatite-gelatin nanocomposite

A nanocomposite of gelatin[GEL]-hydroxyapatite[HAp] was prepared using the biomimetic process. The hydroxyapatite nanocrystals were precipitated in aqueous solution of gelatin at pH 8 and 38 degrees C. The chemical bonding between calcium ions of HAp and carboxyl ions of GEL molecules induced a red-shift of the 1339 cm(-1) band of GEL in FT-IR analysis. TEM images and electron diffraction patterns for the nanocomposite strongly indicate the self-organization of HAp nanocrystals along the GEL fibrils. Electron diffraction for the nanocomposites showed a strong preferred orientation of the (002) plane in HAp nanocrystals. The development of HAp nanocrystals in an aqueous GEL solution was highly influenced by the concentration ratio of GEL to HAp. A higher concentration of GEL induced the formation of tiny crystallites (4 nm x 9 nm size), while a lower concentration of GEL contributed to the development of bigger crystallites (30 nm x 70 nm size). From DT/TGA data, the HAp-GEL nanocomposite showed typically three exothermic temperatures. The increase in decomposition temperatures indicates the formation of a primary chemical bond between HAp and GEL. The higher concentration of GEL supplies abundant reaction sites containing groups such as carboxyl, which can bind with calcium ions. The abundant supply of reaction sites leads to a very large number of HAp nuclei. However, the formation of a large number of nuclei depletes the concentration of calcium ions that available for growth to the extent that the nuclei cannot grow very large. This in turn will lead to the creation of a large number of tiny nanocrystals at this higher GEL concentration.     

Electrospinning biomedical nanocomposite fibers of hydroxyapatite/poly(lactic acid) for bone regeneration

Development of fibrous matrices of bioceramic-biopolymer nanocomposite offers great potential in the field of bone regeneration and tissue engineering. However, in order to produce electrospun fibers with homogeneous structure, it is essential for the ceramic powder to be fine and to remain stable in suspension. Herein, we developed a novel method whereby the bioceramic hydroxyapatite (HA) was kept in suspension in biopolymer poly(lactic acid) (PLA). The strategy was to introduce a surfactant hydroxysteric acid (HSA) between the hydrophilic HA powder and the hydrophobic chloroform-dissolved PLA. The HA nanopowder was dispersed effectively in HSA and mixed homogeneously with PLA. Continuous and uniform fibers were generated successfully with diameters of approximately 1-2 microm, and featured a well-developed nanocomposite structure of HA nanopowder-dispersed PLA. Initial cellular assays showed excellent cell attachment and proliferation and also enhanced expression of alkaline phosphatase at 7 days of culturing. The HA-PLA nanocomposite fibers may be potentially useful in tissue engineering applications, particularly as three-dimensional substrates for bone growth.     

Preparation and characterization of polyurethane optical phantoms

We describe a method for the preparation of a polyurethane phantom to simulate the optical properties of biologic tissues at two wavelengths in the visible and near-infrared spectral range. We characterize the addition of added molecular absorbers with relatively narrow absorption bands [full width at half maximum (FWHM) 32 and 76 nm for Epolight 6084 and 4148, respectively] for independent absorption at 690 nm for absorption up to 5 cm(-1), and 830 nm for absorptions up to 3 cm(-1). Absorption by both dyes is linear with concentration in these respective regions and is consistent in polyurethane both before and after curing. The dyes are stable over long durations with no more than 4% change. The absorption of visible light by polyurethane decreases with time and is stable by one year with a drop of 0.03+/-0.003 cm(-1) from 500 to 830 nm. The scattering properties are selected by the addition of TiO2 particles to the polyurethane, which we functionally describe for the 690- and 830-nm wavelengths as related to the weight per volume. We demonstrate that the variation in absorption and scattering properties for large batch fabrication (12 samples) is +/-3%. The optical properties of the phantoms have not significantly changed in a period of exceeding one year, which makes them suitable for use as a reference standard.     

聚六亚甲基碳酸酯聚氨酯脲的制备与性能评价

目的评价聚六亚甲基碳酸酯聚氨酯脲(PCU)的性能。方法通过拉伸试验、96℃±4℃热水水解试验、次氯酸钠溶液中氧化实验、全血凝血时间实验、测定动态凝血时间实验、血小板黏附实验等,分别测定了PCU的力学性能、耐水解性、耐氧化性和抗凝血性,并与国外产品Chronoflex(Chro)进行对比。结果PCU的强度与Chro相近,但其弹性和柔性逊色于Chro,PCU的耐水解性能和耐氧化性能略优于Chro,PCU和Chro都具有良好的抗凝血性能。结论PCU是一种性能与Chro相近的“生物稳定”聚氨酯。