纳米羟基磷灰石/丝素蛋白/聚己内酯复合超细纤维的制备及表征

通过静电纺丝法制备出纳米羟基磷灰石/丝素蛋白/聚己内酯复合超细纤维,利用扫描电镜、红外光谱仪、X射线衍射仪对纳米羟基磷灰石/丝素蛋白/聚己内酯复合超细纤维形貌和结构进行表征,并进行了拉伸测试。结果表明,随着超细纤维中羟基磷灰石含量的增加,纤维的直径逐渐降低,纤维中聚己内酯的结晶逐渐变差。相比于丝素蛋白/聚己内酯超细纤维,含有质量比为30%羟基磷灰石的复合超细纤维仍具有较好的力学性能。体外小鼠成纤维细胞(L929)培养表明,纳米羟基磷灰石/丝素蛋白/聚己内酯复合超细纤维对细胞没有毒性。     

纳米羟基磷灰石丝素蛋白仿生矿化材料的制备研究

以Ca(NO₃)₂与Na₃PO₄为无机相的前驱体,将丝素蛋白直接溶于Ca(NO₃)₂溶液中,不经过脱盐处理,直接滴入Na₃PO₄溶液中反应,在37℃下丝素蛋白和羟基磷灰石晶体之间相互作用,仿生合成了纳米羟基磷灰石(n-HA)丝素蛋白(SF)生物矿化材料.用FTIR、XRD、XPS和SEM进行表征.结果表明,羟基磷灰石和丝素蛋白两相间具有较强的化学键合,矿化材料中无机相包含少量碳酸根,为缺钙类骨羟基磷灰石并且呈现一定的长轴取向性,说明丝素蛋白大分子对羟基磷灰石晶体的成核和生长起着模板和调控作用.矿化物颗粒尺寸在50~200nm之间,其抗压强度为32.21MPa,可作为非承重部位骨组织缺损修复材料.     

电活性的石墨烯/丝素蛋白取向纳米纤维膜的制备和表征

丝素蛋白和石墨烯由于具有优异性能而被广泛用于生物医学领域。为了更好的仿生天然神经的拓扑结构和电学微环境,将氧化石墨烯(GO)纳米片涂层到丝素蛋白取向纳米纤维膜上,用Vitamin C还原GO制备了具有电活性的石墨烯/丝素蛋白取向纳米纤维膜。扫描电镜(SEM)观察结果显示随着GO涂层时间的增加,纤维的直径逐渐增加,纤维的表面变得越来越粗糙并出现皱褶。电性能测试结果发现用40mmol/L Vitamin C溶液50℃还原2h,取向丝素蛋白纳米纤维膜的电导率可达到1.66×10~(-2)S/cm。傅里叶变换衰减全反射红外光谱(ATR-FT-IR)、拉曼光谱(Raman spectrum)、X射线光电子能谱(XPS)的表征结果表明GO成功地涂层到取向丝素蛋白纳米纤维膜上并且被Vitamin C有效还原。     

纳米羟基磷灰石／丝素蛋白多孔支架材料的制备和表征

采用硝酸钙-丝素蛋白溶液与磷酸钠反应仿生合成纳米羟基磷灰石/丝素蛋白(n-HA/SF)复合材料,并以NaHCO3和NaCl为致孔剂制备了多孔复合支架材料,采用TEM、IR、SEM和EDX对其进行了表征.结果表明,复合材料中HA的粒径在20～50nm之间,是一种CO2-3部分替代型弱结晶类骨针晶,在形貌和尺寸等方面类似于人体骨磷灰石晶体;HA和SF两相间存在强烈的键合作用,复合支架材料呈高度多孔结构,孔壁上富含微孔,孔隙间贯通性高.EDX分析结果表明,HA在有机基体中分布均匀,钙磷元素比为1.66,当复合材料和致孔剂的比例为1:0.5时,其抗压强度可达20.23MPa.     

生物陶瓷及其复合材料表面诱导类骨磷灰石层形成的研究

钙磷生物材料表面类骨磷灰石层的形成对其植入体内诱导新骨生成起非常重要的作用.本实验采用2倍模拟体液(2×SBF)为介质,通过仿生浸泡的方法研究了羟基磷灰石(HA)陶瓷及其复合材料羟基磷灰石/聚乳酸(HA/PLA)、羟基磷灰石/聚己內脂(HA/PCL)、羟基磷灰石/丝素(HA/SF)和羟基磷灰石/壳聚糖(HA/CS)表面诱导类骨磷灰石层形成的差异.实验结果表明,HA陶瓷及其复合材料在2×SBF溶液中仿生浸泡7d后,各样品表面均有一层厚度不同的类骨磷灰石层生成.并且该类骨磷灰石层的结晶度较低,晶粒较细(15～30nm),与人体自然骨无机物的结构非常类似.其中,在这4种复合材料中,HA/CS和HA/SF复合材料中因丝素蛋白和壳聚糖富含多种功能基团,能有效促进类骨磷灰石晶体的形核和生长,因而诱导类骨磷灰石生成的能力最强,显示其良好的生物活性.     

纳米羟基磷灰石／丝素蛋白生物复合材料的制备和表征

采用硝酸钙-丝素蛋白溶液与磷酸钠原位合成纳米羟基磷灰石(HA),在反应过程中丝蛋白(SF)诱导HA晶体生长,仿生合成HA/SF复合材料,用TEM、IR、TGA、XRD和SEM进行表征。结果表明,HA为CO23-部分替代型,粒径为10 nm~40 nm之间的弱结晶类骨晶体,在形貌和结晶度等方面与人体骨磷灰石相似。HA/SF复合材料中丝蛋白的含量约为25%,HA和SF两相存在强烈化学键合作用。SEM观察结果表明,HA微粒被SF完全包裹,两者之间没有明显相分离。     

仿生制备的再生丝素蛋白水溶液的静电纺丝（Ⅲ）-金属离子的影响

通过添加在家蚕丝腺体中含量较多的钾、钙、镁3种金属离子,研究了不同种类的金属离子对丝素蛋白水溶液的流变性能及静电纺丝的影响.研究结果表明,添加K 后溶液的表观粘度较大,而含Ca2 和Mg2 的丝素溶液粘度较小;分别加入0.225mol/L K 、Ca2 和Mg2 后,再生丝素蛋白水溶液在pH为6.0浓度为30%时都可通过静电纺丝法成纤.对于添加了0.225mol/L的混合金属离子的pH为6.0的再生丝素蛋白水溶液而言,丝素溶液的静电纺丝可纺浓度在27%～35%之间,随着丝素浓度的增加,所制纤维的平均直径增大,直径分布变宽.通过拉曼光谱、X衍射、DSC对再生丝素蛋白纤维的结构进行了表征,发现再生丝素蛋白纤维中已含有少量的Silk Ⅱ结构,主要是无定形结构.     

丝素蛋白在室温离子液体中的溶解与再生性能研究

室温离子液体1-烯丙基-3-甲基氯代咪唑和1-(2-羟乙基)-3-甲基氯代咪唑是丝素蛋白的新型良溶剂.丝素蛋白在1-烯丙基-3-甲基氯代咪唑和1-(2-羟乙基)-3-甲基氯代咪唑中的溶解度分别为14.5%(质量分数)(100℃)和8.0%(质量分数)(80℃).向离子液体丝素溶液中加入乙醇或正丁醇可获得再生丝素蛋白.XRD和FT-Raman研究表明再生丝素蛋白膜的构象主要是β-折叠结构.TGA数据表明再生丝素蛋白的热稳定性比天然丝素纤维有所降低,热失重残留物有所增加.同时,机械性能和溶失率分析结果显示从离子液体中再生的丝素蛋白表现出良好的湿态机械性能和优异的稳定性.     

再生丝素/丝胶共混蛋白水溶液的静电纺丝

采用静电纺丝方法制备了再生丝素/丝胶共混纤维,分析了共混配比对再生丝素/丝胶水溶液流变性能和静电纺可纺性的影响,通过扫描电镜、拉曼光谱和DSC等手段研究了所得纤维的形态和微细结构及其力学性能。研究结果表明:随着溶液中丝胶含量的增加,体系的表观粘度增大,静电纺纤维的直径减小且直径分布变窄;并且丝胶的存在有利于丝素蛋白从无规卷曲或α-螺旋结构向β-折叠结构转变,由此可提高静电纺纤维的力学性能。     

非织造多孔丝素蛋白/纳米羟基磷灰石复合支架材料的制备和表征

采用水刺法制备了多孔丝素蛋白(SF)支架,通过原位矿化法制备了纳米羟基磷灰石(HAP)/丝素蛋白复合材料。HAP呈现均匀棒状结构,而没有丝素蛋白支架时HAP仅形成颗粒状结构,这表明蛋白支架可诱导HAP形成均匀棒状结构,这可能是源于丝素蛋白中羧基和溶液中Ca2+的静电作用;丝素蛋白支架起到了复合材料基材和HAP原位生长的高分子模板双重作用。采用红外光谱、X射线衍射和力学性能测试对复合材料的结构性能进行了表征,红外谱图表明,HAP中的PO3-4与SF中的—NH2存在强烈的氢键相互作用,而X射线衍射图谱则表明复合材料中SF的保持了β折叠结构和结晶性能,使得复合材料具有良好的力学性能和多孔性能,有利于成骨细胞在支架上的增殖,同时,该方法制备的复合材料力学性能好,可大面积制备,成本低、制备方便,易于实现工业化,可望广泛用于骨修复支架材料。     

Tissue response and biodegradation of composite scaffolds prepared from Thai silk fibroin,gelatin and hydroxyapatite

This work aimed to investigate tissue responses and biodegradation, both in vitro and in vivo, of four types of Bombyx mori Thai silk fibroin based-scaffolds. Thai silk fibroin (SF), conjugated gelatin/Thai silk fibroin (CGSF), hydroxyapatite/Thai silk fibroin (SF4), and hydroxyapatite/conjugated gelatin/Thai silk fibroin (CGSF4) scaffolds were fabricated using salt-porogen leaching, dehydrothermal/chemical crosslinking and an alternate soaking technique for mineralization. In vitro biodegradation in collagenase showed that CGSF scaffolds had the slowest biodegradability, due to the double crosslinking by dehydrothermal and chemical treatments. The hydroxyapatite deposited from alternate soaking separated from the surface of the protein scaffolds when immersed in collagenase. From in vivo biodegradation studies, all scaffolds could still be observed after 12 weeks of implantation in subcutaneous tissue of Wistar rats and also following ISO10993-6: Biological evaluation of medical devices. At 2 and 4 weeks of implantation the four types of Thai silk fibroin based-scaffolds were classified as “non-irritant” to “slight-irritant”, compared to Gelfoam^® (control samples). These natural Thai silk fibroin-based scaffolds may provide suitable biomaterials for clinical applications.     

HAp/SiO2复合陶瓷材料的制备及体外生物活性

HAp和SiO2的混合粉末压制成型后,在1200℃下烧结得到含5.0%(质量分数)SiO2的HAp/SiO2生物陶瓷复合材料。烧结体的XRD及FT-IR分析结果表明,SiO2的添加促进了HAp发生热分解。其主要物相为α-TCP、HAp、Ca2P2O7以及生物玻璃(bioglass,BG)等。体外生物活性实验结果显示,样品浸泡在模拟体液(SBF)中24h后,表面出现花瓣状磷灰石沉积物,72h后进而生成板状沉积物,120h后在样品表面覆盖了较厚的类骨磷灰石层,经XRD测试分析表明,该层主要为碳酸羟基磷灰石(HCA)。制备的HAp-5.0%(质量分数)SiO2生物陶瓷复合材料具有比纯HAp更加优越的体外生物活性,可期待作为一种新的骨修复材料。     

磷酸氢钙/氨基酸共聚物生物活性复合材料

通过原位聚合法制备了磷酸氢钙/四元氨基酸共聚物(DCP/PAA)复合材料, 并采用XRD和IR方法对其组成结构进行了表征, 用模拟体液和Tris-HCl溶液分别研究了复合材料的体外生物活性和降解性能。结果表明: DCP/PAA复合材料的无机相与有机相存在着一定的相互作用; 聚合反应时间对复合材料的强度有明显的影响, 反应时间越长, PAA的黏度越大, 材料的抗压强度越高; 复合材料具有很好的生物活性, 其表面能够在模拟体液中形成磷灰石层, 新形成磷灰石的Ca与P原子比为1.59, 为缺钙磷灰石; 复合材料能够在Tris-HCl溶液中降解, PAA的黏度对复合材料在Tris-HCl中的降解有一定的影响。复合材料浸泡在Tris-HCl溶液中, 溶液的pH值在浸泡初期略有下降, 但4周后稳定在7.0~7.2, 与人体环境pH值接近。     

磷石膏/聚氨基酸医用多孔复合材料的制备与表征

用化学共沉淀法合成了钙磷原子比(Ca/P)为1.50和1.67的磷石膏(PG), 用熔融聚合法制备四元氨基酸共聚物(PAA4), 用挤出发泡法制备了两种磷石膏/四元氨基酸共聚物(PG/PAA4)多孔复合材料。通过TGA、SEM、XRD、IR、EDS等对两种复合材料的组成结构进行了表征, 并研究了复合材料在磷酸缓冲液(PBS)中的体外降解性能。结果表明: 无机组分PG在两种复合材料中分布均一, 质量分数均为60%左右; PG/PAA4复合材料的孔径为100~400 μm, 孔隙率在60%左右; 多孔复合材料的有机和无机相之间有化学作用。PG/PAA4复合材料具有良好的体外降解性能, 其失重率随PG钙磷比的增加而增加, 降解液的pH值维持在6.9~7.4之间。PG/PAA4复合材料降解后, 其表面沉积了片状钙磷化合物, 推测该复合材料可能具有生物活性。     

渗析分离丝素蛋白盐溶液的研究

采用浓差渗析器进行丝素蛋白盐溶液的脱盐试验，研究了操作参数对脱盐试验的影响，确定了原料液丝素蛋白浓度10％、料液和自来水流量40L／h为其较合理的工艺参数．采用此设备脱盐，脱盐效率较高，脱盐率达到99.36％，丝素蛋白回收率达到89．7％．     

磷灰石包覆金红石型TiO2纳米粉体的制备及表征

将金红石型TiO₂纳米粉体浸泡在模拟体液中, 于37℃经过不同时间, 制备出磷灰石包覆金红石型TiO₂纳米粉体. 用XRD、SEM、TEM、EDX、FTIR、ICP AES和BET方法对复合粉体进行了表征, XRD结果表明,磷灰石的含量可随着金红石型TiO2在模拟体液中浸泡时间的延长而增加. FTIR结果中显示了磷灰石的O-H和PO₄^3-吸收峰, 说明复合粉体中有磷灰石存在. ICP-AES结果表明溶液中Ca和P浓度随浸泡时间延长而下降, 表明时间延长后更多的Ca和P被消耗. TEM和EDX结果证明了金红石型TiO₂表面有磷灰石存在, HRTEM结果显示磷灰石（211）面的晶格间距为0.27nm, 晶粒尺寸约为40nm.     

An effective and green H_2O_2/H_2O/O_3 oxidation method for carbon nanotube to reinforce epoxy resin

Facile green oxidation methods are always desired to functionalize carbon nanotubes(CNTs) in the production of advanced CNT/epoxy composites. In the present work, an optimized H_2O_2/H_2O/O_3 oxidation method was developed, and performances of the H_2O_2/H_2O/O_3 oxidized CNT in epoxy matrix were tested and compared with that of the H_2O/O_3 oxidized CNT and the most commonly used concentrated HNO_3 oxidized CNT. The physical and chemical characteristics of the obtained oxidized CNTs were systematically characterized via transmission electron microscopy(TEM), X-ray photoelectron spectroscopy(XPS) and Raman. Mechanical performances of the obtained composites were explored by tensile tests,impact tests, dynamic mechanical analysis(DMA) and fracture toughness tests. It was found that the H_2O_2/H_2O/O_3 oxidized CNT exhibited all-around overwhelming advantages over the concentrated HNO_3 oxidized CNT on reinforcing the epoxy matrix, while the H_2O/O_3 oxidized CNT only improved the material strength. Reinforcing mechanisms for the different methods oxidized CNTs were studied and compared.The optimized H_2O_2/H_2O/O_3 oxidation method makes scaled production possible, avoids environment pollutions, and holds great potentials to replace the most commonly used concentrated HNO3 oxidation method to oxidize CNT during the preparation of the advanced CNT/epoxy composite.     

Apatite wollastonite–poly methyl methacrylate bio-composites

Bio-composites consisting of sol–gel processed apatite wollastonite (AW) glass ceramics and poly methyl methacrylate (PMMA) were prepared by hot compaction method. Density of the composites decreased with increase in PMMA content, while, biaxial flexural strength (BFS) was 21 MPa for 20 wt.% PMMA and beyond which it decreased. A correlation between phase compositions of AW glass ceramics with BFS was attempted from the XRD results. In vitro bioactivity of the composites in a simulated body fluid (SBF) showed the formation of spherical globules on the surface within 7 days of soaking as observed by environmental SEM. Thin film XRD and EDX measurement confirmed these globules to be bone like apatite with Ca/P ratio 1.53 and FTIR measurement showed the corresponding peaks for phosphates. Results indicated the bone bonding ability of the composites by forming a surface apatite (calcium phosphate) layer in SBF and the growth increased with increase in soaking durations. ICP measurement of the remaining SBF after 7, 14 and 21 days soaking of samples was found to be in good agreement with the EDX analysis results.     

Fabrication and characterization of silk fibroin/bioactive glass composite films

Composite films of silk fibroin (SF) with nano bioactive glass (NBG) were prepared by the solvent casting method, and the structures and properties of the composite films were characterized. Fourier transform infrared (FT-IR) spectroscopy analysis shows that the random coil and β-sheet structure co-exist in the SF films. Results of field emission scanning electron microscope (FESEM) indicate that the NBG particles are uniformly dispersed in the SF films. The measurements of the water contact angles suggest that the incorporation of NBG into SF can improve the hydrophilicity of the composites. The bioactivity of the composite films was evaluated by soaking in 1.5 times simulated body fluid (1.5 × SBF), and formation of a hydroxycarbonate apatite (HCA) layer was determined by XRD and FESEM. The results show that the SF/NBG composite film is bioactive as it induces the formation of HCA on the surface of the composite film after soaking in 1.5 × SBF for 7 days. In vitro osteoblasts attachment and proliferation tests show that the composite film is a good matrix for the growth of osteoblasts. Consequently, the incorporation of NBG into the SF film can enhance both the bioactivity and biocompatibility of the film, which suggests that the SF/NBG composite film may be a potential biomaterial for bone tissue engineering.     

Fabrication of mineralized electrospun PLGA and PLGA/gelatin nanofibers and their potential in bone tissue engineering

Surface mineralization is an effective method to produce calcium phosphate apatite coating on the surface of bone tissue scaffold which could create an osteophilic environment similar to the natural extracellular matrix for bone cells. In this study, we prepared mineralized poly(d,l-lactide-co-glycolide) (PLGA) and PLGA/gelatin electrospun nanofibers via depositing calcium phosphate apatite coating on the surface of these nanofibers to fabricate bone tissue engineering scaffolds by concentrated simulated body fluid method, supersaturated calcification solution method and alternate soaking method. The apatite products were characterized by the scanning electron microscopy (SEM), Fourier transform-infrared spectroscopy (FT-IR), and X-ray diffractometry (XRD) methods. A large amount of calcium phosphate apatite composed of dicalcium phosphate dihydrate (DCPD), hydroxyapatite (HA) and octacalcium phosphate (OCP) was deposited on the surface of resulting nanofibers in short times via three mineralizing methods. A larger amount of calcium phosphate was deposited on the surface of PLGA/gelatin nanofibers rather than PLGA nanofibers because gelatin acted as nucleation center for the formation of calcium phosphate. The cell culture experiments revealed that the difference of morphology and components of calcium phosphate apatite did not show much influence on the cell adhesion, proliferation and activity.