Preparation of 3-D porous fibroin scaffolds by freeze drying with treatment of methanol solutions

In this study, silk scaffolds with appropriate porous structures were prepared by adjusting solution concentrations and providing treatment with methanol solutions in the way of freeze drying. The effects of the preparation conditions on the microstructures and properties of the scaffolds were discussed. Fibroin solutions with different concentrations of 4, 6, 8, 10 wt% were used respectively to prepare the scaffolds. The effects of the addition of 20 vol% methanol before or after freeze drying to the 4 wt% fibroin solution were investigated. As demonstrated by Scanning Electron Microscope (SEM), the fib-roin scaffolds prepared without methanol had porous microstructures composed of thin sheets, and the sizes of the pores decreased with the increase of the fibroin solution concentrations, while the scaffolds prepared in the presence of methanol showed porous microstructures formed by fine-particle aggregates. The porosities and mechanical properties of the prepared fibroin scaffolds under different conditions were tested. The crystalline structures and conformations of the fibroin scaffolds were detected by Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD).     

Fiber-Based Chitosan Tubular Scaffolds for Soft Tissue Engineering： Fabrication and in Vitro Evaluation

Porous, two-ply tubular chitosan conduits for guided tissue regeneration were fabricated by combining the textile technique （inner layer） with the thermally induced phase separation process （outer layer）. A hollow chitosan tube was prepared using an industrial warp knitting process with chitosan yarns. Then, an appropriate diameter mandrel was inserted into the pre-fabricated tube. The tube and the mandrel were dipped into the chitosan solution together, taken out, and freeze-dried. After being neutralized in alkaline solution and dried at room temperature, the mandrel was removed to create the chitosan tubular scaffold. Scanning electron micrographs show that the resulting tubes have a biphasic wall structure, with a fibrous inner layer and a semipermeable outer layer. The swelling properties and the mechanical strength before and after in vitro degradation were investigated. The biocompatibility of the scaffolds was also investigated by co-culturing neuroblastoma cells （N2A, mouse） with the scaffolds. The results suggest that these chitosan tubular scaffolds are useful for the regeneration of tissues requiring a tubular scaffold.     

Electrospun composites of PHBV/pearl powder for bone repairing

Electrospun fi ber has highly structural similarity with natural bone extracelluar matrix(ECM). Many researches about fabricating organic–inorganic composite materials have been carried out in order to mimic the natural composition of bone and enhance the biocompatibility of materials. In this work, pearl powder was added to the poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(PHBV) and the composite nano fi ber scaffold was prepared by electrospinning. Mineralization ability of the composite scaffolds can be evaluated by analyzing hydroxyapatite(HA)formation on the surface of nano fi ber scaffolds. The obtained composite nano fi ber scaffolds showed an enhanced mineralization capacity due to incorporation of pearl powder. The HA formed amount of the composite scaffolds was raised as the increase of pearl powder in composite scaffolds. Therefore, the prepared PHBV/pearl composite nano fi ber scaffolds would be a promising candidate as an osteoconductive composite material for bone repairing.     

3D Nanocomposite Hydrogel Scaffolds Fabricated by Rapid Prototyping for Bone Tissue Engineering

Colloidal gels made of oppositely charged nanoparticles are a novel class of hydrogels and can exhibit pseudoplastic behavior which will enable them to mold easily into specific shapes. These moldable gels can be used as building blocks to self-assemble into integral scaffolds from bottom to up through electrostatic forces. However, they are too weak to maintain scaffold morphology just depending on interparticle interactions such as Van der Waals attraction and electrostatic forces especially for bone tissue engineering. In this study, oppositely charged gelatin nanoparticles were firstly prepared by two-step desolvation method, followed by the mixture with water to form colloid gels. To solve the problem of weak mechanical performance of colloid gels, gelatin macromolecules were introduced into the prepared gels to form blend gels. The blend gels can be easily processed into three-dimensional （3D） porous scaffolds via motor assisted microsyringe （MAM） system, a nozzle-based rapid prototyping technology, under mild conditions. After fabrication the scaffolds were erosslinked by glutaraldehyde （ GA, 25 % solution in water by weight）, then the crosslinked gelatin macromolecules network could form to improve the mechanical properties of colloid gels. The average particle size and zeta potential of gdatin nanoparticles were measured by Nano- ZS instrument. The morphology and microstructures of scaffolds were characterized by macroscopic images. The mechanical properties of the scaffolds were studied by a universal material testing machine.     

Preparation and evaluation of biomimetric nano-hydroxyapatite-based composite scaffolds for bone-tissue engineering

In the present study,novel biomimetic composite scaffolds with a composition similar to that of natural bone were prepared,using nano-hydroxyapatite,collagen,and phosphatidylserine.The scaffolds possess an interconnected porous structure with a porosity of 84%.The pore size ranges from several micrometers up to about 400 m.In-vitro studies in simulated body fluids showed that the morphologies of the products derived from mineralization can be regulated by the extracellular matrix components of the scaffolds;this in turn leads to creation of a large number of hydroxyapatite crystals on the scaffold surface.The regulatory properties of collagen and phosphatidylserine also influenced the cell response to the composite scaffolds.MC3T3-E1 cells attached and spread on the surfaces of the materials and interacted with the substrates;this may be the result of charged groups on the composite materials.Radiological analysis suggested that calluses and bone bridges formed in defects within 12 weeks.These composite scaffolds may therefore be a suitable replacement in bone-tissue engineering.     

Fabrication of Multi-layered Composite Scaffolds by Bi-directional Electrospinning Method

A multi-layered composite scaffolds consisting of poly （ L- ne） （ P （LLA-CL） ）, collagen （COL） and chitosan （CS） were fabricated by a bi-directional electrospinnlng method. Synthetic P （LLA-CL） was used as the middle layer to enhance the strength, while natural COL/CS blending （9： 1, v/v） was used as the bioactive surfaces （inner and outer layers ） to improve the biocompatibility. Each three transitional layers were set between inner/outer layer and middle layer for delamination resistance. Scanning electron microscopy （SEM） was used to observe the fiber morphology. The Fourier transform infrared attenuated total reflectance spectroscopy （FTIR-ATR） spectra, X- ray diffraction （XRD） and thermogravimetry （TG） tests were used to analyze the physical properties of the scaffolds. The results showed that the modified clectrospinning method bad no negative effect on the components, crystal structure and thermostability of the scaffolds, but could effectively combine the mechanical property of synthetic material and biocompatibility of natural materials. Such method could be applied to the fabrication of composite scaffolds for vascular, skin. and nerve tissue engineering.     

Bulk Al–Al_3Zr composite prepared by mechanical alloying and hot extrusion for high-temperature applications

Bulk Al/Al_3Zr composite was prepared by a combination of mechanical alloying(MA) and hot extrusion processes. Elemental Al and Zr powders were milled for up to 10 h and heat treated at 600℃ for 1 h to form stable Al_3Zr. The prepared Al_3Zr powder was then mixed with the pure Al powder to produce an Al–Al_3Zr composite. The composite powder was finally consolidated by hot extrusion at 550℃. The mechanical properties of consolidated samples were evaluated by hardness and tension tests at room and elevated temperatures. The results show that annealing of the 10-h-milled powder at 600℃ for 1 h led to the formation of a stable Al_3Zr phase. Differential scanning calorimetry(DSC) results confirmed that the formation of Al_3Zr began with the nucleation of a metastable phase, which subsequently transformed to the stable tetragonal Al_3Zr structure. The tension yield strength of the Al-10wt%Al_3Zr composite was determined to be 103 MPa, which is approximately twice that for pure Al(53 MPa). The yield stress of the Al/Al_3Zr composite at 300℃ is just 10% lower than that at room temperature, which demonstrates the strong potential for the prepared composite to be used in high-temperature structural applications.     

Preparation of bioactive glasses with controllable degradation behavior and their bioactive characterization

Bioactive glasses and ceramics have been widely investigated for bone repair because of their excel-lent bioactive characteristics. However, these biomaterials undergo incomplete conversion into a bone-like material, which severely limits their biomedical application. In this paper, borosilicate bioac-tive glasses were prepared by traditional melting process. The results showed that borosilicate glasses possessed high biocompatibility and bioactivity. In addition, when immersed in a 0.02 mol/L K2HPO4 solution, particles of a borate glass were fully converted to HA. The desirable conversion rate to HA may be achieved through the adjustment of the B2O3/SiO2 ratio. The results of XRD and FTIR analysis indicated that the degradation product was carbonate-substituted hydroxyapatite, which was similar to the inorganic component of bone.     

Effect of Y_2O_3 content in the pack mixtures on microstructure and oxidation resistance of B–Y modified silicide coating

B–Y modified silicide coatings were prepared on Nb–Si based alloy by pack cementation at 1300 ℃ for 10 h. The effect of Y_2O_3 content in the pack mixtures on microstructure and oxidation resistance of the coatings was investigated. The results show that the four coatings have similar structures, which possess a(Nb,X)Si_2 outer layer and a(Nb,X)_5Si_3 transitional layer. Y_2O_3 content in the pack mixtures has an obvious effect on the Si content in the coating. The mass gains of the coatings prepared with 0.5, 1, 2 and 3 wt% Y_2O_3 in pack mixtures are 2.33, 1.96, 2.05 and 2.86 mg/cm~2 after oxidation at 1250 ℃ for 100 h, respectively. The coating prepared with 1 wt% Y_2O_3 exhibits the best oxidation resistance due to the formation of a dense glass-like borosilicate scale.     

Functionalization of PCL fibrous membrane with RGD peptide by a naturally occurring condensation reaction

Poly(e-caprolactone)(PCL)is widely adopted as an ingredient for tissue engineering scaffolds.To improve its cell affinity,in this study,we developed a new method to introduce bioactive RGD peptides onto the surface of PCL via condensation reaction between 2-cyanobenzothiazole(CBT)and D-cysteine.The PCL fibrous membranes were prepared by electrospinning,and RGD functionalization was characterized by fluorescence microscopy,scanning electron microscopy(SEM),X-ray photoelectron spectroscopy(XPS)and water contact angle(WCA).As expected,our results demonstrated the successful RGD immobilization on the surface of PCL.RGD modification improved the hydrophilicity of PCL,changing their WCA from 112.20°to38.35°.Cell adhesion,spreading and proliferation of 3T3fibroblasts were also enhanced.We therefore believe that the methods reported in this study was facile and effective for functional modification of the hydrophobic PCL scaffolds.The moderate reaction conditions are also suitable for covalent immobilization of bioactive molecules onto PCL.     

Production of Poly(3-Hydroxybutyrate-co-3-Hydroxyhexanoate) Using Aeromonas hydrophila 4AK4 Grown in Mixed Carbon Source

IntroductionPolyhydroxyalkanoates (PHAs) are intracellularenergy storage materials and/or reduction agentsthat are synthesized by many microorganisms[1 ] .Because these bacterial PHAs are trulybiodegradable thermoplastics or elastomers,theyhave attracted much attention in industry[2 ,3 ] .Poly(3 -hydroxybutyrate) (PHB) wasthefirst PHA to beproduced on a large scale and poly (3 -hydroxybutyrate-co-3 -hydroxyvalerate) (PHBV)was laterused in commercial applications.Recently,novel PHA copo…     

Production of Poly （3—Hydroxybutyrate—co—3—Hydroxyhexanoate） by Aeromonas hydrophila and Recombinant Escherichia coli

Aeromonas hydrophila (A. hydrophila) 4AK4 produced poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) with an almost constant 3-hydroxyhexanoate (3HHx) content of 10 % - 15 % from lauric acid and/or soybean oil. Both A. hydrophila 4AK4 and recombinant Escherichia coil (E. coil) JMU193(pBH32) produced PHBHHx with controllable 3HHx content when fed lauric acid and another co-substrate. With glucose or gluconate as the co-substrate, the 3HHx content in the copolyester produced by A. hydrophila 4AK4 was reduced slightly from 12% to 9%. However, the 3HHx content in the copolyester produced by E. coll JMU193 (pBH32) was significantly reduced from 9% to 2% with fructose as the co-substrate. These results show that regulation of 3HHx content in PHBHHx can be achieved using genetically engineered E. coll.     

Dexamethasone-Loaded PLGA Microspheres Incorporated PLLA/PLGA/PCL Composite Scaffold for Bone Tissue Engineering

The combination of micro-carriers and polymer scaffolds as promising bone grafts have attracted considerable interest in recent decades.The poly(L-lactic acid)/poly(lactic-co-glycolic acid)/polycaprolactone(PLLA/PLGA/PCL)composite scaffold with porous structure was fabricated by thermally induced phase separation(TIPS).Dexamethasone(DEX)was incorporated into PLGA microspheres and then loaded on the PLLA/PLGA/PCL scaffoldtopreparethedesiredcompositescaffold.The physicochemical properties of the prepared composite scaffold were characterized.The morphology of rat bone marrow mesenchymal stem cells(BMSCs)grown on scaffolds was observed using scanning electron microscope(SEM)and fluorescence microscope.The resultsshowedthatthePLLA/PLGA/PCLscaffoldhad interconnected macropores and biomimetic nanofibrous structure.In addition,DEX can be released from scaffold in a sustained manner.More importantly,DEX loaded composite scaffold can effectively support the proliferation of BMSCs as indicated by fluorescence observation and cell proliferation assay.The results suggested that the prepared PLLA/PLGA/PCL composite scaffold incorporating drug-loaded PLGA microspheres could hold great potential for bone tissue engineering applications.     

Incorporation of Sphingosine 1-Phosphate Loaded Mesoporous Silica Nanoparticles into Poly( L-lactic acid ) Nanofibrous Scaffolds for Bone Tissue Engineering

Controlled release of the functional factors is the key to improve clinical therapeutic efficacy during the tissue repair and regeneration.The three-dimensional(3D)scaffold can provide not only physical properties such as high strength and porosity but also an optimal environment to enhance tissue regeneration.Sphingosine1-phosphate(S1P),an angiogenic factor,was loaded into mesoporous silica nanoparticles(MSNs)and then incorporated into poly(L-lactic acid)(PLLA)nanofibrous scaffold,which was fabricated by thermally induced phase separation(TIPS)method.The prepared scaffolds were examined by attenuated total reflection Fourier transform infrared spectroscopy(ATR-FTIR),scanning electron microscopy(SEM),and transmission electron microscopy(TEM)and compressive mechanical test.The ATR-FTIR result demonstrated the existence of MSNs in the PLLA nanofibrous scaffold.The SEM images showed that PLLA scaffold had regular pore channel,interconnected pores and nanofibrous structure.The addition of MSNs at appropriate content had no visible effect on the structure of scaffold.The compressive modulus of scaffold containing MSNs was higher than that of the scaffold without MSNs.Furthermore,fluorescein isothiocyanate(FITC)was used as model molecule to investigate the release behavior of S1P from MSNsincorporated PLLA(MSNs/PLLA)nanofibrous scaffold.The result showed that the composite scaffold largely reduced the initial burst release and exhibited prolonged release of FITC than MSNs.Thus,these results indicated that S1P-loaded composite nanofibrous scaffold has potential applications for bone tissue engineering.     

Strontium Substituted Nanohydroxyapatite Incorporated 3D Printing Scaffold for Bone Tissue Engineering

The customized implants which are composed of polycaprolactone( PCL) and strontium substituted nanohydroxyapatite( SrHA) were fabricated successfully by using fused deposition modeling( FDM),which is a simple 3 D printing technology for fabricating personalized products. The physical and chemical properties of composite scaffolds were characterized by transmission electron microscopy( TEM), Fourier transform infrared spectroscopy( FTIR), X-Ray diffraction( XRD) and inductively coupled plasma-atomic emission spectroscopy( ICPAES). The results suggested that strontium element was successfully doped into nanohydroxyapatite and all scaffolds showed the homogeneous network structure. Furthermore, the in vitro biocompatibility of the scaffolds was evaluated by cell counting kit-8( CCK-8) assay. The data indicated that the prepared scaffolds exhibited excellent biocompatibility to bone marrow mesenchymal stem cells( BMSCs). Besides,strontium element can be released from PCL-SrHA scaffolds in a sustained manner. Therefore,the 3 D printing PCL-SrHA scaffolds hold great potential for bone tissue engineering.     

3D打印制备HA/PCL复合材料组织工程支架的研究

应用熔融沉积成型技术（FDM）制备羟基磷灰石（HA）/聚己内酯（PCL）组织工程支架,探讨其内部结构和力学性能。以羟基磷灰石和聚己内酯为原料,采用熔融共混技术分别制备HA质量分数为20%的nano-HA/PCL和micro-HA/PCL复合材料,使用自主研发的熔体微分FDM 3D打印机制备HA/PCL复合材料组织工程支架。通过显微镜观察发现,所制备的nano-HA/PCL和micro-HA/PCL组织工程支架具有均匀分布且相互连通的近似矩形的孔隙。nano-HA/PCL和micro-HA/PCL组织工程支架的断面图分析结果表明,nano-HA/PCL组织工程支架中HA粒子分布均匀,而micro-HA/PCL组织工程支架中HA粒子发生了团聚,导致nano-HA/PCL组织工程支架的拉伸强度和弯曲强度均高于micro-HA/PCL组织工程支架。因此,利用熔体微分FDM 3D打印机打印生物活性nano-HA/PCL复合材料组织工程支架在骨组织工程中具有潜在的应用前景。     

纳米羟基磷灰石/左旋聚乳酸纳米纤维复合支架的制备与表征

采用热致相分离法制备了一系列具有微米级孔隙的纳米纤维状纳米羟基磷灰石/左旋聚乳酸（n-HA/PLLA）复合支架材料。并采用扫描电境和体外降解实验对其进行了表征。扫描电镜观测到复合支架的微观结构为具有微米级孔隙的纳米纤维状骨架,其不规则孔的孔径在几到几百纳米的范围内,纳米纤维直径在200-500nm的范围内;体外降解实验的结果表明,与纯聚乳酸支架相比,n-HA/PLLA复合支架的降解液的pH的变化更加缓慢。     

新型前交叉韧带支架材料的构建及其生物相容性的实验研究

探讨以聚羟基丁酸己酯/聚左旋乳酸(PHB/PLLA1∶1)胶原杂化支架作为前交叉韧带组织工程载体材料的可行性。制备"三明治"样结构PHB/PLLA共聚物并测量其孔隙率等指标。以I型胶原对制备的PHB/PLLA支架进行杂化,获得PHB/PLLA胶原杂化支架。扫描电镜观察其表面结构。将兔皮肤成纤维细胞(SF)接种于PHB/PLLA胶原杂化支架,共培养5d后,扫描电镜下观察其在材料上生长情况。PHB/PLLA支架杂化后胶原填充于纤维空隙,分布比较均匀。体外培养的皮肤成纤维细胞成功种植在支架材料上,在材料上粘附、生长良好。说明构建的支架材料具有良好的三维构型和生物相容性,有望为前交叉韧带损伤的修复提供了一种新型的支架材料。