Preparation and characterization of alpha-amylase-immobilized thermal-responsive composite hydrogel membranes.

Composite hydrogel membranes of crosslinked poly(N-isopropylacrylamide-co-N-acryloxysuccinimide-co-2-hydroxyet hyl methacrylate) [P(NIPAAm-NAS-HEMA)] with starch, as a macropore forming agent, on nonwoven polyester was prepared. The membranes could swell and de-swell around the characteristic lower critical solution temperature (LCST) of poly(N-isopropylacrylamide) (PNIPAAm). It was demonstrated that the presence of macropores in the membranes could improve the immobilization efficiency as well as lead to a short responding time upon temperature change across the LCST. Immobilized alpha-amylase could retain as high as 33% of the activity of the free enzyme with a loading level of 0.60-0.65 mg/cm2 when the membrane preparation and enzyme immobilization conditions were optimized. The half time (T0.5) for the swelling or de-swelling response of the gel phase within the membranes was less than 2 min, and the 90% time (T0.9) was less than 6 min. The permeability for maltose through the membranes could change as much as 4.9-fold when the temperature was raised above or reduced below the LCST.     

Preparation and characterization of nano-hydroxyapatite/chitosan composite scaffolds

A novel nano-hydroxyapatite (HA)/chitosan composite scaffold with high porosity was developed. The nano-HA particles were made in situ through a chemical method and dispersed well on the porous scaffold. They bound to the chitosan scaffolds very well. This method prevents the migration of nano-HA particles into surrounding tissues to a certain extent. The morphologies, components, and biocompatibility of the composite scaffolds were investigated. Scanning electron microscopy, porosity measurement, thermogravimetric analysis, X-ray diffraction, X-ray photoelectron spectroscopy, and Fourier transformed infrared spectroscopy were used to analyze the physical and chemical properties of the composite scaffolds. The biocompatibility was assessed by examining the proliferation and morphology of MC 3T3-E1 cells seeded on the scaffolds. The composite scaffolds showed better biocompatibility than pure chitosan scaffolds. The results suggest that the newly developed nano-HA/chitosan composite scaffolds may serve as a good three-dimensional substrate for cell attachment and migration in bone tissue engineering.     

煅烧骨/壳聚糖复合材料的制备及表征

文题释义：                                                                                                     天然骨：主要由无机的羟基磷灰石和有机的胶原成分构成,并具有一定的力学性能。以羟基磷灰石和磷酸三钙为主的磷酸盐材料拥有良好的骨传导性和部分骨诱导性,能够与宿主的骨直接发生骨结合,已成为目前临床应用最多的骨移植材料。 锻烧骨：是经高温锻烧异体动物骨所获得的无机材料,主要成分是羟基磷灰石,其钙磷比接近于人骨,拥有极好的生物相容性和优越的骨引导性。与人工合成的羟基磷灰石相比,不用考虑煅烧骨材料的结构形貌,而且材料来源广泛、制作成本低。 背景：壳聚糖具备优异的理化性能与良好的生物相容性,但其缺乏骨结合的生物活性,需要与其他材料复合用于骨组织修复中。 目的：将煅烧骨与壳聚糖复合,分析其理化性能和细胞毒性。 方法：采用溶液共混法制备煅烧骨与壳聚糖质量比分别为1/2、1/1、2/1的复合材料,表征3种复合材料的理化性能。在第5代小鼠成纤维细胞 L929中分别加入3种复合材料浸提液,CCK-8法检测复合材料的细胞毒性。 结果与结论：①X射线衍射和红外光谱显示,3种复合材料的主要成分均为羟基磷灰石与β-磷酸三钙,并且随着煅烧骨比例的增加,复合材料中的羟基磷灰石/β-磷酸三钙的特征衍射峰逐渐增强;②扫描电镜显示,煅烧骨颗粒较均匀地分散于壳聚糖介质中;③随着煅烧骨比例的增加,复合材料的抗压强度逐渐降低;④培养7 d时,3种复合材料浸提液中的细胞生长良好,形态无明显变化;培养9 d的时间内,3种复合材料浸提液中的细胞相对增殖率均在90%以上,细胞毒性均为1级,符合生物材料的安全标准;⑤结果表明,煅烧骨/壳聚糖复合材料具备良好的结构特征、理化性能及合适的抗压强度,并且安全无毒。 ORCID: 0000-0003-3519-4485(廖健) 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     

Preparation and characterization of thin film surface coatings for biological environments

An approach to the problem of selecting synthetic materials for use in biological media is presented. Firstly, a surface energetic criterion of biocompatibility of foreign surfaces is suggested. This criterion, which is based on an analysis of the surface interactions between a typical biological fluid (i.e. blood) and synthetic surfaces, is founded on the premise that a sufficiently low (but not very low) solid-bioloical fluid interfacial free energy of the order of 1–3 dyne/cm, is necessary in order to fulfil the dual requirements of maintaining a low thermodynamic driving force for the adsorption of fluid components on the solid surface as well as a mechanically stable solid-fluid interface.

In the second part of this investigation, an experimental approach involving the radio frequency (rf) sputter deposition of thin solid films of tightly adhering polymeric compounds on materials with the desired bulk characteristics, is shown to he a promising method of tailoring the surface properties of many types of synthetic materials for use in biological environments. The preparation and surface characterization of thin, solid films of oxidized fluorocarbon coatings (from a Teflon FEP target) by rf sputtering is illustrated. The deposited polymer films were characterized for their surface morphology, thickness, elemental surface chemical composition and their wetting properties in a biological environment, by the techniques of scanning electron microscopy (SEM), ellipsometry, electron spectroscopy for chemical analysis (ESCA) and contact angle measurements, respectively. Based on the ESCA and contact angle results, it emerges that the surfaces of such polymeric coatings possess sufficient mobility to considerably alter their structures between different environments (such as air and water) and thereby present different wetting characters to these environments. The contact angle procedure developed in this investigation permitted the estimation of the relevant wetting properties of such mobile surfaces in an aqueous environment (which is the environment encountered in most biological fluids).

In the final part of this investigation, the possibility of effecting a drastic reduction in the solid-water interfacial free energy of the sputtered polymer surfaces by physical and or chemical modification of their surfaces and thereby improving their biocompatibility is illustrated.     

Preparation and characterization of nano-hydroxyapatite/chitosan/konjac glucomannan composite

Nano-hydroxyapatite (n-HA)/chitosan (CS)/konjac glucomannan (KGM) composite was prepared by coprecipitation method and investigated by thermal gravitivity/differentiate thermal analysis, Fourier transform infrared spectroscopy, X-ray diffraction, inductively coupled plasma emission spectroscopy, scanning electron microscopy, and energy dispersive X-ray analyzer. The analyses showed that the three phases of n-HA, CS, and KGM combined closely to each other. Further, in vitro tests were conducted to investigate the degradation and bioactivity of the composite. During immersion in simulated body fluid (SBF), pores appeared and a new substance containing Ca and P formed on the surface of the composite. Also, the concentration of Ca and P in SBF changed and weight loss of the composite was observed during time. The composite revealed a high degradation in SBF. Evidently, the new composite has a potential to be used as a carrier of implantable drug delivery system. The biodegradation rate and route could be different from CS and KGM, which will provide an opportunity to control the degradation rate or drug releasing rate by simply adjusting the ratio of CS and KGM.     

Preparation and characterization of bioactive mesoporous calcium silicate-silk fibroin composite films

Composite films of bioactive mesoporous calcium silicate (MCS)/silk fibroin (SF) and conventional calcium silicate (CS)/SF were fabricated by the solvent casting method, and the structures and properties of the composite films were characterized. Results of field emission scanning electron microscope (FESEM) indicated that MCS or CS was uniformly dispersed in the SF films. The measurements of the water contact angles suggested that the incorporation of either MCS or CS into SF could improve the hydrophilicity of the composite films, and the former was more effective than the later. The bioactivity of the composite films was evaluated by soaking in a simulated body fluid (SBF), and the formation of a hydroxycarbonate apatite (HCA) layer was determined by XRD and FT-IR. The results showed that the MCS/SF composite films have significantly enhanced apatite-forming bioactivity compared with the CS/SF composite films owing to the highly specific surface area and pore volume of MCS. In vitro cell attachment and proliferation tests showed that the MCS/SF composite film was a good matrix for the growth of MG63 cells. Consequently, the MCS/SF composite film possessed excellent physicochemical and biological properties, indicating its potential application for bone tissue engineering by designing 3D scaffolds according to its corresponding composition.     

Preparation and characterization of a highly macroporous biodegradable composite tissue engineering scaffold

A unique composite scaffold for bone-tissue engineering applications has been prepared by combining biodegradable poly(lactide-co-glycolide) (PLGA) with bioresorbable calcium phosphate (CaP) cement particles through the process of particle fusion and phase separation/particle leaching. The scaffold is characterized by a highly interconnected macroporosity, with macropores of 0.8-1.8 mm and porosities ranging from 81% to 91%, and improved mechanical properties with respect to the polymer alone, producing excellent dimensional stability. The scaffold properties were controlled by adjusting the processing parameters, including PLGA molar mass and concentration, CaP/PLGA ratio, and porogen size. The differences in mechanical properties between dry, wet/room temperature, and wet/37 degrees C testing conditions, of which the latter are more relevant for materials to be employed in a biological milieu, were investigated. Thus, a scaffold made from PLGA IV 1.13, PLGA concentration 12.5%, and CaP/PLGA ratio 2:1 exhibited significantly different compressive strengths of 0.16 MPa and 0.04 MPa when tested under dry and wet/37 degrees C conditions, respectively. .     

纳米羟基磷灰石/羧甲基壳聚糖-海藻酸钠复合骨水泥的性能

背景：目前普遍使用的黏合剂对粉碎骨折块进行黏合复位或多或少都存在一些缺陷。 目的：研制具有黏接骨骼作用的生物活性骨水泥。 方法：应用共沉淀法制备纳米羟基磷灰石/羧甲基壳聚糖-海藻酸钠复合材料作为骨水泥的固相粉体,将柠檬酸衍生物配制成溶液作为液相。通过优化实验,从骨水泥的固化时间、抗压强度、抗拉强度、抗稀散性等方面确定最佳配比。 结果与结论：纳米羟基磷灰石/羧甲基壳聚糖-海藻酸钠质量比为65/35,其中羧甲基壳聚糖和海藻酸钠质量比为4∶1时复合成粉体,并按固液比为1.0∶0.5(g∶mL)调拌后形成的骨水泥呈膏状,塑形性和抗稀散性能良好,固化时间12~18 min,抗压强度为(4.5±2.1) MPa。体外黏接猪股骨头抗拉强度在不同室温下无显著性差异无显著性意义(P > 0.05),固化后2 h的抗拉强度达到24 h的94%。骨水泥为多孔状结构,孔径为100~300 μm,纳米羟基磷灰石分布较均匀。提示制备的纳米羟基磷灰石/羧甲基壳聚糖-海藻酸钠复合骨水泥具有良好的生物活性、适当的力学强度以及较好的黏合强度。     

Preparation and characterization of collagen-hydroxyapatite composite used for bone tissue engineering scaffold

In this study, highly porous collagen-HA scaffolds were prepared by solid-liquid phase separation method. Microstructure of the composites was characterized by SEM, TEM and XRD. The results show that collagen-HA scaffolds are porous with three-dimension interconnected fiber microstructure, pore sizes are 50-150 microm, and HA particles are dispersed evenly among collagen fiber. Compared with pure collagen, the mechanical property of collagen-HA composite improves significantly. To gain further insight into cell growth throughout 3D scaffolds, the cell proliferation and attachment on the scaffold in vitro was investigated. The collagen-HA composite has good biocompatibility, and adding HA does not affect the histocompatibility of the scaffold materials. The porous collagen-HA composite is suitable as scaffold used for bone tissue engineering.     

Preparation, characterization, and performance of magnetic iron-carbon composite microparticles for chemotherapy

Magnetic microcarrier particles useful for delivering chemotherapeutic drug molecules are described. The particles are formed by joint deformation of iron and carbon in a ball mill. Physical, chemical, and functional characterization has been carried out on the particles. Physical characteristics include microscopy, particle size analysis (0.5-5 microm), surface area (250 m2/g), water vapor adsorption isotherm (hydrophobic surface), and analysis of the iron-carbon interface by M?ssbauer spectroscopy, X-ray diffraction, and differential thermal analysis. Chemical analysis was used to identify elements in the particles other than carbon and iron. Functional characteristics measured included the particles' ability to adsorb and desorb doxorubicin, cytotoxicity, and their magnetic susceptibility.     

Antimonocyte serum. Preparation and characterization

Cell suspensions enriched with monocytes were obtained from the blood of Vietnam normal pig and rabbits injected with avirulent Welshimer strain of L. monocytogenes by velocity sedimentation on BSA gradient and separation on Ficoll-Triosil gradient. They were used for immunization of rabbits and a sheep, respectively. The antisera specific for monocytes (RAMS and SAMS) were obtained by scrupulous absorption with lymphocytes and granulocytes.     

海螵蛸/外消旋聚乳酸复合人工骨材料的制备及表征

BACKGROUND: Cuttlebone which is rich in calcium has a natural affinity with the nature bone tissue. However, when the cuttlebone is applied as supporting material independently, its brittleness is larger and the intensity is lower. OBJECTIVE: To independently research and develop cuttlebone/racemic polylactic acid composite artificial bone tissue scaffolds, and detect their various performance.   METHODS: The composite artificial bone was prepared by combining the cuttlebone and racemic polylactic acid at the weight ratio of 1:2, 2:1 and 1:1 using salting out method. The pure cuttlebone was taken as control. The microstructure, porosity, water binding force, biological mechanics and in vitro degradation performance were determined. RESULTS AND CONCLUSION: The surface of cuttlebone had spine shape, clear texture and cellular distribution. Composite artificial bone was similar to the cancellous bone, whose big pore diameter was 500-1 500 μm and small pore diameter was 150-300 μm. Composite artificial bone had less micropores, and the connectivity between the holes is good. The bending strength, the elastic modulus, in vitro degradation properties of these three kinds of composite artificial bone were better compared with cuttlebone (P < 0.05), porosity was less than that of cuttlebone (P < 0.05), and there were no significant differences between these composite artificial bone groups. There was no significant difference in the water binding force between these four groups. These results demonstrate that cuttlebone/racemic polylactic acid composite artificial bone has good microstructure, mechanical properties and degradation performance.      

Preparation and characterization of a novel tobramycin-containing antibacterial collagen film for corneal tissue engineering

Corneal disease is a major cause of blindness and keratoplasty is an effective treatment method. However, clinical treatment is limited due to a severe shortage of high-quality allogeneic corneal tissues and the bacterial infection after corneal transplantation. In this study, we develop a novel artificial and antibacterial collagen film (called Col-Tob) for corneal repair. In the Col-Tob film, the tobramycin, which is an aminoglycoside antibiotic to treat various types of bacterial infections, was cross-linked by 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide and N-hydroxysuccinimide onto the collagen. Physical properties, antibacterial property and biocompatibility of the films were characterized. The results indicate that the film is basically transparent and has appropriate mechanical properties. Cell experiments show that human corneal epithelial cells could adhere to and proliferate well on the film. Most importantly, the film exhibits excellent antibacterial effect in vitro. Lamellar keratoplasty shows that the Col-Tob film can be sutured in rabbit eyes and are epithelialized completely in15 ± 5 days, and their transparency is restored quickly in the first month. Corneal rejection reaction, neovascularization and keratoconus are not observed within 3 months. This film, which can be prepared in large quantities and at low cost, should have potential application in corneal repair.     

Preparation and characterization of TAM-loaded HPMC/PAN composite fibers for improving drug-release profiles

The present paper reports the preparation and characterization of composite hydroxypropyl methylcellulose/polyacrylonitrile (HPMC/PAN)-medicated fibers via a wet spinning technique. Tamoxifen (TAM) was selected as a model drug. Numerous analyses were conducted to characterize the mechanical, structure and morphology properties of the composite fibers. The drug content and in vitro dissolution behavior were also investigated. SEM images showed that the TAM-loaded HPMC/PAN composite fibers had a finger-like outer skin and a porous structure. FT-IR spectra demonstrated that there was a good compatibility between polymer and drug. Results from X-ray diffraction and DSC suggested that most of the incorporated TAM was evenly distributed in the fiber matrix in an amorphous state, except for a minority that aggregated on the surface of fibers. The drug content in the fibers was lower than that in the spinning solution and about 10% of TAM was lost during spinning process. In vitro dissolution results indicated that, compared to TAM-PAN fibers, HPMC/PAN composite systems had weaker initial burst release effects and more drug-loading. The combination of hydrophilic polymer HPMC with PAN could improve the performance of polymer matrix composite fibers in regulating the drug-release profiles.     

Preparation,characterization and in vitro release of gentamicin from coralline hydroxyapatite-gelatin composite microspheres

Composite microspheres have been prepared from bioactive ceramics such as coralline hydroxyapatite [CHA, Ca10(PO4)6(OH)2] granules, a biodegradable polymer, gelatin and an antibiotic, gentamicin. In our earlier work, we have shown a gentamicin release from CHA granules--chitosan composite microspheres. In the present investigation, an attempt was made to prepare the composite microspheres containing coralline hydroxyapatite and gelatin (CHA-G), which were prepared by the dispersion polymerization technique and the gentamicin was incorporated by the absorption method. The crystal structure of the composite microspheres was analyzed using X-ray powder diffractometer. The Fourier transformed infrared spectrum clearly indicated the presence of amide and hydroxyl groups in the composite microspheres. Scanning electron micrographs and optical micrographs show that the composite microspheres are spherical in shape and porous in nature. The particle size of composite microspheres was analyzed and the average size was found to be 16 microm. The thermal behavior of composite microspheres was studied using thermogravimetric analysis and differential scanning calorimetric analysis. The cumulative in vitro release profile of gentamicin from composite microspheres showed near zero order patterns.     

京尼平交联仿骨结构诱导性骨组织工程支架材料的制备与表征

 背景：目前,尽管利用各种材料制备的组织工程化骨研究取得了一定进展,但均表现出诸如支架材料降解速度与新生骨组织形成速率不匹配、组织生长缓慢、降解代谢产物有毒性等缺陷。目的：构建一种新型的仿骨结构诱导性骨组织工程支架材料,评价其物理化学及生物学性能。方法：以壳聚糖包被淫羊藿苷制备微球,检测其体外缓释效果;将载药微球与胶原蛋白复合构建支架材料的管芯;将羟基磷灰石、聚己内酯与胶原蛋白依次以0∶3∶3、1∶3∶3、2∶3∶3、3∶3∶3的比例混合于六氟异丙醇中,通过静电纺丝技术依次电纺制得具有4层结构的支架材料外管;以1%京尼平将经嵌套的管芯与外管交联在一起。利用万能材料试验机、表面接触角仪、红外光谱、扫描电镜、吸水率、透气性、孔隙率、体外降解实验等对交联前后外管材料的结构和特征进行表征,并评价骨髓间充质干细胞与外管材料的生物相容性;Wistar大鼠皮下埋置实验进一步评价交联前后外管材料的组织相容性。结果与结论：药物在管芯中具有良好的缓释效果;制备的骨组织工程支架材料具有良好的均一性,交联后外管材料的力学性能、吸水率、透气性均高于未交联组(P < 0.05),且体外降解速率显著低于未交联组(P < 0.05)。苏木精-伊红染色显示骨髓间充质干细胞可良好贴附于交联前后的外管材料上;交联的外管材料植入Wistar大鼠皮下后均无炎症反应。表明交联后诱导性骨组织工程支架材料具有良好的生物相容性及力学性能。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     

Preparation and characterization of rabies virus hemagglutinin.

Rabies virus glycoprotein, released by treatment with Triton X-100, was isoelectrically focused in a sucrose gradient containing the nonionic detergent octylglucoside. Removal of the detergent by dialysis resulted in aggregates of variable size and shape. The hemagglutinating activity of this preparation was approximately sixfold higher than that of the intact virus. The protein with hemagglutinating activity and with a buoyant density of 1.237 consisted solely of polypeptide chains of the G-protein and contained 0.38% phospholipids and 16 ng of Triton X-100 per mg of protein. In the National Institutes of Health test the hemagglutinin conferred a significantly higher protective activity than detergent-associated glycoprotein and was as effective as an inactivated virus vaccine. However, after the application of a single dose of hemagglutinin, the onset of protection was delayed by approximately 7 days when compared with inactivated virus vaccine.     

Preparation and characterization of porous PDLLA/HA composite foams by supercritical carbon dioxide technology

A composite poly(D,L)lactic acid (PDLLA)/hydroxyapatite (HA) biomaterial was prepared by in situ polymerization of D,L-lactide monomer and HA. Supercritical CO2 (SC CO2) technology was developed to prepare the biodegradable composite foams for use in tissue regeneration. In this technology, NaCl particles were used as porogen to produce an open-pore structure. Organic solvents were not used and high temperature was not necessary. The problem with pore interconnectivity was resolved. High-porosity composite foams (up to 90% +/- 2% porosity) were obtained with pore sizes ranging from 100 to 300 microm suitable for cell seeding. The microstructure and morphology of the composite foams could be controlled by saturation pressure, saturation time, and temperature as well as amount of NaCl particles. The compressive strength and water absorbability of the composite foams were also determined. With an increase in HA amount, the molecular weight of PDLLA/HA composite foams decreased, but the mechanical strength and hydrophilicity increased slightly.     

Preparation and characterization of macroporous chitosan-gelatin/beta-tricalcium phosphate composite scaffolds for bone tissue engineering

A biodegradable composite scaffold was developed using beta-tricalcium phosphate (beta-TCP) with chitosan (CS) and gelatin (Gel) in the form of a hybrid polymer network (HPN) via co-crosslinking with glutaraldehyde. Various types of scaffolds were prepared by freezing and lyophilizing. These scaffolds were characterized by Fourier transform infrared, X-ray diffractometer, and scanning electron microscopy. The macroporous composite scaffolds exhibited different pore structures. Compressive properties were improved, especially compressive modulus from 3.9-10.9 MPa. Biocompatibility was evaluated subcutaneously on rabbits. A mild inflammatory response was observed over 12 weeks. The results suggest that the scaffolds can be utilized in nonloading bone regeneration.     

Preparation and characterization of a novel bioactive restorative composite based on covalently coupled polyurethane-nanohydroxyapatite fibres

Nanohydroxyapatite (n-HAp) was prepared using a sol-gel method. n-HAp powder was obtained from the gel form by heat treatment followed by grinding using ball milling. A novel polyurethane composite material was prepared by chemically binding the hydroxyapatite to the diisocyanate component in the polyurethane backbone through solvent polymerization. The procedure involved the stepwise addition of monomeric units of the polyurethane and optimizing the reagent concentrations. The resultant composite material was electrospun to form fibre mats. The fibres were less than 1mum in thickness and contained no beads or irregularities. Chemical structural characterization of both the ceramics and the novel polymers were carried out by Fourier transform infrared and Raman spectroscopy. X-ray diffraction, scanning electron microscopy (SEM), transmission electron microscopy and Brunauer-Emmett-Teller surface area analysis were also employed to observe the crystal lattice and size and surface area of the n-HAp. Further characterization (by energy-dispersive X-ray analysis and SEM) of the spun fibres revealed the presence of elements associated with hydroxyapatite and polyurethane without the presence of any loose particles of hydroxyapatite, indicating the formation of the covalent bond between the ceramics and the polymer backbone.