Electrospun chitosan/hydroxyapatite nanofibers for bone tissue engineering

Chitosan (CS) nanofibers were prepared by an electrospinning technique and then treated with simulated body fluid (SBF) to encourage hydroxyapatite (HA) formation on their surface. The CS/HA nanofibers were subjected to scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared spectroscopy, and X-ray diffraction (XRD) to confirm HA formation as well as determine the morphology of the nanofibrous scaffolds. The SEM image indicated that the distribution of HA on the CS nanofibers was homogeneous. The results from EDS and XRD indicated that HA was formed on the nanofibrous surfaces after 6-day incubation in the SBF. The calcium/phosphorus ratio of deposited HA was close to that of natural bone. To determine biocompatibility, the CS/HA scaffolds were applied to the culture of rat osteosarcoma cell lines (UMR-106). The cell densities on the CS/HA nanofibers were higher than those on the CS nanofibers, the CS/HA film, and the CS film, indicating that cell proliferation on CS/HA nanofibers was enhanced. Moreover, the early osteogenic differentiation on CS/HA was also more significant, due to the differences in chemical composition and the surface area of CS/HA nanofibers. The biocompatibility and the cell affinity were enhanced using the CS/HA nanofibers. This indicates that electrospun CS/HA scaffolds would be a potential material in bone tissue engineering.     

Inorganic/organic (SiO₂)/PEO hybrid electrospun nanofibers produced from a modified sol and their surface modification possibilities

Ceramic silica (SiO(2)) hybrid nanofibers were prepared by electrospinning of solutions containing biocompatible polymer and modified silica precursors. The new hybrid nanofibers are based on polyethylene oxide (PEO) and a new solution of modified sol-gel particles of mixture containing tetraethoxysilane (TEOS) and 3-glycidyloxypropyltriethoxysilane (GPTEOS) in a weight ratio of 3:1. Adding high-molecular-weight PEO into the silica sol is found to enhance the formation of the silica nanofibers and leads to reduce the water-soluble carrying polymer down to 1.2%wt. Transmission electron microscopy (TEM) and attenuated total reflection fourier transformation infrared ATR-FTIR measurements are suggested that PEO is encapsulated by the silica component. This excellent formulation renders electrospinning of SiO(2) a robust process for an easy production of controllable silicate nanofibrous matrices. For instance, nanofibers with average diameter down to 400 nm are accessible by varying the weight ratio between the polymer and the inorganic precursor. These are reduced to 120 nm after the pyrolysis process. Moreover, the surface of the nanofibers could be easily modified, either by Al(3+) leading to aluminium silicate coatings, or by incorporation of Ca(2+) ions and subsequent bioactive hydroxyl carbonate apatite (HAP) formation. These hybrid silica nanofibers are possess a unique collective properties can have a great impact either in high-temperature reinforced materials and filtration or in biomedical applications such as in dentistry and bone tissue engineering.     

Bioglass® Coatings on Biodegradable Poly(3‐hydroxybutyrate) (P3HB) Meshes for Tissue Engineering Scaffolds

Osteoconduction and non‐toxic bioresorbability can be achieved by combining Bioglass® particles and Poly (3‐hydroxybutyrate) (P3HB) fibre meshes in novel composites for tissue engineering scaffolds. Bioglass® coatings readily induce hydroxyapatite (HA) formation on fibre surfaces in vitro, while biodegradable P3HB yields non toxic degradation products. In the present investigation, P3HB meshes were used, which were generated by means of an embroidery technology on the basis of yarns with 12 and 24 filaments with diameters of ～ 30 μm. Bioglass® particles of average particle size < 5 μm were used to produce coatings on P3HB meshes by slurry dipping. By varying the concentration of Bioglass® particles in aqueous slurry, coating thickness and homogeneity could be controlled. Optimally coated meshes were incubated in simulated body fluid (SBF) for 3, 7, 14, and 21 days to detect formation of HA, as a qualitative assessment of bioactivity. Scanning electron microscopy (SEM) observations coupled with X‐ray diffraction analyses revealed the presence of HA crystals on mesh surfaces following 3 days of incubation in SBF. The amount of HA crystals was shown to increase with incubation time in SBF. Minimal polymer degradation was seen after 21 days in SBF, suggesting a suitable time frame for tissue replacement. The novel Bioglass® /P3HB composite meshes developed here are potential materials for bone tissue engineering scaffold applications.     

Plasma spraying of zirconia-reinforced hydroxyapatite composite coatings on titanium: Part II Dissolution behaviour in simulated body fluid and bonding degradation

The change of phase, morphology and bond strength of plasma sprayed hydroxyapatite (HA) coating and ZrO2/HA composite coatings immersed in simulated body fluid (SBF) for various periods of time was studied. X-ray diffractometry (XRD) and scanning electron microscopy (SEM) were used to identify the phase and observe the morphology of the coating surface before and after immersion. In addition, inductively coupled plasma emission spectroscopy (ICP) was used to measure the ion release rate of coatings in SBF for various periods of time. Observation of the morphology by SEM shows that the composite coating with the addition of ZrO2 in HA significantly reduced the dissolution rate of impurity phases in simulated body fluid. The argument was supported by measurement of Ca2+ ion concentration in SBF. During plasma spraying, less OH- ions were lost in a ZrO2-containing composite coating. This factor, together with the reduced effective surface of the ZrO2-containing HA coating, were attributed to the reduced dissolution rate of the composite coatings. All the plasma sprayed coatings degraded after immersion in SBF owing to dissolution of constituents in the coating, however, the addition of ZrO2 in HA improved the bonding strength of HA coating after immersion in SBF.     

Release of lysozyme from electrospun PVA/lysozyme-gelatin scaffolds

This article describes an electrospinning process in fabricating ultra fine fibers with core-shell structure. A biodegradable polymer, poly(vinyl alcohol) (PVA), was used as the shell; lysozyme was a kind of antioxidant; and gelatin were used as the core.Morphology and microstructure of the ultra fine fibers were characterized by scanning electron microscope (SEM), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS) analysis. As a comparison, composite nanofiber PVA/lysozyme-gelatin blend was prepared by a normal electrospinning process. In vitro drug release behaviors of the nanofibrous membranes were determined in phosphatebuffered saline (PBS) solution. It was found that core-shell nanofibers PVA/lysozyme-gelatin obviously exhibit higher initial release rates compared to that of PVA/lysozymegelatin blend nanofibers. The current method may find wide application in controlled release of bioactive proteins and tissue engineering.     

Morphology and immersion behavior of plasma-sprayed hydroxyapatite/bioactive glass coatings

A series of hydroxyapatite/bioactive glass (HA/BG) coatings have been plasma-sprayed on Ti6Al-4V substrate using HA/BG powders that were prepared by both sinter-granulation and direct mixing methods. The morphology and immersion behavior of these coatings in a simulated body fluid (SBF) were investigated. The results showed that in-house fabricated BG and sinter-granulated HA powders were irregularly shaped and dense. When 5 wt % or more BG was added in HA, the powder became rough and porous. X-ray diffraction (XRD) patterns showed that the presence of BG enhanced the decomposition of HA structure during fabrication of the powders. Reasonably high bond strengths were obtained from all coatings. The granulated type HA/BG coatings showed no significant differences in bond strength from the mixed type HA/BG coatings. The plasma spray process itself and the presence of BG enhanced the decomposition of apatite. Surface morphology of all sinter-granulated type coatings was similar to that of monolithic HA coating, that was comprised of patches of smooth and shiny glassy film and irregularly-shaped particles on its surface. The dissolution depth of plasma-sprayed coatings immersed in SBF was largely dependent on the type and composition of the coating. Granulated type HA/BG coatings were much less dissolvable than monolithic HA or mixed type HA/BG coatings. It seems that the presently used granulation method for the preparation of HA/BG powders plays a predominant role in determining the dissolution behavior of the plasma-sprayed coatings. ©©2000 Kluwer Academic Publishers     

Wet chemical process to enhance osteoconductivity of electrospun chitosan nanofibers

In this study, chitosan/hydroxyapatite (CS/HA) nanofibers were prepared using a wet chemical method. First, CS nanofibers with uniform diameters were fabricated using electrospinning. Then, a wet chemical process was used to mineralize nanofiber surfaces to form a homogeneous HA deposit. Reactions with three cycles were found to optimize biomimetic properties of the HA. The mineralization process required only approximately 3 h, which corresponded to a saving of 98 % in preparation time compared with that needed by the process using a simulated body fluid (SBF). According to the attachment and spreading of UMR (rat osteosarcoma) cells on the CS/HA composite fibers, the deposited mineralization layer significantly enhanced cell affinity of the CS nanofibers and the HA created by the wet chemical method was as effective as that created by the SBF. The composite nanofibrous scaffolds produced by the wet chemical process also promoted osteogenic differentiation by inducing ossification. Thus, expressions of collagen type I, alkaline phosphatase, osteocalcin, bone sialoprotein, and osterix were all enhanced. These results demonstrated that composite electrospun fibers can be efficiently prepared using wet chemical method and the resulting nanofibrous scaffolds have considerable potential in future bone tissue engineering applications.     

等离子喷涂制备HA/ZrO2复合涂层

采用等离子喷涂技术,在Ｔｉ－６Ａｌ－４Ｖ基体上成功地制备了羟基磷灰石／氧化锆（ＨＡ／ＺｒＯ２）复合涂层,对涂层的微观结构,相组成和结合强度进行了研究,并以模拟体液试验评估涂层的生物活性,结果表明,复合涂层较有较为微观结构,ＨＡ／ＺｒＯ２复合涂层的结合强度明显高于ＨＡ涂层,ＨＡ／６０ｗｔ％ＺｒＯ２涂层的结合强度高达２８．５ＭＰａ,为ＨＡ涂层的２．２倍,复合涂层在模拟体液中浸泡一段时间后,表面覆盖一层     

Preparation and characterization of poly(acrylonitrile-co-acrylic acid) nanofibrous composites with Fe3O4 magnetic nanoparticles

Nanofibrous composites are a new class of polymer materials with controlled and tailored properties. Novel Fe₃O₄/poly(acrylonitrile-co-acrylic acid) nanofibrous composites with magnetic behavior have been prepared by a simple electrospinning process. The nanofibrous composites were characterized by X-ray diffraction, field emission scanning electron microscopy and vibrating sample magnetometer. The distribution of Fe₃O₄ nanoparticles inside the nanofibrous composites was investigated by field emission scanning electron microscopy. X-ray diffraction revealed the presence of Fe₃O₄ nanoparticles in the nanofibrous composites. The maximum saturation magnetization for the composites, measured at 300 K, was 30.51 emu/g.     

聚乳酸复合纳米纤维创面敷料的制备及性能

采用静电纺丝技术制备了聚乳酸(PLLA)纳米纤维毡、壳聚糖/PLLA纳米纤维毡和明胶/PLLA纳米纤维毡。利用扫描电镜(SEM)、图像分析软件等手段研究了纳米纤维微观形貌,并研究各种创面敷料的吸水性、保水性和水蒸汽通透性等性能。结果表明,壳聚糖/PLLA、明胶/PLLA复合纳米纤维毡的吸水性和保水性有显著提高,水蒸汽通透性略有下降,是理想的创面敷料材料。     

Electrophoretic deposition of carbon nanotubes–hydroxyapatite nanocomposites on titanium substrate

Carbon nanotubes–hydroxyapatite (CNTs–HA) composites were synthesized, using an in situ chemical method and characterized by X-ray diffraction (XRD) and transmission electron microscopy (TEM). HA particles were uniformly absorbed on the CNTs, with strong interfacial bonding. The CNTs–HA composites behaved like single composites when deposited on a titanium substrate by electrophoretic deposition (EPD). EPD was carried out at 10, 20 and 40 V, for 0.5 to 8 min at each voltage. Coating efficiency and weight increased with increasing deposition time, while the slope of the curves decreased, indicating a decrease in deposition rate. The CNTs–HA coating morphology was analyzed with scanning electron microscopy (SEM). The results revealed that decreasing the voltage used for deposition coatings could reduce cracking frequency. Potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) studies showed that the deposition coatings protected the titanium substrate from corroding in simulated body fluid (SBF). In addition, in vitro cellular responses to the CNTs–HA coatings were assessed to investigate the proliferation and morphology of osteoblast cell line.     

Synthesis of nanocrystalline hydroxyapatite using surfactant template systems: Role of templates in controlling morphology

Hydroxyapatite (HA) nanopowder was synthesized by reverse microemulsion technique using calcium nitrate and phosphoric acid as starting materials in aqueous phase. Cyclohexane, hexane, and isooctane were used as organic solvents, and Dioctyl sulfosuccinate sodium salt (AOT), dodecyl phosphate (DP), NP5 (poly(oxyethylene)₅ nonylphenol ether), and NP12 (poly(oxyethylene)₁₂ nonylphenol ether) as surfactants to make the emulsion. Effect of synthesis parameters, such as type of surfactant, aqueous to organic ratio (A/O), pH and temperature on powder characteristics were studied. It was found that the surfactant templates played a significant role in regulating the morphology of the nanoparticle. Hydroxyapatite nanoparticle of different morphologies such as spherical, needle shape or rod-like were obtained by adjusting the conditions of the emulsion system. Synthesized powder was characterized using X-ray diffraction (XRD), BET surface area and transmission electron microscopy (TEM). Phase pure HA nanopowder with highest surface area of 121 m²/g were prepared by this technique using NP5 as a surfactant. Densification studies showed that this nanoparticle can give about 98% of their theoretical density. In vitro bioactivity of the dense HA compacts was confirmed by excellent apatite layer formation after 21 days in SBF solution. Cell material interaction study showed good cell attachment and after 5 days cells were proliferated on HA compacts in OPC1 cell culture medium. The results imply this to be a versatile approach for making hydroxyapatite nanocrystals with controlled morphology and excellent biocompatibility.     

Cell adhesion and proliferation on biomimetic calcium-phosphate coatings produced by a sodium silicate gel methodology

The present study describes a methodology to produce bioactive coatings on the surface of starch based biodegradable polymers or other polymeric biomaterials. As an alternative to the more typical bioactive glass percursors, a sodium silicate gel is being employed as a nucleating agent, for inducing the formation of a calcium-phosphate (Ca-P) layer. The method has the advantage of being able to coat efficiently both compact materials and porous 3D architectures aimed at being used on tissue replacement applications and as bone tissue engineering scaffolds. This treatment is also very effective in reducing the incubation periods, being possible to observe the formation of an apatite-like layer, only after 6 h of immersion in a simulated body fluid (SBF). The influence of the SBF concentration on the formation of the apatite coating was also studied. The apatite coatings formed under different conditions were analyzed and compared in terms of morphology, chemical composition and structure. After the first days of SBF immersion, the apatite-like films exhibit the typical cauliflower like morphology. With increasing immersion times, these films exhibited a partially amorphous nature and the Ca/P ratios became very closer to the value attributed to hydroxyapatite (1.67). The obtained results are very promising for pre-calcifying bone tissue engineering scaffolds. Therefore, in order to study cell behavior and response to these apatite coatings, adhesion, morphology, and proliferation of a human osteoblast cell line (SaOS-2) was also analyzed after being cultured in the coatings formed after 15 days of immersion in SBF. Results indicate a good correlation between crystallinity of the apatite like coatings formed in these conditions and respective cell spreading and morphology. In general, higher cell proliferation was observed for higher crystalline Ca-P coatings.     

Hydroxyapatite/Bioactive Glass Films Produced by a Sol–Gel Method: In Vitro Behavior

Hydroxyapatite (HA) and HA/bioactive glass (49S) films were deposited on Si(100) substrates by a sol–gel dip‐coating method. The microstructure and in vitro bioactivity of the films were investigated by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), and X‐ray photoelectron spectroscopy (XPS). Polycrystalline HA and amorphous bioactive glass films were obtained after annealing at 600 and 400 °C, respectively. The crystallization temperature of HA was determined to be around 568 °C. The surfaces of the HA films were covered with an apatite layer consists of spherulites formed by nanosized needle‐like aggregates after the soaking in simulated body fluid (SBF) for 10 days, while amorphous HA/bioactive glass surface was covered with larger spherical crystallites. Both XPS and EDS results obtained from HA/bioactive glass film, after soaking in SBF, showed increasing P amounts on the surface at the expense of Si. The higher density of the newly formed layer on HA/bioactive glass surface than that of the HA surface after 10 days of soaking was evidence of increased reaction rate and apatite forming ability when bioactive glass layer is present on the HA films.     

等离子喷涂制备HA/ZrO2复合涂层

采用等离子喷涂技术,在Ｔｉ－６Ａｌ－４Ｖ基体上成功地制备了羟基磷灰石／氧化锆（ＨＡ／ＺｒＯ_２）复合涂层,对涂层的微观结构、相组成和结合强度进行了研究,并以模拟体液试验评估涂层的生物活性．结果表明,复合涂层具有较为均匀的微观结构．ＨＡ／ＺｒＯ_２复合涂层的结合强度明显高于 ＨＡ涂层, ＨＡ／６０ ｗｔ％ ＺｒＯ_２涂层的结合强度高达 ２８．５ＭＰａ,为 ＨＡ涂层的 ２．２倍．复合涂层在模拟体液中浸泡一段时间后,表面覆盖一层碳酸磷灰石（ｃａｒｂｏｎａｔｅ－ａｐａｔｉｔｅ）,表明涂层具有良好的生物活性．     

Development of a Tamsulosin Hydrochloride Controlled-Release Capsule Consisting of Two Different Coated Pellets

Tamsulosin hydrochloride (TSH) controlled-release capsule (pellets) was successfully prepared using a novel, simple, and flexible multiunit drug delivery system, which consisted of two different coated pellets. The TSH-loaded core pellets consisting of microcrystalline cellulose (MCC), lactose, Carbopol^® 974P, and the active agent, were prepared by extrusion/spheronization method. Eudragit^® NE30D and Eudragit^® L30D-55 were used as the coating materials to prepare sustained-release (SR) pellets and enteric-release (ER) pellets. The coated pellets were prepared using two different equipments: centrifugal coater and fluidized-bed coater. By adjusting the ratio of SR and ER pellets, more than one blend ratios, which meet the in vitro release criterion were obtained. A similarity factor (f₂) was employed to choose the optimum proportion compared with the commercial product (Harnal^® capsule). The morphology of the pellet surfaces was examined by scanning electron microscopy (SEM) before and after dissolution. The release profiles were significantly affected by changing the proportions of SR and ER. The optimum ratio is SR:ER?=?2:1 using a centrifugal coater (f₂?=?61.93) and SR:ER?=?3:1 using a fluidized coater (f₂?=?66.42). This result suggests that blending these two-part pellets (SR and ER) can provide an alternative to preparing a controlled-release dosage form, instead of blending of the coating polymer.     

Corrosion behaviour and bioactivity of electrophoretically deposited hydroxyapatite on titanium in physiological media (Hanks’ solution)

Hydroxyapatite (HA) coatings were developed on titanium by electrophoretic deposition at various deposition potentials from 30 to 60 V and at a constant deposition time of 5 minutes using the synthetic HA (Ca₁₀(PO₄)₆(OH)₂,) powder in a suspension of dimethyleformamide (DMF, HCON(CH₃)₂). The electrochemical corrosion behavior of the HA coatings in simulated body fluid (SBF Hanks?? solution) at 37 °C and pH 7.4 was investigated by means of open-circuit potential (OCP) measurement and potentiodynamic polarization tests. The OCP test showed that the values OCP for the coated samples shifted to more noble potential than for uncoated titanium, especially after addition of dispersants. The polarization test revealed that all HA coated specimens had a corrosion resistance higher than that of the substrate, especially after addition of dispersants such as polyvinyl butyral (PVB), polyethylene glycol (PEG) and triethanolamine (TEA) to the suspension. The coating morphology after polarization, characterized by scanning electron microscopy (SEM), showed penetration of electrolyte into the HA coats. Bone bioactivity of the coatings was also studied by immersion of coated specimens in Hanks?? solution for 3 and 7 days. Apatite granules growth on the surface of the HA layers was observed.     

Bioresorbable and bioactive composite materials based on polylactide foams filled with and coated by Bioglass® particles for tissue engineering applications

Poly(DL-lactide) (PDLLA) foams and bioactive glass (Bioglass®) particles were used to form bioresorbable and bioactive composite scaffolds for applications in bone tissue engineering. A thermally induced phase separation process was applied to prepare highly porous PDLLA foams filled with 10 wt % Bioglass® particles. Stable and homogeneous layers of Bioglass® particles on the surface of the PDLLA/Bioglass® composite foams as well as infiltration of Bioglass® particles throughout the porous network were achieved using a slurry-dipping technique. The quality of the bioactive glass coatings was reproducible in terms of thickness and microstructure. In vitro studies in simulated body fluid (SBF) were performed to study the formation of hydroxyapatite (HA) on the surface of the PDLLA/Bioglass® composites, as an indication of the bioactivity of the materials. Formation of the HA layer after immersion in SBF was confirmed by X-ray diffraction and Raman spectroscopy measurements. The rate of HA formation in Bioglass®-coated samples was higher than that observed in non-coated samples. SEM analysis showed that the HA layer thickness rapidly increased with increasing time in SBF in the Bioglass®-coated samples. The high bioactivity of the developed composites suggests that the materials are attractive for use as bioactive, resorbable scaffolds in bone tissue engineering.     

Poly(D,L-lactide) (PDLLA) foams with TiO2 nanoparticles and PDLLA/TiO2-Bioglass® foam composites for tissue engineering scaffolds

Porous poly(D,L-lactide) PDLLA foams containing 0, 5 and 20 wt% of TiO₂ nanoparticles were fabricated and characterised. The addition of Bioglass^® particles was also studied in a composite containing 5 wt% of Bioglass^® particles and 20 wt% of TiO₂ nanoparticles. The microstructure of the four different foam types was characterised using scanning electron microscopy (SEM) and their mechanical properties assessed by quasi-static compression testing. The in vitro behaviour of the foams was studied in simulated body fluid (SBF) at three different time points: 3, 21 and 28 days. The degradation of the samples was characterised quantitatively by measuring the water absorption and weight loss as a function of immersion time in SBF. The bioactivity of the foams was characterised by observing hydroxyapatite (HA) formation after 21 days of immersion in SBF using SEM and confirmed with X-ray diffraction (XRD) analysis. It was found that the amount of HA was dependent on the distribution of TiO₂ nanoparticles and on the presence of Bioglass^® in the foam samples.     

Bioactivity of calcium phosphate coatings prepared by electrodeposition in a modified simulated body fluid

The purpose of this study was to investigate bioactivity of calcium phosphate coatings prepared by electrodeposition in a modified simulated body fluid (SBF). Calcium phosphates were electrodeposited on commercially pure titanium substrates in the modified SBF at 60 °C for 1 h maintaining the cathodic potentials of − 1.5 V, − 2 V, and − 2.5 V (vs. SCE). Subsequently, the calcium phosphate coatings were transformed into apatites during immersion in the SBF at 36.5 °C for 5 days. The apatites consisted of needle-shaped crystallites distributed irregularly with different grain sizes. As the coatings were electrodeposited at higher cathodic potential, the crystallite of the apatites got denser and the grain sizes of the apatites became bigger during subsequent immersion in the SBF. However, as the coatings were electrodeposited at higher cathodic potential, the coatings were transformed into apatites with lower crystallinity and the Ca/P atomic ratio of the apatites got higher than 1.67, that of stoichiometric hydroxyapatite, after subsequent immersion in the SBF. In addition, CO₃²⁻ ions contained in the modified SBF were incorporated in the calcium phosphate coating during electrodeposition and had an influence on transforming the calcium phosphate into bonelike apatite during subsequent immersion in the SBF showing that CO₃²⁻ incorporated in the apatites disturbed crystallization of the apatites. These results revealed that the coating electrodeposited at − 2.0 V (vs. SCE) in the modified SBF containing CO₃²⁻ ions was the most bioactive showing transformation into carbonate apatite similar to bone apatite.