Synthesis of bioceramic scaffolds for bone tissue engineering by rapid prototyping technique

This study presents a novel rapid prototyping process to fabricate silicate/hydroxyapatite (HA) bone scaffolds for tissue engineering applications. The HA particles are embedded in the gelled silica matrix to form a green part of bioceramic bone scaffold after processing by selective laser sintering. The composition of the bioceramic scaffold is in the series of SiO₂·P₂O₅·CaO. Results indicate that the proposed process could fabricate a multilayer hollow shell structure with brittle property but sufficient integrity for handling prior to post-processing. The fabricated bone scaffold models had a surface finish of 25 μm, a dimensional shrinkage of 16 %, a maximum bending strength of 4.7 MPa, and an apparent porosity of 28 % under the laser energy density of 1.5 J/mm². In vitro bioactivity evaluation by optical density value using a microculture tetrazolium test assay revealed that the bioceramic bone scaffolds were suitable for cell culture, demonstrating their application in tissue engineering.     

Porous poly(glycerol sebacate) (PGS) elastomer scaffolds for skin tissue engineering

Poly (glycerol sebacate) (PGS) elastomer scaffolds with different porosity for skin tissue engineering were fabricated via particulate leaching. The introduction of pores lowers the hydrophilicity but improves the water uptake capability of PGS. The gel content of PGS increases with the increase of salt mass ratio, but the degree of swelling goes the opposite way due to the existence of the porous structure. The degradation rate of PGS can be tailored and controlled by the porous structure, which is of great value for its applications in tissue engineering. The feasibility of these porous PGS scaffolds for skin tissue engineering was evaluated by seeding mouse dermal fibroblasts (MDFs) onto the scaffold. In vitro cell culture results indicate good attachment, proliferation and deep penetration of MDFs into porous PGS scaffolds, which confirms the excellent biocompatibility of these scaffolds.     

Chitosan modified poly(L-lactide) microspheres as cell microcarriers for cartilage tissue engineering

The surfaces of poly(l-lactide) (PLLA) microspheres were modified by chitosan via a method of hydrolysis and grafting-coating to improve their compatibility to chondrocytes. The PLLA microspheres with a diameter of 74-150mum were fabricated by an oil/water emulsion solvent evaporation method, followed by hydrolysis in alkaline solution to produce a larger number of carboxyl groups. Using water-soluble carbodiimide as a coupling reagent, chitosan was covalently grafted onto the microspheres. Due to the physical entanglement and insolubility at neutral pH, unbonded chitosan molecules were stably remained to yield a large amount of coated chitosan. Biological performance of the control PLLA and the chitosan-coated PLLA microspheres were assessed by in vitro culture of rabbit auricular chondrocytes. After 24h and 7d culture, the chitosan-coated PLLA microspheres, especially the ones with larger chitosan amount, exhibited stronger ability to promote cell attachment and proliferation, and maintain the secretion function of the chondrocytes. Therefore, the chitosan-coated PLLA microspheres can be potentially used as the injectable cell microcarriers for chondrogenesis in cartilage tissue engineering.     

A study on the bioactivity of chitosan/nano-hydroxyapatite composite scaffolds for bone tissue engineering

In order to increase the biocompatibility and bioactivity of chitosan, hydroxyapatite had been in situ combined into chitosan scaffolds. The bioactivity of the composite scaffolds was studied by examining the apatite formed on the scaffolds by incubating in simulated body fluid and the activity of preosteoblasts cultured on them. The apatite layer was assessed using scanning electronic microscope (SEM), X-ray diffraction (XRD), Fourier-Transformed Infrared spectroscopy (FTIR) and weight measurement. Composite analysis showed that after incubation in simulated body fluid on both of the scaffolds carbonate hydroxyapatite was formed. With increasing nano-hydroxyapatite content in the composite, the quantity of the apatite formed on the scaffolds increased. Compared with pure chitosan, the composite with nano-hydroxyapatite could form apatite more readily during the biomimetic process, which suggests that the composite possessed better mineralization activity. Furthermore, preosteoblast cells cultured on the apatite-coated scaffolds showed different behavior. On the apatite-coated composite scaffolds cells presented better proliferation than on apatite-coated chitosan scaffolds. In addition, alkaline phosphatase activities of cells cultured on the scaffolds in conditioned medium were assessed. The cells on composite scaffolds showed a higher alkaline phosphatase activity which suggested a higher differentiation level. The results indicated that the addition of nano-hydroxyapatite improved the bioactivity of chitosan/nano-hydroxyapatite composite scaffolds. On the other hand, that is to say composition of substrates could affect the apatite formation on them, and pre-loaded hydroxyapatite can enhance the apatite-coating. It will also be significant in preparation of apatite-coating polymer scaffolds for bone tissue engineering.     

In situ mineralization of hydroxyapatite on poly(vinyl alcohol) monolithic scaffolds for tissue engineering

A poly(vinyl alcohol) (PVA)/hydroxyapatite (HAp) composite monolithic scaffold is prepared via thermally impacted non-solvent induced phase separation method, successively followed by an alternate soaking process. The morphology of the resulting composite monolith is observed by scanning electron microscopy (SEM). The formation of hydroxyapatite is confirmed by X-ray diffraction, SEM in combination with energy dispersive X-ray analysis, and Fourier transform infrared spectroscopy. The effects of soaking cycle and soaking time upon the formation of hydroxyapatite on the monolith surface are systematically investigated. With the increase of soaking cycle and soaking time, the amount of the formed hydroxyapatite increases. As the soaking cycle increases, the water uptake of the composite monolith decreases. The PVA/HAp composite monolith greatly has a promising application as scaffold of bone tissue engineering.     

Preparation and mechanical behavior of PLGA/nano-BCP composite scaffolds during in-vitro degradation for bone tissue engineering

In this paper, the yield strength and elastic modulus of Poly (lactide-co-glycolide) (PLGA) and PLGA/nano-biphasic calcium phosphate (nBCP) composite scaffolds, before and during in-vitro degradation, have been evaluated. Composite scaffolds were made by using PLGA matrix and 10-50 wt.% nBCP powder as the reinforcement material. All scaffolds, with more than 89% porosity, were fabricated by thermally-induced phase separation (TIPS). During in-vitro degradation (0-8 weeks), the PLGA/nBCP scaffolds showed both more weight loss and better mechanical properties as compared to neat PLGA scaffolds. The PLGA/nBCP scaffolds with 30 wt.% nBCP illustrated the highest value of yield strength among the composite scaffolds, before and after degradation, until 6 weeks. After 8 weeks, the yield strength values were very poor and close to each other. The values of elastic modulus for all samples were less than the half of their initial values after 6 weeks. However, after 8 weeks, the elastic moduli of all samples reduced to negligible values.     

Model Compounds for Iron Proteins. Structures and Magnetic, Spectroscopic, and Redox Properties of Fe(III)M(II) and [Co(III)Fe(III)](2)O Complexes with (&mgr;-Carboxylato)bis(&mgr;-phenoxo)dimetalate and (&mgr;-Oxo)diiron(III) Cores

A series of heterobimetallic complexes of the type [Fe(III)M(II)L(&mgr;-OAc)(OAc)(H(2)O)](ClO(4)).nH(2)O (2-5) and [{Fe(III)Co(III)L(&mgr;-OAc)(OAc)}(2)(&mgr;-O)](ClO(4))(2).3H(2)O (6) where H(2)L is a tetraaminodiphenol macrocyclic ligand and M(II) = Zn(2), Ni(3), Co(4), and Mn(5) have been synthesized and characterized. The (1)H NMR spectrum of 6 exhibits all the resonances between 1 and 12 ppm. The IR and UV-vis spectra of 2-5 indicate that in all the cases the metal ions have similar coordination environments. A disordered crystal structure determined for 3 reveals the presence of a (&mgr;-acetate)bis(&mgr;-phenoxide)-Ni(II)Fe(III) core, in which the two metal ions have 6-fold coordination geometry and each have two amino nitrogens and two phenolate oxygens as the in-plane donors; aside from the axial bridging acetate, the sixth coordination site of nickel(II) is occupied by the unidentate acetate and that of iron(III) by a water molecule. The crystal structure determination of 6 shows that the two heterobinuclear Co(III)Fe(III) units are bound by an Fe-O-Fe linkage. 6 crystallizes in the orthorhombic space group Ibca with a = 17.577(4) ?, b = 27.282(7) ?, c = 28.647(6) ?, and Z = 8. The two iron(III) centers in 6 are strongly antiferromagnetically coupled, J = -100 cm(-1) (H = -2JS(1).S(2)), whereas the other two S(1) = S(2) = (5)/(2) systems, viz. [Fe(2)(III)(HL)(2)(&mgr;-OH)(2)](ClO(4))(2) (1) and the Fe(III)Mn(II) complex (5), exhibit weak antiferromagnetic exchange coupling with J = -4.5 cm(-1) (1) and -1.8 cm(-1) (5). The Fe(III)Ni(II) (3) and Fe(III)Co(II) (4) systems, however, exhibit weak ferromagnetic behavior with J = 1.7 cm(-1) (3) and 4.2 cm(-1) (4). The iron(III) center in 2-5 exhibits quasi-reversible redox behavior between -0.44 and -0.48 V vs Ag/AgCl associated with reduction to iron(II). The oxidation of cobalt(II) in 4 occurs quasi-reversibly at 0.74 V, while both nickel(II) and manganese(II) in 3 and 5 undergo irreversible oxidation at 0.85 V. The electrochemical reduction of 6 leads to the generation of 4.     

Adsorption of Hg(II) Ions onto Binary Biopolymeric Beads of Carboxymethyl Cellulose and Alginate

The removal of Hg(II) ions from aqueous solution by adsorption onto cross-linked polymeric beads of carboxymethyl cellulose (CMC) and sodium alginate was studied at fixed pH (6) and room temperature 28 ± 0.2°C. The cross-linked polymeric beads were characterized by FTIR spectra. Sorption capacity of the polymer for the mercury ions was investigated in aqueous media consisting different amounts of mercury ions (2.5 to 100 mg dm^?3) and at different pH values (2 to 8). Adsorption behavior of Hg(II) ions could be modeled using both the Langmuir and Freundlich isotherms. The dynamic nature of adsorption was quantified in terms of several kinetic constants such as rate constants for adsorption (k₁) and Lagergreen rate constant (K_ad). The influence of various experimental parameters such as effect of pH, contact time, solid-to-liquid ratio, salt effect, and temperature effect etc. were investigated on the adsorption of Hg(II) ions.     

Tailoring the porosity and morphology of gelatin-methacrylate polyHIPE scaffolds for tissue engineering applications

Gelatin is a natural protein with many desirable properties for application as a biomaterial, including scaffolding for tissue engineering. In this work gelatin A with a molecular weight in the range 50-100 kg mol-1 was modified with methacrylic anhydride and processed into a concentrated oil-in-water emulsion. Polymerization of the continuous phase gave rise to a polyHIPE, a porous material possessing a highly interconnected, trabecular morphology. In the paper, we focused on the goal of obtaining matrixes characterized by suitable sizes of both voids and interconnects, to allow an in depth colonization from transplanted cells. In this respect, we investigated the role of the volume percentage of the dispersed phase and the effect of additives. It was established that high pore volumes (>or=90%) are to be preferred, because they allow the production of solid foams characterized by average void and interconnect diameters of approximately 20 and 10 microm, respectively. These values are still inadequate for the intended application of these scaffolds but represent a good starting point for further improvements. These were achieved through the use of additives, namely sodium chloride and dimethyl sulfoxide, which partially destabilized the precursor emulsion and allowed a solid foam to be obtained with void and interconnect diameters in the range of 30-150 microm and 10-50 microm, respectively.     

Morphology and characterization of 3D micro-porous structured chitosan scaffolds for tissue engineering

This research studies the morphology and characterization of three-dimensional (3D) micro-porous structures produced from biodegradable chitosan for use as scaffolds for cells culture. The chitosan 3D micro-porous structures were produced by a simple liquid hardening method, which includes the processes of foaming by mechanical stirring without any chemical foaming agent added, and hardening by NaOH cross linking. The pore size and porosity were controlled with mechanical stirring strength. This study includes the morphology of chitosan scaffolds, the characterization of mechanical properties, water absorption properties and in vitro enzymatic degradation of the 3D micro-porous structures. The results show that chitosan 3D micro-porous structures were successfully produced. Better formation samples were obtained when chitosan concentration is at 1–3%, and concentration of NaOH is at 5%. Faster stirring rate would produce samples of smaller pore diameter, but when rotation speed reaches 4000 rpm and higher the changes in pore size is minimal. Water absorption would reduce along with the decrease of chitosan scaffolds’ pore diameter. From stress–strain analysis, chitosan scaffolds’ mechanical properties are improved when it has smaller pore diameter. From in vitro enzymatic degradation results, it shows that the disintegration rate of chitosan scaffolds would increase along with the processing time increase, but approaching equilibrium when the disintegration rate reaches about 20%.     

Novel 3D printed shape-memory PLLA-TMC/GA-TMC scaffolds for bone tissue engineering with the improved mechanical properties and degradability

The biodegradable substitution materials for bone tissue engineering have been a research hotspot. As is known to all, the biodegradability, biocompatibility, mechanical properties and plasticity of the substitution materials are the important indicators for the application of implantation materials. In this article, we reported a novel binary substitution material by blending the poly(lactic-acid)-co-(trimethylene-carbonate) and poly(glycolic-acid)-co-(trimethylene-carbonate), which are both biodegradable polymers with the same segment of flexible trimethylene-carbonate in order to accelerate the degradation rate of poly(lactic-acid)-co-(trimethylene carbonate) substrate and improve its mechanical properties. Besides, we further fabricate the porous poly(lactic-acid)-co-(trimethylene-carbonate)/poly(glycolic-acid)-co-(trimethylene-carbonate) scaffolds with uniform microstructure by the 3D extrusion printing technology in a mild printing condition. The physicochemical properties of the poly(lactic-acid)-co-(trimethylene-carbonate)/poly(glycolic-acid)-co-(trimethylene-carbonate) and the 3D printing scaffolds were investigated by universal tensile dynamometer, fourier transform infrared reflection (FTIR), scanning electron microscope (SEM) and differential scanning calorimeter (DSC). Meanwhile, the degradability of the PLLA-TMC/GA-TMC was performed in vitro degradation assays. Compared with PLLA-TMC group, PLLA-TMC/GA-TMC groups maintained the decreasing T_g, higher degradation rate and initial mechanical performance. Furthermore, the PLLA-TMC/GA-TMC 3D printing scaffolds provided shape-memory ability at 37 ℃. In summary, the PLLA-TMC/GA-TMC can be regarded as an alternative substitution material for bone tissue engineering.     

Surface Performance and Cytocompatibility Evaluation of Acrylic Acid-Mediated Carboxymethyl Chitosan Coating on Poly(tetrafluoroethylene-co-hexafluoropropylene)

To improve the biocompatibility of poly(tetrafluoroethylene-co-hexafluoropropylene) (FEP) film, a technique based on Ar plasma pretreatment and UV-induced grafting polymerization was used to immobilize carboxymethyl chitosan (CMCS) on the FEP film surfaces. Initially Ar plasma was used to treat FEP film. Then, plasma treated FEP film was modified via UV-induced grafting polymerization with hydrophilic acrylic acid (AAc) monomer. The following immobilization of CMCS on the FEP-pAAc surface was carried out via an amidation reaction. The change of chemical composition and surface morphology of FEP film were characterized by attenuated total reflectance Fourier transform infrared (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), scanning electronic microscopy (SEM) and atomic force microscopy (AFM). Results of water contact angles measurement showed that the hydrophilicity of the surface has improved significantly after surface modification. Furthermore, methyl thiazolyl tetrazolium (MTT) assay and cell morphology analysis confirmed that mouse fibroblasts (L929 cells) attachment and proliferation were improved remarkably on the modified FEP surface. These results suggest that CMCS were successfully employed to surface engineering FEP film, and enhanced its cell biocompatibility. The approach presented here may be exploited for surface modification of biomaterials.     

A pH- and Redox-Potentiometric Study of Equilibria between Fe(II), Fe(III), and N-(Carboxymethyl)Aspartic Acid

Complexation in the Fe²⁺–Fe³⁺–N-(carboxymethyl)aspartic acid (H₃L) system in aqueous solutions was studied by pH- and redox-potentiometric titration at 25°C and at an ionic strength of 0.1 (KCl). Depending on the H₃L concentration and pH, neutral, protonated, and hydroxo complexes of iron(III) can be formed in the solutions. The stability constants for all the detected complexes were calculated, and the distribution plots for the fractions of complexes vs. the solution pH were constructed.     

Synthesis of Carboxymethyl Cellulose Based on Short Fibers and Lignified Part of Flax Pedicels (Boon)

The conditions under which carboxymethyl ethers of cellulose with various degrees of substitution can be prepared from wastes formed in production of flax fibers were studied. The structure of the cellulose ethers synthesized was studied by IR Fourier spectroscopy and physicochemical methods.     

Preparation and Kinetic Modeling of Cross-Linked Chitosan Microspheres Immobilized Zn(II) for Urea Adsorption

The preparation of Zn(II)-immobilized glutaraldehyde-crosslinked chitosan microspheres which was modified with epichlorohydrin, tetraethylenepentamine, and bromoacetic acid was presented in this work. The Zn(II)-immobilized value of the microspheres (Ac-TEPA-CS) is 43.6 mg g^?1, which is higher than the blank microspheres. The adsorption of urea onto Zn-Ac-TEPA-CS was studied in a batch system. Langmuir and Freundlich adsorption models were applied to describe the experimental isotherm and isotherm constant, and the kinetic of adsorption process were estimated. These data fits well with Langmuir isotherm and also indicated that the adsorption process is exothermic and follow the pseudo-second-order kinetics. The adsorption capacity depends upon the pH, the temperature and the initial concentration of urea. It observed that Zn-Ac-TEPA-CS could be repeatedly used by elution and regeneration without significant loss of adsorption capacity.     

O-（4-双胍基苯甲酰基）壳聚糖盐酸盐的制备及其抗菌活性

通过壳聚糖与含双胍基结构的芳酰氯反应,得到具有抗菌活性的双胍基化壳聚糖衍生物。利用对双胍基苯甲酸盐酸盐与氯化亚砜的反应,得到盐酸对双胍基苯甲酰氯后,再在甲烷磺酸-二甲基亚砜（MeSO3H-DMSO）介质中与壳聚糖进行缩合反应,得到了O-（4-双胍基苯甲酰基）壳聚糖盐酸盐（p-BGBC）。用FT-IR、1H-NMR、GP...     

Synthesis of multifunctional macroporous-mesoporous TiO<Subscript>2</Subscript>-bioglasses for bone tissue engineering

Hierarchical porous TiO₂-bioglasses (TiO₂-BGs) with the macropore with the size of 30–50 μm and the mesopore with the diameter of 4.4–5.6 nm have been synthesized through the evaporation-induced self-assembly method. The corn stalks were used as the macroporous template and P123 as the mesoporous template in the process. The chemical and physical properties of the hierarchical porous TiO₂-BGs before and after immersion in simulated fluid (SBF) were evaluated by X-ray diffraction, scanning electron microscope, Fourier transform infrared spectroscopy, nitrogen adsorption–desorption, and Energy dispersive spectrometer. The effect of TiO₂ content on the formation of surface hydroxyapatite and drug release profiles of the fabricated TiO₂-BGs in SBF were investigated in detail. It was found that macroporous-mesoporous TiO₂-bioglasses (MM TiO₂-BGs) exhibited a good ability of surface hydroxyapatite formation comparing with macroporous-mesoporous bioglasses. It took only 3 h for the MM TiO₂-BGs to be covered with the hydroxyapatite layer. It can be ascribed to the present of Ti–OH which may improve the spontaneous growth of apatite by consuming the calcium and phosphate ions from SBF. Additionally, MM TiO₂-BGs also showed good drug sustained release profiles. Therefore, the multifunctional MM TiO₂-BGs reported here could be a good candidate for application in bone tissue engineering.     

128例脑性瘫痪血钙、锌、铁、铜、镁元素分析

为探讨小儿脑性瘫痪(CP)体内微量元素的变化,测定了128例脑性瘫痪患者血钙、锌、铁、铜、镁含量,随机选择128例正常儿童进行对照分析。结果表明,两组病例血锌、铜、镁均在正常范围,脑瘫组血钙56例、铁45例低于正常参考值,健康组血钙16例、铁11例低于正常参考值。经统计学处理,两组病例中血钙、铁、铜、镁含量具有显著性差异(P<0.05),血锌无显著性差异(P>0.05)。提示脑瘫儿童血钙、铁、铜、镁含量均低于正常儿童,补充钙、铁、镁等可能有利于脑瘫康复。     

Manganoan rockbridgeite Fe4.32Mn0.62Zn0.06(PO4)3(OH)5: structure analysis and 57Fe Mössbauer spectroscopy

The structure of the basic iron phosphate rockbridgeite [iron manganese zinc tris­(phosphate) penta­hydroxide] was reinvestigated with special emphasis on the cation distribution deduced from new X‐ray and ⁵⁷Fe Mössbauer data. Rockbridgeite is orthorhombic, space group Cmcm, and shows three different Fe sites, one with symmetry, another with m symmetry and the third in a general position. One phosphate group has the P atom on a site with m symmetry, while the other has the P atom at a site with mm symmetry. Two Fe sites are fully occupied by ferric iron, while Mn³⁺ and Fe²⁺ are situated at a third, principally Fe, site. Structural data, bond‐valence sums and polyhedral distortion parameters suggest a new inter­pretation of the rockbridgeite ⁵⁷Fe Mössbauer spectrum.