Fabrication of bioactive composite by developing PLLA onto the framework of sintered HA scaffold

Composite porous scaffolds of hydroxyapatite (HA)/poly-l-lactide (PLLA) were fabricated by a two-step immersing replication method. Structure and mechanical properties of both the single HA scaffold and the composite HA/PLLA scaffold were determined. The bioactivity of the scaffolds was evaluated by soaking in a simulated body fluid (SBF), and the formation of the apatite layer was determined by X-ray diffraction (XRD), Scanning Electron Microscope (SEM) and Energy-Dispersive Spectrometer (EDS). The results showed that without changing the highly interconnected porous structure, the HA/PLLA composite scaffold was mechanically enhanced to a great deal of extent compared with single HA scaffold. On the other hand, it is also suggested that the HA/PLLA scaffold was bioactive as it induced the formation of apatite on the surface of the composite scaffolds after soaking in SBF for 7 days.     

Preparation of poly(3-hydroxybutyrate)/nano-hydroxyapatite composite scaffolds for bone tissue engineering

Poly(3-hydroxybutyrate)/nano-hydroxyapatite (PHB/nHA) composite scaffolds were fabricated via powder mixing, compression moulding, and particle leaching technique. The scaffolds had high porosity with interconnected porous architecture, a favorable structure for cell attachment and new bone tissue ingrowth. A homogeneous dispersion and a uniform distribution of HA nanoparticles in the polymer matrix were obtained. The scaffolds exhibited improved compressive modulus and compressive strength, which were all in the range of compressive modulus and compressive strength of cancellous bone. In addition, the use of toxic organic solvents was eliminated. Thus, the fabricated PHB/nHA composite scaffolds tend to be promising for application in bone tissue engineering.     

Development of polycaprolactone/chitosan blend porous scaffolds

Polycaprolactone (PCL) and chitosan were blended to fabricate porous scaffolds for tissue-engineering applications by employing a concentrated acetic acid solution as solvent and salt particles as porogen. These scaffolds showed well-controlled and interconnected porous structures. The pore size and porosity of the scaffolds could be effectively modulated by selecting appropriate amounts and sizes of porogen. The results obtained from compressive mechanical measurements indicated that PCL/chitosan could basically retain their strength in their dry state compared to individual components. In a hydrated state, their compressive stress and modulus could be still well maintained even though the weight ratio of chitosan reached around 50 wt%.     

Preparation of highly interconnected porous hydroxyapatite scaffolds by chitin gel-casting

In this study, a simple and effective method for producing highly interconnected porous hydroxyapatite (HA) scaffolds was developed by combining gel-casting, particle-leaching and extrusion techniques. Chitin (CT) sol was used to disperse HA particles and wax spheres were introduced as porogens for their excellent deformability. In extrusion process, the accumulated wax spheres in point-to-point contact can transform into surface-to-surface contact by means of the extrusion pressure. Thus, the obtained porous HA scaffolds exhibited an interconnected channel network after leaching out of the porogens. The results showed that the scaffolds prepared by different size of wax spheres exhibited nearly the same volumetric porosity of about 86%, while the compressive strengths decreased as the pore size increased. Therefore, the method developed can be used to effectively tailor the pore size of HA scaffolds while maintaining a high porosity. The highly porous HA scaffolds with excellent interconnectivity are expected to be a promising bone substitute in clinical practice.     

Characterization of poly(3-hydroxybutyrate)/nano-hydroxyapatite composite scaffolds fabricated without the use of organic solvents for bone tissue engineering applications

Poly(3-hydroxybutyrate)/nano-hydroxyapatite (PHB/nHA) composite scaffolds were fabricated without the use of organic solvents at different mass fractions of HA nanoparticles. HA nanoparticles were homogeneously dispersed as primary particles in the polymer matrix of the scaffolds at 10 and 15 wt.% nHA content. Agglomeration of HA nanoparticles occurred when the nHA content of the scaffolds reached 20 wt.%. All the scaffolds had high porosities with interconnected porous structure and optimized pore size ranges. Mechanical properties of all the scaffolds were in the range of mechanical properties of cancellous bone. Scaffolds were biocompatible to MG-63 cells in the indirect method of cytotoxicity evaluation. Also, the morphology of the attached MG-63 cells in direct contact with the scaffolds indicated the appropriate cell-scaffold interaction. Thus, the PHB/nHA composite scaffolds investigated in this study tend to be favorable for bone tissue engineering applications.     

多孔ZnO/羟基磷灰石生物复合材料的制备与性能

利用放电等离子烧结技术制备多孔ZnO/羟基磷灰石（HA）生物复合材料,研究不同纳米ZnO含量对ZnO/HA复合材料微观结构、孔隙特征、力学性能、矿化和降解性能的影响。结果表明:烧结后ZnO/HA复合材料主要由HA相和ZnO相组成;随着ZnO含量提高,多孔ZnO/HA复合材料孔隙率缓慢增大,抗压强度略有减小,弹性模量变化不大;多孔ZnO/HA复合材料的孔隙率>40%,孔径在50～500 μm之间,抗压强度>148 MPa,弹性模量为6.5 GPa左右,能够满足骨修复材料的要求;模拟人工体液中矿化和降解实验表明,多孔ZnO/HA复合材料浸泡7天后表面开始形成大量类骨磷灰石层,且随着ZnO含量增加,磷灰石形成能力明显增强而降解速率加快。      

多孔硅酸钙生物陶瓷的制备及体外活性和降解性研究

以聚乙二醇为造孔剂,制备了多孔硅酸钙生物陶瓷.应用本文提供的方法,可制得孔连通性好、气孔率53.7%-73.6%、抗压强度4.9-48.5MPa、大孔孔径200-500μm的多孔陶瓷.研究表明,多孔硅酸钙生物陶瓷的力学强度随造孔剂含量的增加而明显降低,而气孔率则相反.应用模拟体液浸泡实验研究了多孔陶瓷的体外生物活性和降解性.研究表明,样品在模拟体液中浸泡1天表面就全部被碳酸代羟基磷灰石层覆盖住.气孔率为63.1%的样品7天的降解率达7.14%,具有良好的降解性.研究结果显示多孔硅酸钙生物陶瓷有望作为硬组织修复支架材料用.     

Fabrication and biological characteristics of β-tricalcium phosphate porous ceramic scaffolds reinforced with calcium phosphate glass

The fabrication process, compressive strength and biocompatibility of porous β-tricalcium phosphate (β-TCP) ceramic scaffolds reinforced with 45P₂O₅–22CaO–25Na₂O–8MgO bioglass (β-TCP/BG) were investigated for their suitability as bone engineering materials. Porous β-TCP/BG scaffolds with macropore sizes of 200–500 μm were prepared by coating porous polyurethane template with β-TCP/BG slurry. The β-TCP/BG scaffolds showed interconnected porous structures and exhibited enhanced mechanical properties to those pure β-TCP scaffolds. In order to assess the effects of chemical composition of this bioglass on the behavior of osteoblasts cultured in vitro, porous scaffolds were immersed in simulated body fluid (SBF) for 2 weeks, and original specimens (without soaked in SBF) seeded with MC3T3-E1 were cultured for the same period. The ability of inducing apatite crystals in simulated body fluid and the attachment of osteoblasts were examined. Results suggest that apatite agglomerates are formed on the surface of the β-TCP/BG scaffolds and its Ca/P molar ratio is ~1.42. Controlling the crystallization from the β-TCP/BG matrix could influence the releasing speed of inorganic ions and further adjust the microenvironment of the solution around the β-TCP/BG, which could improve the interaction between osteoblasts and the scaffolds.     

Preparation and in vitro bioactivity of poly(d,l-lactide) composite containing hydroxyapatite nanocrystals

The nano-sized hydroxyapatite (n-HA) was incorporated into poly(d,l-Lactide) (PDLLA) to form a bioactive and biodegradable composite for application in hard tissue replacement and regeneration. Thin film of PDLLA composite containing 20 mass% of n-HA fillers was successfully developed through integration of solvent co-blending and hot pressing techniques. firstly, n-HA and PDLLA were chemically synthesized, respectively, then mixed together and homogeneously dispersed in N,N-dimethyl formamide(DMF) solvent, finally, the dried blended hybrid containing PDLLA matrix and n-HA fillers was put into the mould and compacted by hot-pressing machine under 8 MPa pressure at 110 °C for 15 min. In vitro studies were conducted using the simulated body fluid(SBF). Composite specimens were soaked in SBF from 1 day to 21 days prior to surface analysis. Results obtained from scanning electron microscopy(SEM) examination, Energy dispersive X-ray detector(EDX) analysis and X-ray diffraction (XRD) analysis showed that a layer of non-stoichiometric apatite formed within 7 days on HA/PDLLA composite surface after its immersion in SBF, demonstrating moderate in vitro bioactivity of n-HA/PDLLA composite, though a moderate rate of apatite formation in SBF was found on initial stage of immersion periods for n-HA/PDLLA composite, compared to the other biomaterial composite. This type of composite film exhibited certain desirable bioactive characteristics, and they are promising bone candidates to develop novel bioactive composites for biomedical application.     

Fabrication and characterization of novel hydroxyapatite/porous carbon composite scaffolds

Novel hydroxyapatite (HA)/porous carbon composite scaffolds were prepared by applying sonoelectrodeposition and a subsequent hydrothermal treatment to previous carbonized phenolic resin coated polyurethane sponges. The interconnected pore network and morphology of HA/porous carbon composite scaffolds were determined by scanning electron microscope (SEM), and the whole surface of porous carbons were evenly coated with the deposited HA layer which was confirmed by EDS and XRD. The porosity (83.5 ± 0.3%) and the bulk density (0.297 ± 0.009 g·cm⁻³) of HA/porous carbon scaffolds were detected by the Archimedes method. The compressive and flexural strength of the scaffolds is 1.187 ± 0.064 MPa and 0.607 ± 0.268 MPa, respectively. Compared with the polymeric surface of 24-well cell culture plates, these novel scaffolds significantly promote the proliferation of human osteoblast-like MG-63 cells, indicating that this novel HA/porous carbon composite scaffold could be used for in vitro 3D culture of osteoblasts.     

Low temperature aqueous precipitation of needle-like nanophase hydroxyapatite

The use of tissue engineered biodegradable porous scaffolds has become an important focus of the biomedical research field. The precursor materials used to form these structures play a vital role in their overall performance thus making the study and synthesis of these selected materials imperative. The authors present a comparison and characterisation of hydroxyapatite (HA), a popular calcium phosphate (CaP) biomaterial, synthesised by an aqueous precipitation (AP) method. The influence of various reaction conditions on the phase, crystallinity, particle size as well as morphology, molecular structure, potential in-vivo bioactivity and cell viability were assessed by XRD, SEM and TEM, FTIR, a simulated body fluid (SBF) test and a live/dead assay using MC3T3 osteoblast precursor cells, respectively. Naturally carbonated nanoparticles of HA with typically needle-like morphology were synthesised by the reported AP method. Initial pH was found to influence the crystallisation process and determine the CaP phase formed as well as the resultant particle and crystallite sizes. A marked change in particle morphology was also observed above pH 9. The use of toluene as a replacement solvent for water up to 60 % was found to reduce the crystallinity of as-synthesised HA. This has marked influence on the effect of ethanolamine (5 wt%), which was found to improve HA crystallinity. SEM and EDS were used to confirm the growth of carbonated apatite on the surface of HA pellets immersed in SBF for up to 28 days. Cell culture results revealed viable cells on all samples where pH was controlled and maintained at 10–11 during precipitation, including those that used ethanolamine and toluene in preparation. When the initial alkali pH was not maintained non-viable cells were observed on HA substrates.     

β-CaSiO_3/丝素蛋白复合支架材料结构与性能的研究

利用冷冻干燥法制备了β-CaSiO_3/丝素蛋白复合支架材料,经XRD和FTIR分析表明复合支架中丝素的结构主要以β-折叠为主;SEM分析显示材料孔隙分布均匀,孔连通性较好,孔径尺寸约为100～300μm.对支架的孔隙率和机械强度等性能进行了表征,研究表明复合支架的孔隙率为83%～87%,机械强度有较大提高.应用模拟体液浸泡实验研究了复合支架的体外生物活性,并用XRD、FESEM和EDS对试样表面进行了表征;结果显示,样品经模拟体液浸泡3天后,表面都能沉积出类骨羟基磷灰石(HA)层,β-CaSiO_3的加入能加快复合支架表面沉积类骨HA的速度.研究结果显示β-CaSiO_3/丝素蛋白复合支架材料有望作为强度较好的生物活性硬组织修复材料.     

Fabrication and characterization of interconnected porous biodegradable poly(ε-caprolactone) load bearing scaffolds

In this study, poly(ε-caprolactone) (PCL)/poly(ethylene oxide) (PEO) (50:50 wt%) immiscible blend was used as a model system to investigate the feasibility of a novel solventless fabrication approach that combines cryomilling, compression molding and porogen leaching techniques to prepare interconnected porous scaffolds for tissue engineering. PCL was cryomilled with PEO to form blend powders. Compression molding was used to consolidate and anneal the cryomilled powders. Selective dissolution of the PEO with water resulted in interconnected porous scaffolds. Sodium chloride salt (NaCl) was subsequently added to cryomilled powder to increase the porosity of scaffolds. The prepared scaffolds had homogeneous pore structures, a porosity of ~50% which was increased by mixing salt with the blend (~70% for 60% wt% NaCl), and a compressive modulus and strength (ε = 10%) of 60 and 2.8 MPa, respectively. The results of the study confirm that this novel approach offers a viable alternative to fabricate scaffolds.     

软骨细胞在聚乳酸支架中的体外生长行为

采用明胶和氯化钠颗粒作为致孔剂,使用溶剂浇铸／颗粒沥滤法制备了高孔隙率、孔间连通和高机械性能的聚乳酸支架,采用软骨细胞体外培养研究了这两种多孔支架对细胞生长性能的影响．结果表明,软骨细胞在以明胶颗粒为致孔剂制备的多孔支架中的相对数量和GAG的分泌量更多,细胞的活性更高。     

Biodegradable polylactide/hydroxyapatite nanocomposite foam scaffolds for bone tissue engineering applications

Supercritical carbon dioxide processing of poly-L-lactide (PLLA)/hydroxyapatite (nHA) nanocomposites was investigated as a means to prepare foams suitable as scaffolds in bone tissue engineering applications. For given foaming parameters, addition of nHA to the PLLA gave reduced cell sizes and improved homogeneity in the size distribution, but did not significantly affect the degree of crystallinity, which remained of the order of 50 wt% in all the foams. The compressive modulus and strength were primarily influenced by the porosity and there was no significant reinforcement of the matrix by the nHA. The mechanical properties of the foams were nevertheless comparable with those of trabecular bone, and by adjusting the saturation pressure and depressurization rate it was possible to generate porosities of about 85 %, an interconnected morphology and cell diameters in the range 200-400 μm from PLLA containing 4.17 vol% nHA, satisfying established geometrical requirements for bone replacement scaffolds.     

Fabrication of HA/PHBV composite scaffolds through the emulsion freezing/freeze-drying process and characterisation of the scaffolds

Biodegradable polymer-based scaffolds containing osteoconductive hydroxyapatite (HA) particles can be very useful for bone tissue engineering. In this investigation, HA nanoparticles were incorporated in poly(hydroxybutyrate-co-valerate) (PHBV) polymer to fabricate osteoconductive composite scaffolds. PHBV and HA/PHBV scaffolds were made using an emulsion freezing/freeze-drying technique. The scaffolds produced were subsequently characterized using several techniques. It was found that the scaffolds were highly porous and had interconnected porous structures. The pore size ranged from several microns to around 300 mum. The spherical HA nanoparticles which were produced in-house through a nanoemulsion process could be incorporated into composite scaffolds although some of these nanoparticles existed on the surface of pore walls when a relatively large amount of HA was used for composite scaffolds. The incorporation of HA nanoparticles also enhanced compressive mechanical properties of the scaffolds.     

Preparation and cellular response of porous A-type carbonated hydroxyapatite nanoceramics

Microwave sintering using the activated carbon as embedding material was applied in preparation of porous A-type carbonated hydroxyapatite ceramics with nano(nCHA) and submicron (mCHA) structure. By examining the linear shrinkages and the compressive strengths of samples at different temperatures, a suitable microwave sintering temperature was achieved. The microwave sintering method was successfully used to prepare A-type CHA with nano or submicron structure, and the mechanism of the formation of A-type carbonate groups was discussed also. Compared with the samples prepared by the conventional sintering method (mHA), the nCHA bioceramics synthesized by the microwave sintering approach had smaller grain size and more uniform microstructure, and showed a compressive strength similar to the conventional samples. In vitro dissolution test proved that nCHA exhibits better degradation property in comparison to pure HA. Rat osteoblasts were cultured with nCHA, mCHA and mHA to evaluate their biocompatibility, and nCHA showed significant enhancement of cells in attachment, proliferation and differentiation. In conclusion, carbonate groups can be easily introduced to HA crystal structure using the activated carbon as embedding material, and microwave sintering is an effective and simple method in preparing A-type CHA with a nanostructure. Results from this in vitro biological study suggest that porous A-type carbonated hydroxyapatite nanoceramics may be a much better candidate for clinical use in terms of bioactivity.     

掺杂钠硅磷灰石水泥多孔支架用于骨缺损再生修复

采用磷酸四钙和磷酸氢钙为磷灰石水泥(AC)粉末, 5wt%硅酸钠水溶液为固化液, 氯化钠为致孔剂, 制备了掺杂钠元素的硅磷灰石水泥(s-AC)多孔支架用于骨缺损再生修复. 结果显示: s-AC多孔支架的成分为含钠硅元素的磷灰石, 支架大孔之间互相贯通, 孔径在200~600 μm, 孔隙率在58%~75%, 抗压强度在1.6~3.8MPa的范围内. 与AC相比, 掺杂一定量的钠硅元素提高了s-AC支架在Tris-HCl溶液中的降解性. 将s-AC多孔支架植入兔股骨缺损处, 组织学分析结果显示: 新生骨在支架材料的表面直接形成, 并长入其中, 相互贯通的多孔支架促进了新骨长入其内部. 结果表明: s-AC支架具有优良的生物相容性、降解性和成骨性, 将是一种优良的骨修复生物材料.     

In vitro mineralization kinetics of poly(<Emphasis Type="SmallCaps">l</Emphasis>-lactic acid)/hydroxyapatite nanocomposite material by attenuated total reflection Fourier transform infrared mapping coupled with principal component analysis

Poly(l-lactic acid)/hydroxyapatite (PLLA/HA) nanocomposite, which combines the properties of PLLA and HA, is suitable to construct scaffold for bone tissue engineering. Its mineralization behavior plays a key role in composite’s property. In this present work, two PLLA/HA composites with porous and compact architecture were fabricated and soaked into simulated body fluid (SBF) at 37 °C for in vitro mineralization, respectively. An attenuated total reflection Fourier transform infrared (ATR FTIR) mapping coupled with principal component analysis was developed to investigate the mineralization kinetics. The FTIR images with an area of 300 × 300 μm² were collected every 7 days. The results suggest that the mineralization of PLLA/HA composites in SBF follows a zero-order kinetic model, no matter what the architecture is. However, it follows a second-order model when the composite is degraded in phosphate-buffered saline solution based on our previous work. The mechanisms of the in vitro mineralization kinetics in different submersion solutions are discussed. Our results alert researchers that they should choose the mineralization medium cautiously.     

Highly porous titanium (Ti) scaffolds with bioactive microporous hydroxyapatite/TiO2 hybrid coating layer

Highly porous Ti scaffolds with a bioactive microporous hydroxyapatite (HA)/TiO₂ hybrid coating layer were fabricated using the sponge replication process and micro-arc oxidation (MAO) treatment to produce the porous Ti scaffold and hybrid coating layer, respectively. In particular, the morphology and chemical composition of the hybrid coating layer were controlled by carrying out the MAO treatment in electrolyte solutions containing various concentrations of HA, ranging from 0 to 30 wt.%. The fabricated sample showed high porosity of approximately 70 vol.% with interconnected pores and reasonably high compressive strength of 18 ± 0.3 MPa. Furthermore, the surfaces could be coated successfully with a bioactive microporous HA/TiO₂ hybrid layer. The amount of HA particles in the hybrid coating layer increased with increasing HA content in the electrolyte solution, while preserving the microporous morphology. This hybrid coating improved the osteoblastic activity of the porous Ti scaffolds significantly.