蜡球造孔法制备多孔HA陶瓷支架及其性能优化

组织工程支架的贯通性对其体内生物学表现具有重要影响。采用甲壳素溶胶体系和蜡球造孔剂制备多孔羟基磷灰石(HA)陶瓷支架, 考查在相同模压条件下, 不同浆料/造孔剂比例对多孔HA陶瓷支架的孔隙率、收缩率、贯通性、多孔结构以及抗压强度的影响。结果表明: 该方法可以制备具有高孔隙的多孔HA陶瓷支架, 随着造孔剂比例的增大, 支架的贯通性更好, 当浆料/造孔剂比例为1:1.2时可以得到孔隙率、贯通性、力学性能最优的多孔HA陶瓷支架。     

纳米羟基磷灰石/聚己内酯复合生物活性多孔支架研究

采用水热法制备了纳米羟基磷灰石（n-HA）及其与聚己内酯（PCL）的复合材料. 用熔融浇铸/食盐微粒浸出法制备了孔径在200～400μｍ、大孔互相贯通的复合材料支架. 通过细胞培养和体内动物实验研究了该支架的生物学性能. 结果表明,复合支架的孔隙率随致孔剂用量的增加而增加,而抗压强度随之而减小;支架的最大孔隙率可达86%,相应的抗压强度为2.4MPa. 成骨细胞在支架上的细胞粘附率和增殖随磷灰石含量增加而提高,复合材料明显高于单纯的PCL支架. 组织学观察显示,新生骨长入多孔支架和复合材料形成了直接的骨性结合. n-HA/PCL复合材料支架有很好的生物相容性和生物活性.     

Nanostructured bioactive glass coating on porous hydroxyapatite scaffold for strength enhancement

Many attempts have been focused on preparing highly porous scaffolds with appropriate mechanical strength. This paper has developed a new route to enhance the compressive strength of porous HA (hydroxyapatite) scaffold (porosity: ∼ 83%, mean pore size:∼ 800 μm). Briefly this route included nanostructure coating of bioactive glass on struts of porous HA. Coating microstructure consisted of the grains with the range between 91 and 320 nm and micron size pores that could be detected by SEM observation. This simple method improved the compressive strength of highly porous HA from 0.22 to 1.49 MPa. The obtained scaffolds provided good mechanical support while maintaining bioactivity so they could be used as tissue engineering scaffolds for low-load bearing applications.     

Nanoindentation on porous bioceramic scaffolds for bone tissue engineering

We report nanoindentation mechanical properties measurements on porous ceramic scaffolds made for tissue engineering applications. The scaffolds have been made from tricalcium phosphate (TCP), hydroxyapatite (HA) nanopowder and mixed powders of HA (50 wt%) and TCP (50 wt%) using the polyurethane sponge method, which produces open porous ceramic scaffolds through replication of a porous polymer template. The scaffolds prepared by this method have a controllable pore size and interconnected pore structure. The crystal structures and morphology of porous scaffolds were determined by X-ray diffraction (XRD) and atomic force microscopy (AFM) respectively. Nanoindentation measurements to a depth of 600 nm showed a Young's modulus value of 10.3 GPa for HA+TCP composite scaffolds and 1.5 GPa for TCP scaffolds. The hardness values were 240 MPa for HA+TCP composites and 21 MPa for TCP sample respectively. The results showed that the mechanical properties of the biodegradable scaffolds can be considerably enhanced with the addition of HA while maintaining the interconnected open pores and pore geometry desirable for bone tissue engineering.     

Biodegradable poly(lactic acid)/hydroxyl apatite 3D porous scaffolds using high-pressure molding and salt leaching

Scaffolds comprising poly(lactic acid) (PLA) and hydroxyl apatite (HA) were fabricated by combination of the high-pressure compression-molding plus salt-leaching techniques. The optimized HA content was determined in terms of the pore morphology, porosity, storage modulus, degradation behavior, hydrophilicity as well as the cell growth ability of the scaffolds. At HA content of 20 wt%, the scaffolds exhibited an interconnected open pore structure with the high porosity of 82.2 %. More importantly, the storage modulus of PLA/HA scaffolds (87.6 MPa) achieved almost three times higher compared with pure PLA scaffolds, while under low-pressure condition, the increase of modulus caused by HA does not reach 150 %. The obvious contrast indicated that HA and high pressure had a synergistic effect on enhancing mechanical properties of porous scaffolds. It was truly interesting that the hydrophilicity of PLA/HA scaffolds was significantly improved by alkaline hydrolysis treatment, which eventually led to the excellent cellular biocompatibility of the scaffolds, as revealed from the morphology and spreading of the cells cultured in our scaffolds. On the whole, the resultant PLA/HA scaffolds are well-suited candidates for the design of tailor-made matrices in tissue engineering.     

Fabrication of three dimensional polymeric scaffolds with spherical pores

This paper reports polymeric scaffolds with spherical internal macropores and relatively large external dimension. Paraffin spheres with the diameter of several hundred microns were prepared by a suspension technique. Particulate leaching technique based on this kind of spherical porogens was combined with room-temperature compression molding technique to fabricate biodegradable poly(D,L-lactic-co-glycolic acid) (PLGA) porous scaffolds potentially for tissue engineering or in situ tissue induction. The scaffolds exhibited ordered macropores with good pore interconnectivity. The porosity ranged from 80 to 97% adjusted simply by varying porogen content. The foams with porosity around 90% have compressive modulus over 3 MPa and compressive strength over 0.2 MPa. As preliminary cell experiments with 3T3 fibroblasts cultured on the porous scaffolds indicate, the processing procedure of the scaffolds has not brought with problem in cytotoxicity.     

基于Micro-CT不同多孔结构陶瓷支架的建模及其贯通性与液流分布分析

贯通性是骨组织工程支架的重要参数, 决定包含蛋白和细胞的体液渗入和组织生长。本研究采用Micro-CT技术对三种不同工艺(球粒堆积、蜡球造孔、纤维堆积)构建的羟基磷灰石多孔支架进行断层扫描, 并从三方面研究支架的贯通性: (1)通过影像重建定量分析支架孔隙的三维贯通结构; (2)统计分析比较三种支架在贯流方向上的孔隙率变化; (3)有限元模拟支架的内部液流分布情况。结果表明, 球粒堆积支架与蜡球造孔支架孔隙率分布较均匀, 而纤维堆积支架孔隙分布较杂乱。液流模拟(流速分布)发现, 球粒堆积支架与蜡球造孔支架中液体流动均匀, 但是蜡球造孔支架孔壁近表面区域存在大量“漩涡流”, 不利于支架内细胞与液流之间的物质交换, 该结果有可能解释球粒堆积支架体内成骨性优于蜡球造孔支架的动物体内实验结果。     

Morphology, mechanical properties, and mineralization of rigid thermoplastic polyurethane/hydroxyapatite scaffolds for bone tissue applications: effects of fabrication approaches and hydroxyapatite size

Rigid thermoplastic polyurethane (TPU)/hydroxyapatite (HA) scaffolds were prepared with micro HA (mHA) and nano HA (nHA) particles, respectively, via the thermally induced phase separation method. The effects of solvent and co-solvent, addition of sodium chloride (NaCl) porogen, and HA particle size were studied together with the morphology, compressive properties, and mineralization behavior of the scaffolds. Depending on the solvent, co-solvent, or porogen used, different porous structures were produced. In particular, a ladder-like morphology was obtained when dioxane (Di) was used as the solvent, whereas an interconnected porous structure was obtained by using dioxane and deionized water (DiW) as co-solvents. Rectangular pores with interconnected channels on the pore walls were achieved by using NaCl crystals as porogens. The TPU/nHA scaffolds showed stronger compressive properties than the TPU/mHA scaffolds and the pure TPU scaffolds. The scaffolds prepared using dioxane and water as co-solvents exhibit the greatest compressive modulus. Furthermore, TPU scaffolds with nHA particles had the ability to form bone apatite when soaked in simulated body fluid (SBF). After being soaked in SBF for 3 weeks, the weight percentage of formed apatite in the TPU/nHA-DiW scaffold was 9.2 %wt of the initial TPU content. Preliminary cytotoxicity tests were conducted using NIH 3T3 fibroblast cells. The high survival rate of these cells and the mineralization behavior suggest biocompatibility and high potential of these composites being used in bone tissue engineering applications.     

Structural characterization and strengthening mechanism of forsterite nanostructured scaffolds synthesized by multistep sintering method

In this study, highly porous forsterite scaffolds with interconnected porosities were synthesized using multi-step sintering (MSS) method. The starting powder was nanosized forsterite, which was synthesized from talc and magnesium carbonate powders. The phase composition, average particle size and morphology of the produced forsterite powder were characterized by X-ray diffraction technique (XRD) and transition electron microscopy (TEM). Forsterite scaffolds were produced by foamy method using polymeric sponges. MSS process including three steps was used to efficiently sinter the forsterite nanopowders without destroying the initial porous structure of polymeric sponges. The results showed that MSS technique is an efficient and appropriate procedure to produce highly porous forsterite scaffolds with pore size in the range of 100–300?μm. The compressive strength, compressive modulus and porosity of C12 specimen (sintered at 1650?°C for 1?h with subsequent annealing at 1000?°C for 1000?min) was 1.88?MPa, 29.2?MPa, and 72.4%, respectively, which is very close to that of cancellous bone. The approach studied in this research can be developed for other nanostructure ceramics to produce highly porous scaffolds with interconnected porosities for load bearing applications.     

Fabrication and characterization of chitosan microspheres agglomerated scaffolds for bone tissue engineering

In the presented paper authors describe a method for bone scaffolds fabrication. The technique is based on the agglomeration of chitosan microspheres. The fabrication process is complex and consists of a few steps: chitosan spheres extrusion, scaffold formation by compression followed by the spheres agglomeration and bonding with cross-linking agent (STPP, sodium tripolyphosphate). The described method allows manufacturing of porous materials with controllable shape, pore size distribution and their interconnectivity. In this technique 3D scaffold porosity can be regulated by altering spheres diameter. Authors studied influence of cross-linker concentrations and time of cross-linking process on the scaffold morphology, mechanical properties, enzymatic degradation rate (in the presence of lysozyme) and human osteoblasts response. Surface morphology and topography were evaluated by SEM. Porosity and pore interconnectivity were observed via μCT scanning. Mechanical tests showed that chitosan scaffolds perform compression characteristic (Young Modulus) similar to natural bone. Cytotoxicity established by XTT assay confirmed that most of the developed composite materials do not show toxic properties. Osteoblast adhesion and morphology were analyzed by SEM and optical microscopy.     

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.     

凝胶注模成型法制备多孔羟基磷灰石陶瓷

采用凝胶注模成型工艺制备了多孔羟基磷灰石陶瓷,并通过X射线分析了多孔陶瓷的相成分,采用扫描电镜观测了孔隙结构和形貌.结果表明,所制备的多孔羟基磷灰石陶瓷的孔隙率均大于80%;孔隙尺寸主要分布在350～600μm,孔壁上存在孔径为60～190μm的贯通孔;X射线衍射证明烧结过程未引入异质成分.所制备的多孔羟基磷灰石陶瓷具有适宜的孔隙直径和孔隙率,且孔隙间具有良好的贯通性.     

Hydroxyapatite (HA) bone scaffolds with controlled macrochannel pores

Hydroxyapatite (HA) macrochanneled porous scaffolds, with a controlled pore structure, were fabricated via a combination of the extrusion and lamination processes. The scaffold was architectured by aligning and laminating the extruded HA and carbon filaments. The macrochannel pores were formed by removing the carbon filaments after thermal treatments (binder removal and sintering). The porosity of the scaffolds was varied between 48 and 73% with a controlled pore size of ∼450 μm, by adjusting the fractions of HA and carbon filaments. As the porosity was increased from 48 to 73%, the compressive strength decreased from 11.5 to 3.2 MPa. However, the osteoblast-like cell responses on the scaffold, such as the proliferation rate and alkaline phosphatase (ALP) activity, were significantly enhanced as the porosity was increased.     

聚乳酸涂层改善网状羟基磷灰石陶瓷支架力学性能的研究

采用有机泡沫浸渍法制备出高孔隙率的三维网状羟基磷灰石(HA)生物陶瓷支架,并采用不同浓度的聚乳酸(PDLLA)溶液涂覆网状HA支架,以达到增强补韧网化陶瓷支架的目的.通过扫描电镜观察PDLLA涂覆网状HA陶瓷支架的显微结构和测试力学强度,研究了不同浓度PDLLA溶液涂覆对网状HA支架的涂层厚度、显微结构和力学性能的影响.实验结果表明:涂覆一定量的PDLLA涂层可以显著提高网状HA生物陶瓷支架的抗压强度,同时保持网状陶瓷支架的贯通性和孔隙率.     

Preparation of 3-D regenerated fibroin scaffolds with freeze drying method and freeze drying/foaming technique

Although three-dimensional fibroin scaffolds have been prepared with freeze drying method, the porosity and pore sizes still can not satisfy the requirement of tissue engineering. In this article, fibroin porous scaffold with high porosity and > 100μm diameter interconnected pores was firstly prepared with freeze drying method through adjusting fibroin concentration. The morphology of different scaffolds lyophilized from different fibroin concentration was observed by SEM. A novel freeze drying improved method, freeze drying/foaming technique, was also devised to prepare fibroin scaffolds at different fibroin concentrations. Using the said method, the porosity and pore size of fibroin scaffolds prepared from 12% concentration were 85.8 ± 4% and 109 ± 20 μm respectively with yield strength up to 450 ± 6 KPa while the porosity and pore size of fibroin scaffolds prepared from 8% concentration were 96.9 ± 3.6% and 120 ± 30 μm respectively with yield strength up to 30 ± 1 KPa. The freeze drying/foaming technique produced scaffolds with a useful combination of high yield strength, interconnected pores, and pore sizes greater than 100 μm in diameter. Through adjusting fibroin concentration and thawing time, the porosity, pore sizes and mechanical properties could be controlled to satisfy the different requirements of tissue engineering. The results suggested that fibroin scaffolds prepared with the above methods could be formed for utility in biomaterial application.     

具有良好贯通性的颗粒造孔支架的制备及表征

支架孔隙贯通性的研究一直是多孔生物陶瓷的研究重点之一.采用石蜡球作为造孔剂, 在常规的颗粒造孔法制备多孔陶瓷支架的基础上,通过二甲苯处理以便在石蜡球间形成桥联结构, 以扩大颗粒间的接触面积,从而提高多孔陶瓷支架的孔隙贯通性. 借助扫描电镜(SEM)观察陶瓷支架的多孔结构,评价二甲苯处理石蜡球对陶瓷支架孔隙贯通性的改善效果; 采用密度法测定了陶瓷支架的孔隙率并计算其收缩率,并用成骨细胞评价陶瓷支架的细胞相容性. 结果表明,通过二甲苯的处理, 不仅改善了陶瓷支架孔隙的贯通性,而且提高了其孔隙率, 但孔隙率对陶瓷支架的收缩率无明显影响.细胞培养实验显示成骨细胞可进入多孔陶瓷支架内部, 并在材料表面正常生长,贯通性好的多孔陶瓷支架可为成骨细胞生长提供更充分的空间.     

具有复杂形状的聚ε-己内酯多孔支架的模压制备方法

通过聚ε-己内酯(PCL)支架的制备，尝试了一种制备具有复杂形状的组织工程三雏多孔支架的新方法一改进的模压／粒子浸出法，并对所得外耳状多孔支架的形态、孔结构和孔隙率进行了表征。模压针对聚ε-己内酯熔体和大量盐粒的混合物进行。该方法所得支架孔隙率高达90％以上。可望用于各种不同复杂形状的三雏多孔支架的制备。     

Chitosan-Based Nanocomposite Scaffolds for Tissue Engineering Applications

This article reports the fabrication of three-dimensional porous chitosan and hydroxyapatite (HA)/chitosan composite scaffolds by the thermally induced phase separation (TIPS) technique, for bone tissue engineering. Different amounts of HA nanoparticles (10%, 20%, and 30% g/g) were added to the chitosan solution to produce HA/chitosan composite scaffolds of varying compositions. The morphology and pore structure of the scaffolds vis-à-vis composition were characterized using scanning electron microscopy (SEM) and an energy dispersive X-ray (EDX). Both pure chitosan and HA/chitosan composite scaffolds were highly porous and had interconnected pores. The pore sizes ranged from several micrometers to a few hundred micrometers. The HA nanoparticles were well dispersed and physically coexisted with chitosan in the composite scaffolds. However, some agglomeration of HA nanoparticles was observed on the surface of pore walls when a relatively large amount of HA was used. The composite 3D scaffolds are very promising for use in bone tissue engineering 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.     

Porous scaffolds of polycaprolactone reinforced with in situ generated hydroxyapatite for bone tissue engineering

Polycaprolactone/hydroxyapatite (PCL/HA) composites were prepared by in situ generation of HA in the polymer solution starting from the precursors calcium nitrate tetrahydrate and ammonium dihydrogen phosphate via sol–gel process. Highly interconnected porosity was achieved by means of the salt-leaching technique using a mixture of sodium chloride and sodium bicarbonate as porogens. Structure and morphology of the PCL/HA composites were investigated by scanning electron microscopy, and mechanical properties were determined by means of tensile and compression tests. The possibility to employ the developed composites as scaffolds for bone tissue regeneration was assessed by cytotoxicity test of the PCL/HA composites extracts and cell adhesion and proliferation in vitro studies.