Study on β-TCP Coated Porous Mg as a Bone Tissue Engineering Scaffold Material

Three-dimensional honeycomb-structured magnesium (Mg) scaffolds with interconnected pores of accurately controlled pore size and porosity were fabricated by laser perforation technique. Biodegradable and bioactive β-tricalcium phosphate (β-TCP) coatings were prepared on the porous Mg to further improve its biocompatibility, and the biodegradation mechanism was simply evaluated in vitro. It was found that the mechanical properties of this type of porous Mg significantly depended on its porosity. Elastic modulus and compressive strength similar to human bones could be obtained for the porous Mg with porosity of 42.6%-51%. It was observed that the human osteosarcoma cells (UMR106) were well adhered and proliferated on the surface of the β-TCP coated porous Mg, which indicates that the β-TCP coated porous Mg is promising to be a bone tissue engineering scaffold material.     

The preparation, cytocompatibility, and in vitro biodegradation study of pure β-TCP on magnesium

Biodegradable and bioactive β-tricalcium phosphate (β-TCP) coatings were prepared on magnesium (Mg) in order to improve its biocompatibility by a chemical method. The tensile bonding strength of β-TCP coating and Mg substrate was measured by the standard adhesion test (ISO 13779-4). And the cytocompatibility of β-TCP coated Mg was studied by using human osteoblast-like MG63 cells. It was found that the MG63 cells could grow well on the surface of β-TCP coated Mg and the cell viability on β-TCP coated Mg was above 80% during the cocultivation of MG63 cells and β-TCP coated Mg for 10 days, indicating no cytotoxicity. It was concluded that the β-TCP coated Mg had good cytocompatibility. The degradation of Mg substrate with β-TCP coating in vitro was studied in detail by XRD, EDX, SEM, and ICP. The results showed that a bone-like apatite continually formed on the surface of the sample with the degradation of both Mg substrate and β-TCP coating in Hank’s solution (a simulated body fluid). The biodegradation mechanism was preliminarily analyzed in the paper.     

Synthesis and degradation properties of b\boldsymbol{\beta} -TCP/BG porous composite materials

b\boldsymbol{\beta}-TCP/BG porous composite materials were successfully fabricated by foaming technology. X-ray diffraction was used to determine the crystal structure of powders. The pore size and distribution of the resulting materials were characterized using scanning electron microscopy. The porosity and degradation performance of materials were also investigated. The results showed that the porous composite materials possessed the pore size ranging from 100 to 500 m\boldsymbol{\mu} m in diameter, whereas the interconnection among macrospores was poor. The porosity in materials increased from 58·7% to 63·47% with BG content ranging from 0 to 3 wt%, further increasing of BG content results in a decrease in porosity. The degradation rate of composite materials can be adjusted by varying the BG content.     

A novel bioactive porous bredigite (Ca<Subscript>7</Subscript>MgSi<Subscript>4</Subscript>O<Subscript>16</Subscript>) scaffold with biomimetic apatite layer for bone tissue engineering

The aim of this study was to develop a novel bioactive, degradable and cytocompatible bredigite (Ca₇MgSi₄O₁₆) scaffold with biomimetic apatite layer for bone tissue engineering. Porous bredigite scaffolds were prepared using polymer sponge method. The bredigite scaffolds with biomimetic apatite layer (BTAP) were obtained by soaking bredigite scaffolds in simulated body fluid (SBF) for 10 days. The porosity and in vitro degradability of BTAP scaffolds were investigated. In addition, osteoblast-like cell morphology, proliferation and differentiation on BTAP scaffolds were evaluated and compared with β-tricalcium phosphate (β-TCP) scaffolds. The results showed that BTAP scaffolds possessed 90% of porosity. The degradation of BTAP scaffolds was comparable to that of β-TCP scaffolds. Cells on BTAP scaffolds spread well and presented a higher proliferation rate and differentiation level as compared with those on β-TCP scaffolds. Our results indicated that BTAP scaffolds were degradable and possessed the function to enhance cell proliferation and differentiation, and might be used as bone tissue engineering materials.     

β-磷酸三钙/聚L-乳酸与大鼠骨膜成骨细胞复合骨修复材料的研究

采用溶液浇铸-模压成型-沥滤方法制备了β-TCP/PLLA多孔支架材料, 将支架材料与大鼠骨膜成骨细胞复合获得新型组织工程骨修复材料. 通过抗压强度及压缩模量的表征研究了支架材料的力学性能; 采用SEM观测、MTT法、碱性磷酸酶活性及骨钙素分泌量检测细胞复合材料的体外成骨特性; 通过裸鼠肌袋种植, 以组织学方法评价细胞复合材料的异位成骨能力. 结果表明: β-TCP/PLLA多孔支架材料孔隙率可调, 孔径为100～00μm, 孔道相互贯通; 材料抗压强度和压缩模量随孔隙率的增大而降低, β-TCP复合PLLA后材料的力学性能高于同孔隙率的纯PLLA多孔材料; 复合支架材料适宜骨膜成骨细胞粘附和生长, 无细胞毒性; 骨膜成骨细胞复合β-TCP/PLLA支架材料的体外成骨特性良好, 且具有体内异位成骨能力.     

Microstructure,mechanical property and corrosion behavior of co-continuous β-TCP/MgCa composite manufactured by suction casting

The co-continuous β-TCP/MgCa composite was fabricated by infiltrating MgCa alloy into porous β-TCP using suction casting technique. The microstructure, mechanical property and corrosion behaviors of the composite have been evaluated by means of scanning electron microscopy (SEM), X-ray diffraction (XRD), mechanical testing, electrochemical and immersion tests. It was shown that the composite structure was compact and the interfacial combination between MgCa alloy and β-TCP scaffold was very well. The composite had an ultimate compressive strength of (147 ± 13) MPa, which was near with the natural bone (2–180 MPa) and about 1000-fold higher than that of the original porous β-TCP scaffold, but it still retained over half of the strength of the MgCa bulk alloy. The electrochemical and immersion tests indicated that the corrosion resistance of the composite was better than that of the MgCa bulk alloy, and the corrosion rate of the MgCa matrix alloy was quicker than that of the porous scaffold for the composite. The corrosion products of the composite surface were mainly Mg(OH)₂, hydroxyapatite (HA) and Ca₃(PO₄)₂.     

In Vitro Characterizations of PLLA/β-TCP Porous Matrix Materials and RMSC-PLLA-β-TCP Composite Scaffolds

To develop a novel degradable poly (L-lactic acid)/β-tricalcium phosphate (PLLA/β-TCP) bioactive materials for bone tissueengineering, β-TCP powder was produced by a new wet process. Porous scaffolds were prepared by three steps, i.e. solventcasting, compression molding and leaching stage. Factors influencing the compressive strength and the degradation behaviorof the porous scaffold, e.g. weight fraction of pore forming agent-sodium chloride (NaCl), weight ratio of PLLA: β-TCP,the particle size of β-TCP and the porosity, were discussed in details. Rat marrow stromal cells (RMSC) were incorporatedinto the composite by tissue engineering approach. Biological and osteogenesis potential of the composite scaffold weredetermined with MTT assay, alkaline phosphatase (ALP) activity and bone osteocalcin (OCN) content evaluation. Resultsshow that PLLA/β-TCP bioactive porous scaffold has good mechanical and pore structure with adjustable compressive strengthneeded for surgery. RMSCs seeding on porous PLLA/     

The effect of loading icariin on biocompatibility and bioactivity of porous β-TCP ceramic

In order to enhance the ability of calcium phosphate-based biomaterials for bone defect repair, icariin (Ica), one natural product with ability of promoting osteoblasts differentiation in vitro and enhancing bone formation in vivo, was loaded into porous β-tricalcium phosphate ceramic (β-TCP) disks. The obtained Ica-loaded porous β-TCP ceramic (Ica/β-TCP) disks were characterized by SEM. The SEM photos indicated that the disks had porous structure and the surface morphology of the porous β-TCP ceramic (β-PTCP) disks had no obvious difference from the Ica/β-TCP disks. The Ica release curve of Ica/β-TCP disks showed a burst release during the first 1 day and the concentration of released Ica during the first 3 days had low cytotoxicity. The loading Ica in Ica/β-TCP disks hardly affected the attachment and morphology of Ros17/28 cells, however, the Ica/β-TCP disks were favorable to supporting the proliferation and differentiation of Ros17/28 cells better compared with the β-PTCP disks. There was plenty of bone-like apatite formed on the surface of Ica/β-TCP disks soaked in SBF solution for three days. After back intramuscular implantation of rats for three months, no obvious osteogenic evidence was detected in β-PTCP disks, but new bone formation was observed in Ica/β-TCP disks. Fibrous tissues and slight inflammatory reaction was also found in the Ica/β-TCP disks and β-TCP disks. Therefore, the loading Ica did not change the biocompatibility of β-TCP ceramic, but enhanced the bioactivity of β-TCP ceramic in vivo. The Ica/β-TCP ceramic had potential to be used for bone defect repair.     

Development of porous HAp and β-TCP scaffolds by starch consolidation with foaming method and drug-chitosan bilayered scaffold based drug delivery system

The inability to maintain high concentrations of antibiotic at the site of infection for an extended period of time along with dead space management is still the driving challenge in treatment of osteomyelitis. Porous bioactive ceramics such as hydroxyapatite (HAp) and beta-tri calcium phosphate (β-TCP) were some of the alternatives to be used as local drug delivery system. However, high porosity and high interconnectivity of pores in the scaffolds play a pivotal role in the drug release and bone resorption. Ceftriaxone is a cephalosporin that has lost its clinical popularity. But has recently been reported to exhibit better bactericidal activity in vitro and reduced probability of resistance development, in combination with sulbactam, a β-lactamase inhibitor. In this article, a novel approach of forming HAp and pure β-TCP based porous scaffolds by applying together starch consolidation with foaming method was used. For the purpose, pure HAp and β-TCP were prepared in the laboratory and after thorough characterization (including XRD, FTIR, particle size distribution, etc.) the powders were used for scaffold fabrication. The ability of these scaffolds to release drugs suitably for osteomyelitis was studied in vitro. The results of the study indicated that HAp exhibited better drug release profile than β-TCP when drug was used alone indicating the high influence of the carrier material. However, this restriction got relaxed when a bilayered scaffold was formed using chitosan along with the drug. SEM studies along with EDAX on the drug-chitosan bilayered scaffold showed closest apposition of this combination to the calcium phosphate surface.     

Low temperature fabrication of high strength porous calcium phosphate and the evaluation of the osteoconductivity

Porous NaO₂–MgO–CaO–P₂O₅ bioglass doped beta-tri-calcium phosphate (β-TCP) bioceramic possessing high mechanical properties and well pore structure with high porosity and high pore connectivity has been prepared through dipping method with the porous polyurethane as the pore forming template. The sintering mechanism and the mechanical properties of the bioglass doped β-TCP scaffold have been investigated by the X-ray diffraction (XRD) analysis, Scanning electron microscope (SEM) and thermal differential analysis (DTA). The scaffold’s in vivo osteoconductivity has been evaluated by implantation of scaffolds into the femurs of New Zealand rabbits. The results show that the porous structure can achieve the densification process at a low temperature about 950°C by a solid solution sintering mechanism and hence dense macropore scaffold with a compressive strength of 4.32 MPa when the porosity is 75% has been obtained. The in vivo test shows that the Na₂O–MgO–CaO–P₂O₅ bioglass doped porous β-TCP bioceramic has a relatively fast bone formation after implantation; after 1 month implantation new deposited bone tissue has been detected on the strut of the porous scaffold and degraded particles also has been found on the surface of the new formed bone. After 6 months implantation the porous scaffold has been thoroughly covered with new formed bone. Results show that the Na₂O–MgO–CaO–P₂O₅ bioglass doped porous β-TCP bioceramic is potential bone tissue engineering scaffold for orthopedic use.     

Effects of Mg doping on sol–gel derived nanocrystalline hydroxyapatite thin films

Abstract

Both pure and Mg doped thin films were fabricated by sol–gel dip coating. The films were sintered either at 800 or 1000°C. The average grain size of the films was significantly affected by Mg substitution in the hydroxyapatite (HA) structure and change in the sintering temperature. The grains were considerably larger in the films sintered at higher temperatures. In addition, Mg doped films contained significantly larger grains compared to undoped HA films. Mg doping also caused rodlike grains at 800°C, and led to whitlockite (β-TCP) formation at 1000°C. The ratio of the existing phases was estimated as β-TCP/HA=27 : 73. All the films had rough surfaces with high porosity. It was also observed that undoped films had higher surface roughness than Mg doped ones.     

Hydroxyapatite formation on porous ceramics of alpha-tricalcium phosphate in a simulated body fluid

Alpha-tricalcium phosphate (α-TCP) ceramic is a bioresorbable material that degrades in bone tissue after implantation, since it exhibits higher solubility than beta-tricalcium phosphate (β-TCP) ceramics. The high solubility of α-TCP in an aqueous solution causes its transformation into hydroxyapatite (HAp) through hydrolysis. While one expects the formation of hydroxyapatite after exposure to an aqueous solution mimicking a body environment, we occasionally find variation in HAp formation in the simulated body fluid (SBF). In the present study, HAp formation resulting from exposure to SBF was investigated for some types of α-TCP ceramics with different porosities and specific surface area. Reduced porosity and large surface area of porous specimens may increase the local density of Ca²⁺ in the surrounding SBF to increase the degree of supersaturation with respect to HAp. Thus, the porosity and specific surface area are significant parameters for determining not only bioabsorbability but also the ability to form HAp.     

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.     

磷灰石-硅灰石/β-磷酸三钙复合多孔支架材料的制备和表征

以磷灰石-硅灰石玻璃陶瓷(AW)粉和β-磷酸三钙(β-TCP)粉为原料. 以硬脂酸为致孔剂. 经模压成型、1170℃烧结制备磷灰石-硅灰石/β-磷酸三钙复合多孔支架材料(AW/βTCP). 采用X射线衍射(XRD)、扫描电镜(SEM)、能谱(EDS)、诱导耦合等离子体原子发射光谱(ICP-AES)等方法分析支架的晶相组成、显微结构、物理性能、生物活性和降解性. 将大鼠骨髓间充质干细胞(rMSCs)与支架体外复合培养评价支架的生物相容性. 结果表明: 所制备的AW/β-TCP支架材料的抗压强度达14.3MPa. 孔隙率达66.9%. 孔径为100～700μm. 具有良好的生物相容性、生物活性和降解性. 可作为骨组织工程支架的候选材料.     

负载淫羊藿苷的丝蛋白/β-磷酸三钙复合骨修复材料制备及性能

在多孔β-Ca_3(PO_4)_2(β-TCP)表面沉积含有淫羊藿苷(ICA)的丝蛋白(SF)层,制备可缓释ICA的SFICA/β-TCP骨修复复合材料,研究SF-ICA/β-TCP复合材料的相关性能。结果表明,SF-ICA/β-TCP复合材料中ICA的引入并未改变基体材料的微观形貌与孔隙率;体外释放实验表明,通过负载量的调控,可以实现SF-ICA/β-TCP复合材料中ICA的高浓度释放(2.80×10~(-4) mg/mL至7.00×10~(-4) mg/mL)和低浓度释放(5×10~(-6) mg/mL至1.0×10~(-5) mg/mL),累计释放量分别达到约5.2×10~(-3) mg和7.0×10~(-5) mg;细胞增殖实验与电镜观察表明,SF-ICA/β-TCP复合材料中ICA的负载对小鼠颅顶前骨细胞的增殖无显著性影响;但碱性磷酸酶活性检测实验表明,负载高含量ICA的SF-ICA/β-TCP复合材料中的细胞具有较高的碱性磷酸酶表达。所制备的负载ICA的SFICA/β-TCP复合材料在体内骨修复领域具有潜在的应用前景。     

Microstructure,mechanical property and corrosion behavior of interpenetrating (HA + β-TCP)/MgCa composite fabricated by suction casting

The novel interpenetrating (HA + β-TCP)/MgCa composites were fabricated by infiltrating MgCa alloy into porous HA + β-TCP using suction casting technique. The microstructure, mechanical properties and corrosion behaviors of the composites have been evaluated by means of scanning electron microscopy (SEM), X-ray diffraction (XRD), mechanical testing, electrochemical and immersion tests. It was shown that the composites had compact structure and the interfacial bonding between MgCa alloy and HA + β-TCP scaffolds was very well. The ultimate compressive strength of the composites was about 500–1000 fold higher than that of the original porous scaffolds, and it still retained quarter-half of the strength of the bulk MgCa alloy. The electrochemical and immersion tests indicated that the corrosion resistance of the composites was better than that of the MgCa matrix alloy, and the corrosion products of the composite surface were mainly Mg(OH)₂, HA and Ca₃(PO₄)₂. Meanwhile, the mechanical and corrosive properties of the (HA + β-TCP)/MgCa composites were adjustable by the choice of HA content.     

Processing and characterization of porous alumina scaffolds

Bioceramic materials are used for the reconstruction or replacement of the damaged parts of the human body. In this study an improved procedure is described for producing ceramic scaffolds with controlled porosity. Bioinert alumina ceramic was used to make porous scaffolds by using indirect fused deposition modeling (FDM), a commercially available rapid prototyping (RP) technique. Porous alumina samples were coated with hydroxyapatite (HAp) to increase the biocompatibility of the scaffolds. Initial biological responses of the porous alumina scaffolds were assessed in vitro using rat pituitary tumor cells (PR1). Both porous alumina and HAp coated alumina ceramics provided favorable sites for cell attachments in a physiological solution at 37 °C, which suggests that these materials would promote good bonding while used as bone implants in vivo. Based on these preliminary studies, similar tests were performed with human osteosarcoma cells. Cell proliferation studies show that both the ceramic materials can potentially provide a non-toxic surface for bone bonding when implanted in vivo.     

Preparation of PLGA/β-TCP composite scaffolds with supercritical CO 2 foaming technique

A high porosity scaffold with suitable compressive strength prepared by a gentle method has become a pressing need. To meet this demand, poly(DL-lactide-co-glycolide) (PLGA) and β-tricalcium phosphate (β-TCP) were designed to prepare composite scaffolds by the supercritical technique. The preparation process consisted of three units: the mixing of PLGA and β-TCP, compression molding of the mixture, and the foaming process. Six influencing factors — temperature, pressure of the scCO₂ system, maintaining time of scCO₂, the ratio of β-TCP to PLGA, the rate of depressurization, and the molecular weight — were investigated. The results collectively indicated that the optimized conditions for the foaming process were that CO₂ pressure and temperature be 8MPa and 39°C, respectively, which should be kept for 8h; the content of β-TCP in the mixture should be 25% and the depressurizing rate be 0.1 MPa/s, using PLGA of an 80kDa molecular weight. Scaffolds with a porosity of 65.47% and a compressive strength of 4.76 MPa could be obtained. The pore size ranged around 100 µm. The material’s use as tissue engineering scaffolding is expected.     

多孔胶原-β-磷酸三钙-硫酸软骨素复合膜的制备与表征

通过碳化二亚胺(EDC)改性、二次冻干制备多孔胶原-β-磷酸三钙-硫酸软骨素复合膜材料.通过扫描电镜(SEM)、X射线衍射分析仪(XRD)与原子力显微镜(AFM)考察了组分变化与制备过程中复合材料的微观形貌变化,并进一步利用红外、孔隙率、MTT细胞毒性实验等分析手段对复合材料的结构与性能进行了表征.实验结果表明,当胶原盐酸溶解液pH=2,胶原与β-磷酸三钙质量比为1∶2(m(Col)∶m(β-TCP)=1∶2)时,复合材料中β-TCP晶相保持较好,其与胶原之间的排列结合最为均匀紧密.经EDC改性后,SEM与AFM实验均显示了交联后的胶原束明显变大变粗,以一定的方向紧密地排列在一起.XRD图谱显示复合材料中β-TCP特征衍射峰明显.复合材料的孔径为80～90 μm,三元膜孔隙率为(90.76士1.28)％,大于纯胶原冻干膜(85.88士0.92)％;红外光谱证实β-TCP中的钙离子与Col上的羧基发生了化学键合,AFM显示β-TCP颗粒能与胶原发生直接联结.复合材料的MTT实验结果为1级,是一种潜在的口腔修复膜材料.     

Preparation and in vitro evaluation of mesoporous hydroxyapatite coated β-TCP porous scaffolds

A mesoporous hydroxyapatite (HA) coating was prepared on a β-tricalcium phosphate (β-TCP) porous scaffold by a sol-gel dip-coating method using the block copolymer Pluronic F127 (EO₁₀₆PO₇₀EO₁₀₆) as the template. For application as a bone graft, in vitro cell response and bone-related protein expression of mesoporous HA coated β-TCP scaffold were investigated, using the non-mesoporous HA coated scaffold as the control group, to evaluate the influence of the mesoporous structure on the biological properties of HA coating. It was found that the increased surface area of the mesoporous HA coating greatly affected the response of MC3T3-E1 osteoblasts and the expression of proteins. An enzyme-linked immunosorbent assay recorded a significantly higher expression of alkaline phosphatase (ALP) and bone sialoprotein (BSP) in the mesoporous group than those in the control group (*p < 0.05) after different incubation periods. The introduction of mesopores enhanced the expression of ALP and BSP in the cells grown on the mesoporous HA coatings, on the premise of maintaining the protein expression in a sequence to ensure the correct temporo-spatial expression in osteogenesis. These results indicated that the mesoporous HA coating would provide a good environment for cell growth, suggesting that it could be used as the coating material for the surface modification of the tissue engineering scaffolds.