改进粒子沥滤法制备PLA/HA支架及其性能

采用溶剂浇铸/真空挥发/粒子沥滤法(SC/VV/PL)制备了聚乳酸(PLA)和PLA/羟基磷灰石(HA)多孔支架,研究了支架的结构、力学性能、亲水性能等.从扫描电镜结果可以看出支架孔径与所用的致孔剂氯化钠(NaCl)的粒径符合良好,PLA和PLA/HA支架的孔隙率均大于79%,压缩模量、接触角、吸水率的测试结果表明,HA的加入显著改善了PLA支架的力学性能和亲水性能.     

A novel fabrication procedure for producing high strength hydroxyapatite ceramic scaffolds with high porosity

Hydroxyapatite (HA) scaffolds were fabricated using the space holder method with a pressureless sintering process in a systematically developed manner at different fabrication stages to increase the strength of the scaffold at high porosity. Polyvinyl alcohol (PVA) and Polymethyl methacrylate (PMMA) were used as binders and space holder agents, respectively. The physical properties of the HA scaffolds were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM), linear shrinkage test, and porosity measurements. The mechanical properties of the HA scaffolds were analyzed using compressive strength measurements. The results revealed that the HA scaffold met the expected quality requirements with a compressive strength of 2.2 MPa at a porosity of 65.6% with pore sizes distributed in the range of 126–385 μm. The shrinkage of the scaffold diameter occurred by 20.27%, this diameter shrinkage predominantly to the shrinkage of the HA scaffold caused by sintering. Besides, suspect that a higher PMMA concentration causes pore size shrinkage upon sintering. The formation of pore interconnections was evidenced by SEM observations and the ‘translucent light method’ developed in this study. The results of the scaffold phase test using XRD showed that the final scaffold consisted only of the HA phase, as the PVA and PMMA phases burned out during the sintering process.     

Formation of porous PLGA scaffolds by a combining method of thermally induced phase separation and porogen leaching

A novel method for the fabrication of porous poly(L -lactide-co-glycolide) (PLGA) scaffolds by combining thermally induced phase separation and porogen leaching is presented in this article. Big pores with about 75–400 μm diameters in the obtained scaffolds were generated by the porogen, sucrose particles, while small pores with diameters less than 20 μm induced via phase separation. Extraction of the solvent, chloroform by ethanol at cool temperatures could reduce the scaffold toxicity. Effects of PLGA concentration, freezing temperature, volume fraction of porogen, and introduction of β-tricalcium phosphate (β-TCP) on morphology, porosity, and compressive properties of the scaffolds were systematically discussed. Results showed that the size of small pores decreased by decreasing the polymer concentration and reducing the freezing temperature, whereas the interconnectivity of the scaffolds was improved by increasing the porogen fraction. The compressive modulus and strength were significantly lowered by increasing the scaffold porosity, that is, by increasing porogen fraction, or decreasing the polymer concentration, or reducing the freezing temperature. Addition of β-TCP into the scaffolds did not influence the compressive modulus significantly but tended to decrease the compressive strength. The obtained scaffolds with diverse pore sizes would be potentially used in bone tissue engineering. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

骨组织工程用PLGA多孔支架的制备及细胞毒性研究

制备能在骨组织工程研究中应用,并具有良好孔隙结构的块状聚（D,L-乳酸-CO-乙醇酸）（PLGA）多孔支架,探索出以冰粒子作为致孔剂,采用粒子滤出方法结合冷冻干燥工艺制备多孔支架的方法.首先将冰颗粒加入预冻的PLGA氯仿溶液中混合均匀,然后把混合物置于液氮中深度冷冻后冷冻干燥,制得多孔支架.对支架孔隙结构分析表明,该工艺制备的多孔支架无致孔剂残留、三维结构良好、孔径与孔隙可通过改变冰粒子的粒径和质量分数来控制;细胞毒性实验表明该多孔支架毒性在0～1级,可作为骨组织工程研究用多孔支架.     

Morphology,wettability, and mechanical properties of polycaprolactone/hydroxyapatite composite scaffolds with interconnected pore structures fabricated by a mini‐deposition system

A new mini‐deposition system (MDS) was developed to fabricate scaffolds with interconnected pore structures and anatomical geometry for bone tissue engineering. Polycaprolactone/hydroxyapatite (PCL/HA) composites with varying hydroxyapatite (HA) content were adopted to manufacture scaffolds by using MDS with a porosity of 54.6%, a pore size of 716 μm in the x‐y plane, and 116 μm in the z direction. The water uptake ratio and compressive modulus of PCL/HA composite scaffold increase from 8 to 39% and from 26.5 to 49.8 MPa, respectively, as the HA content increases from 0 to 40%. PCL/HA composite scaffolds have better wettability and mechanical properties than pure PCL scaffold. A PCL/HA composite scaffold for mandible bone repair was successfully fabricated with both interconnected pore structures and anatomical shape to demonstrate the versatility of MDS. POLYM. ENG. SCI., 2012. © 2012 Society of Plastics Engineers     

Three-Dimensional Microstructural Characterization of Porous Hydroxyapatite Using Confocal Laser Scanning Microscopy

The characterization of porosity is crucial in the development and commercialization of ceramic bone replacement technology, since the pore size and interconnectivity play a central role in both biological function (bone ingrowth and nutrient flow) as well as mechanical properties of bone scaffolds. The ability of confocal laser scanning microscopy (CLSM) to image three-dimensional (3-D) structures with large vertical depths (∼2 mm) and fine vertical resolution (∼1 μm) is utilized in this article to characterize the 3-D microstructures of hydroxyapatite (HA) bone scaffold specimens with porosity ranging from roughly 60–70 vol%. Various CLSM techniques are applied to image and interpret the HA pore structure, including Z -series stacking, topographic profiling, and Phi- Z scanning and contour mapping.     

Polymer template fabrication of porous hydroxyapatite scaffolds with interconnected spherical pores

Porous hydroxyapatite (HA) scaffolds with interconnected spherical pores were fabricated by slip casting using a polymer template. Templates were produced using polymer beads, NaCl, and adhesive (N100). Effects of the preparation process on the pore structures and mechanical properties of the porous HA scaffolds were investigated. Pore interconnectivity was improved by adding NaCl particles with appropriate diameters to the polymer template. The size of the adhesive area could be controlled by adjusting the concentration of N100. The pore size could be controlled between 200 ± 42 and 400 ± 81 μm, and the porosity between 50.2 and 73.1%, by changing the size of the polymer beads and the volume of the NaCl particles. The compressive strength decreased as the porosity or pore size increased.     

Fabrication of Hydroxyapatite/Ethylene-Vinyl Acetate/Polyamide 66 Composite Scaffolds by the Injection-Molding Method

This research investigated the injection-molding techniques to produce hydroxyapatite (HA)/ethylene-vinyl acetate (EVA)/polyamide 66 (PA66) composite scaffolds. The effects of HA, EVA, azodicarbonamide (AC) content and shot size on the mechanical properties, pore morphology, porosity and crystallization behavior of the composite scaffolds were analyzed by XRD, DSC, SEM and mechanical test. The compressive modulus and strength of the HA/EVA/PA66 scaffolds with a pore size of 200–600 µm are close to the cancellous bone. Compared with common methods to fabricate scaffolds, this process makes the fabrication of composite scaffolds come true in a rapid and convenient manner.     

Fabrication of polylactic acid/polyethylene glycol (PLA/PEG) porous scaffold by supercritical CO2 foaming and particle leaching

PLA/PEG/NaCl blends were melt‐blended followed by gas foaming and particle leaching process to fabricate porous scaffold with high porosity and interconnectivity. A home‐made triple‐screw compounding extruder was used to intensify the mixability and dispersion of NaCl and PEG in the PLA matrix. Supercritical carbon dioxide was used as physical blowing agent for the microcellular foaming process. Sodium chloride (NaCl) was used as the porogen to further improve the porosity of PLA scaffold. This study investigated the effects of PEG and NaCl on the structure and properties of the PLA‐based blend, as well as the porosity, pore size, interconnectivity, and hydrophilicity of porous scaffolds. It was found that the incorporation of PEG and NaCl significantly improved the crystallization rate and reduced viscoelasticity of PLA. Moreover, scaffolds obtained from PLA/PEG/NaCl blends had an interconnected bimodal porous structure with the open‐pore content about 86% and the highest porosity of 80%. And the presence of PEG in PLA/NaCl composite improved the extraction ability of NaCl particles during leaching process, which resulted in a well‐interconnected structure. The biocompatibility of the porous scaffolds fabricated was verified by culturing fibroblast cells for 10 days. POLYM. ENG. SCI., 55:1339–1348, 2015. © 2015 Society of Plastics Engineers     

Injection‐molded porous hydroxyapatite/polyamide‐66 scaffold for bone repair and investigations on the experimental conditions

Hydroxyapatite/polyamide‐66 (HA/PA66) composite scaffolds were prepared using injection‐molding technique and also analyzed by means of scanning electron microscopy, X‐ray diffraction, differential scanning calorimetry, Fourier transform infrared spectroscopy, and mechanical testing. Compared with common methods to fabricate scaffolds, this process can fabricate composite scaffolds in a rapid and convenient manner by adjusting the experimental conditions of foaming agent and shot size. The interactions between PA66 and HA particles affect the crystallization behavior of PA66 and the pore structure of scaffolds. HA particles can increase the stiffness of composite scaffolds accompanied by the reduction of impact strength, pore size and porosity. The obtained 40 wt% HA/PA66 composite scaffolds with a pore size ranging from 100–500 μm and a porosity more than 65% can simultaneously meet the requirements of porous structure and mechanical performance. POLYM. ENG. SCI., 54:1003–1012, 2014. © 2013 Society of Plastics Engineers     

Ultrasonication‐Induced Modification of Hydroxyapatite Nanoparticles onto a 3D Porous Poly(lactic acid) Scaffold with Improved Mechanical Properties and Biocompatibility

A 3D porous poly(lactic acid) (PLA) scaffold with high porosity and well‐connected pores is fabricated using a vacuum‐assisted solvent casting technique. Its surface is modified with hydroxyapatite (HA) nanoparticles using ultrasonication to prepare an HA‐modified PLA/HA scaffold. For reference, an HA‐blended (b‐PLA‐HA) scaffold is fabricated via the solution blending method. The morphology, porosity, hydrophilicity, swelling ratio, mechanical properties, and cell viability of the PLA, b‐PLA‐HA, and PLA/HA scaffolds are systematically studied. The results show that HA nanoparticles are successfully introduced onto the surface of the PLA/HA scaffold, and strong interactions occur between the HA nanoparticles and the PLA matrix. The PLA/HA scaffold still has a high porosity of more than 85% after ultrasonication. The hydrophilicity and mechanical properties of the PLA/HA scaffold are significantly higher than those of the PLA and b‐PLA‐HA scaffolds. Compared with the PLA and b‐PLA‐HA scaffolds, the attachment and growth of mouse embryonic osteoblasts cells (MC3T3‐E1) cultured on the PLA/HA scaffold significantly improve, due to most HA nanoparticles on the surface, resulting in a good and direct interaction between the cells and the scaffold. Therefore, the PLA/HA scaffold possesses great potential to be used as a tissue engineering scaffold.     

Biocompatibility and strengthening of porous hydroxyapatite scaffolds using poly(l-lactic acid) coating

Hydroxyapatite (HA) is a well-known biocompatible bone substitute. Porous HA is more resorbable and osteoconductive compared with non-porous HA, and has been studied both experimentally and clinically. However, the mechanical strength of porous HA scaffolds is known to be weak. In this study, we developed a porous HA scaffold coated with a synthetic biodegradable polymer, poly(l-lactic acid) (PLLA), to strengthen the scaffold. PLLA-coated HA pellets were used to investigate the in vitro proliferation and alkaline phosphatase (ALP) activity of osteoblasts. PLLA-coated porous HA scaffolds were observed using scanning electron microscopy to investigate surface characteristics, porosity, and mechanical strength. PLLA coating concentration varied from 2 to 10 wt%. Osteoblast proliferation was higher in HA samples coated with PLLA compared with non-coated. ALP activity was highest on 8 wt% PLLA-coating after 3 days and on 4 wt% and 6 wt% PLLA after 9 and 12 days. Porous HA scaffolds with higher concentrations of PLLA were found to have a smoother, flatter surface. This enhanced proliferation and attachment of osteoblasts onto the porous HA scaffold. PLLA solution at a concentration of 10 wt% decreased scaffold porosity to half that of HA scaffolds with no PLLA coating. Scaffold mechanical strength was increased two-fold with a PLLA concentration of 2 wt%. Based on in vitro experimentation, it can be concluded that PLLA-coating on porous HA scaffolds enhances both the biocompatibility and the mechanical strength.     

Modification of hydroxyapatite (HA) powder by carboxymethyl chitosan (CMCS) for 3D printing bioceramic bone scaffolds

The poor mechanical properties of 3D printed HA bone scaffold is always a challenge in tissue engineering, to address this issue, carboxymethyl chitosan (CMCS) was proposed to modify HA bone scaffolds by a physical blending method in this research. A series of HA and HA/CMCS composite ceramic scaffolds were printed by using piezoelectric inkjet 3D printing technology, and their properties were investigated in terms of forming quality, structural morphology, mechanical properties, degradability, cytotoxicity, and cell adhesion growth. The results of forming quality and structural morphology show that with the increase of CMCS content, the forming quality of the samples deteriorated, the pore size and porosity increased. However, when the content of CMCS reached 5 wt%, obvious cracks appeared on the surface of the sample, and the forming quality was relatively poor. The mechanical testing results indicated the toughness of composites could be enhanced by incorporating CMCS into HA, which was attributed to the higher strength connections of the CMCS polymer network between HA particles and the stronger interaction between HA and CMCS molecules. FTIR spectra further revealed the strong hydrogen bonding interaction between CMCS and HA. Moreover, the degradation rate and mineralization ability of the sample increased with the content of CMCS, but the compressive strength during degradation increased with the CMCS content, indicating that incorporating CMCS into HA cannot only improve the mechanical property and biological activity of the scaffold but also makes up the defect of slow degradation of pure HA scaffold. Finally, the cytotoxicity, cell adhesion, and cell proliferation tests show that HA and HA/CMCS composite samples had good cytocompatibility, HA/CMCS sample with 3 wt% CMCS possessed the best bioactivity. In summary, HA/CMCS composite powder with 3 wt% CMCS content is the optimal matrix material for 3D printing bone scaffolds.     

Porous hydroxyapatite scaffolds produced by the combination of the gel-casting and freeze drying techniques

A technique combining gel-casting and freeze drying methods is introduced to prepare porous hydroxyapatite scaffolds which allow for better control of the scaffold microstructure and have improved mechanical properties. A monomeric system which is known to be a suitable gelling agent for setting ceramic suspensions into dense forms was selected to produce ceramic foams. Different concentrations of sodium lauryl sulphate solution were added into the hydroxyapatite gel suspension as a pore former. The effect of the solid content on the mechanical properties of the scaffold was also investigated. Rapid freezing with liquid nitrogen was performed according to the freeze drying technique and the porous structure and morphology of the scaffolds were analyzed by scanning electron microscopy. The mechanical properties of the hydroxyapatite scaffolds were determined by testing compressive strength using a universal testing machine. The prepared scaffolds were characterized by well-defined pore connectivity along with directional, uniform and completely open porosity. The maximum compressive strength of about 17?MPa obtained from the suspension consisted of 50% solid content with 20% concentration of sodium lauryl sulphate solution. The results show that sodium lauryl sulphate solution plays a significant role in changing the pore structure of hydroxyapaite scaffolds in systems having high solid content.     

β-TCP scaffold coated with PCL as biodegradable materials for dental applications

Requirements for an ideal scaffold include biocompatibility, biodegradability, mechanical strength and sufficient porosity and pore dimensions. Beta tricalcium phosphate (β-TCP) has competent biocompatibility and biodegradability, but has low mechanical strength because of its porous structure. Polycaprolactone (PCL) is a biodegradable polymer with elastic characteristics and good biocompatibility. In this study, β-TCP/PCL composites were prepared in different ratio and their morphology, phase content, mechanical properties, biodegradation and biocompatibility were investigated. After coating, surfaces of β-TCP scaffolds were covered with the PCL while some of the pores were partially clogged. The compression and bending strength of β-TCP scaffolds were significantly enhanced by PCL coating. The degradation rate of the scaffold in Tris buffer was reduced with higher content of the PCL coating. MTT and ALP assays showed that the osteoblast cells could proliferate and differentiate on PCL coated scaffolds as well as on bare β-TCP scaffolds. Based on the comprehensive analysis achieved in this study, it is concluded that the β-TCP/PCL composite scaffold fabricated with 40% β-TCP and 5% PCL exhibits optimum properties suitable for dental applications.     

Two-step modification process to improve mechanical properties and bioactivity of hydroxyfluorapatite scaffolds

Porous ceramic scaffolds are synthetic implants, which support cell migration and establish sufficient extracellular matrix (ECM) and cell-cell interactions to heal bone defects. Hydroxyapatite (HA) scaffolds is one of the most suitable synthetic scaffolds for hard tissue replacement due to their bioactivity, biocompatibility and biomimetic features. However, the major disadvantages of HA is poor mechanical properties as well as low degradability rate and apatite formation ability. In this study, we developed a new method to improve the bioactivity, biodegradability and mechanical properties of natural hydroxyfluorapatite (HFA) by applying two-step coating process including ceramic and polymer coats. The structure, morphology and bioactivity potential of the modified and unmodified nanocomposite scaffolds were evaluated using transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and energy dispersive spectroscopy (EDS). The scaffold with optimized mechanical properties was HFA-30?wt%HT (HT stands for hardystonite) with a total porosity and pore size of 89?±?1 and 900–1000?µm, respectively. The compressive modulus and strength of HFA (porosity ~ 93?±?1) were improved from 108.81?±?11.12–251.45?±?12.2?MPa and 0.46?±?0.1–1.7?±?0.3?MPa in HFA-30?wt%HT sample, respectively. After applying poly(ε-caprolactone fumarate) (PCLF) polymer coating, the compressive strength and modules increased to 2.8?±?0.15 and 426.1?±?15.14?MPa, respectively. The apatite formation ability of scaffolds was investigated using simulated body fluid (SBF). The results showed that applying the hardystonite coating improve the apatite formation ability; however, the release of ions increased the pH. Whereas, modified scaffolds with PCLF could control the release of ions and improve the apatite formation ability as well.     

Three dimensional polyurethane/ hydroxyapatite bioactive scaffolds: The role of hydroxyapatite on pore generation

Fabrication of a suitable scaffold with highly interconnected and well-distributed pores for cell proliferation and growth in the field of bone tissue engineering is of high importance. In this study, three-dimensional porous polyurethane (PU) scaffolds, with 0, 15, 25, and 32 wt% hydroxyapatite (HA), were fabricated. In this regard, HA was incorporated into PU constituents prior to starting in-situ polymerization of PU. Porosity and density measurement of the scaffolds revealed that higher amount of HA in the scaffolds led to increasing the former and decreasing the latter quantity. The field emission scanning electron microscopy (FESEM) images revealed that by increasing HA content, the pore size showed a descending trend while the number of pores increased. This would be attributed to the type of interactions between HA and PU, and the role of HA in pore formation. Mechanical test revealed that Young's Modulus of the samples was reduced by increasing scaffold porosity caused by the increase of HA content. Bioactivity tests in the simulated body fluid (SBF) showed the ability of scaffolds forming apatite precipitates. MTT assay showed that by increasing HA content, MG63 osteoblast cell viability increased and FESEM images revealed proper attachment of the cells to the scaffold surface.     

Preparation and characterization of aliphatic polyurethane and hydroxyapatite composite scaffold

Novel porous composite scaffolds for tissue engineering were prepared from aliphatic biodegradable polyurethane (PU) elastomer and hydroxyapatite (HA). It was found that the aliphatic PU was possible to load up to 50 wt % HA. The morphology and properties of the scaffolds were characterized by scanning electron microscope, X‐ray diffraction, infrared absorption spectra, mechanical testing, dynamic mechanical analysis, and in vitro degradation measurement. The results indicated that the HA/PU scaffolds had an interconnected porous structure with a pore size mainly ranging from 300 to 900 μm, and 50–200 μm micropores existed on the pores' walls. The average pore size of macropores and micropores are 510 and 100 μm, respectively. The compressive strength of the composite scaffolds showed higher enhancement with increasing HA content. In addition, the polymer matrix was completely composed of aliphatic component and exhibited progressive mass loss in vitro degradation, and the degradation rate depended on the HA content in PU matrix. The porous HA/PU composite may have a good prospect to be used as scaffold for tissue engineering. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

可注射nHA/PU复合多孔骨修复支架制备及表征

兼具良好孔隙率和原位任意塑形固化的可注射复合多孔骨修复材料在临床不规则骨缺损的治疗方面显示出巨大的优势。本研究通过优化双组分设计,以水为发泡剂制备可注射纳米羟基磷灰石/聚氨酯（nHA/PU）复合多孔支架。利用扫描电子显微镜（SEM）、傅里叶变换红外光谱（FTIR）、X射线衍射（XRD）、力学测试及Gillmore针测试等手段对制备的支架进行结构形貌、化学组成、力学性能和固化时间表征。结果表明,本研究制备的可注射nHA/PU复合多孔支架孔隙率高、孔隙贯通性好,孔径分布在100~700μm,适宜细胞和组织向孔内生长;添加20% nHA显著提高了PU支架的力学强度,但降低了支架的孔隙率;可注射支架在8h固化,适宜临床操作。本研究制备的可注射nHA/PU复合多孔支架在不规则骨缺损修复领域具有较大的应用潜力。     

Three-dimensional fibroin/collagen scaffolds derived from aqueous solution and the use for HepG2 culture

Although three-dimensional fibroin scaffolds have been prepared with freeze–drying method, these scaffolds still cannot meet the requirements of tissue engineering. In this article, a new process is described to form fibroin-based porous scaffolds with controllable structure and morphological features. When collagen was added to fibroin solution, the viscosity of the blend solution increased because of the interaction between fibroin and collagen, and then it restrained the unwanted fibroin leaf formation in freezing process that generally appeared in the previous fibroin scaffold preparation. With methanol treatment, the fibroin/collagen scaffolds became water-stable, following the transition from random and -helix to β-sheet conformation. The aqueous-fibroin porous scaffolds had highly homogeneous and interconnected pores with pore sizes ranging from 127 to 833 μm, depending on the fibroin concentration. The porosity of scaffolds was >90%, and the yield strength and modulus were up to 354±25 kPa and 30±0.1 MPa, respectively, when the blend solution, containing 20% collagen, maintained 4% fibroin concentration. Adhesion, spreading and proliferation of HepG2 cells on fibroin and fibroin/collagen blend scaffolds were also observed to investigate the biocompatibility. Scanning electron microscopy (SEM) and MTT analyses demonstrated that the adding of collagen evidently facilitated HepG2 attachment and proliferation in vitro. These new fibroin based three-dimensional scaffolds provided much more excellent properties due to the greatly improved control of pore size, the uniform pore distribution, the hydrophility, the mechanical properties and the biocompatibility compared with those of reported three-dimensional fibroin scaffolds.