Manufacture and characterization of a new Si-Ca-P biphasic ceramic

A ceramic of eutectoid composition based on a silicocarnotite (S)- tricalcium phosphate (TCP) sub-system was prepared, and the morphology of the structure investigated. The ceramic material, obtained by a solid-state reaction and slow solidification (6?°C/min) through the eutectoid temperature region (1158?±?2?°C), presented a typical eutectoid microstructure of alternate layers of both silicocarnotite and α-TCP_ss phases. The microstructure of the two phases was homogeneous and free of crystalline and structural defects. High-resolution electron microscopy revealed that the interfaces between the α-TCP_ss and silicocarnotite lamellae within the eutectoid grains were faultless and no sign of an intermediate region between both phases. Moreover in both phases, the lattice planes resolved at the interface were free of defects. The orientation relation between the lamellae of the eutectoid grains was determined as follows: {130} α-TCP_ss // {302} silicocarnotite. This eutectoid lamellae structure presents a new approach for the bioengineering application for hard tissue replacement as the material combines unique structural and bioactive properties and provides a platform for direct integrations with natural tissue.     

Study of biodegradable ceramic bone graft substitute

Abstract

The aim of the present study was to obtain a biodegradable porous calcium phosphate implants as a synthetic bone graft substitute. The calcium phosphate used in the present study consisted of hydroxyapatite (HA) and dicalcium phosphate anhydrous (DCP). Porous bioceramic was fabricated by a foam casting method. By polyurethane foam and slurry containing HA/DCP (3 : 1 weight ratio) powder, water and additives a high porous structure with ～70% was created. The X-ray diffractometry revealed that the β-tricalcium phosphate (β-TCP) formation is major phase. Surface morphology analysis and porosity evaluations were performed. The variation in the compressive strength, elastic modulus and dissolution behaviour of immersed synthetic bone graft in simulated physiological solution investigated.     

Gelatin-layered and multi-sized porous β-tricalcium phosphate for tissue engineering scaffold

The multi-sized porous β-tricalcium phosphate scaffolds were fabricated by freeze drying followed by slurry coating using a multi-sized porous sponge as a template. Then, gelatin was dip coated on the multi-sized porous β-tricalcium phosphate scaffolds under vacuum. The mechanical and biological properties of the fabricated scaffolds were evaluated and compared to the uniformly sized porous scaffolds and scaffolds that were not coated by gelatin. The compressive strength was tested by a universal testing machine, and the cell viability and differentiation behavior were measured using a cell counting kit and alkaline phosphatase activity using the MC3T3-E1 cells. In comparison, the gelatin-coated multi-sized porous β-tricalcium phosphate scaffold showed enhanced compressive strength. After 14 days, the multi-sized pores were shown to affect cell differentiation, and gelatin coatings were shown to affect the cell viability and differentiation. The results of this study demonstrated that the multi-sized porous β-tricalcium phosphate scaffold coated by gelatin enhanced the mechanical and biological strengths.     

3D Composite scaffolds using strontium containing bioactive glasses

In this study, it was aimed to fabricate and characterize three-dimensional composite scaffolds derived from Sr-doped bioactive glass for bone tissue engineering applications. The scaffolds were fabricated by using polymer foam replication technique and coated with gelatin to be able to improve the properties of them. The porous scaffolds were successfully synthesized using optimized process parameters. Both coated and uncoated scaffolds favored precipitation of calcium phosphate layer when they were soaked in simulated body fluid (SBF). Gelatin coating improved the mechanical properties of the scaffold and also it did not change the bioactive behavior of the scaffold. It was observed that there was a good pore interconnectivity maintained in the scaffold microstructure. Results indicated that scaffolds can deliver controlled doses of strontium toward the SBF medium. That is the determinant for bone tissue regeneration, as far as strontium is known to positively act on bone remodeling.     

Wollastonite/hydroxyapatite scaffolds with improved mechanical,bioactive and biodegradable properties for bone tissue engineering

Wollastonite/hydroxyapatite composite scaffolds are proposed as bone graft. An investigation on scaffold with varying reinforcing wollastonite content fabricated by polymeric sponge replica is reported. The composition, sintering behavior, morphology, porosity and mechanical strength were characterized. All the scaffolds had a highly porous well-interconnected structure. A significant increase in mechanical strength is achieved by adding a 50% wollastonite phase. The most mechanically resistant (50/50) wollastonite/hydroxyapatite scaffolds were soaked in both simulated body fluid (SBF) and Tris–HCl solution in order to assess bioactivity and biodegradability. A carbo-hydroxyapatite layer formed on their surfaces when immersed in SBF. The biodegradability tests reveals that the composite scaffold shows a higher degradation rate compared to pure hydroxyapatite used as comparison. These results demonstrate that the incorporation of a 50% of wollastonite phase in hydroxyapatite matrix is effective in improving the strength and the bioactive and biodegradable properties of the porous scaffolds.     

Evaluation of direct light processing for the fabrication of bioactive ceramic scaffolds: Effect of pore/strut size on manufacturability and mechanical performance

Bioactive ceramic scaffolds for bone regeneration consisting of a three-dimensional mesh of interpenetrating struts with square section were fabricated via Digital Light Processing (DLP). The ability of the technique to manufacture 3D porous structures from β-tricalcium phosphate (β-TCP) powders with different dimensions of struts and pores was evaluated, identifying the possibilities and limitations of the manufacturing process. Small pore sizes were found to seriously complicate the elimination of excess slurry from the scaffold’s innermost pores. The effect of the strut/pore size on the mechanical performance of the scaffolds under compressive stresses was also evaluated, but no significant influence was found. Under compressive stresses, the structures resulted weaker when tested perpendicularly to the printing plane due to interlayer shear failure. Interlayer superficial grooves are proposed as potential failure-controlling defects, which could also explain the lack of a Weibull size effect on the mechanical strength of the fabricated DLP scaffolds.     

DLP 3D printing porous β-tricalcium phosphate scaffold by the use of acrylate/ceramic composite slurry

Digital light processing (DLP) 3D printing has been utilized to fabricate controlled porous β-tricalcium phosphate (β-TCP) scaffolds, which promote cell adhesion and angiogenesis during bone regeneration. However, the current limitation of DLP 3D printing for the fabrication of β-TCP scaffold is how to prepare a low viscosity ceramic slurry and remove the toxicity of residual non-polymerized slurry. The present study has developed a low viscosity ceramic slurry system by mixing β-TCP with photosensitive acrylate resin, and the viscosity of slurry is about 3 Pa s and the solid content of β-TCP can be as high as 60 wt%. After optimizing the ratio of slurry, printing, degreasing and sintering processes, the maximum compressive strength of the DLP printed scaffolds reaches up to 9.89 MPa, while the porosity keeps ca. 40%. According to the proliferation of cells, it confirms the preserved biocompatibility of DLP-fabricated β-TCP scaffolds. These porous scaffolds made by DLP 3D printing technology is of great significance for bone regeneration, and will also help to expand the application of DLP technology in biomedical field.     

层状结构磷酸钙骨水泥组织工程支架材料的制备与表征

采用可溶粒子造孔法结合冷等静压成型技术,模拟扁骨的结构,制备了一种新型层状结构的多孔磷酸钙骨水泥组织工程支架材料,并用XRD和SEM等手段对其组成和结构进行了表征,用万能材料试验机测定了支架的抗压强度.结果表明,材料由致密层和多孔层构成,具有与扁骨类似的结构.其中致密层起到了增强作用,可以显著提高支架的强度.支架多孔层的孔隙率(77.26±1.99)%,孔隙直径在100～400 μm,决定于可溶盐晶粒的大小;致密层的孔隙率(20.78±0.56)%,主要是磷酸钙骨水泥固化过程中产生的微孔.     

Bioactive and Biocompatible Macroporous Scaffolds with Tunable Performances Prepared Based on 3D Printing of the Pre‐Crosslinked Sodium Alginate/Hydroxyapatite Hydrogel Ink

Bioactive and biocompatible porous scaffold materials with adjustable pore structures and drug delivery capability are one of the key elements in bone tissue engineering. In this work, bioactive and biocompatible sodium alginate (SA)/hydroxyapatite (HAP) macroporous scaffolds are facilely and effectively fabricated based on 3D printing of the pre‐crosslinked SA/HAP hydrogels followed by further crosslinking to improve the mechanical properties of scaffolds. The pore structures and porosity (>80%) of the porous scaffolds can be readily tailored by varying the formation conditions. Furthermore, the in vitro biomineralization tests show that the bioactivity of the porous scaffolds is effectively enhanced by the addition of HAP nanoparticles into the scaffold matrix. Furthermore, the anti‐inflammatory drug curcumin is loaded into the porous scaffolds and the in vitro release study shows the sustainable drug release function of the porous scaffolds. Moreover, mouse bone mesenchymal stem cells (mBMSCs) are cultured on the porous scaffolds, and the results of the in vitro biocompatibility experiment show that the mBMSCs can be adhered well on the porous scaffolds. All of the results suggest that the bioactive and biocompatible SA/HAP porous scaffolds have great application potential in bone tissue engineering.     

Physicochemical properties and biomimetic behaviour of α-TCP-chitosan based materials

In this paper we present a complex study on the preparation and characterization of the cement-type implant material composed of α-tricalcium phosphate and chitosan. α-TCP has already been used as an initial component of bone cement formulations. In most cases, however, it was synthesized using a solid state reaction. In our studies, highly reactive α-TCP powder was obtained by a wet chemical method. Chitosan is well known as a safe and biocompatible polymer. In our studies it was used to improve the characteristic of the cements. Chitosan was introduced into the bone cements in the form of acetic acid solution and acted as a binder for agglomerates of calcium phosphate grains but at the same time the impeded creation of interconnections inside the agglomerates. The concentration of acetic acid influenced the pore size distribution. In vitro investigations proved the high bioactive potential of the examined materials.     

Preparation of chitosan-sodium alginate/bioactive glass composite cartilage scaffolds with high cell activity and bioactivity

Chitosan-sodium alginate/bioactive glass (CSB) composite cartilage scaffold with outstanding in vitro mineralization property and cytocompatibility is synthesized by freeze drying method. The effect of bioactive glass (BG) addition on the microstructure, porosity, swelling/degradation ratio, in vitro mineralization property and cytocompatibility of CSB scaffold is investigated by the characterization techniques of SEM, XRD, FTIR and BET. Results showed that CSB composite cartilage scaffold had a three-dimensional (3D) porous structure, and both porosity and average pore size met the requirements of cartilage tissue repair. Among, the typical CSB-1.0 had the largest overall pore size and lowest compressive modulus (1.083 ± 0.002 MPa). As the amount of BG increased, pore volume and porosity of CSB scaffolds gradually decreased, and the swelling and degradation ratios gradually reduced. After immersing in SBF for 3 d, cauliflower like hydroxyapatite (HA) was formed on CSB surface, indicating that the scaffold had good in vitro mineralization property. Moreover, the introduction of BG into the composite scaffold can improve the relative cell viability of MC3T3-E1 cells, and CSB-1.0 has the strongest ability to promote the proliferation of cells. Therefore, the as-obtained CSB scaffold can be used as a strong candidate for cartilage tissue engineering scaffold to meet clinical needs.     

Fabrication,characterization and cellular biocompatibility of porous biphasic calcium phosphate bioceramic scaffolds with different pore sizes

Facile wet-chemical methods are applied to synthesize hydroxyapatite and β-tricalcium phosphate nanoparticles, respectively. Porous biphasic calcium phosphate (BCP) bioceramic scaffolds are then fabricated using as-prepared HA and β-tricalcium phosphate nanoparticle powders. The macro pore diameter of BCP bioceramic scaffolds can be controlled by adjusting the amount of surfactants. The average diameter of the macro pores in BCP bioceramic scaffolds increases from 100 to 600 µm with the decrease amount of sodium dodecyl sulfate from 0.8 to 0.5 g, respectively. The BCP bioceramic scaffolds gradually degrade and the calcium-phosphate compounds fully deposit when soaking in simulated body fluid solution. Moreover, The BCP bioceramic scaffolds have outstanding biocompatibility to promote the cellular growth and proliferation of human dental pulp stem cells (hDPSCs). The hDPSCs also demonstrate favorable cellular adhering capacity on the pore surface of scaffolds, especially on the scaffolds with 100–200 µm pore diameter. The porous BCP bioceramic scaffold with inter-connected pore structure, outstanding in vitro cellular biocompatibility, favorable cell viability and adhesion ability will be a promising biomaterial for bone or dentin tissue regeneration.     

Synthesis and in vitro biodegradation of pure octacalcium phosphate spheres

Octacalcium phosphate (OCP) is a key precursor of biological apatite in hard tissues with excellent osteoconductive and biodegradable properties for bone regeneration. OCP spherical granules are expected to be useful as drug delivery carriers, since OCP has high specific surface area. Although there have been some reports of OCP sphere preparation, methods for preparing pure OCP spheres are limited. The objective of this study is the preparation of spherical granules of pure OCP and assessment of their in vitro biodegradation in physiological conditions. We successfully prepared spherical pure OCP granules with a size of ~500 µm without any organic additives by simple immersion of α-tricalcium phosphate spherical granules in pH 5.0 acetate buffered solutions at 60°C. The granules had core-shell structure composed of OCP crystals different particle size. The spherical granules showed 20%-40% in vitro degradation in physiological conditions; however, the phase transition of OCP was not significantly observed.     

β-Tricalcium phosphate silica aerogel as an alternative bioactive ceramic for the potential use in dentistry

ABSTRACT

In this study, a mesoporous silica aerogel with β-tricalcium phosphate (β-TCP-AE) was manufactured. The effect of β-TCP-AE on gene expressions (BMP2, BMP7, Runx2 and OSX) of SAOS-2 cells was tested. For the in vivo evaluation, the ‘calvaria critical-size defect’ model was used: following 1 and 3 months of the artificial surgical bone defects filled with β-TCP-AE, histopathological analyses were performed. Gene expression studies demonstrated a mild osteoblastic differentiation of the SAOS-2 cells triggered after seven days of β-TCP-AE treatment. Digital histology of rat’s calvarial bone defects reconstructed with β-TCP-AE showed that after 1 month, calcifications and early ossifications developed with the presence of capillary-rich fibrous inflammation and remnants of exogenous compounds which nearly disappeared by the third month, and replaced with multiple newly formed bone islets mediated by osteoblasts. Based on our results, this bioceramic compound appears to have favourable properties for the use as a scaffold in the reconstructive medical practice.     

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.     

Effect of particle size and freezing conditions on freeze-casted scaffold

Freeze casting is one of the emerging and novel manufacturing routes to fabricate porous scaffolds for various applications including orthopedic implants, drug delivery, energy storing devices etc. Thus, it becomes important to understand this process in a deeper sense. Present work was focused to study the effect/influence of basic parameters, particle sizes, and freezing conditions on the mechanical properties and microstructures of porous scaffold fabricated by freeze casting. β-tricalcium phosphate (β-TCP) and hydroxyapatite (HAp) powder with particle sizes of 10?μm and 20?nm were used. Prepared slurries were freeze casted at constant freezing temperature (5?°C) and constant freezing rate (1.86?°C/min) to study the effect of freezing conditions on mechanical and microstructural properties of the porous scaffold. It was observed that porous scaffold fabricated by nanoparticles has given better porosity (63.22–76.16%), than scaffold fabricated by microparticles (13–43.05%) at given solid loading of both freezing conditions. Although, the range of pore size of the scaffold fabricated by nanoparticles (CFR: 2.60–0.84?μm; CFT: 1.66–0.46?μm) was lower than that of scaffold fabricated by microparticles (CFR: 9.45–4.83?μm; CFT: 4.72–2.84?μm). The compressive strength of scaffolds prepared by nanoparticles was in the range of trabecular bone. Moreover, the results of present work will pave the way for the fabrication of porous scaffold with desired pore size and porosity for various implants, energy, and drug delivery applications.     

Bone Fracture-Treatment Method: Fixing 3D-Printed Polycaprolactone Scaffolds with Hydrogel Type Bone-Derived Extracellular Matrix and β-Tricalcium Phosphate as an Osteogenic Promoter

Bone formation and growth are crucial for treating bone fractures. Improving bone-reconstruction methods using autologous bone and synthetic implants can reduce the recovery time. Here, we investigated three treatments using two different materials, a bone-derived decellularized extracellular matrix (bdECM) and β-tricalcium phosphate (β-TCP), individually and in combination, as osteogenic promoter between bone and 3D-printed polycaprolactone scaffold (6-mm diameter) in rat calvarial defects (8-mm critical diameter). The materials were tested with a human pre-osteoblast cell line (MG63) to determine the effects of the osteogenic promoter on bone formation in vitro. A polycaprolactone (PCL) scaffold with a porous structure was placed at the center of the in vivo rat calvarial defects. The gap between the defective bone and PCL scaffold was filled with each material. Animals were sacrificed four weeks post-implantation, and skull samples were preserved for analysis. The preserved samples were scanned by micro-computed tomography and analyzed histologically to examine the clinical benefits of the materials. The bdECM–β-TCP mixture showed faster bone formation and a lower inflammatory response in the rats. Therefore, our results imply that a bdECM–β-TCP mixture is an ideal osteogenic promoter for treating fractures.     

Temperature-dependent morphology changes of noble metal tricalcium phosphate-nanocomposites

Calcium phosphates, functionalized with nano-sized metal particles, are a promising material class for the treatment of bone defects. However, a sintering process is required in principle to achieve sufficient strength of calcium phosphate scaffolds. In this work laser-generated nano-sized silver, gold and platinum particles were adsorbed on micro-sized β-tricalcium phosphate particles and further heat treated at temperatures between 600 and 1200 °C. Gold and platinum nanoparticles underwent exponential growth starting at about 600 °C, while sintering of β-tricalcium phosphate started at 800 °C. We hypothesise that this phenomenon is caused by a heat-induced evaporation and growth process where the decrease of the particle number is directly correlated with the size increase. The silver nanoparticles on the other hand formed a new phase with the calcium phosphate (AgCa₁₀(PO₄)₇) during the heat treatments and could not be observed within the ceramic scaffold anymore. Addressing the lack of information in nanoparticle-combined calcium phosphate scaffolds, this study contributes to the further modification of bone replacement materials with biologically relevant functions and molecules.     

Brushite Cements from Polyphosphoric Acid, Calcium Phosphate Systems

Brushite cement is more soluble than apatitic cement under physiological conditions. Thus, brushite cement may be resorbable in vivo when used as a bone substitute material. Our group has previously reported the formation of brushite cement from orthophosphoric acid and several calcium phosphates such as β-tricalcium phosphate or nanocrystalline hydroxyapatite. In this study, polyphosphoric acid was investigated as a calcium phosphate cement component. We found that brushite cement was not formed when polyphosphoric acid was mixed with β-tricalcium phosphate but instead monocalcium phosphate monohydrate (MCPM) was formed. Although stronger in compression when tested dry, MCPM is readily soluble under physiological conditions and cement loses structural integrity within hours of aging in water. However, by varying the solid component phases, we have discovered a new route to the formation of brushite cement. The mixture of polyphosphoric acid, water, and tetracalcium phosphate yielded cement composed predominantly of brushite. The mechanical performance, microstructure, and setting time of this cement appeared to be dependent on the composition of the set cement, which in turn was determined by the composition of cement reactants in the cement slurry.     

Synthesis,Characterization and <Emphasis Type="Italic">in-vitro</Emphasis> Study of Chitosan/Gelatin/Calcium Phosphate Hybrid Scaffolds Fabricated Via Ion Diffusion Mechanism for Bone Tissue Engineering

In this study, biomimetic scaffolds were designed to investigate calcium phosphate formation via a double diffusion mechanism within a gelatin/chitosan hydrogel in biological pH and temperature. Three types of samples with initial percentages of chitosan (20, 30 and 40 wt. %) were prepared. Diffusion of calcium and phosphate ions through the hydrogel formed a precipitation layer. Samples were freeze dried to form porous scaffolds and soaked in glutaraldehyde to increase their mechanical properties. X-ray diffraction (XRD), Fourier transform infra-red (FTIR) spectroscopy and scanning electron microscopy (SEM) were employed to investigate the microstructure and to characterize the prepared scaffolds. Analysis of precipitation indicated the presence of brushite and hydroxyapatite. The amorphous calcium phosphate phase converted into crystalline hydroxyapatite after immersion in simulated body fluid which mimics the formation of hydroxyapatite in the human body. FTIR results suggested the presence of structural hydroxyl and phosphate bonds in the structure of the prepared scaffolds which could be due to the formation of hydroxyapatite. With increasing amount of chitosan in the composite scaffold, the water up-take ability was increased from 380 to 660 %, yield strength and Young’s modulus slightly decreased and the crystalinity of the precipitated phase increased. Mechanical properties obtained from the samples were in the range of cancellous bone. MTT assay results and alkaline phosphatase activity showed prepared scaffolds had proper biocompatibility.