Human Cancellous Bone Research Articles

3D gel printing of porous calcium silicate scaffold for bone tissue engineering

Potential porous calcium silicate (CaSiO3) scaffolds for bone tissue engineering were successfully prepared by 3D gel printing technology. The average particle size of CaSiO3 powder is about 5.119 μm. The CaSiO3 slurry prepared for printing had been exhibited shear-thinning properties, and the maximum solid loading of the slurry was 72 wt%. The green scaffolds were sintered at 1150 °C for 4 h after degreasing at 250 °C for 3 h, 420 °C for 9 h, and 660 °C for 3 h, respectively. The sintered scaffolds have well-distributed designed voids and microporous. The shrinkage in length, width, and height is 12.29 ± 0.20%, 12.43 ± 0.17%, 13.15 ± 0.19%, respectively. The porosity of the sintered scaffold is approximately 62%, and the compressive strength is about 16.52 MPa, which is equivalent to the strength of human cancellous bone.

Fabrication, Morphology Analysis, and Mechanical Properties of Ti Foams Manufactured Using the Space Holder Method for Bone Substitute Materials

Porous titanium (Ti) offers several key attributes as a biomedical material. Among the known characteristics of Ti relevant to biomedical applications, the mechanical performance and effects of a pore structure on the deformation characteristics under compressive loading were examined. The space holder method was employed to generate Ti foams with target porosities of 60%, 70%, and 80%. A micro-computed to mography analysis and light and scanning electron microscopy were performed to examine the pore morphology and microstructure. The mechanical properties along with the elastic modulus and compressive strength were evaluated via uniaxial compression testing. Ti foam samples with three porosity levels displayed average elastic moduli and compressive strengths comparable with those of human cancellous and cortical bone. All the Ti foam samples had elastic moduli similar to those of cancellous bone with their open porous structures. Although the foam samples with ~60% porosity had compressive strength comparable to that of cortical bone, the samples with ~80% porosity displayed compressive strength similar to that of cancellous bone. The results indicate that Ti foam scaffolds produced using the space holder method have great potential for applications in hard tissue engineering, as their mechanical properties and pore structures are similar to those of bone.

Synthesis and characterization of a novel open cellular Mg-based scaffold for tissue engineering application

Tissue engineering is a field which aims to regenerate damaged tissues by enhancing tissue growth through the porous architecture of the scaffolds which is desired to mimic the human cancellous bone. Mg-based scaffolds are gaining importance in the field of tissue engineering owing to its potential application as a biomaterial. However, fabrication of porous Mg remains a daunting task due to its highly reactive nature. In the present work, a novel Mg-based open cell porous structure with pore interconnectivity and significant strength is successfully fabricated using powder metallurgy approach and Ti-woven wire mesh as a space holding material. Pore morphology and percentage porosity can be easily altered by adjusting the Ti-wire diameter and shape of construct. SEM, EDX and µ-CT analysis were performed to assess the microstructural properties of the fabricated scaffold which revealed a uniform distribution of pores with porosity varying in range 50–60%. The measured values of ultimate compressive strength and elastic modulus using quasi static compression test were found to be 101 MPa and 2 GPa, respectively. Further to improve corrosion resistance of fabricated scaffold, alloying and coating were carried out. Preliminary degradation study as well as cytocompatibility studies using L929 cells was carried out to validate the potential of fabricated scaffold for bone healing/repair applications. Fabricated porous structures showed improved corrosion resistance as well as cell viability of more than 90%, suggesting it as a promising development for bone scaffolding applications in future.

Robocasting of Cu2+ & La3+ doped sol–gel glass scaffolds with greatly enhanced mechanical properties: Compressive strength up to 14 MPa

This research details the successful fabrication of scaffolds by robocasting from high silica sol–gel glass doped with Cu2+ or La3+. The parent HSSGG composition within the system SiO2–CaO–Na2O–P2O5 [67% Si – 24% Ca – 5% Na – 4% P (mol%)] was doped with 5 wt% Cu2+ or La3+ (Cu5 and La5). The paper sheds light on the importance of copper and lanthanum in improving the mechanical properties of the 3–D printed scaffolds. 1 h wet milling was sufficient to obtain a bioglass powder ready to be used in the preparation of a 40 vol% solid loading paste suitable for printing. Moreover, Cu addition showed a small reduction in the mean particle size, while La exhibited a greater reduction, compared with the parent glass. Scaffolds with macroporosity between 300 and 500 µm were successfully printed by robocasting, and then sintered at 800 °C. A small improvement in the compressive strength (7–18%) over the parent glass accompanied the addition of La. However, a much greater improvement in the compressive strength was observed with Cu addition, up to 221% greater than the parent glass, with compressive strength values of up to ∼14 MPa. This enhancement in compressive strength, around the upper limit registered for human cancellous bones, supports the potential use of this material in biomedical applications. Statement of Significance3D porous bioactive glass scaffolds with greatly improved compressive strength were fabricated by robocasting from a high silica sol–gel glasses doped with Cu2+ or La3+. In comparison to the parent glass, the mechanical performance of scaffolds was greatly improved by copper-doping (>220%), while a modest increase of ∼9% was registered for lanthanum-doping. Doping ions (particularly La3+) acted as glass modifiers leading to less extents of silica polymerisation. This favoured the milling of the glass powders and the obtaining of smaller mean particle sizes. Pastes with a high solid loading (40 vol%) and with suitable rheological properties for robocasting were prepared from all glass powders. Scaffolds with dimensions of 3 × 3 × 4 mm and macro-pore sizes between 300 and 500 µm were fabricated.

Cancellous-Bone-like Porous Iron Scaffold Coated with Strontium Incorporated Octacalcium Phosphate Nanowhiskers for Bone Regeneration.

The repair of large bone defects poses a grand challenge in tissue engineering. Thus, developing biocompatible scaffolds with mechanical and structural similarity to human cancellous bone is in great demand. Herein, we fabricated a three-dimensional (3D) porous iron (Fe) scaffold with interconnected pores via a template-assisted electrodeposition method. The porous Fe scaffold with a skeleton diameter of 143 μm had the porosity >90%, an average pore size of 345 μm, and a yield strength of 3.5 MPa. Such structure and mechanical strength were close to those of cancellous bone. In order to enhance the biocompatibility of the scaffold, strontium incorporated octacalcium phosphate (Sr-OCP) was coated on the skeletons of the porous Fe scaffold. The coated Sr-OCP was in the form of nanowhiskers with a mean diameter of 300 nm and length of 30 μm. Such Sr-OCP coating could effectively reduce the release rate of the Fe ions to a level which was safe for the human body. Both in vitro cytotoxicity tests by extraction method and direct contact assay confirmed that the Sr-OCP coating could promote the cell adhesion and substantially enhance the biocompatibility of the porous Fe scaffolds. Thus, the cancellous-bone-like porous structure with compatible mechanical properties and excellent biocompatibility enables the present Sr-OCP coated porous Fe scaffold to be a promising candidate for bone repair and regeneration.

Improving the anti-washout property of calcium phosphate cement by introducing konjac glucomannan/κ-carrageenan blend.

Anti-washout calcium phosphate cement (CPC) was prepared by dissolving water-soluble konjac glucomannan (KGM) and κ-carrageenan (KC) blend in the cement liquid. The anti-washout property, setting time, compressive strength and in vitro cytocompatibility of the CPC modified with KGM/KC blend were evaluated. The results indicated that the CPC pastes modified with KGM/KC blend exhibited excellent anti-washout property. The addition of KGM/KC blend shortened the setting time and increased the injectability of CPC. Although the introduction of KGM/KC blend reduced the compressive strength of CPC, the compressive strength still surpassed that of human cancellous bone. The optimal KGM/KC mass ratio was 2:8, with which the modified cement exhibited the most efficient washout resistance and the highest compressive strength. The introduction of KGM/KC blend obviously promoted the proliferation of mouse bone marrow mesenchymal stem cells. This anti-washout CPC modified by KGM/KC blend with excellent in vitro cytocompatibility will have good prospects for application in bone defect repair.

Mechanical characterization of pore-graded bioactive glass scaffolds produced by robocasting

Abstract Since the discovery of 45S5 Bioglass® by Larry Hench, bioactive glasses have been widely studied as bone substitute materials and, in more recent years, have also shown great promise for producing three-dimensional scaffolds. The development of additive manufacturing techniques and their application in bone tissue engineering allows the design and fabrication of complex structures with controlled porosity. However, achieving strong and mechanically-reliable bioactive glass scaffolds is still a great challenge. Furthermore, there is a relative paucity of studies reporting an exhaustive assessment of other mechanical properties than compressive strength of glass-derived scaffolds. This research work aimed at determining key mechanical properties of silicate SiO2-Na2O-K2OMgO-CaO-P2O5 glass scaffolds fabricated by robocasting and exhibiting a porosity gradient. When tested in compression, these scaffolds had a strength of 6 MPa, a Young’s modulus around 340 MPa, a fracture energy of 93 kJ/m3 and a Weibull modulus of 3, which provides a quantification of the scaffold reliability and reproducibility. Robocasting was a suitable manufacturing method to obtain structures with favorable porosity and mechanical properties comparable to those of the human cancellous bone, which is fundamental regarding osteointegration of bone implants.

Improvement of anti-washout property of calcium phosphate cement by addition of konjac glucomannan and guar gum.

The inferior anti-washout property of injectable calcium phosphate cement (CPC) limits its wider application in clinic. In this study, the improvement of anti-washout performance of CPC by addition of konjac glucomannan or guar gum, which was dissolved in the CPC liquid, was first studied. The influence of KGM/GG blend with different mass ratios on the anti-washout property, compressive strength and in vitro cytocompatibility of CPC was estimated. The results revealed that small amount of KGM or GG could obviously enhance the anti-washout property of CPC. Moreover, the washout resistance efficiency of KGM/GG blend was better than KGM or GG alone. The addition of KGM/GG blend slightly shortened the final setting time of CPC. Although the introduction of KGM/GG blend reduced the compressive strength of CPC, the compressive strength still reached or surpassed that of human cancellous bone. The best KGM/GG mass ratio was 5:5, which was most efficient at not only reducing CPC disintegration, but also increasing compressive strength. The addition of KGM/GG blend obviously promoted the cells proliferation on the CPC. In short, the CPC modified by KGM/GG blend exhibited excellent anti-washout property, appropriate setting time, adequate compressive strength, and good cytocompatibility, and has the potential to be used in bone defect repair. The addition of KGM/GG blend significantly improved the anti-washout property of CPC. The best KGM/GG mass ratio was 5:5, which was most efficient in reducing the CPC disintegration.

A biocompatible thermoset polymer binder for Direct Ink Writing of porous titanium scaffolds for bone tissue engineering

There is increasing demand for synthetic bone scaffolds for bone tissue engineering as they can counter issues such as potential harvesting morbidity and restrictions in donor sites which hamper autologous bone grafts and address the potential for disease transmission in the case of allografts. Due to their excellent biocompatibility, titanium scaffolds have great potential as bone graft substitutes as they mimic the structure and properties of human cancellous bone. Here we report on a new thermoset bio-polymer which can act as a binder for Direct Ink Writing (DIW) of titanium artificial bone scaffolds. We demonstrate the use of the binder to manufacture porous titanium scaffolds with evenly distributed and highly interconnected porosity ideal for orthopaedic applications. Due to their porous structure, the scaffolds exhibit an effective Young's modulus similar to human cortical bone, alleviating undesirable stress-shielding effects, and possess superior strength. The biocompatibility of the scaffolds was investigated in vitro by cell viability and proliferation assays using human bone-marrow-derived Mesenchymal stem cells (hMSCs). The hMSCs displayed well-spread morphologies, well-organized F-actin and large vinculin complexes confirming their excellent biocompatibility. The vinculin regions had significantly larger Focal Adhesion (FA) area and equivalent FA numbers compared to that of tissue culture plate controls, showing that the scaffolds support cell viability and promote attachment. In conclusion, we have demonstrated the excellent potential of the thermoset bio-polymer as a Direct Ink Writing ready binder for manufacture of porous titanium scaffolds for hard tissue engineering.

Morphological and mechanical characterization of topologically ordered open cell porous iron foam fabricated using 3D printing and pressureless microwave sintering

Topologically ordered porous structures (TOPSs) have shown great potential in biomedical application. For biodegradable application, TOPS can prove to be advantageous especially for iron-based biodegradable materials. However, limited work has been reported which discusses the fabrication and characterization of iron-based TOPS. Hence, in the present work, topologically ordered open cell porous iron foam (TOPIF) was developed using a novel fabrication procedure consisting of 3D printing and pressureless microwave sintering. Different unit cell structures namely cubic, truncated octahedron and pyramid were used. Porosities in the range of 45.6–86.9% and 5–22% variation in dimensions were obtained. Compressive modulus of elasticity, plateau stress and ultimate compressive strength of TOPIF with 45.6–86.9% porosity were found in the range of 218.67–854.04 MPa, 4.24–21.60 MPa and 13.16–52.06 MPa respectively. Moreover, flexural modulus of elasticity and ultimate flexural strength in the range of 161–753 MPa and 9–38 MPa respectively were obtained. Analysis based on Gibson-Ashby model was performed and good agreement with experimental results was obtained. A comparative study showed that the fabricated TOPIF samples possessed ideal properties as required for an augmentation procedure of a human cancellous bone.

T cell immunomodulation by clinically used allogeneic human cancellous bone fragments: a potential novel immunotherapy tool

Multipotential stromal cells (MSCs) demonstrate strong immunomodulation capabilities following culture expansion. We have previously demonstrated that human cancellous bone fragments (CBFs) clinically used as viable allografts for spinal fusion have resident MSCs that exhibit T cell immunomodulation after monolayer expansion. This study investigated the immunomodulatory ability of these CBFs without MSC culture-expansion. CD4 positive T cells were induced to proliferate using CD3/CD28 stimulation and added to CBFs at different ratios of T cells per gram of CBF. A dose-dependent suppressive effect on T cell proliferation was evident and correlated with increased culture supernatant levels of TGF-ß1, but not PGE2. CBF-driven immunosuppression was reduced in co-cultures with TGF-ß neutralising antibodies and was higher in cell contact compared to non-contact cultures. CBF gene expression profile identified vascular cell adhesion molecule-1, bone marrow stromal antigen 2/CD317 and other interferon signalling pathway members as potential immunomodulatory mediators. The CD317 molecule was detected on the surface of CBF-resident cells confirming the gene expression data. Taken together, these data demonstrate that human clinically used CBFs are inherently immunomodulatory and suggest that these viable allografts may be used to deliver therapeutic immunomodulation for immune-related diseases.

Replication and bioactivation of Ti-based alloy scaffold macroscopically identical to cancellous bone from polymeric template with TiNbZr powders

In the present work, a new type of porous Ti-based alloy scaffold with high porosity (about 75%) and interconnected pores in the range of 300–1000 µm was fabricated by polymeric foam replication method with TiNbZr powders. This porous scaffold, which is consisted with major β phase Ti and minor α Ti phase, exhibits a compressive strength of 14.9 MPa and an elastic modulus of 0.21 GPa, resembling the mechanical properties of nature human cancellous bone (σ = 10–50 MPa, E = 0.01–3.0 GPa). To improve its osteogenic potential, a bioactive nanostructural titanate network coating was applied to the scaffold surface using hydrothermal treatment. The bone-like apatite inducing ability of the treated scaffold was systemically assessed using SBF immersion during 3–28 days. The nanostructural titanate network coated on porous TiNbZr scaffold is favorable for apatite nucleation and subsequent growth due to the hydrolysis of titanate. The results suggest that highly porous TiNbZr scaffolds with an appropriate bioactive coating, which was fabricated in this study, could be potentially used for bone tissue engineering application.

The effect of charge density on the velocity and attenuation of ultrasound waves in human cancellous bone

Cancellous bone is a highly porous material, and two types of waves, fast and slow, are observed when ultrasound is used for detecting bone diseases. There are several possible stimuli for bone remodelling processes, including bone fluid flow, streaming potential, and piezoelectricity. Poroelasticity has been widely used for elucidating the bone fluid flow phenomenon, but the combination of poroelasticity with charge density has not been introduced. Theoretically, general poroelasticity with a varying charge density is employed for determining the relationship between wave velocity and attenuation with charge density. Fast wave velocity and attenuation are affected by porosity as well as charge density; however, for a slow wave, both slow wave velocity and attenuation are not as sensitive to the effect of charge density as they are for a fast wave. Thus, employing human femoral data, we conclude that charged ions gather on trabecular struts, and the fast wave, which moves along the trabecular struts, is significantly affected by charge density.

Synthesis and Characterization of Akermanite by Mechanical Milling and Subsequent Heat Treatment

In this study, nano-structured akermanite was synthesized by mechanical milling and mixing using calcium oxide, magnesium oxide, cobalt oxide and silicon dioxide in a planetary ball milling with the speed of 500 rpm for 3 h. The milled powder was pelletized and sintered at 1200°C subsequently were characterized by X-ray diffraction (XRD), transmission Fourier transform spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM) confirming FESEM a dense sintered akermanite. The diametral tensile strength (DTS) value of akermanite was found to be 5.71 MPa, which was in the range of human cancellous bone i.e. 1.5-38 MPa. Finally, akermanite induced obvious apatite formation on the surface after 7 days of soaking. All the results postulated that the akermanite ceramics might be potentially used as bone repair biomaterials for non-load bearing applications.

Ultrasonic backscatter difference measurements of cancellous bone from the human femur: Relation to bone mineral density and microstructure.

Ultrasonic backscatter techniques are being developed to detect changes in cancellous bone caused by osteoporosis. One technique, called the backscatter difference technique, measures the power difference between two portions of a backscatter signal. The goal of the present study is to investigate how bone mineral density (BMD) and the microstructure of human cancellous bone influence four backscatter difference parameters: the normalized mean of the backscatter difference (nMBD) spectrum, the normalized slope of the backscatter difference spectrum, the normalized intercept of the backscatter difference spectrum, and the normalized backscatter amplitude ratio (nBAR). Ultrasonic measurements were performed with a 3.5 MHz broadband transducer on 54 specimens of human cancellous bone from the proximal femur. Volumetric BMD and the microstructural characteristics of the specimens were measured using x-ray micro-computed tomography. Of the four ultrasonic parameters studied, nMBD and nBAR demonstrated the strongest univariate correlations with density and microstructure. Multivariate analyses indicated that nMBD and nBAR depended on trabecular separation and possibly other microstructural characteristics of the specimens independently of BMD. These findings suggest that nMBD and nBAR may be sensitive to changes in the density and microstructure of bone caused by osteoporosis.

Non-linearly viscoelastic constitutive equation of cancellous bone tissue and identification of material constants

In the paper a new constitutive equation for human cancellous bone tissue was formulated. A strain energy function was postulate, which describes the elastic response of the tissue to the load. The nonlinear viscoelasticity was also taken into account in the constitutive model. The material constants in the constitutive equation were identified on the basis of the results of stress relaxation tests and monotonic compression tests using the curve fitting method. The Levenberg-Marquardt algorithm of the least square method was used here. On the basis of the relaxation tests, the values of relaxation times were identified, while the elastic and viscoelastic constants were determined on the basis of monotonic compression tests. The tests were performed at room temperature. On the cuboidal samples of cancellous bone obtained from femoral heads during the surgeries of hip joint prosthesis implantation.

Effects of Moisture Content and Loading Profile on Changing Properties of Bone Micro-Biomechanical Characteristics.

BackgroundOur study explored the influences of hydration conditions and loading methods on the mechanical properties of cortical bones and cancellous bones.Material/MethodsElastic modulus and hardness of human cortical bones and cancellous bones that contained different moisture levels (20%, 30%, 40%, 50%, and 60%) were measured with nanoindentation with different peak loads and loading rates. Cortical bones with 20% and 60% moisture were tested with 30 nm, 40 nm, and 50 nm peak loads at 6 nm/s, 8 nm/s, and 10 nm/s loading rates, respectively. Cancellous bones with 5% or 40% moisture percentages were tested with 600 μN, 750 μN, and 1000 μN peak loads at 200 μN/s, 250 μN/s, and 333 μN/s loading rates, respectively.ResultsUnder the same loading condition, specimens with higher moisture contents showed decreased elastic modulus and hardness. Under different loading conditions, the loading modes had little influence on elastic modulus and hardness of cortical bone and cancellous bone with low moisture, but had significant influence on specimens with higher moistures.ConclusionsThe elastic modulus and bone hardness were affected by the moisture content and the loading conditions in cortical and cancellous bones with high hydration condition but not in those with low hydration condition.

Microstructure and fracture properties of open-cell porous Ti-6Al-4V with high porosity fabricated by electron beam melting

The open-cell porous Ti-6Al-4V structure with high porosity, intended to be applied as replacement for human cancellous bone, were fabricated by electron beam melting (EBM). Computer aided design (CAD) was applied to design porous structures using the same unit cell with different unit cell sizes from 2.5 to 4 mm, different ligament widths from 600 to 900 μm, and different pore sizes from 1200 to 1800 μm, in order to achieve high porosity of 80% in avoiding the stress shielding effect. In comparison with the CAD designs and the EBM samples, there were minor discrepancies in terms of pore size and ligament width, mainly a result of melting pool. The measured data on the Young's modulus and yield strength of the EBM porous samples can be predicted by the Gibson and Ashby model. All samples with high porosity were found to match well with cancellous bone, with Young's modulus of 2 GPa and yield stress of 31 MPa, effective to diminish the risk of stress shielding effect. For porous EBM sample with high 80% porosity, the work of fracture can increase from 15.9 to 47.6 kJ/m2 with increasing ligament width from 600 to 900 μm.

Quick x-ray microtomography using a laser-driven betatron source

Laser-driven X-ray sources are an emerging alternative to conventional X-ray tubes and synchrotron sources. We present results on microtomographic X-ray imaging of a cancellous human bone sample using synchrotron-like betatron radiation. The source is driven by a 100-TW-class titanium-sapphire laser system and delivers over $10^8$ X-ray photons per second. Compared to earlier studies, the acquisition time for an entire tomographic dataset has been reduced by more than an order of magnitude. Additionally, the reconstruction quality benefits from the use of statistical iterative reconstruction techniques. Depending on the desired resolution, tomographies are thereby acquired within minutes, which is an important milestone towards real-life applications of laser-plasma X-ray sources.

Comparison of the Pullout Strength of Different Pedicle Screw Designs and Augmentation Techniques in an Osteoporotic Bone Model.

Study DesignMechanical study.PurposeTo compare the pullout strength of different screw designs and augmentation techniques in an osteoporotic bone model.Overview of LiteratureAdequate bone screw pullout strength is a common problem among osteoporotic patients. Various screw designs and augmentation techniques have been developed to improve the biomechanical characteristics of the bone–screw interface.MethodsPolyurethane blocks were used to mimic human osteoporotic cancellous bone, and six different screw designs were tested. Five standard and expandable screws without augmentation, eight expandable screws with polymethylmethacrylate (PMMA) or calcium phosphate augmentation, and distal cannulated screws with PMMA and calcium phosphate augmentation were tested. Mechanical tests were performed on 10 unused new screws of each group. Screws with or without augmentation were inserted in a block that was held in a fixture frame, and a longitudinal extraction force was applied to the screw head at a loading rate of 5 mm/min. Maximum load was recorded in a load displacement curve.ResultsThe peak pullout force of all tested screws with or without augmentation was significantly greater than that of the standard pedicle screw. The greatest pullout force was observed with 40-mm expandable pedicle screws with four fins and PMMA augmentation. Augmented distal cannulated screws did not have a greater peak pullout force than nonaugmented expandable screws. PMMA augmentation provided a greater peak pullout force than calcium phosphate augmentation.ConclusionsExpandable pedicle screws had greater peak pullout forces than standard pedicle screws and had the advantage of augmentation with either PMMA or calcium phosphate cement. Although calcium phosphate cement is biodegradable, osteoconductive, and nonexothermic, PMMA provided a significantly greater peak pullout force. PMMA-augmented expandable 40-mm four-fin pedicle screws had the greatest peak pullout force.