3D printed agglomerates for granule breakage tests

In the research into agglomeration, a long term barrier is the lack of a universally accepted method to evaluate the breakage propensity of agglomerates. Computer simulation is often used but is limited by the lack of identical, controlled agglomerates to test and validate simple models, let alone replicate the complex structure of real industrial agglomerates. This paper presents work on the characterisation of strength of model test agglomerates prepared by a 3D printing production method enabling fully reproducible structures. Agglomerates were designed using Solidworks 2014 software and printed by an Objet500 Connex 3D printer. Materials with different mechanical properties were used to print the particles and the inter particle bonds, allowing a series of combinations of bond strength, particle strength and agglomerate structure to be tested. Compression and impact tests were performed to investigate the breakage behaviour of the printed agglomerates in terms of agglomerate orientations, bond properties and strain rates. This method will allow more rigorous testing of agglomerate breakage models

Bioactivation of 3D Cell-Imprinted Polydimethylsiloxane Surfaces by Bone Protein Nanocoating for Bone Tissue Engineering

Physical and chemical parameters that mimic the physiological niche of the human body have an influence on stem cell fate by creating directional signals to cells. Micro/nano cell-patterned polydimethylsiloxane (PDMS) substrates, due to their ability to mimic the physiological niche, have been widely used in surface modification. Integration of other factors such as the biochemical coating on the surface can achieve more similar microenvironmental conditions and promote stem cell differentiation to the target cell line. Herein, we investigated the effect of physical topography, chemical functionalization by acid bone lysate (ABL) nanocoating, and the combined functionalization of the bone proteins' nanocoated surface and the topographically modified surface. We prepared four distinguishing surfaces: plain PDMS, physically modified PDMS by 3D cell topography patterning, chemically modified PDMS with bone protein nanocoating, and chemically modified nano 3D cell-imprinted PDMS by bone proteins (ABL). Characterization of extracted ABL was carried out by Bradford staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis, followed by the MTT assay for evaluation of cell viability on ABL-coated PDMS. Moreover, field emission scanning electron microscopy and profilometry were used for the determination of optimal coating thickness, and the appropriate coating concentration was identified and used in the study. The binding and retention of ABL to PDMS were confirmed by Fourier transform infrared spectroscopy and bicinchoninic acid assay. Sessile drop static water contact angle measurements on substrates showed that the combined chemical functionalization and nano 3D cell-imprinting on the PDMS surface improved surface wettability by 66% compared to plain PDMS. The results of ALP measurement, alizarin red S staining, immunofluorescence staining, and real-time PCR showed that the nano 3D cell-imprinted PDMS surface functionalized by extracted bone proteins, ABL, is able to guide the fate of adipose derived stem cellss toward osteogenic differentiation. Eventually, chemical modification of the cell-imprinted PDMS substrate by bone protein extraction not only improved the cell adhesion and proliferation but also contributed to the topographical effect itself and caused a significant synergistic influence on the process of osteogenic differentiation

Analysis of pin milling of pharmaceutical materials

Milling is an important process for tailoring the particle size distribution for enhanced dissolution, content uniformity, tableting, etc., specially for active pharmaceutical ingredients and excipient in pharmaceutical industries. Milling performance of particulate solids depends on the equipment operating conditions (geometry, process conditions and input energy etc.) as well as material properties (particle size, shape, and mechanical properties, such as Young’s modulus, hardness and fracture toughness). In this work, a newly developed approach to assess the breakability of pharmaceutical materials using an aerodynamic dispersion method has been combined with the Discrete Element Method (DEM) to simulate the dynamic behaviour of a number of pharmaceutical materials in a pin mill. A sensitivity analysis is carried out addressing the effect of the milling conditions (rotational speed of the mill and feed particle flow rate) and feed properties on the milled products in terms of the shift in the specific surface area of the milled particles. The outcome of the work is used as a method to predict the breakage of the particles for the milling conditions where chipping takes place

3D Traction Forces in Cancer Cell Invasion

Cell invasion through a dense three-dimensional (3D) matrix is believed to depend on the ability of cells to generate traction forces. To quantify the role of cell tractions during invasion in 3D, we present a technique to measure the elastic strain energy stored in the matrix due to traction-induced deformations. The matrix deformations around a cell were measured by tracking the 3D positions of fluorescent beads tightly embedded in the matrix. The bead positions served as nodes for a finite element tessellation. From the strain in each element and the known matrix elasticity, we computed the local strain energy in the matrix surrounding the cell. We applied the technique to measure the strain energy of highly invasive MDA-MB-231 breast carcinoma and A-125 lung carcinoma cells in collagen gels. The results were compared to the strain energy generated by non-invasive MCF-7 breast and A-549 lung carcinoma cells. In all cases, cells locally contracted the matrix. Invasive breast and lung carcinoma cells showed a significantly higher contractility compared to non-invasive cells. Higher contractility, however, was not universally associated with higher invasiveness. For instance, non-invasive A-431 vulva carcinoma cells were the most contractile cells among all cell lines tested. As a universal feature, however, we found that invasive cells assumed an elongated spindle-like morphology as opposed to a more spherical shape of non-invasive cells. Accordingly, the distribution of strain energy density around invasive cells followed patterns of increased complexity and anisotropy. These results suggest that not so much the magnitude of traction generation but their directionality is important for cancer cell invasion

Biological Evaluation of a Novel Tissue Engineering Scaffold of Layered Double Hydroxides (LDHs)

Bone Tissue Engineering (BTE) Composed of Three Main Parts: Scaffold, Cells and Signaling Factors. Several Materials and Composites Are Suggested as a Scaffold for BTE. Biocompatibility is One of the Most Important Property of a BTE Scaffold. in This Work Synthesis of a Novel Nanocomposite Including Layered Double Hydroxides (LDH) and Gelatin is Carried Out and its Biological Properties Were Studied. the Co-Precipitation (PH=11) Method Was Used to Prepare the LDH Powder, using Calcium Nitrate, Magesium Nitrate and Aluminum Nitrate Salts as Starting Materials. the Resulted Precipitates Were Dried. X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM) Analyses Were Used to Characterize the Synthesized Powders. the Results Demonstrated the Presence of Nanocrystals of Ca-LDH and Mg-LDH as Hexagonal and Layered Morphology. the Obtained Powders Were Composed to Gelatin Via Solvent Casting Method Then Freez Dried. the Scaffold Was Prepared Via Membrane Lamination Method from the Resulted Layers that Linked Together with Gelatin as Binder. in Order to Investigate the Scaffold Cytotoxicity MTT Assay Was Done with a Osteosarcoma Cell Line. No Toxic Response Was Observed in Specimens. as a Major Result, It Was Demonstrated that the Specimen Showed a Significant Cellular Response. Then Osteosarcoma Cells Were Cultured for 7-Day and 14-Day Extract of Powders. the Composites Osteoconductivity Was Investigate with Cells Alkaline Phosphatase Extraction. the Results Demonstrated that the Ca-LDH/gelatin Composite Scaffold Has a Good Potential for Bone Tissue Engineering Applications and Mg-LDH Specimen Has a Better Osteconductivity. © (2012) Trans Tech Publications

NEDD9 Stabilizes Focal Adhesions, Increases Binding to the Extra-Cellular Matrix and Differentially Effects 2D versus 3D Cell Migration

The speed of cell migration on 2-dimensional (2D) surfaces is determined by the rate of assembly and disassembly of clustered integrin receptors known as focal adhesions. Different modes of cell migration that have been described in 3D environments are distinguished by their dependence on integrin-mediated interactions with the extra-cellular matrix. In particular, the mesenchymal invasion mode is the most dependent on focal adhesion dynamics. The focal adhesion protein NEDD9 is a key signalling intermediary in mesenchymal cell migration, however whether NEDD9 plays a role in regulating focal adhesion dynamics has not previously been reported. As NEDD9 effects on 2D migration speed appear to depend on the cell type examined, in the present study we have used mouse embryo fibroblasts (MEFs) from mice in which the NEDD9 gene has been depleted (NEDD9 −/− MEFs). This allows comparison with effects of other focal adhesion proteins that have previously been demonstrated using MEFs. We show that focal adhesion disassembly rates are increased in the absence of NEDD9 expression and this is correlated with increased paxillin phosphorylation at focal adhesions. NEDD9−/− MEFs have increased rates of migration on 2D surfaces, but conversely, migration of these cells is significantly reduced in 3D collagen gels. Importantly we show that myosin light chain kinase is activated in 3D in the absence of NEDD9 and is conversely inhibited in 2D cultures. Measurement of adhesion strength reveals that NEDD9−/− MEFs have decreased adhesion to fibronectin, despite upregulated α5β1 fibronectin receptor expression. We find that β1 integrin activation is significantly suppressed in the NEDD9−/−, suggesting that in the absence of NEDD9 there is decreased integrin receptor activation. Collectively our data suggest that NEDD9 may promote 3D cell migration by slowing focal adhesion disassembly, promoting integrin receptor activation and increasing adhesion force to the ECM

Jamming during particle spreading in additive manufacturing

Additive manufacturing (AM) is going through an exponential growth, due to its enormous potential for rapid manufacturing of complex shapes. One of the manufacturing methods is based on powder processing, but its major bottleneck is associated with powder spreading, as mechanical arching adversely affects both product quality and speed of production. Here we analyse transient jamming of gas-atomised metal powders during spreading. These particles are highly frictional, as they have asperities and multiple spheres and are prone to jamming in narrow gaps. Therefore their detailed characterisations of mechanical properties are critical to be able to reliably predict the jamming frequency as influenced by powder properties and process conditions. Special methods have been used to determine the physical and mechanical properties of gas-atomised stainless steel powders. These properties are then used in numerical simulations of powder spreading by the Discrete Element Method. Particle shape is reconstructed for the simulations as a function of particle size. The characteristic size D₉₀ by number (i.e. the particle size, based on the projected-area diameter, for which 90% of particles by number are smaller than this value) is used as the particle dimension accountable for jamming. Jamming is manifested by empty patches over the work surface. Its frequency and period have been characterised as a function of the spreader gap height, expressed as multiple of D₉₀. The probability of formation of empty patches and their mean length, the latter indicating jamming duration, increase sharply with the decrease of the gap height. The collapse of the mechanical arches leads to particle bursts after the blade. The frequency of jamming for a given survival time decreases exponentially as the survival time increases

Experimental study of the deformation and breakage of 3D printed agglomerates: Effects of packing density and inter-particle bond strength

Characterization of the mechanical properties of agglomerates is important in order to understand their deformation and breakage. However, research progress has been hampered by limitations in our ability to manufacture reproducible agglomerates with well-controlled and fully characterised mechanical properties. In this paper, we report on the preparation and testing of agglomerates with tuneable properties using 3D printing technology. Two typical agglomerate structures with different packing densities were designed and printed using a PolyJet 3D printer. Each agglomerate consisted of rigid primary particles connected by either rigid or rubber-like inter-particle cylindrical bonds. Compression tests (using speeds in the range 0.02–0.5 mm/s) and drop weight impact tests were carried out to investigate the effect of bond material and strain rate on mechanical properties of the agglomerates. The results show that strain rate affects their deformation and breakage significantly, and breakage patterns of the two structures are different under uniaxial compression and impact test conditions. These results demonstrate the broad utility of 3D printed agglomerates as ideal “test” agglomerates for a range of breakage studies, including validating computer simulations of DEM breakage