429 research outputs found
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The influence of silanisation on the mechanical and degradation behaviour of PLGA/HA composites.
This study investigates the influence of silanisation on the mechanical and degradation behaviour of PLGA/HA composites. Three different silanes (mercaptopropyl trimethoxy silane (MPTMS), aminopropyl trimethoxy silane (APTMS) and aminopropyltriethoxy silane (APTES)) were applied to HA substrates in order to study the effect of head group (which binds to the polymer) and tail group (which binds to the surface hydroxyl groups in HA). A composite of hydroxyapatite (HA) and poly(d,l lactide-co-glycolide (50:50)) (PLGA) was investigated. The influence of concentration, the reaction time, drying temperature and substrate surface on silanisation was examined. TGA was used to detect the degree of silanisation. HA with MPTMS (1wt.% MPTMS with reaction time of 1h) was used as filler in PLGA-30wt.% HA composites for an in-vitro degradation study carried out in PBS. In addition, the mechanical properties of the composites were studied. Silanisation affects the properties of the composite by improving the bonding at the interface and hence it was found to influence the plastic mechanical properties rather than the elastic mechanical properties or the degradation profile of the composite.The authors are grateful to Riverside Medical Group for the funding.This is the accepted manuscript for a paper published in Materials Science and Engineering: C Volume 48, 1 March 2015, Pages 642–650, doi: 10.1016/j.msec.2014.12.05
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Crosslinking Collagen Constructs: Achieving Cellular Selectivity Through Modifications of Physical and Chemical Properties
Collagen-based constructs have emerged in recent years as ideal candidates for tissue engineering implants. For many biomedical applications, collagen is crosslinked in order to improve the strength, stiffness and stability of the construct. However, the crosslinking process may also result in unintended changes to cell viability, adhesion or proliferation on the treated structures. This review provides a brief overview of some of both the most commonly used and novel crosslinkers used with collagen, and suggests a framework by which crosslinking methods can be compared and selected for a given tissue engineering application
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Investigating the morphological, mechanical and degradation properties of scaffolds comprising collagen, gelatin and elastin for use in soft tissue engineering.
Collagen-based scaffolds can be used to mimic the extracellular matrix (ECM) of soft tissues and provide support during tissue regeneration. To better match the native ECM composition and mechanical properties as well as tailor the degradation resistance and available cell binding motifs, other proteins or different collagen types may be added. The present study has explored the use of components such as gelatin or elastin and investigated their effect on the bulk physical properties of the resulting scaffolds compared to those made from pure collagen type I. The effect of altering the composition and crosslinking was evaluated in terms of the scaffold structure, mechanical properties, swelling, degradation and cell attachment. Results demonstrate that scaffolds based on gelatin had reduced tensile stiffness and degradation time compared with collagen. The addition of elastin reduced the overall strength and stiffness of the scaffolds, with electron microscopy results suggesting that insoluble elastin interacts best with collagen and soluble elastin interacts best with gelatin. Carbodiimide crosslinking was essential for structural stability, strength and degradation resistance for scaffolds of all compositions. In addition, preliminary cell adhesion studies showed these highly porous structures (pore size 130-160 μm) to be able to support HT1080 cell infiltration and growth. Therefore, this study suggests that the use of gelatin in place of collagen, with additions of elastin, can tailor the physical properties of scaffolds and could be a design strategy for reducing the overall material costs
Targeted protein delivery: carbodiimide crosslinking influences protein release from microparticles incorporated within collagen scaffolds
open access articleTissue engineering response may be tailored via controlled, sustained release of active agents from protein-loaded degradable microparticles incorporated directly within three-dimensional (3D) ice-templated collagen scaffolds. However, the effects of covalent crosslinking during scaffold preparation on the availability and release of protein from the incorporated microparticles have not been explored. Here, we load 3D ice-templated collagen scaffolds with controlled additions of poly-(DL-lactide-co-glycolide) microparticles. We probe the effects of subsequent N-(3-dimethylaminopropyl)- N0-ethylcarbodiimide hydrochloride crosslinking on protein release, using microparticles with different internal protein distributions. Fluorescein isothiocyanate labelled bovine serum albumin is used as a model protein drug. The scaffolds display a homogeneous microparticle distribution,
and a reduction in pore size and percolation diameter with increased microparticle addition, although these values did not fall below those reported as necessary for cell invasion. The protein distribution within the microparticles, near the surface or more deeply located within the microparticles, was important in determining the release profile and effect of crosslinking, as the surface
was affected by the carbodiimide crosslinking reaction applied to the scaffold. Crosslinking of microparticles with a high proportion of protein at the surface caused both a reduction and delay in protein release. Protein located within the bulk of the microparticles, was protected from the crosslinking reaction and no delay in the overall release profile was seen
Feature importance in multi-dimensional tissue-engineering datasets: random forest assisted optimization of experimental variables for collagen scaffolds
Ice-templated collagen-based tissue-engineering scaffolds are ideal for controlled tissue regeneration since they mimic the micro-environment experienced in vivo. The structure and properties of scaffolds are fine-tuned during fabrication by controlling a number of experimental parameters. However, this parameter space is large and complex, rendering the interpretation of results and selection of optimal parameters to be challenging in practice. This paper investigates the impact of a cross section of this parameter space (drying conditions and solute environment) on the scaffold microstructure. Qualitative assessment revealed the previously unreported impact of drying temperature and pressure on pore wall roughness, and confirmed the influence of collagen concentration, solvent type, and solute addition on pore morphology. For quantitative comparison, we demonstrate the novel application of random forest regression to analyze multi-dimensional biomaterials datasets, and predict microstructural attributes for a scaffold. Using these regression models, we assessed the relative importance of the input experimental parameters on quantitative pore measurements. Collagen concentration and pH were found to be the largest factors in determining pore size and connectivity. Furthermore, circular dichroism peak intensities were also revealed to be a good predictor for structural variations, which is a parameter that has not previously been investigated for its effect on a scaffold microstructure. Thus, this paper demonstrates the potential for predictive models such as random forest regressors to discover novel relationships in biomaterials datasets. These relationships between parameters (such as circular dichroism spectra and pore connectivity) can therefore also be used to identify and design further avenues of investigation within biomaterials
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MicroCT analysis of connectivity in porous structures: optimizing data acquisition and analytical methods in the context of tissue engineering.
Micro-computed X-ray tomography (MicroCT) is one of the most powerful techniques available for the three-dimensional characterization of complex multi-phase or porous microarchitectures. The imaging and analysis of porous networks are of particular interest in tissue engineering due to the ability to predict various large-scale cellular phenomena through the micro-scale characterization of the structure. However, optimizing the parameters for MicroCT data capture and analyses requires a careful balance of feature resolution and computational constraints while ensuring that a structurally representative section is imaged and analysed. In this work, artificial datasets were used to evaluate the validity of current analytical methods by considering the effect of noise and pixel size arising from the data capture, and intrinsic structural anisotropy and heterogeneity. A novel 'segmented percolation method' was developed to exclude the effect of anomalous, non-representative features within the datasets, allowing for scale-invariant structural parameters to be obtained consistently and without manual intervention for the first time. Finally, an in-depth assessment of the imaging and analytical procedures are presented by considering percolation events such as micro-particle filtration and cell sieving within the context of tissue engineering. Along with the novel guidelines established for general pixel size selection for MicroCT, we also report our determination of 3 μm as the definitive pixel size for use in analysing connectivity for tissue engineering applications
Multi-scale mechanical response of freeze-dried collagen scaffolds for tissue engineering applications.
Tissue engineering has grown in the past two decades as a promising solution to unresolved clinical problems such as osteoarthritis. The mechanical response of tissue engineering scaffolds is one of the factors determining their use in applications such as cartilage and bone repair. The relationship between the structural and intrinsic mechanical properties of the scaffolds was the object of this study, with the ultimate aim of understanding the stiffness of the substrate that adhered cells experience, and its link to the bulk mechanical properties. Freeze-dried type I collagen porous scaffolds made with varying slurry concentrations and pore sizes were tested in a viscoelastic framework by macroindentation. Membranes made up of stacks of pore walls were indented using colloidal probe atomic force microscopy. It was found that the bulk scaffold mechanical response varied with collagen concentration in the slurry consistent with previous studies on these materials. Hydration of the scaffolds resulted in a more compliant response, yet lesser viscoelastic relaxation. Indentation of the membranes suggested that the material making up the pore walls remains unchanged between conditions, so that the stiffness of the scaffolds at the scale of seeded cells is unchanged; rather, it is suggested that thicker pore walls or more of these result in the increased moduli for the greater slurry concentration conditions.The authors are grateful to the Nano Doctoral Training Centre (NanoDTC), University of Cambridge, and the EPSRC who supported this work through the EP/G037221/1 grant.This is the final published version. It originally appeared at http://www.sciencedirect.com/science/article/pii/S1751616114003397#
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Fabrication of free standing collagen membranes by pulsed-electrophoretic deposition.
This work reports an important new development in the production of collagen membranes, based on pulsed electrophoretic deposition (P-EPD), suitable for a wide range of biomedical applications. Collagen membranes are of great interest as a biomaterial and in a range of other industries, though current production techniques suffer from limitations with scaling up, homogeneity, and complex shapes. P-EPD can be used to rapidly create detachable, large-area, homogeneous products with controlled thickness in a wide variety of shapes. We provide a new understanding of the influence of a range of parameters (pulse width, voltage, duty cycle, solvent additions) and their effects on membrane structure. Characterisation by AFM, SEM, and cryoSEM revealed the ability to produce dense, structurally defect-free membranes, and significantly, we show and discuss the ability to produce thicker membranes by sequential deposition without seeing a corresponding increase in cell electrical resistance. We anticipate this novel, rapid, and controllable method for the production of collagen membranes to be of interest for a wide range of fields.The authors wish to acknowledge the support of theEngineering and Physical Sciences Research Council(EPSRC)grants EP/K503009/1, EP/J500380/1, EP/L504920/1, EP/M506485/1, and EP/M508007/1,Geistlich Pharma AG, and the European ResearchCouncil(ERC)Advanced Grant 320598 3D-
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Self-assembly of collagen bundles and enhanced piezoelectricity induced by chemical crosslinking.
The piezoelectricity of collagen is purported to be linked to many biological processes including bone formation and wound healing. Although the piezoelectricity of tissue-derived collagen has been documented across the length scales, little work has been undertaken to characterise the local electromechanical properties of processed collagen, which is used as a base for tissue-engineering implants. In this work, three chemically distinct treatments used to form structurally and mechanically stable scaffolds-EDC-NHS, genipin and tissue transglutaminase-are investigated for their effect on collagen piezolectricity. Crosslinking with EDC-NHS is noted to produce a distinct self-assembly of the fibres into bundles roughly 300 nm in width regardless of the collagen origin. These fibre bundles also show a localised piezoelectric response, with enhanced vertical piezoelectricity of collagen. Such topographical features are not observed with the other two chemical treatments, although the shear piezoelectric response is significantly enhanced upon crosslinking. These observations are reconciled by a proposed effect of the crosslinking mechanisms on the molecular and nanostructure of collagen. These results highlight the ability to modify the electromechanical properties of collagen using chemical crosslinking methods.ERC, Bill and Melinda Gates Foundation, Geistlich Pharma A
Cell Invasion in Collagen Scaffold Architectures Characterized by Percolation Theory.
The relationship between biological scaffold interconnectivity and cell migration is an important but poorly understood factor in tissue regeneration. Here a scale-independent technique for characterization of collagen scaffold interconnectivity is presented, using a combination of X-ray microcomputed tomography and percolation theory. Confocal microscopy of connective tissue cells reveals this technique as highly relevant for determining the extent of cell invasion.The authors acknowledge financial support from EPSRC, Geistlich Pharma AG and ERC Advanced Grant 320598 3D-E.This is the final version of the article. It first appeared from Wiley at http://dx.doi.org/10.1002/adhm.201500197
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