Spectroscopy on the wing: Investigating possible differences in protein secondary structures in feather shafts of birds using Raman spectroscopy.

The central shaft of a bird's flight feather bears most of the aerodynamic load during flight and exhibits some remarkable mechanical properties. The shaft comprises two parts, the calamus and the rachis. The calamus is at the base of the shaft, while the rachis is the longer upper part which supports the vanes. The shaft is composed of a fibrous outer cortex, and an inner foam-like core. Recent nanoindentation experiments have indicated that reduced modulus values, Er, for the inner and outer regions of the cortex can vary, with the Er values of the inner region slightly greater than those of the outer region. In this work, Raman spectroscopy is used to investigate the protein secondary structures in the inner and outer regions of the feather cortex. Analysis of the Amide I region of Raman spectra taken from four birds (Swan, Gull, Mallard and Kestrel) shows that the ß-sheet structural component decreases between the inner and outer region, relative to the protein side-chain components. This finding is consistent with the proposal that Er values are greater in the inner region than the outer region. This work has shown that Raman spectroscopy can be used effectively to study the change in protein secondary structure between the inner and outer regions of a feather shaft.

Predicting the Fracture Toughness of Human Cancellous Bone in Fractured Neck of Femur Patients Using Bone Volume and Micro-Architecture.

The current protocol used to determine if an individual is osteoporotic relies on assessment of the individual's bone mineral density (BMD), which allows clinicians to judge the condition of a patient with respect to their peers. This, in essence, evaluates a person's fracture risk, because BMD is a good surrogate measure for strength and stiffness. In recent studies, the authors were the first to produce fracture toughness (FT) data from osteoporotic (OP) and osteoarthritic (OA) patients, by using a testing technique which basically analyzes the prerequisite stress conditions for the onset of growth of a major crack through cancellous bone tissue. FT depends mainly on bone quantity (BV/TV, bone volume/tissue volume), but also on bone micro-architecture (mArch), the inner trabecular design of the bone. The working research hypothesis of the present study is that mArch offers added prediction power to BV/TV in determining FT parameters. Consequently, our aim was to investigate the use of predictive models for fracture toughness and also to investigate if there are any significant differences between the models produced from samples loaded across (AC, transverse to) the main trabecular orientation and along (AL, in parallel) the trabeculae. In multilinear regression analysis, we found that the strength of the relationships varied for a crack growing in these two orthogonal directions. Adding mArch variables in the Ac direction helped to increase the R2 to 0.798. However, in the AL direction, adding the mArch parameters did not add any predictive power to using BV/TV alone; BV/TV on its own could produce R2 = 0.730. The present results also imply that the anisotropic layout of the trabeculae makes it more difficult for a major crack to grow transversely across them. Cancellous bone models and remodels itself in a certain way to resist fracture in a specific direction, and thus, we should be mindful that architectural quality as well as bone quantity are needed to understand the resistance to fracture.

Effect of hormone replacement therapy in matrix metalloproteinase expression and intimal hyperplasia development after vascular injury.

BACKGROUND: Postmenopausal women taking hormone replacement therapy (HRT) require secondary intervention after vascular reconstruction more frequently than women not taking HRT, often due to increased development of intimal hyperplasia (IH). Matrix metalloproteinases (MMPs) play a role in IH by degradation and remodeling of components of the vascular basement membrane. The MMP pathway is regulated by a balance between MMPs, membrane-type MMPs (MT-MMPs), and tissue inhibitor of MMPs (TIMPs). We have recently provided evidence for unbalanced regulation of the MT1-MMP/MMP-2 pathway in vascular smooth muscle cells (VSMCs) exposed to hormones in vitro. Herein we study the role of HRT in the development of IH in a postmenopausal rodent model of vascular injury and in the modulation of this MMP regulatory pathway in vivo. METHODS: Female rats were aged to 12 months. Animals were ovariectomized (OVX) and 4 weeks later hormones or placebo was delivered via a 90-day slow-release pellet. After 6 weeks of HRT each rat underwent balloon angioplasty of the left common carotid artery. At 14 days postinjury tissue samples were collected and stained with trichrome elastin and for isoform-specific MMPs. RESULTS: After vascular injury, the intima:media (I:M) ratio was decreased in OVX rats receiving placebos as compared with non-OVX controls (P < 0.05). In OVX animals receiving HRT, estrogen with and without progesterone and progesterone alone slightly increased I:M ratio compared with placebo, although no significant difference was found in any HRT group. Injury-induced intimal expression of MMP-2 and -9 was decreased in OVX placebo animals compared with non-OVX controls (P < 0.05). MMP-2 and -9 levels were subsequently increased by each type of hormone therapy compared with placebo, with a significant increase in MMP-9 in response to estrogen with and without progesterone (P < 0.05). Conversely, TIMP-2 was decreased by estrogen compared with placebo (P < 0.05). There was no effect on intimal MT1-MMP in any group. CONCLUSIONS: In this study we detected a statistically significant decrease in IH as a result of OVX. Subsequent HRT exposure resulted in increased I:M ratios compared with OVX animals given placebo, although significance was not reached with the doses given. Long-term exogenous exposure may have a more deleterious effect compared with acute exposure and should be examined further. We also demonstrated a significant reduction in MMP-2 and -9 and TIMP-2 in response to OVX. Subsequent hormone exposure resulted in the upregulation of MMP-2 and -9 without a counterregulatory increase in TIMP, indicating that HRT modulates the MMP regulatory pathway in vivo. The data suggest that the lack of hormones after OVX protects against pathologic remodeling in our aged model of disease and that exposure to both natural and exogenous hormones could be a negative risk factor resulting in an exaggerated vascular response to injury. Future studies should focus on in vivo manipulation of unbalanced MMP regulation for prevention of IH in response to HRT and in general. Furthermore, the age-associated difference in response to the presence of natural hormones in young vs aged models should be investigated.

Pseudotumour formation due to tribocorrosion at the taper interface of large diameter metal on polymer modular total hip replacements.

We present an in-depth failure analysis of two large diameter bearing metal-on-polymer (MoP) modular total hip replacements, which have required revision surgery due to pseudotumour formation. The failure analysis showed a discrete pattern of material loss from the distal end of the head taper/stem trunnion interface. We postulate that the use of a proximal contacting taper design had provided insufficient mechanical locking between the head and the stem, enabling the head to toggle on the trunnion. In addition, the difference in angle between the taper and the trunnion formed a crevice between the two components. Through a combination of crevice environment, mechanically assisted corrosion, mechanical wear and erosion; debris and metal-ions have been released resulting in the adverse local tissue reactions (ALTR).

Depth profiling via nanoindentation for characterisation of the elastic modulus and hydraulic properties of thin hydrogel layers.

The accurate determination of the mechanical properties of hydrogels is of fundamental importance for a range of applications, including in assessing the effect of stiffness on cell behaviour. This is a particular issue when using thin hydrogel layers adherent to stiff substrate supports, as the apparent stiffness can be significantly influenced by the constraint of the underlying impermeable substrate, leading to inaccurate measurements of the elastic modulus and permeability of thin hydrogel layers. This study used depth profiling nanoindentation and a poroelastic model for spherical indentation to identify the elastic moduli and hydraulic conductivity of thin polyacrylamide (PAAm) hydrogel layers (∼27 µm-782 µm thick) on impermeable substrates. The apparent stiffness of thin PAAm layers increased with indentation depth and was significantly greater than those of thicker hydrogels, which showed no influence of indentation depth. The hydraulic conductivity decreased as the geometrical confinement of hydrogels increased, indicating that the fluid became more constrained within the confinement areas. The impact of geometrical confinement on the apparent modulus and hydraulic conductivity of thin PAAm hydrogel layers was then established, and their elastic moduli and intrinsic permeability were determined in relation to this effect. This study offers valuable insights into the mechanical characterisation of thin PAAm hydrogel layers used for the fundamental study of cell mechanobiology.

Measuring the elastic modulus of soft biomaterials using nanoindentation.

The measurement of the elastic modulus of soft biomaterials via nanoindentation relies on the accurate determination of the zero-point of the tip-sample interaction on which the depth of penetration into the sample is based. Non-cantilever based nanoindentation systems were originally designed for hard materials, and therefore monitoring the zero-point contact presents a significant challenge for the characterisation of very soft biomaterials. This study investigates the ability of non-cantilever based nanoindentation to differentiate between hydrogels with elastic moduli on the order of single kiloPascals (kPa) using a bespoke soft contact protocol and low flexural stiffness of instrument. Polyethylene glycol (PEG) hydrogels were fabricated as a model system with a range of elastic moduli by varying the polymer concentration and degree of crosslinking. Elastic modulus values were calculated using the Oliver-Pharr method, Hertzian contact model, as well as a viscoelastic model to account for the time-dependent behaviour of the gels. The stiffness measurements were validated by measuring cantilever beams with the equivalent flexural stiffness to that of the PEG hydrogels being tested. The results demonstrated a high repeatability of the measurements, enabling differentiation between hydrogels with elastic moduli in the single kPa to hundreds of kPa range.

Influence of food consistency on growth and morphology of the mandibular condyle.

The objective of this study was to determine if variation in the shape and mineralization of the mandibular condyle are the result of natural adaptation in response to different functional loading demands. Eight female Kuni Kuni piglets were randomly assigned to two groups of four, receiving either a soft or hard diet. Each animal was given three separate doses of vital stains intravenously at set time points during the study. At 8.5 months, animals were euthanized and temporomandibular joints (TMJs) were excised. Histological analysis was used to measure the amount of new bone deposition in the anterior, central, and posterior regions of the mandibular condyle. Backscatter electron (BSE) imaging was used as a semiquantitative estimate of bone mineralization in these two diet groups. Histology revealed that the degree of new bone deposition in the hard-diet group was significantly (n = 4, P < 0.001, paired t-test) higher than that of the soft-diet group. Also, the majority (87%) of animals fed a hard diet tended to show greater new bone deposition on the leftside in comparison to the right, indicating a chewing preference for the left side. In both groups, the degree of new bone deposition was significantly (P < 0.01) higher in the posterior area than in other regions. BSE imaging corroborated basic histology results, with significantly (P < 0.01) higher mineralization levels detected in the hard-diet group. These findings indicate that diet consistency has a small but significant effect on the rate of bone deposition in the mandibular condyle.

Quantitative assessment of dentine mineralization and tubule occlusion by NovaMin and stannous fluoride using serial block face scanning electron microscopy.

Dentine hypersensitivity (DH) is one of the most common dental conditions affecting most adults during their lifetime. Tubule occlusion is a widely accepted method for treating DH. Current in-vitro techniques such as focused ion beam, scanning electron microscopy (SEM), or hydraulic conductance that are used to determine tubule occlusion do not provide the depth of occlusion, are time-consuming, expensive and the volume of dentine tested is limited. The presented study aimed to assess the ability of serial block-face SEM (SBF-SEM) to section dentine, to quantify the number of occluded tubules including the depth of penetration by NovaMin and stannous fluoride (SnF2 ) and to compare mineral density between the control and treated dentine. Results demonstrated that NovaMin provided a better occlusion with 100% of the tubules blocked at the surface compared to 83% for SnF2 . The grayscale value (230.42) was significantly higher (p ≤ 0.05) after treatment with NovaMin compared to SnF2 (222.06) and the control (196.37), indicating increased mineral density and dentine mineralization. SBF-SEM has the potential to be used for large volume analysis of bone-like materials at high resolution with minimal sample preparation over a short period. It can be significantly useful in the development and research of new biomaterials.

It's worth cleaning - The examination of the female taper could identify a particular cause of trunnionosis at revision 16 years after total hip arthroplasty.

Adverse reaction to metal debris (ARMD) is an issue in metal-on-metal (MoM) total hip replacements (THR). It mainly affects large-head MoM THR, whereas 28-32 mm MoM pairings are associated with low long-term revision rates. However, the bearing surface is not necessarily the only cause of metal debris. This report documents with advanced analysis of the retrievals a particular cause of trunnionosis in late failure of a small diameter MoM THR and illustrates the importance of cleaning of the taper when seating the head in THR. A 65-year-old patient was revised due to ARMD 16 years after small diameter MoM THR. Debridement and exchange of the inlay and the head had been performed through an anterior approach. While the cup and the outer surface of the head were accessible to direct analysis by an optical coordinate measuring machine, the female taper had to be analysed indirectly by measuring an imprint. Wear from the cup and the head was within expected low ranges. The analysis of the female taper identified bone fragments, which contributed to trunnionosis. Failure due to ARMD after MoM THR is not necessarily caused by the bearing, but can be due to trunnionosis. Bone fragments within the taper contact in this case highlight the importance of meticulous cleaning of the taper before seating the head, to avoid trunnionosis.

Structure and properties of baleen in the Southern right (Eubalaena australis) and Pygmy right whales (Caperea marginata).

Baleen is a resilient and keratinised filter-feeding structure attached to the maxilla of mysticete whales. It is strong and tough, yet a pliant and resilient material, that withstands extreme pressures in the oral cavity during feeding. We investigated the structure, water content, wettability and mechanical properties of baleen of the Southern right (SRW) and Pygmy right whales (PRW), to understand the effects of hydration on the physical and mechanical properties of baleen. Sixty 25 × 15mm baleen subsamples were prepared from one individual of SRW and PRW. Half were hydrated in circulated natural seawater for 21 days and half were dry. Water content analysis showed that SRW baleen was 21.2% water weight and PRW was 26.1%. Wettability testing indicated that surfaces of both hydrated and dried SRW and PRW baleen were hydrophilic, with hydrated samples of both species having lower contact angle values. For the SRW, the average contact angle of hydrated baleen was 40° ± 13.2 and 73° ± 6 for dried samples. Hydrated PRW baleen had an average contact angle of 44° ± 15.3, which was lower than in dried samples (74° ± 2.9). Three-point bending mechanical tests showed that the average maximum flexural stress of dried SRW (134.1 ± 34.3 MPa) and PRW samples (117.8 ± 22.3 MPa) were significantly higher than those of hydrated SRW (25.7 ± 6.3 MPa) and PRW (19.7 ± 4.8 MPa) baleen. Scanning electron microscope images showed the stratification of the outer cortical layer, with cross-linked keratin fibres observed within and between baleen keratin sheets. Hydrated baleen, as in its natural and functional behaviour, has greater flexibility and strength, attributes necessary for the complex filter feeding mechanism characteristic of whales. Hydration must be considered when addressing the physical and mechanical properties of baleen, especially when using dried museum specimens.

A comparative study on the physicochemical characteristics of nanoparticles released in vivo from CoCrMo tapers and cement-stem interfaces of total hip replacements.

The good biocompatibility and corrosion resistance of the bulk CoCrMo alloy has resulted in it being used in the manufacture of implants and load bearing medical devices. These devices, however, can release wear and corrosion products which differ from the composition of the bulk CoCrMo alloy. The physicochemical characteristics of the particles and the associated in vivo reactivity are dictated by the wear mechanisms and electrochemical conditions at the sites of material loss. Debris released from CoCrMo hip bearings, taper junctions, or cement-stem interfaces can, therefore, have different chemical and morphological characteristics, which provide them with different in vivo toxicities. Here, we propose to assess and compare the characteristics of the particles released in vivo from CoCrMo tapers and cement-stem interfaces which have received less attention compared to debris originating from the hip bearings. The study uses state-of-art characterization techniques to provide a detailed understanding of the size, morphology, composition, and chemistry of the particles liberated from the wear and corrosion flakes from revised hip replacements, with an enzymatic treatment. The phase analyses identified Cr2 O3 nanoparticles released from tapers and cement-stem interfaces, whose composition did not vary with origin or particle morphology. The size distributions showed significantly smaller particles were released from the stems, compared to the particles originating from the corresponding tapers. The investigation demonstrates that the tribocorrosive processes occurring at the taper and stem interfaces both result in Cr2 O3 nanoparticle formation.

Upregulation of epithelial metallothioneins by metal-rich ultrafine particulate matter from an underground railway.

Airborne particulate matter (PM) is a leading cause of mortality and morbidity. However, understanding of the range and mechanisms of effects of PM components is poor. PM generated in underground railways is rich in metals, especially iron. In the ultrafine (UFPM; <0.1 µm diameter) fraction, the combination of small size and metal enrichment poses an unknown health risk. This study aimed to analyse transcriptomic responses to underground UFPM in primary bronchial epithelial cells (PBECs), a key site of PM deposition. The oxidation state of iron in UFPM from an underground station was determined by X-ray absorption near edge structure (XANES) spectroscopy. Antioxidant response was assayed using a reporter cell line transfected with an antioxidant response element (ARE)-luciferase construct. Differentiated PBECs were exposed to UFPM for 6 h or 24 h for RNA-Seq and RT-qPCR analysis. XANES showed predominance of redox-active Fe3O4, with ROS generation confirmed by induction of ARE-luciferase expression. 6 h exposure of PBECs to UFPM identified 52 differentially expressed genes (DEGs), especially associated with epithelial maintenance, whereas 24 h exposure yielded 23 DEGs, particularly involved with redox homeostasis and metal binding. At both timepoints, there was upregulation of members of the metallothionein family, low molecular weight proteins with antioxidant activity whose main function is binding and homeostasis of zinc and copper ions, but not iron ions. This upregulation was partially inhibited by metal chelation or ROS scavenging. These data suggest differential regulation of responses to metal-rich UFPM depending on exposure period, and highlight novel pathways and markers of PM exposure, with the role of metallothioneins warranting further investigation.

Investigation of nano-sized debris released from CoCrMo secondary interfaces in total hip replacements: Digestion of the flakes.

The in vivo release of wear debris and corrosion products from the metallic interfaces of total hip replacements is associated with a wide spectrum of adverse body reactions and systemic manifestations. The origin of debris and the electrochemical conditions at the sites of material loss both play a role in determining the physicochemical characteristics of the particles, and thus influence their in vivo reactivity. Debris retrieved from revised CoCrMo tapers and cement-stem interfaces consists of heterogeneous flakes that comprise mechanically mixed metal particles, corrosion products and organic material. Detailed investigation of the size and composition of the metal debris contained within these composites requires the digestion of the flakes to release the small metal particles. Here, we compare alkaline and enzymatic digestion methods that both aim to fragment the flakes and reveal their smallest building blocks. The characterization of debris cleaned with both methods revealed crystalline Cr oxide nanoparticles and clusters. Comparison between the treatments showed that the alkaline method is more efficient in fragmenting the flakes and provided cleaner and generally smaller nanoparticles than exhibited in debris released with the enzymatic treatment. The provision of cleaner nanoparticles from the alkaline method also allows the physicochemical properties of the particles to be more clearly identified. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 107B: 424-434, 2019.

Some basic relationships between density values in cancellous and cortical bone.

Density is a salient property of bone and plays a crucial role in determining the mechanical properties of both its cancellous and cortical structural forms. Density is defined in a number of ways at either the bone tissue (D(app), apparent) or the bone material level (D(mat), material). The concept of density is relatively simple, but measuring it in the context of bone is a complex issue. The third dimension of the problem is the concept of porosity, or BV/TV (ratio of bone material volume over tissue volume). Recent investigations from our laboratory have revealed an interdependence of D(app) and D(mat) in the cancellous bone of at least four different cohorts of human patients. To clarify the underlying causes of this behaviour, we produced here equivalent relationships from specimens originating from cortical and cancellous areas of the same bone. Plots of D(app) vs. D(mat) showed that D(mat) was not a monotonic function of increasing D(app), but instead showed a 'boomerang'-like pattern. By empirically dissecting the data in two regions for D(app) above and below a value equal to 1.3gcm(-3), we were able to objectively isolate the bone in trabecular and compact forms. Our findings may have implications not only for the segregation of bone in these two structural forms, but also for the mechanobiological and physiological processes that govern the regulation of compact and trabecular bone areas.

Human Skeletal Stem Cell Response to Multiscale Topography Induced by Large Area Electron Beam Irradiation Surface Treatment.

The healthcare socio-economic environment is irreversibly changing as a consequence of an increasing aging population, consequent functional impairment, and patient quality of life expectations. The increasing complexity of ensuing clinical scenarios compels a critical search for novel musculoskeletal regenerative and replacement strategies. While joint arthroplasty is a highly effective treatment for arthritis and osteoporosis, further innovation and refinement of uncemented implants are essential in order to improve implant integration and reduce implant revision rate. This is critical given financial restraints and the drive to improve cost-effectiveness and quality of life outcomes. Multi-scale modulation of implant surfaces, offers an innovative approach to enhancement in implant performance. In the current study, we have examined the potential of large area electron beam melting to alter the surface nanotopography in titanium alloy (Ti6Al4V). We evaluated the in vitro osteogenic response of human skeletal stem cells to the resultant nanotopography, providing evidence of the relationship between the biological response, particularly Collagen type I and Osteocalcin gene activation, and surface nanoroughness. The current studies demonstrate osteogenic gene induction and morphological cell changes to be significantly enhanced on a topography Ra of ~40 nm with clinical implications therein for implant surface treatment and generation.

Acoustically modulated biomechanical stimulation for human cartilage tissue engineering.

Bioacoustofluidics can be used to trap and levitate cells within a fluid channel, thereby facilitating scaffold-free tissue engineering in a 3D environment. In the present study, we have designed and characterised an acoustofluidic bioreactor platform, which applies acoustic forces to mechanically stimulate aggregates of human articular chondrocytes in long-term levitated culture. By varying the acoustic parameters (amplitude, frequency sweep, and sweep repetition rate), cells were stimulated by oscillatory fluid shear stresses, which were dynamically modulated at different sweep repetition rates (1-50 Hz). Furthermore, in combination with appropriate biochemical cues, the acoustic stimulation was tuned to engineer human cartilage constructs with structural and mechanical properties comparable to those of native human cartilage, as assessed by immunohistology and nano-indentation, respectively. The findings of this study demonstrate the capability of acoustofluidics to provide a tuneable biomechanical force for the culture and development of hyaline-like human cartilage constructs in vitro.

Localized tissue mineralization regulated by bone remodelling: A computational approach.

Bone is a living tissue whose main mechanical function is to provide stiffness, strength and protection to the body. Both stiffness and strength depend on the mineralization of the organic matrix, which is constantly being remodelled by the coordinated action of the bone multicellular units (BMUs). Due to the dynamics of both remodelling and mineralization, each sample of bone is composed of structural units (osteons in cortical and packets in cancellous bone) created at different times, therefore presenting different levels of mineral content. In this work, a computational model is used to understand the feedback between the remodelling and the mineralization processes under different load conditions and bone porosities. This model considers that osteoclasts primarily resorb those parts of bone closer to the surface, which are younger and less mineralized than older inner ones. Under equilibrium loads, results show that bone volumes with both the highest and the lowest levels of porosity (cancellous and cortical respectively) tend to develop higher levels of mineral content compared to volumes with intermediate porosity, thus presenting higher material densities. In good agreement with recent experimental measurements, a boomerang-like pattern emerges when plotting apparent density at the tissue level versus material density at the bone material level. Overload and disuse states are studied too, resulting in a translation of the apparent-material density curve. Numerical results are discussed pointing to potential clinical applications.

Damage mechanisms at the cement-implant interface of polished cemented femoral stems.

The occurrence of damage on polished femoral stems has been widely reported in the literature, and bone cement has been implicated in a tribocorrosive failure process. However, the mechanisms of cement-mediated damage and the impact of cement formulation on this process are not well understood. In this study, 13 Zimmer CPT polished femoral stems, and the corresponding cement specimens were retrieved at revision surgery and analyzed using high-resolution imaging techniques. Surface damage attributed to tribocorrosion was observed on all stems. Corrosion product, in the form of black flaky surface debris, was observed on the surface of cement specimens; both energy-dispersive X-ray spectroscopy and inductively coupled plasma mass spectrometry(ICP-MS) confirmed the presence of cobalt and chromium, with the ICP-MS showing much higher levels of Cr compared to Co when compared to the original stem material. Agglomerates of ZrO2 radiopacifier were also identified on the cement surface and, in some cases, showed evidence of abrasive wear; the size of these particles correlated well with elliptical pitting evident on the surfaces of the corresponding stems. This evidence supports the hypothesis that agglomerates of hard radiopacifier particles within the cement may induce a wear-dominated tribocorrosive interaction at the stem-cement interface that damages the surface of polished CoCr femoral stems. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2027-2033, 2017.

Large animal in vivo evaluation of a binary blend polymer scaffold for skeletal tissue-engineering strategies; translational issues.

Binary blend polymers offer the opportunity to combine different desirable properties into a single scaffold, to enhance function within the field of tissue engineering. Previous in vitro and murine in vivo analysis identified a polymer blend of poly(l-lactic acid)-poly(ε-caprolactone) (PLLA:PCL 20:80) to have characteristics desirable for bone regeneration. Polymer scaffolds in combination with marrow-derived skeletal stem cells (SSCs) were implanted into mid-shaft ovine 3.5 cm tibial defects, and indices of bone regeneration were compared to groups implanted with scaffolds alone and with empty defects after 12 weeks, including micro-CT, mechanical testing and histological analysis. The critical nature of the defect was confirmed via all modalities. Both the scaffold and scaffold/SSC groups showed enhanced quantitative bone regeneration; however, this was only found to be significant in the scaffold/SSCs group (p = 0.04) and complete defect bridging was not achieved in any group. The mechanical strength was significantly less than that of contralateral control tibiae (p < 0.01) and would not be appropriate for full functional loading in a clinical setting. This study explored the hypothesis that cell therapy would enhance bone formation in a critical-sized defect compared to scaffold alone, using an external fixation construct, to bridge the scale-up gap between small animal studies and potential clinical translation. The model has proved a successful critical defect and analytical techniques have been found to be both valid and reproducible. Further work is required with both scaffold production techniques and cellular protocols in order to successfully scale-up this stem cell/binary blend polymer scaffold. © 2015 The Authors. Journal of Tissue Engineering and Regenerative Medicine published by John Wiley & Sons, Ltd.

Nanomechanical properties of bird feather rachises: exploring naturally occurring fibre reinforced laminar composites.

Flight feathers have evolved under selective pressures to be sufficiently light and strong enough to cope with the stresses of flight. The feather shaft (rachis) must resist these stresses and is fundamental to this mode of locomotion. Relatively little work has been done on rachis morphology, especially from a mechanical perspective and never at the nanoscale. Nano-indentation is a cornerstone technique in materials testing. Here we use this technique to make use of differentially oriented fibres and their resulting mechanical anisotropy. The rachis is established as a multi-layered fibrous composite material with varying laminar properties in three feathers of birds with markedly different flight styles; the Mute Swan (Cygnus olor), the Bald Eagle (Haliaeetus leucocephalus) and the partridge (Perdix perdix). These birds were chosen not just because they are from different clades and have different flight styles, but because they have feathers large enough to gain meaningful results from nano-indentation. Results from our initial datasets indicate that the proportions and orientation of the laminae are not fixed and may vary either in order to cope with the stresses of flight particular to the bird or with phylogenetic lineage.