A definition of bioinks and their distinction from biomaterial inks.

Biofabrication aims to fabricate biologically functional products through bioprinting or bioassembly (Groll et al 2016 Biofabrication 8 013001). In biofabrication processes, cells are positioned at defined coordinates in three-dimensional space using automated and computer controlled techniques (Moroni et al 2018 Trends Biotechnol. 36 384-402), usually with the aid of biomaterials that are either (i) directly processed with the cells as suspensions/dispersions, (ii) deposited simultaneously in a separate printing process, or (iii) used as a transient support material. Materials that are suited for biofabrication are often referred to as bioinks and have become an important area of research within the field. In view of this special issue on bioinks, we aim herein to briefly summarize the historic evolution of this term within the field of biofabrication. Furthermore, we propose a simple but general definition of bioinks, and clarify its distinction from biomaterial inks.

Fully printed high performance humidity sensors based on two-dimensional materials.

Fully printed humidity sensors based on two-dimensional (2D) materials are described. Monolayer graphene oxide (GO) and few-layered black phosphorus (BP) flakes were dispersed in low boiling point solvents suitable for inkjet printing. The humidity sensors were fabricated by printing GO and BP sensing layers on printed silver nanoparticle electrodes. The electrical response of the GO and BP sensors to humidity levels ranges from 11 to 97% relative humidity, which revealed a high capacitance sensitivity of 4.45 × 104 times for the GO sensor and 5.08 × 103 times for the BP sensor at 10 Hz operation frequency. Response/recovery times of the GO and BP sensor were found to be 2.7/4.6 s and 4.7/3.0 s respectively. These sensors also showed sensitive and fast response to a proximal human fingertip, showing potential applications in contactless switching.

Peptide hydrogel in vitro non-inflammatory potential.

Peptide-based hydrogels have attracted significant interest in recent years as these soft, highly hydrated materials can be engineered to mimic the cell niche with significant potential applications in the biomedical field. Their potential use in vivo in particular is dependent on their biocompatibility, including their potential to cause an inflammatory response. In this work, we investigated in vitro the inflammatory potential of a ß-sheet forming peptide (FEFEFKFK; F: phenylalanine, E: glutamic acid; K: lysine) hydrogel by encapsulating murine monocytes within it (3D culture) and using the production of cytokines, IL-ß, IL-6 and TNFα, as markers of inflammatory response. No statistically significant release of cytokines in our test sample (media + gel + cells) was observed after 48 or 72 h of culture showing that our hydrogels do not incite a pro-inflammatory response in vitro. These results show the potential biocompatibility of these hydrogels and therefore their potential for in vivo use. The work also highlighted the difference in monocyte behaviour, proliferation and morphology changes when cultured in 2D vs. 3D. © 2016 The Authors Journal of Peptide Science published by European Peptide Society and John Wiley & Sons Ltd.

Frequency-modulated atomic force microscopy localises viscoelastic remodelling in the ageing sheep aorta.

Age-related aortic stiffening is associated with cardiovascular diseases such as heart failure. The mechanical functions of the main structural components of the aorta, such as collagen and elastin, are determined in part by their organisation at the micrometer length scale. With age and disease both components undergo aberrant remodelling, hence, there is a need for accurate characterisation of the biomechanical properties at this length scale. In this study we used a frequency-modulated atomic force microscopy (FM-AFM) technique on a model of ageing in female sheep aorta (young: ~18 months, old: >8 years) to measure the micromechanical properties of the medial layer of the ascending aorta. The novelty of our FM-AFM method, operated at 30kHz, is that it is non-contact and can be performed on a conventional AFM using the ×³cantilever tune' mode, with a spatial (areal) resolution of around 1.6µm(2). We found significant changes in the elastic and viscoelastic properties within the medial lamellar unit (elastic lamellae and adjacent inter-lamellar space) with age. In particular, there was an increase in elastic modulus (Young; geometric mean (geometric SD)=42.9 (2.26)kPa, Old=113.9 (2.57)kPa, P<0.0001), G' and G″ (storage and loss modulus respectively) (Young; G'=14.3 (2.26)kPa, Old G'=38.0 (2.57)kPa, P<0.0001; Young; G″=14.5 (2.56)kPa, Old G″=32.8 (2.52)kPa, P<0.0001). The trends observed in the elastic properties with FM-AFM matched those we have previously found using scanning acoustic microscopy (SAM). The utility of the FM-AFM method is that it does not require custom AFM hardware and can be used to simultaneously determine the elastic and viscoelastic behaviour of a biological sample.

Localized micro- and nano-scale remodelling in the diabetic aorta.

Diabetes is strongly associated with cardiovascular disease, but the mechanisms, structural and biomechanical consequences of aberrant blood vessel remodelling remain poorly defined. Using an experimental (streptozotocin, STZ) rat model of diabetes, we hypothesized that diabetes enhances extracellular protease activity in the aorta and induces morphological, compositional and localized micromechanical tissue remodelling. We found that the medial aortic layer underwent significant thickening in diabetic animals but without significant changes in collagen or elastin (abundance). Scanning acoustic microscopy demonstrated that such tissue remodelling was associated with a significant decrease in acoustic wave speed (an indicator of reduced material stiffness) in the inter-lamellar spaces of the vessel wall. This index of decreased stiffness was also linked to increased extracellular protease activity (assessed by semi-quantitative in situ gelatin zymography). Such a proteolytically active environment may affect the macromolecular structure of long-lived extracellular matrix molecules. To test this hypothesis, we also characterized the effects of diabetes on the ultrastructure of an important elastic fibre component: the fibrillin microfibril. Using size exclusion chromatography and atomic force microscopy, we isolated and imaged microfibrils from both healthy and diabetic aortas. Microfibrils derived from diabetic tissues were fragmented, morphologically disrupted and weakened (as assessed following molecular combing). These structural and functional abnormalities were not replicated by in vitro glycation. Our data suggest that proteolysis may be a key driver of localized mechanical change in the inter-lamellar space of diabetic rat aortas and that structural proteins (such as fibrillin microfbrils) may be biomarkers of diabetes induced damage.

Current concepts and advances in the application of tissue engineering in otorhinolaryngology and head and neck surgery.

OBJECTIVE: This paper reviews the progress in the rapidly expanding scientific discipline of tissue engineering, which may have an integral role in the future of otorhinolaryngology. This article seeks to inform on the current concepts and principles of tissue engineering, and describe the state of the art research and developments in this exciting field as applied to ENT and head and neck surgery. METHOD: In order to carry out a comprehensive review of the literature spanning the past 30 years, a search of relevant publications was performed using the Web of Knowledge, Medline and PubMed databases. RESULTS: This search identified 85 scholarly articles, which were utilised as the basis of this review. CONCLUSION: Given the current rate of development of tissue engineering research, it is likely that tissue-engineered implants will be widely used in surgical practice, including ENT and head and neck surgery.

Gel-cast glass-ceramic tissue scaffolds of controlled architecture produced via stereolithography of moulds.

Two glass-ceramic scaffolds with a simple cubic structure of 500 µm square ligaments and square channels of width 400 or 600 µm have been fabricated by gel-casting into moulds produced by stereolithography, followed by mould removal, polymer burnout and sintering. The scaffolds have crushing strengths of 41 ± 14 and 17 ± 5 Mpa, respectively. Using a method of assembling discrete slices of scaffold, we are able to study cell behaviour within a scaffold by disassembly. Both scaffold structures were seeded with primary human osteoblasts and these penetrate, adhere, spread and proliferate on the scaffold structure. The larger channel diameter scaffold shows a greater cell population (despite its smaller surface area) and more pronounced production of ECM components (collagen and mineralization) with increased time in culture. Studies of sectioned scaffolds show that cell density and ECM production decrease with depth and that the difference between the two scaffold architectures is maintained.

Conical tungsten stamps for the replication of pore arrays in anodic aluminium oxide films.

A tungsten master stamp has been generated by applying a novel procedure that includes two-step anodizing, followed by sequential anodizing and pore widening to develop nominally funnelled pores. These conical-shaped pores were filled with tungsten by sputter coating to manufacture a master stamp. Under a pressure of 65 MPa, the master stamp successfully embossed the surface of annealed and electropolished aluminium. The embossed surface was then used to control the position of pores created by anodizing under the conditions used to produce the original pore array.

Nanoindentation of histological specimens: Mapping the elastic properties of soft tissues.

Although alterations in the gross mechanical properties of dynamic and compliant tissues have a major impact on human health and morbidity, there are no well-established techniques to characterize the micromechanical properties of tissues such as blood vessels and lungs. We have used nanoindentation to spatially map the micromechanical properties of 5-mum-thick sections of ferret aorta and vena cava and to relate these mechanical properties to the histological distribution of fluorescent elastic fibers. To decouple the effect of the glass substrate on our analysis of the nanoindentation data, we have used the extended Oliver and Pharr method. The elastic modulus of the aorta decreased progressively from 35 MPa in the adventitial (outermost) layer to 8 MPa at the intimal (innermost) layer. In contrast, the vena cava was relatively stiff, with an elastic modulus >30 MPa in both the extracellular matrix-rich adventitial and intimal regions of the vessel. The central, highly cellularized, medial layer of the vena cava, however, had an invariant elastic modulus of ~20 MPa. In extracellular matrix-rich regions of the tissue, the elastic modulus, as determined by nanoindentation, was inversely correlated with elastic fiber density. Thus, we show it is possible to distinguish and spatially resolve differences in the micromechanical properties of large arteries and veins, which are related to the tissue microstructure.

Novel collagen scaffolds with predefined internal morphology made by solid freeform fabrication.

Novel collagen scaffolds possessing predefined and reproducible internal channels with widths of 135 microm and greater have been produced. The process employed to make the collagen scaffold utilises a sacrificial mould, manufactured using solid freeform fabrication technology, and critical point drying technique. A computer aided design (CAD) file of the mould to be produced is created. This mould is manufactured using a phase change ink-jet printer. A dispersion of collagen is then cast into the mould and frozen. The mould is dissolved away with ethanol and the collagen scaffold is then critical point dried with liquid carbon dioxide. The effect of processing on the tertiary structure of collagen is assessed by monitoring the wavenumber of the N-H stretching vibration peak using Fourier transform infra-red spectroscopy and it is found that processing does not denature the collagen. Ultraviolet-visual spectroscopy was used to detect the presence of any contamination from the sacrificial mould on the collagen. The ability to use computer aided design and manufacture (CAD/CAM) provides a route to optimise scaffold designs using collagen in tissue engineering applications.

Manufacture of biomaterials by a novel printing process.

Tricalcium phosphate (TCP) scaffolds with controlled internal porosity were fabricated with a suspension of TCP in diacrylate cross-linking monomers using a mold prepared by ink-jet printing. Scaffolds were removed by selective dissolution of the mold. They were heat treated for removal of the acrylic binder followed by sintering. Despite a considerable linear shrinkage, scaffold porosity was retained after sintering. Composite scaffolds were fabricated from TCP in poly(ethylene glycol) diacrylate using an identical gel casting route.

Pathology of olivopontocerebellar atrophy with glutamate dehydrogenase deficiency.

We report the neuropathologic findings in the first patient with recognized glutamate dehydrogenase (GDH) deficiency to come to postmortem examination. He had progressive cerebellar ataxia beginning at age 21. He died at age 47 of pulmonary emboli. Postmortem examination revealed pancerebellar, olivary, and mild pontine atrophy, demyelination of the posterior columns, degeneration of anterior horn and dorsal root ganglion cells, and reduction of myelinated fibers in the sural nerve. In addition, there was neuronal storage of lipopigment diffusely throughout the CNS and the autonomic neurons, with cell distention, atrophy, and loss in selected areas.

Spectrum of symptomatic brain-stem metastasis.

Three cases are presented which depict the spectrum of neurological disability attendant on intra-axial brain-stem metastases, ranging from fulminant decline to a more leisurely and less disabling course. The rarity of primary glioma of brain-stem compared with brain-stem metastases in a general hospital population in the age group from 50 years and over is emphasized. Clinical deficit, certain ancillary findings, and response to therapy do not serve to separate brain-stem glioma from secondary brain-stem metastasis. The primary tumour may not be apparent when central nervous system symptoms appear or even for as long as two years after. It is recommended that the diagnosis of primary brain-stem glioma in the middle-aged adult be provisional and increasingly tentative over the age of 50 years.

Cardiobacterium hominis endocarditis with cerebral mycotic aneurysm.

Cardiobacterium hominis, a recently recognized Gram-negative pathogen, was recovered in blood cultures from a 65-year-old man with indolent endocarditis of previously normal heart valves. Despite the low virulence of the organism, major cardiac damage required valvular replacement, and there were multiple cerebral emboli with development of a mycotic aneurysm. After bacteriological cure, he died of a ruptured aneurysm.