Centrifugally-spun polyhydroxybutyrate fibres: Effect of process solvent on structure,morphology and cell response

The structure and morphology of a novel form of poly(β-hydroxybutyrate) produced by gel-spinning is described. The entangled fibrous nature of the material, which resembles 'cotton wool' suggests possible functions in wound scaffolding devices. The surface structure and fibre diameter distribution of the fibres have been investigated using phase contrast and scanning electron microscopy. Fibres were found to possess a variety of surface irregularities, such as pores and indentations, with diameters mainly in the range 1-15 μm. Additionally, individual fibres were occasionally found to be fused or forked together with neighbours. The effects of blending with various polysaccharides and of altering the process solvent on fibre morphology were also investigated. Under hydrolytic degradation conditions (pH 10.6, 70°C) the fibres degraded by gradual fragmentation and erosion to fibre fragments, particulate matter and eventually to monomer. Altering the production process influenced both the fibre diameter distributions and surface morphology of the constituent fibres. Mammalian and human epithelial cells were used to study the cellular interaction with the spun fibres. SEM studies show that there is little or no cell adhesion to the unmodified fibres, but surface treatment by means of acid and alkali washes promoted cell proliferation on the materials, probably as a result of the introduction of hydroxyl and carboxyl at the surface. Fabrication of non-woven mats, which were subsequently acid or alkali treated, provided a conventional way of forming a celladhesive matrix which may have potential value as a wound scaffold. Neither cell line exhibited any cytotoxic response to these polymers.     

Scanning electron microscopic studies of the myelinated nerve fibres of the mouse sciatic nerve with special reference to the Schwann cell cytoplasmic network external to the myelin sheath

Summary The three-dimensional morphology of the surface of myelinated nerve fibres in the mouse sciatic nerve was studied by scanning electron microscopy after combined potassium hydroxide treatment and collagenase digestion (to remove the surrounding collagen fibrils and basal laminae from nerve fibres) as well as by transmission electron microscopy.The myelinated nerve fibre appeared as a long cylinder with sporadic annular constrictions corresponding to the nodes of Ranvier. Slight swellings of the surface due to Schwann cell nuclei were usually found at the middle of each internode. The surface of the nerve fibre clearly exhibited a network of bulges, which consisted of longitudinal bands extending from the nuclear swelling to the nodes of Ranvier through the internode, and transverse trabeculae bridging between these longitudinal bands. These bulges on the surface of nerve fibres were the site of the retained Schwann cell cytoplasm external to the myelin lamellae. These cytoplasmic networks on myelinated fibres presumably corresponded to the networks described by Cajal following silver impregnation. In addition, other thin elevations and focal round swellings were also found associated with these longitudinal bands and transverse trabeculae. These networks of Schwann cell cytoplasm are considered to be cytoplasmic channels for nutrition.The two apposing paranodal bulbs of nodes of Ranvier were often asymmetrical in their structure. The networks of the paranodal region were more complicated than those in the internode. The networks of Schwann cell cytoplasm converged into a continuous circumferential collar toward the node, which in turn gave rise to finger-like projections into the nodal gap.     

Gravity spun polycaprolactone fibres for soft tissue engineering: interaction with fibroblasts and myoblasts in cell culture

Poly(epsilon-caprolactone) (PCL) fibres were produced by wet spinning from solutions in acetone under low shear (gravity flow) conditions. As-spun PCL fibres exhibited a mean strength and stiffness of 7.9 MPa and 0.1 GPa, respectively and a rough, porous surface morphology. Cold drawing to an extension of 500% resulted in increases in fibre strength (43 MPa) and stiffness (0.3 GPa) and development of an oriented, fibrillar surface texture. The proliferation rate of Swiss 3T3 mouse fibroblasts and C2C12 mouse myoblasts on as-spun, 500% cold-drawn and gelatin-modified PCL fibres was determined in cell culture to provide a basic measure of the biocompatibility of the fibres. Proliferation of both cell types was consistently higher on gelatin-coated fibres relative to as-spun fibres at time points below 7 days. Fibroblast growth rates on cold-drawn PCL fibres exceeded those on as-spun fibres but myoblast proliferation was similar on both substrates. After 1 day in culture, both cell types had spread and coalesced on the fibres to form a cell layer, which conformed closely to the underlying topography. The high fibre compliance combined with a potential for modifying the fibre surface chemistry with cell adhesion molecules and the surface architecture by cold drawing to enhance proliferation of fibroblasts and myoblasts, recommends further investigation of gravity-spun PCL fibres for 3-D scaffold production in soft tissue engineering.     

Electrospinning and Evaluation of PHBV-Based Tissue Engineering Scaffolds with Different Fibre Diameters,Surface Topography and Compositions

While electrospinning is an effective technology for producing poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) micrometre-scale fibrous scaffolds for tissue regeneration, electrospinning of PHBV fibrous scaffolds composed of sub-micrometre fibres, surface-porous fibres or nanocomposite fibres is rarely explored. In this study, the average PHBV fibre diameter was successfully reduced to the sub-micrometre scale by dissolving a conductivity-enhancing salt in the polymer solution for electrospinning. Surface-porous fibres were made using a mixture of solvents, and carbonated hydroxyapatite (CHA) nanoparticles were incorporated into the fibres with the aid of an ultrasonic power source. Water contact angle measurements demonstrated that both fibre diameter reduction and CHA incorporation enhanced the wettability of the fibrous scaffolds. Tensile properties of the scaffolds were not undermined by the reduction of fibre diameter and the presence of surface pores. In vitro biological evaluation using a human osteoblast-like cell line (SaOS-2) demonstrated that all types of fibrous scaffolds supported cell attachment, spreading and proliferation. Analysis of cell morphology revealed similar projected cell areas on all types of scaffolds. However, cells on sub-micrometre fibres possessed a lower cell aspect ratio than cells on microfibres. The reduction of fibre diameter to the sub-micrometre scale enhanced cell proliferation after 14 days cell culture, while the incorporation of CHA nanoparticles in microfibres significantly enhanced the alkaline phosphatase activity of SaOS-2 cells. The control of fibre diameter, surface topography and composition is important in developing electrospun PHBV-based scaffolds for specific tissue-engineering applications.     

Modified alkali digestion method for pulmonary asbestos fibre counts

In using the standard alkali digestion method for pulmonary asbestos fibre count, it was found that carbonaceous particles often obscured the presence of asbestos bodies (coated fibres) rendering their quantification inaccurate, particularly in lungs with a high soot particle content and a low fibre count. We applied the chloroform-ethanol separation technique from the bleaching agent digestion method to the alkali digestion method to remove the carbonaceous particles. It was found that coated fibres are much easier to visualize and count. The uncoated fibre count was nevertheless more accurate with the original method. The coated and uncoated fibre counts from 21 lung specimens were compared using both methods. The total fibre count was considered better derived from using both techniques.     

Reactive fibre reinforced glass ionomer cements

The mechanical properties of glass ionomer cements used in restorative dentistry reinforced by chopped glass fibres were investigated. Reactive glass fibres with a composition in the system SiO(2)-Al(2)O(3)-CaF(2)-Na(3)AlF(6) and a thickness of 26 microm were drawn by a bushing process. The manufacturing parameters were optimized with respect to maximum strength of the glass fibre reinforced ionomer cements. Powder to liquid ratio, pre-treatment of the glass, grain size distribution and fibre volume fraction were varied. Glass fibre and cement were characterized by X-ray diffraction, transmission electron microscopy and energy dispersive spectroscopy techniques, respectively. The highest flexural strength of the reinforced cement (15.6 MPa) was found by compounding 20 vol% reactive fibres and extending the initial dry gelation period up to 30 min. Microscopic examination of the fractured cements indicated a distinct reactive layer at the fibre surface. A pronounced fibre pull out mode gives rise to an additional work-of-fracture contributed by pulling the fibres out of the fracture surface.     

Fibre type proportion and fibre size in trapezius muscle biopsies from cleaners with and without myalgia and its correlation with ragged red fibres, cytochrome-c-oxidase-negative fibres, biomechanical output, perception of fatigue, and surface electromyography during repetitive forward flexions

In the literature enlarged/increased cross-sectional area (CSA) of type I muscle fibres has been reported as a morphological mark of work-related localised myalgia in the descending part of the trapezius muscle of women. These studies did not use enough subjects or lacked an adequate control group. The recording of surface electromyograms (EMG) is central to the research field of work-related myalgia. However, the influence of intrinsic muscle properties – such as the effect of muscle fibre distribution upon surface EMG – has to be better understood in order properly to evaluate this method as a possible diagnostic and preventive tool. This study had two aims. Firstly, it investigated the muscle fibre distribution and CSA in work-related myalgia in trapezius muscles. Secondly, the multivariate relationships among muscle morphology and histochemistry [ragged-red (RR) fibres, and cytochrome-c-oxidase-negative-fibre changes] EMG, perceived fatigue, and biomechanical output of shoulder flexions were analysed. The raw data have been presented in an earlier study. The participants in this study were 25 female cleaners with work-related myalgia of the trapezius muscle and 25 female cleaners not experiencing work-related myalgia of this muscle. The control group comprised 21 healthy female teachers who had not been exposed to highly repetitive work or static muscle work. Smaller CSA of type II fibres were found in cleaners compared to teachers. In this study the CSA of type I fibres of the trapezius muscles associated with myalgia were no greater than in muscles without myalgia. The prevalence of RR fibres together with age, fibre type proportions, CSA and working as a cleaner correlated with the ability to relax as recorded electromyographically. The relative mean frequency of the EMG of the trapezius muscle correlated with the prevalence of RR fibres, but it did not correlate with the proportions and CSA of different fibre types. Low biomechanical outputs and low signal amplitude increases of the EMG during the test were associated with high proportions of type IIB fibres. The smaller CSA of type II fibres in cleaners might have reflected a different muscle activation pattern due to different occupational demands in cleaners than in teachers. Morphological or histopathological variables can influence the three EMG variables investigated. Electronic Publication     

New biotextiles for tissue engineering: Development,characterization and in vitro cellular viability

This work proposes biodegradable textile-based structures for tissue engineering applications. We describe the use of two polymers, polybutylene succinate (PBS) proposed as a viable multifilamentand silk fibroin (SF), to produce fibre-based finely tuned porous architectures by weft knitting. PBS is here proposed as a viable extruded multifilament fibre to be processed by a textile-based technology. A comparative study was undertaken using a SF fibre with a similar linear density. The knitted constructs obtained are described in terms of their morphology, mechanical properties, swelling capability, degradation behaviour and cytotoxicity. The weft knitting technology used offers superior control over the scaffold design (e.g. size, shape, porosity and fibre alignment), manufacturing and reproducibility. The presented fibres allow the processing of a very reproducible intra-architectural scaffold geometry which is fully interconnected, thus providing a high surface area for cell attachment and tissue in-growth. The two types of polymer fibre allow the generation of constructs with distinct characteristics in terms of the surface physico-chemistry, mechanical performance and degradation capability, which has an impact on the resulting cell behaviour at the surface of the respective biotextiles. Preliminary cytotoxicity screening showed that both materials can support cell adhesion and proliferation. These results constitute a first validation of the two biotextiles as viable matrices for tissue engineering prior to the development of more complex systems. Given the processing efficacy and versatility of the knitting technology and the interesting structural and surface properties of the proposed polymer fibres it is foreseen that the developed systems could be attractive for the functional engineering of tissues such as skin, ligament, bone or cartilage.     

Muscle morphology,self‐reported physical activity and insulin resistance syndrome

In a population‐based sample of 475 men the associations between muscle morphology, self‐reported physical activity (PA) and insulin resistance (IR) syndrome were investigated. Also, we studied to what degree muscle morphology contributes to the association between PA and IR syndrome. Muscle morphology and the components of IR syndrome were compared in four groups categorized according to self‐reported habitual PA data. We found a significantly higher percentage of type I fibres, fibre area and number of capillaries around the fibres and a lower proportion of type IIB fibres with higher level of PA. The relative distribution of type I fibres and capillarization were positively related to high density lipoprotein (HDL) cholesterol and negatively to serum triglycerides (TG) and plasminogen activator inhibitor‐1 (PAI‐1) activity. The percentage of type IIB fibres was were inversely related to HDL cholesterol and positively to serum TG, PAI‐1 activity and resting heart rate. Insulin sensitivity was positively and independently related to PA level (P < 0.001). Regression analysis including all relevant variables regarding insulin sensitivity indicated that the significant explanatory variables left in the equation were body mass index (BMI), glucose intolerance, PAI‐1 activity, serum free fatty acid concentration, proportion of type IIB fibres, HDL cholesterol level, drug treatment, PA level, and waist‐to‐hip ratio, which together explained 55% of the variation in the insulin sensitivity index. In conclusion, both fibre type distribution and muscle capillary density might contribute to the beneficial effect of PA on IR syndrome.     

Muscle morphology,self-reported physical activity and insulin resistance syndrome

In a population-based sample of 475 men the associations between muscle morphology, self-reported physical activity (PA) and insulin resistance (IR) syndrome were investigated. Also, we studied to what degree muscle morphology contributes to the association between PA and IR syndrome. Muscle morphology and the components of IR syndrome were compared in four groups categorized according to self-reported habitual PA data. We found a significantly higher percentage of type I fibres, fibre area and number of capillaries around the fibres and a lower proportion of type IIB fibres with higher level of PA. The relative distribution of type I fibres and capillarization were positively related to high density lipoprotein (HDL) cholesterol and negatively to serum triglycerides (TG) and plasminogen activator inhibitor-1 (PAI-1) activity. The percentage of type IIB fibres was were inversely related to HDL cholesterol and positively to serum TG, PAI-1 activity and resting heart rate. Insulin sensitivity was positively and independently related to PA level (P < 0.001). Regression analysis including all relevant variables regarding insulin sensitivity indicated that the significant explanatory variables left in the equation were body mass index (BMI), glucose intolerance, PAI-1 activity, serum free fatty acid concentration, proportion of type IIB fibres, HDL cholesterol level, drug treatment, PA level, and waist-to-hip ratio, which together explained 55% of the variation in the insulin sensitivity index. In conclusion, both fibre type distribution and muscle capillary density might contribute to the beneficial effect of PA on IR syndrome.     

Dependence of cross-bridge kinetics on myosin light chain isoforms in rabbit and rat skeletal muscle fibres

Cross-bridge kinetics underlying stretch-induced force transients was studied in fibres with different myosin light chain (MLC) isoforms from skeletal muscles of rabbit and rat. The force transients were induced by stepwise stretches (< 0.3% of fibre length) applied on maximally Ca²⁺-activated skinned fibres. Fast fibre types IIB, IID (or IIX) and IIA and the slow fibre type I containing the myosin heavy chain isoforms MHC-IIb, MHC-IId (or MHC-IIx), MHC-IIa and MHC-I, respectively, were investigated. The MLC isoform content varied within fibre types. Fast fibre types contained the fast regulatory MLC isoform MLC2f and different proportions of the fast alkali MLC isoforms MLC1f and MLC3f. Type I fibres contained the slow regulatory MLC isoform MLC2s and the slow alkali MLC isoform MLC1s. Slow MLC isoforms were also present in several type IIA fibres. The kinetics of force transients differed by a factor of about 30 between fibre types (order from fastest to slowest kinetics: IIB > IID > IIA ≫ I). The kinetics of the force transients was not dependent on the relative content of MLC1f and MLC3f. Type IIA fibres containing fast and slow MLC isoforms were about 1.2 times slower than type IIA fibres containing only fast MLC isoforms. We conclude that while the cross-bridge kinetics is mainly determined by the MHC isoforms present, it is affected by fast and slow MLC isoforms but not by the relative content of MLC1f and MLC3f. Thus, the physiological role of fast and slow MLC isoforms in type IIA fibres is a fine-tuning of the cross-bridge kinetics.     

The elastin network: its relationship with collagen and cells in articular cartilage as visualized by multiphoton microscopy

A combination of two‐photon fluorescence (TPF), second harmonic generation (SHG) and coherent anti‐Stokes Raman scattering (CARS) imaging has been used to investigate the elastin fibre network in healthy equine articular cartilage from the metacarpophalangeal joint. The elastin fibres were identified using their intrinsic two‐photon fluorescence and immuno‐staining was used to confirm the identity of these fibres. SHG was used to reveal the collagen matrix and the collagen fibre orientations were determined from their SHG polarization sensitivity, while CARS was used to clearly delineate the cell boundaries. Extensive elastin fibre networks were found in all the joint regions investigated. The elastin was found predominantly in the superficial zone (upper 50 μm) and was aligned parallel to the articular surface. Elastin was also detected in the pericellular matrix surrounding the superficial chondrocytes; however, individual fibres could not be resolved in this region. Variations in the density and organization of the fibres were observed in different regions on the joint surface.     

A comparison between the physiological and histochemical characterisation of urethral striated muscle in the guinea pig

In this in vitro study comparison has been made between the actomyosin ATPase activity and the contractile properties of the external urethral sphincter in the guinea pig. Histochemical analysis showed the external urethral sphincter to contain a mixture of alkali and acid stable actomyosin ATPase positive fibres in the ratio of 3:1. External urethral sphincter isometric contraction measurements were undertaken using specimens mounted transversely or longitudinally with respect to the urethral lumen. These contraction parameters have shown that the majority of fibres which constitute the external urethral sphincter correspond to the fast twitch type. In order to determine whether the relatively small fibre diameter of urethral striated muscle influenced the contraction results, when compared with controls, corrections for different volume ratios of slow and fast fibres were applied to the results. These modified values revealed close correlation between muscle fibre actomyosin ATPase content and isometric contraction responses.     

Gravity spinning of polycaprolactone fibres for applications in tissue engineering

Poly(epsilon-caprolactone) (PCL) fibres have been produced by wet spinning from solutions in acetone under low shear (gravity flow) conditions. The tensile strength and stiffness of as-spun fibres were highly dependent on the concentration of the spinning solution. Use of a 6% w/v solution resulted in fibres having strength and stiffness of 1.8 MPa and 0.01 GPa, respectively, whereas these values increased to 9.9 MPa and 0.1 GPa when fibres were produced from 20% w/v solutions. Cold drawing to an extension of 500% resulted in further increases in fibre strength (up to 50 MPa) and stiffness (0.3 GPa). The surface morphology of as-spun fibres was modified, to yield a directional grooved pattern by drying in contact with a mandrel having a machined topography characterised by a peak-peak separation of 91 microm and a peak height of 30 microm. Limited in vitro studies of cell behaviour in contact with the fibres were performed using cell culture. The number of attached fibroblasts and myoblasts on as-spun PCL fibres after 5 days in cell culture was lower than on tissue culture plastic by a factor 2 and 1.5, respectively, but higher than on Dacron monofilament by a factor of 4 and 11, respectively. The high fibre compliance and the potential for controlling the fibre surface architecture to promote contact guidance effects together with the maintained proliferation of fibroblasts and myoblasts on as-spun PCL fibres in vitro recommends their use for 3-D scaffold production in soft tissue engineering.     

Morphology of cardiac muscle in septic shock. Observations with a porcine septic shock model

The morphology of cardiac muscle was investigated in a porcine model of septic shock, created by intermitted application of Escherichia coli-endotoxin. The earliest lesions, found after 18 h of septic shock, were endothelial cell swelling, marked leucostasis and slight ischaemic alterations of the muscle fibres. At the end point of the experiments, after 48 h, some fibrin thrombi were found associated with more pronounced ischaemic alterations of cardiac muscle cells and some necrotic fibres. Comparing these findings with the severe endothelial and muscle fibre lesions found in skeletal muscle, the endothelial cells of the heart microvasculature, are clearly more resistant to the attack of the endotoxins and mediators liberated in septic shock.     

Muscle fibre damage and regeneration resulting from surgical limb distraction

Using an animal model of limb distraction, the extent of muscle fibre damage and atrophy resulting from distraction at two different rates (1.3 or 3.0 mm day(-1)) was investigated. It was found that at the high rate of distraction there was a significantly greater loss of range of joint movement and more muscle fibre atrophy and fibre damage than at the low rate. Muscle fibre damage is usually followed by regeneration. This involves the expression of the neonatal form of myosin heavy chain, which can therefore be used as an indicator of regeneration. It was found that whilst many more fibres showed evidence of damage at the high compared to the low rate, the number of fibres expressing neonatal myosin was significantly reduced, indicating the presence of a population of fibres which was undergoing degeneration without subsequent regeneration. Thus it would appear that beyond a certain rate of distraction, regeneration may be insufficient to replace contractile material damaged by overstretching. It is suggested that these fibres are replaced with connective tissue. This process may contribute to the muscle weakness and loss of range of joint movement which sometimes accompanies limb distraction procedures.     

Transformations of the retinal topography along the visual pathway of the chicken

Summary It is still unclear how the retinotectal map of the chick is formed during development. In particular, it is not yet known whether or not the organization of fibres plays a role in the formation of this map. In order to contribute to the solution of this problem, we analysed the representation of the retinal topography at closely spaced intervals along the fibre pathway. We injected HRP into various sites of the tectal surface and traced the labelled fibre bundles back to the retina. The retinal topography was reconstructed at ten different levels, i.e. in the retina, the optic nerve head, the middle of the optic nerve, the chiasm (three levels), the optic tract (three levels), and the optic tectum. We obtained the following results: (1) The labelled fibre bundles as well as the fields of labelled retinal ganglion cells were always well delimited and coherent. (2) The reconstructions show that transformations of the retinal topography occur in the fibre pathway. The first and most important transformation is found in the optic nerve head where the retinal image is mirrored across an axis extending from dorsotemporal to ventronasal retina. In addition, the retinal representation is split in its temporal periphery. Thus, central and centrotemporal fibres are no longer in the centre of the image but close to the dorsal border of the nerve. Peripheral fibres are found along the medial, ventral and lateral circumference of the nerve. In the optic tract a second transformation occurs. The retinal topography is rotated clockwise by about 90 degrees and flattened to a band. The flattening is accompanied by a segregation of fibre bundles so that eventually central and centrotemporal retinal fibres are located centrally, ventral fibres dorsally and dorsal retinal fibres ventrally in the tract. By these two transformations an organization of fibres is produced in the optic tract which can be projected onto the tectal surface without major changes given that dorsal and ventral fibres remain in their relative positions, and that deep lying fibres project to the rostral and central tectum, superficial fibres to the caudal tectum.The transformations which we have observed follow specific rules and thus maintain order in the pathway although retinotopy is lost. In conjunction with our earlier studies on the development of the retinotectal system we conclude that fibres are laid down in a chronotopic order. The transformations take place under particular structural constraints. Thus, an organization of fibres is provided in the optic tract which results in a retinotopic map when projected onto the tectal surface. This is stated for the order of magnitude of fibre bundles as investigated in this study. At the level of individual fibres additional factors may play an important role.     

Post-self-assembly experimentation on extruded collagen fibres for tissue engineering applications

Extruded collagen fibres have been shown to constitute a biomimetic three-dimensional scaffold with numerous tissue engineering applications. The multi-step fabrication process of this material provides opportunities for further advancements to improve the properties of the final product. Herein we investigated the influence of the post-self-assembly washing baths on the structural, mechanical and thermal properties of these fibres. The surface morphology and the inter-fibre packing were similar for every treatment. The overnight incubation in isopropanol yielded fibres with the highest temperature and energy of denaturation (p<0.013). Typical s- and j-shape stress-strain curves were obtained for all treatments in the dry and wet state respectively. Rehydration of the fibres resulted in increased fibre diameter (p<0.006) and reduced stress (p<0.001), force (p<0.001) and modulus (p<0.002) values for every treatment. In the dry state, the alcohol-treated fibres were characterized by the highest stress (p<0.002) values; whilst in the wet state the Tris-HCl-treated fibres were the weakest (p<0.006). For every treatment, in both dry and wet state, a strong and inverse relationship between the fibre diameter and the stress at break was observed. Overall, the fibres produced were characterized by properties similar to those of native tissues.     

Effects of fibre reinforcement on the mechanical properties of brushite cement

In this study the effect of structure and amount of polyglactin fibre incorporation into a brushite forming calcium phosphate cement system and the effect of mechanical compaction on the fibre modified system were investigated. In comparison the effect of resorbable polycaprolactone surface coating of cement specimens was investigated. The results showed that, apart from the mechanical properties of the reinforcing material, the structure of the incorporated fibres, regular or random, is crucial for the resulting flexural strength and modulus of elasticity. Fibre reinforcement could also be combined with mechanical compaction of the cement/fibre composite paste leading to a possible 7-fold increase in flexural strength or an almost 5-fold increase in modulus of elasticity. Reinforcement of the tensile surface of cement grafts may ultimately improve strength where required, especially in conjunction with bone fixation devices.     

Creating polymer hydrogel microfibres with internal alignment via electrical and mechanical stretching

Hydrogels have been widely used for 3-dimensional (3D) cell culture and tissue regeneration due to their tunable biochemical and physicochemical properties as well as their high water content, which resembles the aqueous microenvironment of the natural extracellular matrix. While many properties of natural hydrogel matrices are modifiable, their intrinsic isotropic structure limits the control over cellular organization, which is critical to restore tissue function. Here we report a generic approach to incorporate alignment topography inside the hydrogel matrix using a combination of electrical and mechanical stretching. Hydrogel fibres with uniaxial alignment were prepared from aqueous solutions of natural polymers such as alginate, fibrin, gelatin, and hyaluronic acid under ambient conditions. The unique internal alignment feature drastically enhances the mechanical properties of the hydrogel microfibres. Furthermore, the facile, organic solvent-free processing conditions are amenable to the incorporation of live cells within the hydrogel fibre or on the fibre surface; both approaches effectively induce cellular alignment. This work demonstrates a versatile and scalable strategy to create aligned hydrogel microfibres from various natural polymers.