Full-field strain distribution in hierarchical electrospun nanofibrous poly-L(lactic) acid/collagen scaffolds for tendon and ligament regeneration: A multiscale study.

Regeneration of injured tendons and ligaments (T/L) is a worldwide need. In this study electrospun hierarchical scaffolds made of a poly-L (lactic) acid/collagen blend were developed reproducing all the multiscale levels of aggregation of these tissues. Scanning electron microscopy, microCT and tensile mechanical tests were carried out, including a multiscale digital volume correlation analysis to measure the full-field strain distribution of electrospun structures. The principal strains (εp1 and εp3) described the pattern of strains caused by the nanofibers rearrangement, while the deviatoric strains (εD) revealed the related internal sliding of nanofibers and bundles. The results of this study confirmed the biomimicry of such electrospun hierarchical scaffolds, paving the way to further tissue engineering and clinical applications.

Self-Sensing Soft Skin Based on Piezoelectric Nanofibers.

The development of electronic skins and wearable devices is rapidly growing due to their broad application fields, such as for biomedical, health monitoring, or robotic purposes. In particular, tactile sensors based on piezoelectric polymers, which feature self-powering capability, have been widely used thanks to their flexibility and light weight. Among these, poly(vinylidenefluoride-trifluoroethylene) (PVDF-TrFE) presents enhanced piezoelectric properties, especially if manufactured in a nanofiber shape. In this work, the enhanced piezoelectric performances of PVDF-TrFE nanofibers were exploited to manufacture a flexible sensor which can be used for wearable applications or e-skin. The piezoelectric signal was collected by carbon black (CB)-based electrodes, which were added to the active layer in a sandwich-like structure. The sensor was electromechanically characterized in a frequency range between 0.25 Hz and 20 Hz-which is consistent with human activities (i.e., gait cycle or accidental bumps)-showing a sensitivity of up to 4 mV/N. The parameters of the signal acquisition circuit were tuned to enable the sensor to work at the required frequency. The proposed electrical model of the nanofibrous piezoelectric sensor was validated by the experimental results. The sensitivity of the sensor remained above 77.5% of its original value after 106 cycles of fatigue testing with a 1 kN compressive force.

Rubber-enhanced polyamide nanofibers for a significant improvement of CFRP interlaminar fracture toughness.

Nanofibrous mats provide substantial delamination hindering in composite laminates, especially if the polymer (as rubbers) can directly toughen the composite resin. Here, the well-known Nylon 66 nanofibers were impregnated with Nitrile Butadiene Rubber (NBR) for producing rubber/thermoplastic membranes for hampering the delamination of epoxy Carbon Fiber Reinforced Polymers (CFRPs). The starting polyamide mats were electrospun using two different solvent systems, and their effect on the mat's thermal and mechanical properties was investigated, as well as the laminate Mode I delamination resistance via Double Cantilever Beam (DCB) tests. Plain Nylon 66 mats electrospun from formic acid/chloroform perform better than the ones obtained from a solvent system containing trifluoroacetic acid, showing up to + 64% vs + 53% in interlaminar fracture toughness (GI), respectively. The effect of NBR coating benefits both nanofiber types, significantly raising the GI. The best results are obtained when interleaving medium-thickness and lightweight mats (20 µm, 9-10 g/m2) with 70-80 wt% of loaded rubber, achieving up to + 180% in GI. The work demonstrates the ability of NBR at improving the delamination hindering of common polyamide nonwovens, paving the way to the use of NBR-coated Nylon 66 nanofibers as effective interleaves for GI enhancement and overall composite safety improvement.

Is Graphene Always Effective in Reinforcing Composites? The Case of Highly Graphene-Modified Thermoplastic Nanofibers and Their Unfortunate Application in CFRP Laminates.

Graphene (G) can effectively enhance polymers' and polymer composites' electric, thermal, and mechanical properties. Nanofibrous mats have been demonstrated to significantly increase the interlaminar fracture toughness of composite laminates, hindering delamination and, consequently, making such materials safer and more sustainable thanks to increased service life. In the present paper, poly(ethylene oxide) (PEO), polycaprolactone (PCL), and Nylon 66 nanofibers, plain or reinforced with G, were integrated into epoxy-matrix Carbon Fiber Reinforced Polymers (CFRPs) to evaluate the effect of polymers and polymers + G on the laminate mechanical properties. The main aim of this work is to compare the reinforcing action of the different nanofibers (polyether, polyester, and polyamide) and to disclose the effect of G addition. The polymers were chosen considering their thermal properties and, consequently, their mechanism of action against delamination. PEO and PCL, displaying a low melting temperature, melt, and mix during the curing cycle, act via matrix toughening; in this context, they are also used as tools to deploy G specifically in the interlaminar region when melting and mixing with epoxy resin. The high extent of modification stems from an attempt to deploy it in the interlaminar layer, thus diluting further in the resin. In contrast, Nylon 66 does not melt and maintain the nanostructure, allowing laminate toughening via nanofiber bridging. The flexural properties of the nanomodifed CFRPs were determined via a three-point bending (3PB) test, while delamination behavior in Mode I and Mode II was carried out using Double Cantilever Beam (DCB) and End-Notched Flexture (ENF) tests, respectively. The lack of a positive contribution of G in this context is an interesting point to raise in the field of nanoreinforced CFRP.

New Application Field of Polyethylene Oxide: PEO Nanofibers as Epoxy Toughener for Effective CFRP Delamination Resistance Improvement.

Delamination is the most severe weakness affecting all composite materials with a laminar structure. Nanofibrous mat interleaving is a smart way to increase the interlaminar fracture toughness: the use of thermoplastic polymers, such as poly(ε-caprolactone) and polyamides (Nylons), as nonwovens is common and well established. Here, electrospun polyethylene oxide (PEO) nanofibers are proposed as reinforcing layers for hindering delamination in epoxy-based carbon fiber-reinforced polymer (CFRP) laminates. While PEO nanofibers are well known and successfully applied in medicine and healthcare, to date, their use as composite tougheners is undiscovered, resulting in the first investigation in this application field. The PEO-modified CFRP laminate shows a significant improvement in the interlaminar fracture toughness under Mode I loading: +60% and +221% in G I,C and G I,R, respectively. The high matrix toughening is confirmed by the crack path analysis, showing multiple crack planes, and by the delamination surfaces, revealing that extensive phase separation phenomena occur. Under Mode II loading, the G II enhancement is almost 20%. Despite a widespread phase separation occurring upon composite curing, washings in water do not affect the surface delamination morphology, suggesting a sufficient humidity resistance of the PEO-modified laminate. Moreover, it almost maintains both the original stiffness and glass transition temperature (T g), as assessed via three-point bending and dynamic mechanical analysis tests. The achieved results pave the way for using PEO nanofibrous membranes as a new effective solution for hindering delamination in epoxy-based composite laminates.

High-resolution X-ray tomographic workflow to investigate the stress distribution in vitreous enamel steels.

Vitreous enamel steels (VES) are a class of metal-ceramic composite materials realised with a low carbon steel basement coated by an enamel layer. During the firing phase to adhere the enamel to the metal, several gas bubbles remain entrapped inside the enamel volume modifying its internal structure. In this work high-resolution X-ray computed tomography (micro-CT) was used to investigate these composite materials. The micro-CT reconstructions enabled a detailed investigation of VES minimising the metal artefacts. The tomograms were used to develop finite element models (FEM) of VES by means of a representative volume element (RVE) to evaluate the thermal residual stresses caused by the manufacturing process, as well as the effect of the 3D bubbles distribution on the internal stress patterns after the thermic gradient. The promising results from this study have the potential to inform further research on such composite materials by optimising manufacturing processes for targeted applications.

Vitreous enamel steels are a particular class of composite materials composed by a low carbon steel basement coated by a vitreous enamel layer. Throughout the firing process applied to fix the enamel on the steel substrate, several gas bubbles remain entrapped inside the internal volume of the enamel modifying its internal microstructure. The presence of these bubbles substantially modifies the internal mechanical state of the structure developing residual stresses both among the bubbles and between the enamel-metal surface. However, to date no methods are still available to properly investigate the 3D bubbles morphology, distribution and stress patterns inside these materials. For this reason, in the present study we developed for the first time a high-resolution X-ray computed tomography (micro-CT) protocol able to investigate the vitreous enamel steels full field structure and numerically study their mechanics when the thermal gradient is applied. The micro-CT scans reconstructions allowed the visualisation of the enamel coating structure minimising metal artefacts. Moreover, the scans were postprocessed developing unpreceded 3D reconstructions with which the distribution, the volume and the mean diameter of the bubbles were analysed and defined. Subsequently, full field finite element computational models able to evaluate the thermal residual stresses produced inside the enamel volume were developed. They permitted to investigate the effect of the bubbles distribution on the internal residual stress patterns due to the thermal gradient generated throughout the cooling phase. The promising results from this study have the potential to inform further research on such composite materials by optimising manufacturing processes for targeted applications.

Self-Assembled NBR/Nomex Nanofibers as Lightweight Rubbery Nonwovens for Hindering Delamination in Epoxy CFRPs.

Still today, concerns regarding delamination limit the widespread use of high-performance composite laminates, such as carbon fiber-reinforced polymers (CFRPs), to replace metals. Nanofibrous mat interleaving is a well-established approach to reduce delamination. However, nanomodifications may strongly affect other laminate thermomechanical properties, especially if achieved by integrating soft materials. Here, this limitation is entirely avoided by using rubbery nitrile butadiene rubber (NBR)/Nomex mixed nanofibers: neither laminate stiffness nor glass-transition temperature (Tg) lowering occurs upon CFRP nanomodification. Stable noncrosslinked nanofibers with up to 60% wt of NBR were produced via single-needle electrospinning, which were then morphologically, thermally, spectroscopically, and mechanically characterized. NBR and Nomex disposition in the nanofiber was investigated via selective removal of the sole rubber fraction, revealing the formation of particular self-assembled structures resembling quasi-core-shell nanofibers or fibril-like hierarchical structures, depending on the applied electrospinning conditions (1.10 and 0.20 mL/h, respectively). Mode I and Mode II loading tests show a significant improvement of the interlaminar fracture toughness of rubbery nanofiber-modified CFRPs, especially GI (up to +180%), while GII enhancement is less pronounced but still significant (+40% in the best case). The two nanofibrous morphologies (quasi-core-shell and fibril-like ones) improve the delamination resistance differently, also suggesting that the way the rubber is located in the nanofibers plays a role in the toughening action. The quasi-core-shell nanofiber morphology provides the best reinforcing action, besides the highest productivity. By contrast, pure Nomex nanofibers dramatically worsen the interlaminar fracture toughness (up to -70% in GI), acting as a release film. The achieved delamination resistance improvements, combined with the retention of both the original laminate stiffness and Tg, pave the way to the extensive and reliable application of NBR/Nomex rubbery nanofibrous mats in composite laminates.

Rubbery-Modified CFRPs with Improved Mode I Fracture Toughness: Effect of Nanofibrous Mat Grammage and Positioning on Tanδ Behaviour.

Carbon Fiber Reinforced Polymers (CFRPs) are widely used where high mechanical performance and lightweight are required. However, they suffer from delamination and low damping, severely affecting laminate reliability during the service life of components. CFRP laminates modified by rubbery nanofibers interleaving is a recently introduced way to increase material damping and to improve delamination resistance. In this work, nitrile butadiene rubber/poly(ε-caprolactone) (NBR/PCL) blend rubbery nanofibrous mats with 60 wt% NBR were produced in three different mat grammages (5, 10 and 20 g/m2) via single-needle electrospinning and integrated into epoxy CFRP laminates. The investigation demonstrated that both mat grammage and positioning affect CFRP tanδ behaviour, evaluated by dynamic mechanical analysis (DMA) tests, as well as the number of nano-modified interleaves. Double cantilever beam (DCB) tests were carried out to assess the mat grammage effect on the interlaminar fracture toughness. Results show an outstanding improvement of GI,R for all the tested reinforced laminates regardless of the mat grammage (from +140% to +238%), while the effect on GI,C is more dependent on it (up to +140%). The obtained results disclose the great capability of NBR/PCL rubbery nanofibrous mats at improving CFRP damping and interlaminar fracture toughness. Moreover, CFRP damping can be tailored by choosing the number and positioning of the nano-modified interleaves, besides choosing the mat grammage.

Functionalisable Epoxy-rich Electrospun Fibres Based on Renewable Terpene for Multi-Purpose Applications.

New bio-based polymers capable of either outperforming fossil-based alternatives or possessing new properties and functionalities are of relevant interest in the framework of the circular economy. In this work, a novel bio-based polycarvone acrylate di-epoxide (PCADE) was used as an additive in a one-step straightforward electrospinning process to endow the fibres with functionalisable epoxy groups at their surface. To demonstrate the feasibility of the approach, poly(vinylidene fluoride) (PVDF) fibres loaded with different amounts of PCADE were prepared. A thorough characterisation by TGA, DSC, DMTA and XPS showed that the two polymers are immiscible and that PCADE preferentially segregates at the fibre surface, thus developing a very simple one-step approach to the preparation of ready-to-use surface functionalisable fibres. We demonstrated this by exploiting the epoxy groups at the PVDF fibre surface in two very different applications, namely in epoxy-based carbon fibre reinforced composites and membranes for ω-transaminase enzyme immobilisation for heterogeneous catalysis.

Tuning the Structure of Nylon 6,6 Electrospun Bundles to Mimic the Mechanical Performance of Tendon Fascicles.

Tendon and ligament injuries are triggered by mechanical loading, but the specific mechanisms are not yet clearly identified. It is well established however, that the inflection and transition points in tendon stress-strain curves represent thresholds that may signal the onset of irreversible fibrillar sliding. This phenomenon often results in a progressive macroscopic failure of these tissues. With the aim to simulate and replace tendons, electrospinning has been demonstrated to be a suitable technology to produce nanofibers similar to the collagen fibrils in a mat form. These nanofibrous mats can be easily assembled in higher hierarchical levels to reproduce the whole tissue structure. Despite the fact that several groups have developed electrospun tendon-inspired structures, an investigation of the inflection and transition point mechanics is missing. Comparing their behavior with that of the natural counterpart is important to adequately replicate their behavior at physiological strain levels. To fill this gap, in this work fascicle-inspired electrospun nylon 6,6 bundles were produced with different collector peripheral speeds (i.e., 19.7 m s-1; 13.7 m s-1; 7.9 m s-1), obtaining different patterns of nanofibers alignment. The scanning electron microcopy revealed a fibril-inspired structure of the nanofibers with an orientation at the higher speed similar to those in tendons and ligaments (T/L). A tensile mechanical characterization was carried out showing an elastic-brittle biomimetic behavior for the higher speed bundles with a progressively more ductile behavior at slower speeds. Moreover, for each sample category the transition and the inflection points were defined to study how these points can shift with the nanofiber arrangement and to compare their values with those of tendons. The results of this study will be of extreme interest for the material scientists working in the field, to model and improve the design of their electrospun structures and scaffolds and enable building a new generation of artificial tendons and ligaments.

Tissue Engineering for the Insertions of Tendons and Ligaments: An Overview of Electrospun Biomaterials and Structures.

The musculoskeletal system is composed by hard and soft tissue. These tissues are characterized by a wide range of mechanical properties that cause a progressive transition from one to the other. These material gradients are mandatory to reduce stress concentrations at the junction site. Nature has answered to this topic developing optimized interfaces, which enable a physiological transmission of load in a wide area over the junction. The interfaces connecting tendons and ligaments to bones are called entheses, while the ones between tendons and muscles are named myotendinous junctions. Several injuries can affect muscles, bones, tendons, or ligaments, and they often occur at the junction sites. For this reason, the main aim of the innovative field of the interfacial tissue engineering is to produce scaffolds with biomaterial gradients and mechanical properties to guide the cell growth and differentiation. Among the several strategies explored to mimic these tissues, the electrospinning technique is one of the most promising, allowing to generate polymeric nanofibers similar to the musculoskeletal extracellular matrix. Thanks to its extreme versatility, electrospinning has allowed the production of sophisticated scaffolds suitable for the regeneration of both the entheses and the myotendinous junctions. The aim of this review is to analyze the most relevant studies that applied electrospinning to produce scaffolds for the regeneration of the enthesis and the myotendinous junction, giving a comprehensive overview on the progress made in the field, in particular focusing on the electrospinning strategies to produce these scaffolds and their mechanical, in vitro, and in vivo outcomes.

Towards Poly(vinylidene fluoride-trifluoroethylene-chlorotrifluoroethylene)-Based Soft Actuators: Films and Electrospun Aligned Nanofiber Mats.

In the pursuit of designing a linear soft actuator with a high force-to-weight ratio and a stiffening behavior, this paper analyzes the electrostrictive effect of the poly(vinylidene fluoride-trifluoroethylene-chlorotrifluoroethylene) polymer in the form of film and aligned electrospun nanofiber mat. An experimental setup is realized to evaluate the electrostrictive effect of the specimens disjointly from the Maxwell stress. In particular, an uniaxial load test is designed to evaluate the specimens' forces produced by their axial contraction (i.e., the electrostrictive effect) when an external electric field is applied, while an uniaxial tensile load test is designed to show the specimens' stiffening properties. This electro-mechanical analysis demonstrates that both the film and the nanofiber mat are electrostrictive, and that the nanofiber mat exhibits a force-to-weight ratio ∼65% higher than the film and, therefore, a larger electrostrictive effect. Moreover, both the film and the nanofiber mat show a stiffening behavior, which is more evident for the nanofiber mat than the film and is proportional to the weight of the material. This study concludes that, thanks to its electro-mechanical properties, the poly(vinylidene fluoride-trifluoroethylene-chlorotrifluoroethylene), especially in the form of aligned electrospun nanofiber mat, has high potential to be used as electro-active polymer for soft actuators in biomedical and biorobotics applications.

Biomimetic Hierarchically Arranged Nanofibrous Structures Resembling the Architecture and the Passive Mechanical Properties of Skeletal Muscles: A Step Forward Toward Artificial Muscle.

Skeletal muscles are considered to date the best existing actuator in nature thanks to their hierarchical multiscale fibrous structure capable to enhance their strength and contractile performances. In recent years, driven by the growing of the soft robotics and tissue-engineering research field, many biomimetic soft actuators and scaffolds were designed by taking inspiration from the biological skeletal muscle. In this work we used the electrospinning technique to develop a hierarchically arranged nanofibrous structure resembling the morphology and passive biomechanical properties of skeletal muscles. To mimic the passive properties of muscle, a low-modulus polyurethane was used. Several electrospun structures (mats, bundles, and a muscle-like assembly) were produced with different internal 3D arrangements of the nanofibers. A thermal characterization through thermogravimetric and differential scanning calorimetry analysis investigated the physico-chemical properties of the material. The multiscale morphological similarities with the biological counterpart were verified by means of scanning electron microscopy investigation. The tensile tests on the different electrospun samples revealed that the muscle-like assembly presented slightly higher strength and stiffness compared to the skeletal muscle ones. Moreover, mathematical models of the mechanical behavior of the nanofibrous structures were successfully developed, allowing to better investigate the relationships between structure and mechanics of the samples. The promising results suggest the suitability of this hierarchical electrospun nanofibrous structure for applications in regenerative medicine and, if combined with active materials, in soft actuators for robotic.

Morphologically bioinspired hierarchical nylon 6,6 electrospun assembly recreating the structure and performance of tendons and ligaments.

Reconstructions of ruptured tendons and ligaments currently have dissatisfactory failure rate. Failures are mainly due to the mechanical mismatch of commercial implants with respect to the host tissue. In fact, it is crucial to replicate the morphology (hierarchical in nature) and mechanical response (highly-nonlinear) of natural tendons and ligaments. The aim of this study was to develop morphologically bioinspired hierarchical Nylon 6,6 electrospun assemblies recreating the structure and performance of tendons and ligaments. First, we built different electrospun bundles to find the optimal orientation of the nanofibers. A 2nd-level hierarchical assembly was fabricated with a dedicated process that allowed tightly joining the bundles one next to the other with an electrospun sheath, so as to improve the mechanical performance. Finally, a further hierarchical 3rd-level assembly was constructed by grouping several 2nd-level assemblies. The morphology of the different structures was assessed with scanning electron microscopy and high-resolution X-ray tomography, which allowed measuring the directionality of the nanofibers in the bundles and in the sheaths. The mechanical properties of the single bundles and of the 2nd-level assemblies were measured with tensile tests. The single bundles and the hierarchical assemblies showed morphology and directionality of the nanofibers similar to the tendons and ligaments. The strength and stiffness were comparable to that of tendons and ligaments. In conclusion, this work showed an innovative electrospinning production process to build nanofibrous Nylon 6,6 hierarchical assemblies which are suitable as future implantable devices and able to mimic the multiscale morphology and the biomechanical properties of tendons and ligaments.

Multiscale hierarchical bioresorbable scaffolds for the regeneration of tendons and ligaments.

Lesions of tendons and ligaments account for over 40% of the musculoskeletal lesions. Surgical techniques and materials for repair and regeneration are currently not satisfactory. The high rate of post-operative complications and failures mainly relates to the technical difficulties in replicating the complex multiscale hierarchical structure and the mechanical properties of the native tendons and ligaments. With the aim of overcoming the limitations of non-biomimetic devices, we developed a hierarchical structure replicating the organization of tendons and ligaments. The scaffold consists of multiple bundles made of resorbable electrospun nanofibers of Poly-L-Lactic acid (PLLA) having tailored dimensions, wrapped in a sheath of nanofibers able to compact the construct. The bundles in turn consist of electrospun nanofibers with a preferential direction. High-resolution x-ray tomographic investigation at nanometer resolution confirmed that the morphology of the single bundles and of the entire scaffold replicated the hierarchical arrangement in the natural tendons and ligaments. To confirm that these structures could adequately restore tendons and ligaments, we measured the tensile stiffness, strength and toughness. The mechanical properties were in the range required to replace and repair tendons and ligaments. Furthermore, human fibroblasts were able to attach to the scaffolds and showed an increase in cell number, indicated by an increase in metabolic activity over time. Fibroblasts were preferentially aligned along the electrospun nanofibers. These encouraging in vitro results open the way for the next steps towards in vivo regeneration of tendons and ligaments.

Tendon Fascicle-Inspired Nanofibrous Scaffold of Polylactic acid/Collagen with Enhanced 3D-Structure and Biomechanical Properties.

Surgical treatment of tendon lesions still yields unsatisfactory clinical outcomes. The use of bioresorbable scaffolds represents a way forward to improve tissue repair. Scaffolds for tendon reconstruction should have a structure mimicking that of the natural tendon, while providing adequate mechanical strength and stiffness. In this paper, electrospun nanofibers of two crosslinked PLLA/Collagen blends (PLLA/Coll-75/25, PLLA/Coll-50/50) were developed and then wrapped in bundles, where the nanofibers are predominantly aligned along the bundles. Bundle morphology was assessed via SEM and high-resolution x-ray computed tomography (XCT). The 0.4-micron resolution in XCT demonstrated a biomimetic morphology of the bundles for all compositions, with a predominant nanofiber alignment and some scatter (50-60% were within 12° from the axis of the bundle), similar to the tendon microstructure. Human fibroblasts seeded on the bundles had increased metabolic activity from day 7 to day 21 of culture. The stiffness, strength and toughness of the bundles are comparable to tendon fascicles, both in the as-spun condition and after crosslinking, with moderate loss of mechanical properties after ageing in PBS (7 and 14 days). PLLA/Coll-75/25 has more desirable mechanical properties such as stiffness and ductility, compared to the PLLA/Coll-50/50. This study confirms the potential to bioengineer tendon fascicles with enhanced 3D structure and biomechanical properties.

Nanovascularization of polymer matrix: generation of nanochannels and nanotubes by sacrificial electrospun fibers.

Several methods for creating vascular structures, made of either discrete or interconnected channels have been developed. The currently employed methods enable the formation of channels with diameters in the millimetric and micrometric scale. However, the formation of an interconnected three-dimensional (3D) vasculature by using a rapid and scalable process is a challenge and largely limits the fields of applicability of these innovative materials. Here, we propose the use of electrospun nonwoven mats as sacrificial fibers to easily generate 3D macroscale vascularized composites containing interconnected networks with channels and tubes having submicrometric and nanometric diameters. The novel approach has the potentialities to give rise to a novel generation of composites potentially displaying new and enhanced functionalities thanks to the nanoscale features of the cavities.