Micro- and Nanostructured Fibrous Composites via Electro-Fluid Dynamics: Design and Applications for Brain.

The brain consists of an interconnected network of neurons tightly packed in the extracellular matrix (ECM) to form complex and heterogeneous composite tissue. According to recent biomimicry approaches that consider biological features as active components of biomaterials, designing a highly reproducible microenvironment for brain cells can represent a key tool for tissue repair and regeneration. Indeed, this is crucial to support cell growth, mitigate inflammation phenomena and provide adequate structural properties needed to support the damaged tissue, corroborating the activity of the vascular network and ultimately the functionality of neurons. In this context, electro-fluid dynamic techniques (EFDTs), i.e., electrospinning, electrospraying and related techniques, offer the opportunity to engineer a wide variety of composite substrates by integrating fibers, particles, and hydrogels at different scales-from several hundred microns down to tens of nanometers-for the generation of countless patterns of physical and biochemical cues suitable for influencing the in vitro response of coexistent brain cell populations mediated by the surrounding microenvironment. In this review, an overview of the different technological approaches-based on EFDTs-for engineering fibrous and/or particle-loaded composite substrates will be proposed. The second section of this review will primarily focus on describing current and future approaches to the use of composites for brain applications, ranging from therapeutic to diagnostic/theranostic use and from repair to regeneration, with the ultimate goal of providing insightful information to guide future research efforts toward the development of more efficient and reliable solutions.

Highly Reinforced Acrylic Resins for Hard Tissue Engineering and Their Suitability to Be Additively Manufactured through Nozzle-Based Photo-Printing.

Mineralized connective tissues represent the hardest materials of human tissues, and polymer based composite materials are widely used to restore damaged tissues. In particular, light activated resins and composites are generally considered as the most popular choice in the restorative dental practice. The first purpose of this study is to investigate novel highly reinforced light activated particulate dental composites. An innovative additive manufacturing technique, based on the extrusion of particle reinforced photo-polymers, has been recently developed for processing composites with a filler fraction (w/w) only up to 10%. The second purpose of this study is to explore the feasibility of 3D printing highly reinforced composites. A variety of composites based on 2,2-bis(acryloyloxymethyl)butyl acrylate and trimethylolpropane triacrylate reinforced with silica, titanium dioxide, and zirconia nanoparticles were designed and investigated through compression tests. The composite showing the highest mechanical properties was processed through the 3D bioplotter AK12 equipped with the Enfis Uno Air LED Engine. The composite showing the highest stiffness and strength was successfully processed through 3D printing, and a four-layer composite scaffold was realized. Mechanical properties of particulate composites can be tailored by modifying the type and amount of the filler fraction. It is possible to process highly reinforced photopolymerizable composite materials using additive manufacturing technologies consisting of 3D fiber deposition through extrusion in conjunction with photo-polymerization.

Fracture Strength and Failure Modes of Endodontically Treated Premolars Restored with Compact and Hollow Composite Posts Subjected to Cyclic Fatigue.

Physical and mechanical properties of continuous carbon or glass fiber reinforced endodontic posts are relevant to increase the retention and resistance of the tooth-restoration system. Hollow posts have been recently designed for delivering the luting cement through the post hole, thus enhancing the post-dentin interface by reducing the risk of air bubbles formation. Methods: Three type of endodontic posts, a carbon fiber hollow post, a glass fiber hollow post and a compact glass fiber post were investigated. Mechanical properties of these posts were assessed through bending tests. Teeth were subjected to fatigue cycling and the strength of restored teeth was detected through static tests. Failure modes were investigated through optical and scanning electron microscopy. Results show that composite posts increase the mechanical stability by more than 100% compared to premolars restored with particulate composite. Carbon fiber posts retain the highest strength (1467 N ± 304 N) among the investigated post and core restoration, but an unfavorable type of fracture has been observed, preventing the tooth re-treatment. Instead, more compliant posts (i.e., glass fiber reinforced composite, providing a strength of 1336 N ± 221 N), show a favorable mode of fracture that allows the re-treatment of teeth in the case that failure occurs. Glass fiber hollow posts show a good trade-off between strength and a favorable type of fracture.

Design of 3D Additively Manufactured Hybrid Structures for Cranioplasty.

A wide range of materials has been considered to repair cranial defects. In the field of cranioplasty, poly(methyl methacrylate) (PMMA)-based bone cements and modifications through the inclusion of copper doped tricalcium phosphate (Cu-TCP) particles have been already investigated. On the other hand, aliphatic polyesters such as poly(ε-caprolactone) (PCL) and polylactic acid (PLA) have been frequently investigated to make scaffolds for cranial bone regeneration. Accordingly, the aim of the current research was to design and fabricate customized hybrid devices for the repair of large cranial defects integrating the reverse engineering approach with additive manufacturing, The hybrid device consisted of a 3D additive manufactured polyester porous structures infiltrated with PMMA/Cu-TCP (97.5/2.5 w/w) bone cement. Temperature profiles were first evaluated for 3D hybrid devices (PCL/PMMA, PLA/PMMA, PCL/PMMA/Cu-TCP and PLA/PMMA/Cu-TCP). Peak temperatures recorded for hybrid PCL/PMMA and PCL/PMMA/Cu-TCP were significantly lower than those found for the PLA-based ones. Virtual and physical models of customized devices for large cranial defect were developed to assess the feasibility of the proposed technical solutions. A theoretical analysis was preliminarily performed on the entire head model trying to simulate severe impact conditions for people with the customized hybrid device (PCL/PMMA/Cu-TCP) (i.e., a rigid sphere impacting the implant region of the head). Results from finite element analysis (FEA) provided information on the different components of the model.

Impact of Magnetic Stimulation on Periodontal Ligament Stem Cells.

The aim of this study was to evaluate the effect of a time-dependent magnetic field on the biological performance of periodontal ligament stem cells (PDLSCs). A Western blot analysis and Alamar Blue assay were performed to investigate the proliferative capacity of magnetically stimulated PDLSCs (PDLSCs MAG) through the study of the MAPK cascade (p-ERK1/2). The observation of ALP levels allowed the evaluation of the effect of the magnetic field on osteogenic differentiation. Metabolomics data, such as oxygen consumption rate (OCR), extracellular acidification rate (ECAR) and ATP production provided an overview of the PDLSCs MAG metabolic state. Moreover, the mitochondrial state was investigated through confocal laser scanning microscopy. Results showed a good viability for PDLSCs MAG. Magnetic stimulation can activate the ERK phosphorylation more than the FGF factor alone by promoting a better cell proliferation. Osteogenic differentiation was more effectively induced by magnetic stimulation. The metabolic panel indicated significant changes in the mitochondrial cellular respiration of PDLSCs MAG. The results suggested that periodontal ligament stem cells (PDLSCs) can respond to biophysical stimuli such as a time-dependent magnetic field, which is able to induce changes in cell proliferation and differentiation. Moreover, the magnetic stimulation also produced an effect on the cell metabolic profile. Therefore, the current study demonstrated that a time-dependent magnetic stimulation may improve the regenerative properties of PDLSCs.

Combination Design of Time-Dependent Magnetic Field and Magnetic Nanocomposites to Guide Cell Behavior.

The concept of magnetic guidance is still challenging and has opened a wide range of perspectives in the field of tissue engineering. In this context, magnetic nanocomposites consisting of a poly(ε-caprolactone) (PCL) matrix and iron oxide (Fe3O4) nanoparticles were designed and manufactured for bone tissue engineering. The mechanical properties of PCL/Fe3O4 (80/20 w/w) nanocomposites were first assessed through small punch tests. The inclusion of Fe3O4 nanoparticles improved the punching properties as the values of peak load were higher than those obtained for the neat PCL without significantly affecting the work to failure. The effect of a time-dependent magnetic field on the adhesion, proliferation, and differentiation of human mesenchymal stem cells (hMSCs) was analyzed. The Alamar Blue assay, confocal laser scanning microscopy, and image analysis (i.e., shape factor) provided information on cell adhesion and viability over time, whereas the normalized alkaline phosphatase activity (ALP/DNA) demonstrated that the combination of a time-dependent field with magnetic nanocomposites (PCL/Fe3O4 Mag) influenced cell differentiation. Furthermore, in terms of extracellular signal-regulated kinase (ERK)1/2 phosphorylation, an insight into the role of the magnetic stimulation was reported, also demonstrating a strong effect due the combination of the magnetic field with PCL/Fe3O4 nanocomposites (PCL/Fe3O4 Mag).

Modification of PMMA Cements for Cranioplasty with Bioactive Glass and Copper Doped Tricalcium Phosphate Particles.

Cranioplasty represents the surgical repair of bone defects or deformities in the cranium arising from traumatic skull bone fracture, cranial bone deformities, bone cancer, and infections. The actual gold standard in surgery procedures for cranioplasty involves the use of biocompatible materials, and repair or regeneration of large cranial defects is particularly challenging from both a functional and aesthetic point of view. PMMA-based bone cement are the most widely biomaterials adopted in the field, with at least four different surgical approaches. Modifications for improving biological and mechanical functions of PMMA-based bone cement have been suggested. To this aim, the inclusion of antibiotics to prevent infection has been shown to provide a reduction of mechanical properties in bending. Therefore, the development of novel antibacterial active agents to overcome issues related to mechanical properties and bacterial resistance to antibiotics is still encouraged. In this context, mechanical, biological, and antibacterial feature against P. aeruginosa and S. aureus bacterial strains of surgical PMMA cement modified with BG and recently developed Cu-TCP bioactive particles have been highlighted.

A Further Analysis on Ti6Al4V Lattice Structures Manufactured by Selective Laser Melting.

Mechanical and architectural features play an important role in designing biomedical devices. The use of materials (i.e., Ti6Al4V) with Young's modulus higher than those of natural tissues generally cause stress shielding effects, bone atrophy, and implant loosening. However, porous devices may be designed to reduce the implant stiffness and, consequently, to improve its stability by promoting tissue ingrowth. If porosity increases, mass transport properties, which are crucial for cell behavior and tissue ingrowth, increase, whereas mechanical properties decrease. As reported in the literature, it is always possible to tailor mass transport and mechanical properties of additively manufactured structures by varying the architectural features, as well as pore shape and size. Even though many studies have already been made on different porous structures with controlled morphology, the aim of current study was to provide only a further analysis on Ti6Al4V lattice structures manufactured by selective laser melting. Experimental and theoretical analyses also demonstrated the possibility to vary the architectural features, pore size, and geometry, without dramatically altering the mechanical performance of the structure.

Bioactive chitosan-based scaffolds with improved properties induced by dextran-grafted nano-maghemite and l-arginine amino acid.

Over the past years, fundamentals of magnetism opened a wide research area of interest, in the field of tissue engineering and regenerative medicine. The integration of magnetic nanoarchitectures into synthetic/natural scaffold formulations allowed obtaining "on demand" responsive structures able to guide the regeneration process. The aim of this work was the design and characterization of three-dimensional (3D) chitosan-based scaffolds containing dextran-grafted maghemite nanoarchitectures (DM) and functionalized with l-arginine (l-Arg) amino acid as bioactive agent. A homogeneous pore distribution and a high degree of interconnection were obtained for all the structures with DMs, which resulted well distributed inside the polymer matrix. All the results suggest that the simultaneous presence of DMs and l-Arg conferred interesting mechano-structural and bioactive properties toward osteoblast-like and human mesenchymal stem cells, differentially stimulating their proliferation both in the absence and in the presence of a time-dependent magnetic field. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1244-1252, 2019.

An analysis on the potential of diode-pumped solid-state lasers for dental materials.

Material structure-property relationship is strongly related to the employed process technology. Over the past years, laser processing of engineering materials has been proposed in many fields and different uses for diode lasers have been found in dentistry. In this contest, the potential of GaN- and InGaN-based laser diodes to cure dental materials was analysed. Two wavelengths of 405 nm and 445 nm were used as heat or light sources for warm condensation of gutta-percha, light transmission in dental posts and brackets or light curing of dental composites. Additive manufacturing approach was considered to fabricate 3D root analogues, suitable supports, positioning systems and moulds for optical measurements. A three-axis CAD/CAM system was implemented for positioning and aligning the laser beam. The ability of diode-pumped solid-state lasers to cure dental materials or to transmit light was compared to that of a traditional instrument. Temperature profile at the apex of an additive manufactured root canal sealed with gutta-percha, light transmission through translucent quartz fiber post or through aesthetic ceramic bracket, bending properties and morphological features of light cured dental composites (Gradia Direct - GC Corporation and Venus Diamond - Heraeus Kulzer) were measured. Results showed a very high potential of diode-pumped solid-state lasers to be used in endodontics, orthodontics and restorative dentistry.

3D laser scanning in conjunction with surface texturing to evaluate shift and reduction of the tibiofemoral contact area after meniscectomy.

Meniscectomy significantly change the kinematics of the knee joint by reducing the contact area between femoral condyles and the tibial plateau, but the shift in the contact area has been poorly described. The aim of our investigation was to measure the shift of the tibiofemoral contact area occurring after meniscectomy. We used laser scans combined to surface texturing for measuring the 3D position and area of the femoral and tibial surfaces involved in the joint. In particular, natural condyles (porcine model) were analysed and the reverse engineering approach was used for the interpretation of the results from compression tests and local force measurements in conjunction with staining techniques. The results suggested that laser scans combined to surface texturing may be considered as a powerful tool to investigate the stained contours of the contact area. Beside the largely documented reduction of contact area and local pressure increase, a shift of the centroid of the contact area toward the intercondylar notch was measured after meniscectomy. As a consequence of the contact area shift and pressure increase, cartilage degeneration close to the intercondylar notch may occur.

Functional levels and MRI patterns of muscle involvement in upper limbs in Duchenne muscular dystrophy.

The aim of the study was to evaluate the spectrum of upper limb functional activities and imaging finding in a cohort of patients affected by Duchenne muscular dystrophy. Thirty-one patients of age between 5 and 29 years were included in the study (17 ambulant and 14 non-ambulant). They were all assessed using the Performance of Upper Limb (PUL) test and muscle MRI of shoulder, arm and forearm in order to establish if the functional scores obtained at shoulder, mid and distal level related to specific patterns of involvement in each upper limb segment on muscle MRI. At shoulder level, latissimus dorsi, serratus anterior, infraspinatus and subscapularis were always involved, even in patients with full functional scores at shoulder level. Diffuse and severe involvement of all muscles was found in the patients with a PUL shoulder functional score of ≤ 5. At arm level biceps brachii, brachialis and triceps were generally concordantly involved or spared. Some degree of involvement could already be detected in patients with reduced scores on the PUL mid domain. They were generally severely involved in patients with functional scores less than 6 at mid-level. At distal level supinator and pronator muscles were often involved, followed by brachioradialis and, less frequently, by the muscles of the flexor compartment. The extensor muscles were generally completely spared. A diffuse and severe involvement was found only in patients who had very low scores (8 or below) on the PUL distal domain. The integrated use of functional scales and imaging allowed to establish patterns of involvement at each level, and the functional scores that were more frequently associated with diffuse and severe involvement.

Theoretical Design of Multilayer Dental Posts Using CAD-Based Approach and Sol-Gel Chemistry.

A computer-aided design (CAD)-based approach and sol-gel chemistry were used to design a multilayer dental post with a compositional gradient and a Young’s modulus varying from 12.4 to 2.3 GPa in the coronal-apical direction. Specifically, we propose a theoretical multilayer post design, consisting of titanium dioxide (TiO2) and TiO2/poly(ε-caprolactone) (PCL) hybrid materials containing PCL up to 24% by weight obtained using the sol-gel method. The current study aimed to analyze the effect of the designed multilayer dental post in endodontically treated anterior teeth. Stress distribution was investigated along and between the post and the surrounding structures. In comparison to a metal post, the most uniform distributions with lower stress values and no significant stress concentration were found when using the multilayer post.

Mechanical and Thermal Properties of Dental Composites Cured with CAD/CAM Assisted Solid-State Laser.

Over the last three decades, it has been frequently reported that the properties of dental restorative composites cured with argon laser are similar or superior to those achieved with conventional halogen and light emitting diode (LED) curing units. Whereas laser curing is not dependent on the distance between the curing unit and the material, such distance represents a drawback for conventional curing units. However, a widespread clinical application of this kind of laser remains difficult due to cost, heavy weight, and bulky size. Recently, with regard to the radiation in the blue region of the spectrum, powerful solid-state lasers have been commercialized. In the current research, CAD (computer-aided design)/CAM (computer-aided manufacturing) assisted solid-state lasers were employed for curing of different dental restorative composites consisting of micro- and nanoparticle-reinforced materials based on acrylic resins. Commercial LED curing units were used as a control. Temperature rise during the photopolymerisation process and bending properties were measured. By providing similar light energy dose, no significant difference in temperature rise was observed when the two light sources provided similar intensity. In addition, after 7 days since curing, bending properties of composites cured with laser and LED were similar. The results suggested that this kind of laser would be suitable for curing dental composites, and the curing process does not suffer from the tip-to-tooth distance.

Collagen density gradient on three-dimensional printed poly(ε-caprolactone) scaffolds for interface tissue engineering.

The ability to engineer scaffolds that resemble the transition between tissues would be beneficial to improve repair of complex organs, but has yet to be achieved. In order to mimic tissue organization, such constructs should present continuous gradients of geometry, stiffness and biochemical composition. Although the introduction of rapid prototyping or additive manufacturing techniques allows deposition of heterogeneous layers and shape control, the creation of surface chemical gradients has not been explored on three-dimensional (3D) scaffolds obtained through fused deposition modelling technique. Thus, the goal of this study was to introduce a gradient functionalization method in which a poly(ε-caprolactone) surface was first aminolysed and subsequently covered with collagen via carbodiimide reaction. The 2D constructs were characterized for their amine and collagen contents, wettability, surface topography and biofunctionality. Finally, chemical gradients were created in 3D printed scaffolds with controlled geometry and porosity. The combination of additive manufacturing and surface modification is a viable tool for the fabrication of 3D constructs with controlled structural and chemical gradients. These constructs can be employed for mimicking continuous tissue gradients for interface tissue engineering.

Synthesis and characterization of divinyl-fumarate poly-ε-caprolactone for scaffolds with controlled architectures.

A vinyl-terminated polycaprolactone has been developed for tissue engineering applications using a one-step synthesis and functionalization method based on ring opening polymerization (ROP) of ε-Caprolactone, with hydroxyl ethyl vinyl ether (HEVE) acting both as the initiator of ROP and as photo-curable functional group. The proposed method employs a catalyst based on aluminium, instead of the most popular Tin(II) 2-ethylhexanoate, to reduce the cytotoxicity. Following the synthesis of the vinyl-terminated polycaprolactone, its reaction with fumaryl chloride (FuCl) results in a divinyl-fumarate polycaprolactone (VPCLF). The polymers obtained were thoroughly characterized using Fourier transform infrared spectroscopy (FTIR) and gel permeation chromatography (GPC) techniques. The polymer has been successfully employed, in combination with N-vinyl pyrrolidone (NVP), to fabricate films and computer-designed porous scaffolds by micro-stereolithography (µ-SL) with gyroid and diamond architectures. Characterization of the networks indicated the influence of NVP content on the network properties. Human mesenchymal stem cells adhered and spread onto VPCLF/NVP networks showing good biological properties and no cytotoxic effect. Copyright © 2016 John Wiley & Sons, Ltd.

Effect of topical antiinflammatory drugs on mechanical behavior of rabbit cornea.

BACKGROUND: A variety of antiinflammatory therapies are employed to promote corneal wound healing. The effects of steroidal and nonsteroidal antiinflammatory drugs on the biomechanical properties of rabbit cornea were investigated over time using tensile tests. METHODS: Full-thickness incisions were made and used to analyze the effects of dexamethasone sodium phosphate 0.1% and diclofenac sodium 0.1% on corneal biomechanical properties during wound healing at 7, 14 and 21 days after surgery. RESULTS: The full-thickness incision deeply modified all of the mechanical properties. At 3 weeks after incision, regardless of the drug therapy, the tensile modulus was about 70% of the value for the intact cornea. CONCLUSIONS: Topical treatment with dexamethasone was particularly effective during the first week after surgery; the second week after surgery, a similar result was observed in the corneas treated with diclofenac. Low doses of steroidal and nonsteroidal antiinflammatory drugs would seem to have the potential to improve biomechanical properties only during the early stage of the healing process of the cornea.

3D additive-manufactured nanocomposite magnetic scaffolds: Effect of the application mode of a time-dependent magnetic field on hMSCs behavior.

Over the past few years, the influence of static or dynamic magnetic fields on biological systems has become a topic of considerable interest. Magnetism has recently been implicated to play significant roles in the regulation of cell responses and, for this reason, it is revolutionizing many aspects of healthcare, also suggesting new opportunities in tissue engineering. The aim of the present study was to analyze the effect of the application mode of a time-dependent magnetic field on the behavior of human mesenchymal stem cells (hMSCs) seeded on 3D additive-manufactured poly(ɛ-caprolactone)/iron-doped hydroxyapatite (PCL/FeHA) nanocomposite scaffolds.

3D fibre deposition and stereolithography techniques for the design of multifunctional nanocomposite magnetic scaffolds.

Magnetic nanocomposite scaffolds based on poly(ε-caprolactone) and poly(ethylene glycol) were fabricated by 3D fibre deposition modelling (FDM) and stereolithography techniques. In addition, hybrid coaxial and bilayer magnetic scaffolds were produced by combining such techniques. The aim of the current research was to analyse some structural and functional features of 3D magnetic scaffolds obtained by the 3D fibre deposition technique and by stereolithography as well as features of multimaterial scaffolds in the form of coaxial and bilayer structures obtained by the proper integration of such methods. The compressive mechanical behaviour of these scaffolds was investigated in a wet environment at 37 °C, and the morphological features were analysed through scanning electron microscopy (SEM) and X-ray micro-computed tomography. The capability of a magnetic scaffold to absorb magnetic nanoparticles (MNPs) in water solution was also assessed. confocal laser scanning microscopy was used to assess the in vitro biological behaviour of human mesenchymal stem cells (hMSCs) seeded on 3D structures. Results showed that a wide range of mechanical properties, covering those spanning hard and soft tissues, can be obtained by 3D FDM and stereolithography techniques. 3D virtual reconstruction and SEM showed the precision with which the scaffolds were fabricated, and a good-quality interface between poly(ε-caprolactone) and poly(ethylene glycol) based scaffolds was observed for bilayer and coaxial scaffolds. Magnetised scaffolds are capable of absorbing water solution of MNPs, and a preliminary information on cell adhesion and spreading of hMSCs was obtained without the application of an external magnetic field.