Nanostructured scaffolds based on bioresorbable polymers and graphene oxide induce the aligned migration and accelerate the neuronal differentiation of neural stem cells.

Within the field of neural tissue engineering, there is a huge need for the development of materials that promote the adhesion, aligned migration and differentiation of stem cells into neuronal and supportive glial cells. In this study, we have fabricated bioresorbable elastomeric scaffolds combining an ordered nanopatterned topography together with a surface functionalization with graphene oxide (GO) in mild conditions. These scaffolds allowed the attachment of murine neural stem cells (NSCs) without the need of any further coating of its surface with extracellular matrix adhesion proteins. The NSCs were able to give rise to both immature neurons and supporting glial cells over the nanostructured scaffolds in vitro, promoting their aligned migration in cell clusters following the nanostructured grooves. This system has the potential to reestablish spatially oriented neural precursor cell connectivity, constituting a promising tool for future cellular therapy including nerve tissue regeneration.

Advances and Perspectives in Dental Pulp Stem Cell Based Neuroregeneration Therapies.

Human dental pulp stem cells (hDPSCs) are some of the most promising stem cell types for regenerative therapies given their ability to grow in the absence of serum and their realistic possibility to be used in autologous grafts. In this review, we describe the particular advantages of hDPSCs for neuroregenerative cell therapies. We thoroughly discuss the knowledge about their embryonic origin and characteristics of their postnatal niche, as well as the current status of cell culture protocols to maximize their multilineage differentiation potential, highlighting some common issues when assessing neuronal differentiation fates of hDPSCs. We also review the recent progress on neuroprotective and immunomodulatory capacity of hDPSCs and their secreted extracellular vesicles, as well as their combination with scaffold materials to improve their functional integration on the injured central nervous system (CNS) and peripheral nervous system (PNS). Finally, we offer some perspectives on the current and possible future applications of hDPSCs in neuroregenerative cell therapies.

Benefits of Polydopamine as Particle/Matrix Interface in Polylactide/PD-BaSO4 Scaffolds.

This work reports the versatility of polydopamine (PD) when applied as a particle coating in a composite of polylactide (PLA). Polydopamine was observed to increase the particle-matrix interface strength and facilitate the adsorption of drugs to the material surface. Here, barium sulfate radiopaque particles were functionalized with polydopamine and integrated into a polylactide matrix, leading to the formulation of a biodegradable and X-ray opaque material with enhanced mechanical properties. Polydopamine functionalized barium sulfate particles also facilitated the adsorption and release of the antibiotic levofloxacin. Analysis of the antibacterial capacity of these composites and the metabolic activity and proliferation of human dermal fibroblasts in vitro demonstrated that these materials are non-cytotoxic and can be 3D printed to formulate complex biocompatible materials for bone fixation devices.

Functionalised collagen spheres reduce H2O2 mediated apoptosis by scavenging overexpressed ROS.

Excess reactive oxygen species (ROS) has been implicated in numerous diseases including cancer, cardiovascular and neurodegenerative diseases. Overexpression of ROS can lead to oxidative stress and subsequently to H2O2-mediated cell apoptosis. In this study, it was demonstrated that biodegradable PLGA microspheres coated with collagen type I and decorated with MnO2 nanoparticles acted as ROS scavengers controlling the H2O2-mediated apoptosis of cells undergoing oxidative stress. The results showed that the functionalized collagen spheres can protect cells even under very harsh conditions of oxidative stress.

Effects of Polydopamine Functionalization on Boron Nitride Nanotube Dispersion and Cytocompatibility.

Boron nitride nanotubes (BNNTs) have unique physical properties, of value in biomedical applications; however, their dispersion and functionalization represent a critical challenge in their successful employment as biomaterials. In the present study, we report a process for the efficient disentanglement of BNNTs via a dual surfactant/polydopamine (PD) process. High-resolution transmission electron microscopy (HR-TEM) indicated that individual BNNTs become coated with a uniform PD nanocoating, which significantly enhanced dispersion of BNNTs in aqueous solutions. Furthermore, the cytocompatibility of PD-coated BNNTs was assessed in vitro with cultured human osteoblasts (HOBs) at concentrations of 1, 10, and 30 µg/mL and over three time-points (24, 48, and 72 h). In this study it was demonstrated that PD-functionalized BNNTs become individually localized within the cytoplasm by endosomal escape and that concentrations of up to 30 µg/mL of PD-BNNTs were cytocompatible in HOBs cells following 72 h of exposure.

From implantation to degradation - are poly (l-lactide)/multiwall carbon nanotube composite materials really cytocompatible?

Poly (l-lactide)'s (PLLA) biodegradable properties are of special value in orthopaedic applications, but its mechanical strength limits its usage. To overcome this PLLA can be reinforced by multiwall carbon nanotubes (MWCNT). In this study the PLLA and MWCNT were combined to prepare nanostructured composites (nanocomposite) at 0, 0.1, 0.5 and 1wt.% reinforcement. The in vitro biocompatibility of these PLLA/MWCNT nanocomposites was evaluated taking into account the various stages of implantation including nanocomposite degradation. PLLA/MWCNT nanocomposites were highly biocompatible with human bone marrow stromal cells (HBMC). The potential surface degradation product, MWCNT, did not induce toxic responses on HBMC. However, the combination of MWCNT with lactic acid, resembling release after bulk degradation, significantly inhibited HBMC proliferation and activity. This study demonstrates the importance of comprehensive evaluations of novel materials for medical applications in predicting possible adverse effects during nanocomposite degradation. FROM THE CLINICAL EDITOR: This study scrutinizes the cytocompatibility of poly-L-lactide reinforced by multiwall carbon nanotubes, and concludes that the combination of MWCNT with lactic acid significantly inhibited human bone marrow stromal cell proliferation and activity, highlighting the importance of comprehensive evaluations of novel materials.

Physicochemical Characterization and In Vitro Activity of Poly(ε-Caprolactone)/Mycophenolic Acid Amorphous Solid Dispersions.

The obtention of amorphous solid dispersions (ASDs) of mycophenolic acid (MPA) in poly(ε-caprolactone) (PCL) is reported in this paper. An improvement in the bioavailability of the drug is possible thanks to the favorable specific interactions occurring in this system. Differential scanning calorimetry (DSC) was used to investigate the miscibility of PCL/MPA blends, measuring glass transition temperature (Tg) and analyzing melting point depression to obtain a negative interaction parameter, which indicates the development of favorable inter-association interactions. Fourier transform infrared spectroscopy (FTIR) was used to analyze the specific interaction occurring in the blends. Drug release measurements showed that at least 70% of the drug was released by the third day in vitro in all compositions. Finally, preliminary in vitro cell culture experiments showed a decreased number of cancerous cells over the scaffolds containing MPA, presumably arising from the anti-cancer activity attributable to MPA.

Self-assembled three-dimensional hydrogels based on graphene derivatives and cerium oxide nanoparticles: scaffolds for co-culture of oligodendrocytes and neurons derived from neural stem cells.

Stem cell-based therapies have shown promising results for the regeneration of the nervous system. However, the survival and integration of the stem cells in the neural circuitry is suboptimal and might compromise the therapeutic outcomes of this approach. The development of functional scaffolds capable of actively interacting with stem cells may overcome the current limitations of stem cell-based therapies. In this study, three-dimensional hydrogels based on graphene derivatives and cerium oxide (CeO2) nanoparticles are presented as prospective supports allowing neural stem cell adhesion, migration and differentiation. The morphological, mechanical and electrical properties of the resulting hydrogels can be finely tuned by controlling several parameters of the self-assembly of graphene oxide sheets, namely the amount of incorporated reducing agent (ascorbic acid) and CeO2 nanoparticles. The intrinsic properties of the hydrogels, as well as the presence of CeO2 nanoparticles, clearly influence the cell fate. Thus, stiffer adhesion substrates promote differentiation to glial cell lineages, while softer substrates enhance mature neuronal differentiation. Remarkably, CeO2 nanoparticle-containing hydrogels support the differentiation of neural stem cells to neuronal, astroglial and oligodendroglial lineage cells, promoting the in vitro generation of nerve tissue grafts that might be employed in neuroregenerative cell therapies.

Lactide and Ethylene Brassylate-Based Thermoplastic Elastomers and Their Nanocomposites with Carbon Nanotubes: Synthesis, Mechanical Properties and Interaction with Astrocytes.

Polylactide (PLA) is among the most commonly used polymers for biomedical applications thanks to its biodegradability and cytocompatibility. However, its inherent stiffness and brittleness are clearly inappropriate for the regeneration of soft tissues (e.g., neural tissue), which demands biomaterials with soft and elastomeric behavior capable of resembling the mechanical properties of the native tissue. In this work, both L- and D,L-lactide were copolymerized with ethylene brassylate, a macrolactone that represents a promising alternative to previously studied comonomers (e.g., caprolactone) due to its natural origin. The resulting copolymers showed an elastomeric behavior characterized by relatively low Young's modulus, high elongation at break and high strain recovery capacity. The thermoplastic nature of the resulting copolymers allows the incorporation of nanofillers (i.e., carbon nanotubes) that further enable the modulation of their mechanical properties. Additionally, nanostructured scaffolds were easily fabricated through a thermo-pressing process with the aid of a commercially available silicon stamp, providing geometrical cues for the adhesion and elongation of cells representative of the nervous system (i.e., astrocytes). Accordingly, the lactide and ethylene brassylate-based copolymers synthesized herein represent an interesting formulation for the development of polymeric scaffolds intended to be used in the regeneration of soft tissues, thanks to their adjustable mechanical properties, thermoplastic nature and observed cytocompatibility.

In vitro preparation of human Dental Pulp Stem Cell grafts with biodegradable polymer scaffolds for nerve tissue engineering.

Human Dental Pulp Stem Cells (hDPSCs) are one of the most promising stem cell sources for tissue engineering and regeneration, due to their extraordinary multi-lineage differentiation ability, ease of extraction from biological waste in dental clinics, safe non-tumorigenic phenotype, immune-tolerance upon in vivo transplantation, and great possibilities of application in autologous tissue reconstruction. The in vitro manipulation of hDPSCs paves the way for drug screening and tailor-made regeneration of damaged tissues, in the context of personalized medicine. The neural crest phenotype of these stem cells gives them the capacity to differentiate to a large variety of cell types, including neural-lineage cells. In this chapter, we describe various culture methods to generate different cellular phenotypes from hDPSCs, which can not only grow as mesenchymal-like plastic adherent cells, but also as non-adherent neurogenic dentospheres in serum-free medium. Floating dentospheres can be used to generate large populations of mature neuronal and glial marker expressing cells, which may be cultured over a substrate of nanopatterned scaffold based on biodegradable poly(lactide-co-caprolactone) (PLCL) to induce a controlled alignment of neurites and cell migration, to generate in vivo biocompatible constructs for nerve tissue bioengineering.

Poly(ethylene oxide)-b-poly(L-lactide) diblock copolymer/carbon nanotube-based nanocomposites: LiCl as supramolecular structure-directing agent.

This work relies on the CNT dispersion in either solution or a polymer matrix through the formation of a three-component supramolecular system composed of PEO-b-PLLA diblock copolymer, carbon nanotubes (CNTs), and lithium chloride. According to a one-pot procedure in solution, the "self-assembly" concept has demonstrated its efficiency using suspension tests of CNTs. Characterizations of the supramolecular system by photon correlation spectroscopy, Raman spectroscopy, and molecular dynamics simulations highlight the charge transfer interaction from the CNTs toward the PEO-b-PLLA/LiCl complex. Finally, this concept was successfully extended in bulk (absence of solvent) via melt-processing techniques by dispersing these complexes in a commercial polylactide (PLA) matrix. Electrical conductivity measurements and transmission electron microscopy attested for the remarkable dispersion of CNTs, confirming the design of high-performance PLA-based materials.

Amorphous solid dispersions in poly(ε-caprolactone)/xanthohumol bioactive blends: physicochemical and mechanical characterization.

This paper reports the obtention of amorphous solid dispersions (ASDs) of xanthohumol (XH) in PCL containing up to 50 wt% of the bioactive compound in the amorphous form thanks to the advantageous specific interactions established in this system. The miscibility of the PCL/XH blends was investigated using DSC. Melting point depression analysis yielded a negative interaction parameter indicating the occurrence of favorable inter-association interactions. XRD analyses performed at room temperature agree with the crystallinity results obtained on the heating runs performed by DSC. FTIR spectroscopy reveals strong C[double bond, length as m-dash]OO-H specific interactions between the hydroxyl groups of XH and the carbonyl groups of PCL. The AFM analysis of the blends obtained by spin-coating shows the variation of crystalline morphology with composition. Finally, tensile tests reveal high toughness retention for the blends in which XH can be dispersed in the amorphous form (containing up to 50 wt% XH). In summary, PCL is a convenient matrix to disperse XH in the amorphous form, bringing the possibility of obtaining completely amorphous bioactive materials suitable for the development of non-stiff biomedical devices.

Smart Layer-by-Layer Polymeric Microreactors: pH-Triggered Drug Release and Attenuation of Cellular Oxidative Stress as Prospective Combination Therapy.

Polymer capsules fabricated via the layer-by-layer (LbL) approach have emerged as promising biomedical systems for the release of a wide variety of therapeutic agents, owing to their tunable and controllable structure and the possibility to include several functionalities in the polymeric membrane during the fabrication process. However, the limitation of the capsules with a single functionality to overcome the challenges involved in the treatment of complex pathologies denotes the need to develop multifunctional capsules capable of targeting several mediators and/or mechanisms. Oxidative stress is caused by the accumulation of reactive oxygen species [e.g., hydrogen peroxide (H2O2), hydroxyl radicals (•OH), and superoxide anion radicals (•O2-)] in the cellular microenvironment and is a key modulator in the pathology of a broad range of inflammatory diseases. The disease microenvironment is also characterized by the presence of proinflammatory cytokines, increased levels of matrix metalloproteinases, and acidic pH, all of which could be exploited to trigger the release of therapeutic agents. In the present work, multifunctional capsules were fabricated via the LbL approach. Capsules were loaded with an antioxidant enzyme (catalase) and functionalized with a model drug (doxorubicin), which was conjugated to an amine-containing dendritic polyglycerol through a pH-responsive linker. These capsules efficiently scavenge H2O2 from solution, protecting cells from oxidative stress, and release the model drug in acidic microenvironments. Accordingly, in this work, a polymeric microplatform is presented as an unexplored combinatorial approach applicable for multiple targets of inflammatory diseases, in order to perform controlled spatiotemporal enzymatic reactions and drug release in response to biologically relevant stimuli.

Crystallization Behavior and Mechanical Properties of Poly(ε-caprolactone) Reinforced with Barium Sulfate Submicron Particles.

Poly(ε-caprolactone) (PCL) was mixed with submicron particles of barium sulfate to obtain biodegradable radiopaque composites. X-ray images comparing with aluminum samples show that 15 wt.% barium sulfate (BaSO4) is sufficient to present radiopacity. Thermal studies by differential scanning calorimetry (DSC) show a statistically significant increase in PCL degree of crystallinity from 46% to 52% for 25 wt.% BaSO4. Non-isothermal crystallization tests were performed at different cooling rates to evaluate crystallization kinetics. The nucleation effect of BaSO4 was found to change the morphology and quantity of the primary crystals of PCL, which was also corroborated by the use of a polarized light optical microscope (PLOM). These results fit well with Avrami-Ozawa-Jeziorny model and show a secondary crystallization that contributes to an increase in crystal fraction with internal structure reorganization. The addition of barium sulfate particles in composite formulations with PCL improves stiffness but not strength for all compositions due to possible cavitation effects induced by debonding of reinforcement interphase.

Novel Hydrogels of Chitosan and Poly(vinyl alcohol) Reinforced with Inorganic Particles of Bioactive Glass.

Chitosan (CS) and poly(vinyl alcohol) (PVA) hydrogels, a polymeric system that shows a broad potential in biomedical applications, were developed. Despite the advantages they present, their mechanical properties are insufficient to support the loads that appear on the body. Thus, it was proposed to reinforce these gels with inorganic glass particles (BG) in order to improve mechanical properties and bioactivity and to see how this reinforcement affects levofloxacin drug release kinetics. Scanning electron microscopy (SEM), X-ray diffraction (XRD), swelling tests, rheology and drug release studies characterized the resulting hydrogels. The experimental results verified the bioactivity of these gels, showed an improvement of the mechanical properties and proved that the added bioactive glass does affect the release kinetics.

Lactide-Valerolactone Copolymers for Packaging Applications.

Lactide-valerolactone copolymers have potential application in the packaging sector. Different copolymers were synthesized, and the kinetics of the copolymerization reactions and the microstructure of the copolymers were analysed. Lactide showed higher reactivity than valerolactone which leads to composition drift through the reaction. Thermal, mechanical and barrier properties of the selected copolymers were studied. Overall, the incorporation of valerolactone results in copolymers with higher ductility than poly(lactide) with intermediate water and oxygen permeability which makes these materials appropriate candidates for use in the packaging sector.

Electrical percolation in extrinsically conducting, poly(ε-decalactone) composite neural interface materials.

By providing a bidirectional communication channel between neural tissues and a biomedical device, it is envisaged that neural interfaces will be fundamental in the future diagnosis and treatment of neurological disorders. Due to the mechanical mismatch between neural tissue and metallic neural electrodes, soft electrically conducting materials are of great benefit in promoting chronic device functionality. In this study, carbon nanotubes (CNT), silver nanowires (AgNW) and poly(hydroxymethyl 3,4-ethylenedioxythiophene) microspheres (MSP) were employed as conducting fillers within a poly(ε-decalactone) (EDL) matrix, to form a soft and electrically conducting composite. The effect of a filler type on the electrical percolation threshold, and composite biocompatibility was investigated in vitro. EDL-based composites exhibited favourable electrochemical characteristics: EDL/CNT-the lowest film resistance (1.2 ± 0.3 kΩ), EDL/AgNW-the highest charge storage capacity (10.7 ± 0.3 mC cm- 2), and EDL/MSP-the highest interphase capacitance (1478.4 ± 92.4 µF cm-2). All investigated composite surfaces were found to be biocompatible, and to reduce the presence of reactive astrocytes relative to control electrodes. The results of this work clearly demonstrated the ability of high aspect ratio structures to form an extended percolation network within a polyester matrix, resulting in the formulation of composites with advantageous mechanical, electrochemical and biocompatibility properties.

The conformation of chloramphenicol in the ordered and disordered phases.

The conformational behavior of chloramphenicol (CHL) in the solid, liquid and vapor phases is revisited here by means of FTIR spectroscopy and QM methods. In the crystalline phase, both the IR analysis and QM computations discard the conformer proposed by Acharya et al. (Acta Cryst., 1979, B35:1360-1363) and support the one proposed by Chatterjee et al. (J. Cryst. Mol. Struct., 1979, 9:295-304), characterized by an intramolecular OH⋯O hydrogen bond in which the primary hydroxyl group acts as hydrogen bond donor. The conformational behavior of CHL in the liquid and gas phases has been analyzed using QM calculations. The Self-Consistent Reaction Field (SCRF) method with the Onsager solvation model has been used for the initial optimizations in solution, and the lowest energy conformers have been refined using the Solvation Model based on Density (SMD). In solution environment the intramolecular OH⋯O hydrogen bond in CHL is reversed so that the secondary hydroxyl group acts as hydrogen bond donor. In addition, the dichloroacetamide group folds back further over the phenyl ring to form an intramolecular CCl⋯π halogen bond. Two different halogen bonds are actually observed (each one with a different chlorine atom) resulting in two different stable conformers, that can be detected by FTIR spectroscopy due to the conformational sensitivity of the CO group to the conformation of the dichloroacetyl group. Finally, the stability of the conformers with the polarity of the medium is also discussed.

Morphology and mechanical properties of poly(ethylene brassylate)/cellulose nanocrystal composites.

Poly(ethylene brassylate), a novel inexpensive biodegradable polyester, has been reinforced with cellulose nanocrystals (CNCs) with the aim of improving its thermal stability and mechanical properties. The composites have been characterized through calorimetry, tensile tests, thermogravimetry and electron microscopy. The addition of small amounts of CNCs improves both the stiffness and the ductility of the composites, suggesting the existence of some compatibilizing effect. Adding large CNC amounts increases the Young modulus (e.g., 150% for 50 wt% CNCs), but now the material shows brittle behavior. Degradation of the CNCs starts at lower temperature suggesting mutual reactivity. The SEM analysis of the composites with ductile behavior reveals the formation of a percolating network crossing through the interconnected domains that conform a PEB-rich continuous phase. Processing consisting on reinforcement dispersion by sonication followed by melt processing results in composites in which the improvement of mechanical properties does not involve any trade-off.

High Throughput Manufacturing of Bio-Resorbable Micro-Porous Scaffolds Made of Poly(L-lactide-co-ε-caprolactone) by Micro-Extrusion for Soft Tissue Engineering Applications.

Porous scaffolds made of elastomeric materials are of great interest for soft tissue engineering. Poly(L-lactide-co-ε-caprolactone) (PLCL) is a bio-resorbable elastomeric copolymer with tailorable properties, which make this material an appropriate candidate to be used as scaffold for vascular, tendon, and nerve healing applications. Here, extrusion was applied to produce porous scaffolds of PLCL, using NaCl particles as a leachable agent. The effects of the particle proportion and size on leaching performance, dimensional stability, mechanical properties, and ageing of the scaffolds were analyzed. The efficiency of the particle leaching and scaffold swelling when wet were observed to be dependent on the porogenerator proportion, while the secant moduli and ultimate tensile strengths were dependent on the pore size. Porosity, swelling, and mechanical properties of the extruded scaffolds were tailorable, varying with the proportion and size of porogenerator particles and showed similar values to human soft tissues like nerves and veins (E = 7-15 MPa, σu = 7 MPa). Up to 300-mm length micro-porous PLCL tube with 400-µm thickness wall was extruded, proving extrusion as a high-throughput manufacturing process to produce tubular elastomeric bio-resorbable porous scaffolds of unrestricted length with tunable mechanical properties.