Unpatterned Bioactive Poly(Butylene 1,4-Cyclohexanedicarboxylate)-Based Film Fast Induced Neuronal-Like Differentiation of Human Bone Marrow-Mesenchymal Stem Cells.

Herein, we present poly(butylene 1,4-cyclohexanedicarboxylate) (PBCE) films characterized by an unpatterned microstructure and a specific hydrophobicity, capable of boosting a drastic cytoskeleton architecture remodeling, culminating with the neuronal-like differentiation of human bone marrow-mesenchymal stem cells (hBM-MSCs). We have used two different filming procedures to prepare the films, solvent casting (PBCE) and compression-moulding (PBCE*). PBCE film had a rough and porous surface with spherulite-like aggregations (Ø = 10-20 µm) and was characterized by a water contact angle = 100°. PBCE* showed a smooth and continuous surface without voids and visible spherulite-like aggregations and was more hydrophobic (WCA = 110°). Both surface characteristics were modulated through the copolymerization of different amounts of ether-oxygen-containing co-units into PBCE chemical structure. We showed that only the surface characteristics of PBCE-solvent-casted films steered hBM-MSCs toward a neuronal-like differentiation. hBM-MSCs lost their canonical mesenchymal morphology, acquired a neuronal polarized shape with a long cell protrusion (≥150 µm), expressed neuron-specific class III ß-tubulin and microtubule-associated protein 2 neuronal markers, while nestin, a marker of uncommitted stem cells, was drastically silenced. These events were observed as early as 2-days after cell seeding. Of note, the phenomenon was totally absent on PBCE* film, as hBM-MSCs maintained the mesenchymal shape and behavior and did not express neuronal/glial markers.

Metal Nanoparticles Embedded in Cellulose Nanocrystal Based Films: Material Properties and Post-use Analysis.

The dispersion of nanoparticles having different size-, shape-, and composition-dependent properties is an exciting approach to design and synthesize multifunctional materials and devices. This work shows a detailed investigation of the preparation and properties of free-standing nanocomposite films based on cellulose nanocrystals (CNC) loaded with three different types of metal nanoparticles. CNC-based nanocomposites having zinc oxide (ZnO), titanium dioxide (TiO2), and silver oxide (Ag2O) have been obtained through evaporation-induced self-assembly (EISA) in acqueous solution. Morphological and optical characteristics, chemical properties, wettability, and antimicrobial assays of the produced films were conducted. Furthermore, disintegrability in composting condition of CNC based nanocomposites was here investigated for the first time. The morphological observations revealed the formation of a chiral nematic structure with uniformly distributed nanoparticles. The bionanocomposite films based on the metal nanoparticles had effective antimicrobial activity, killing both Escherichia coli RB ( E. coli RB) and Staphylococcus aureus 8325-4 ( S. aureus 8325-4). The simplicity method of film preparation, the large quantity of cellulose in the world, and the free-standing nature of the nanocomposite films offer highly advantageous characteristics that can for the new development of multifunctional materials.

Adipose Stem Cell Translational Applications: From Bench-to-Bedside.

During the last five years, there has been a significantly increasing interest in adult adipose stem cells (ASCs) as a suitable tool for translational medicine applications. The abundant and renewable source of ASCs and the relatively simple procedure for cell isolation are only some of the reasons for this success. Here, we document the advances in the biology and in the innovative biotechnological applications of ASCs. We discuss how the multipotential property boosts ASCs toward mesenchymal and non-mesenchymal differentiation cell lineages and how their character is maintained even if they are combined with gene delivery systems and/or biomaterials, both in vitro and in vivo.

The interaction of bacteria with engineered nanostructured polymeric materials: a review.

Bacterial infections are a leading cause of morbidity and mortality worldwide. In spite of great advances in biomaterials research and development, a significant proportion of medical devices undergo bacterial colonization and become the target of an implant-related infection. We present a review of the two major classes of antibacterial nanostructured materials: polymeric nanocomposites and surface-engineered materials. The paper describes antibacterial effects due to the induced material properties, along with the principles of bacterial adhesion and the biofilm formation process. Methods for antimicrobial modifications of polymers using a nanocomposite approach as well as surface modification procedures are surveyed and discussed, followed by a concise examination of techniques used in estimating bacteria/material interactions. Finally, we present an outline of future sceneries and perspectives on antibacterial applications of nanostructured materials to resist or counteract implant infections.

Combined effects of Ag nanoparticles and oxygen plasma treatment on PLGA morphological, chemical, and antibacterial properties.

The purpose of this study is to investigate the combined effects of oxygen plasma treatments and silver nanoparticles (Ag) on PLGA in order to modulate the surface antimicrobial properties through tunable bacteria adhesion mechanisms. PLGA nanocomposite films, produced by solvent casting with 1 wt % and 7 wt % of Ag nanoparticles were investigated. The PLGA and PLGA/Ag nanocomposite surfaces were treated with oxygen plasma. Surface properties of PLGA were investigated by field emission scanning electron microscopy (FESEM), atomic force microscopy (AFM), static contact angle (CA), and high resolution X-ray photoelectron spectroscopy (XPS). Antibacterial tests were performed using an Escherichia coli RB (a Gram negative) and Staphylococcus aureus 8325-4 (a Gram positive). The PLGA surface becomes hydrophilic after the oxygen treatment and its roughness increases with the treatment time. The surface treatment and the Ag nanoparticle introduction have a dominant influence on the bacteria adhesion and growth. Oxygen-treated PLGA/Ag systems promote higher reduction of the bacteria viability in comparison to the untreated samples and neat PLGA. The combination of Ag nanoparticles with the oxygen plasma treatment opens new perspectives for the studied biodegradable systems in biomedical applications.

Novel Nanostructured Scaffolds of Poly(butylene trans-1,4-cyclohexanedicarboxylate)-Based Copolymers with Tailored Hydrophilicity and Stiffness: Implication for Tissue Engineering Modeling.

Here, we present novel biocompatible poly(butylene trans-1,4-cyclohexanedicarboxylate) (PBCE)-based random copolymer nanostructured scaffolds with tailored stiffness and hydrophilicity. The introduction of a butylene diglycolate (BDG) co-unit, containing ether oxygen atoms, along the PBCE chain remarkably improved the hydrophilicity and chain flexibility. The copolymer containing 50 mol% BDG co-units (BDG50) and the parent homopolymer (PBCE) were synthesized and processed as electrospun scaffolds and compression-molded films, added for the sake of comparison. We performed thermal, wettability, and stress-strain measures on the PBCE-derived scaffolds and films. We also conducted biocompatibility studies by evaluating the adhesion and proliferation of multipotent mesenchymal/stromal cells (hBM-MSCs) on each polymeric film and scaffold. We demonstrated that solid-state properties can be tailored by altering sample morphology besides chemical structure. Thus, scaffolds were characterized by a higher hydrophobicity and a lower elastic modulus than the corresponding films. The three-dimensional nanostructure conferred a higher adsorption protein capability to the scaffolds compared to their film counterparts. Finally, the PBCE and BDG50 scaffolds were suitable for the long-term culture of hBM-MSCs. Collectively, the PBCE homopolymer and copolymer are good candidates for tissue engineering applications.

Tuning multi/pluri-potent stem cell fate by electrospun poly(L-lactic acid)-calcium-deficient hydroxyapatite nanocomposite mats.

In this study, we investigated whether multipotent (human-bone-marrow-derived mesenchymal stem cells [hBM-MSCs]) and pluripotent stem cells (murine-induced pluripotent stem cells [iPSCs] and murine embryonic stem cells [ESCs]) respond to nanocomposite fibrous mats of poly(L-lactic acid) (PLLA) loaded with 1 or 8 wt % of calcium-deficient nanohydroxyapatite (d-HAp). Remarkably, the dispersion of different amounts of d-HAp to PLLA produced a set of materials (PLLA/d-HAp) with similar architectures and tunable mechanical properties. After 3 weeks of culture in the absence of soluble osteogenic factors, we observed the expression of osteogenic markers, including the deposition of bone matrix proteins, in multi/pluripotent cells only grown on PLLA/d-HAp nanocomposites, whereas the osteogenic differentiation was absent on stem-cell-neat PLLA cultures. Interestingly, this phenomenon was confined only in hBM-MSCs, murine iPSCs, and ESCs grown on direct contact with the PLLA/d-HAp mats. Altogether, these results indicate that the osteogenic differentiation effect of these electrospun PLLA/d-HAp nanocomposites was independent of the stem cell type and highlight the direct interaction of stem cell-polymeric nanocomposite and the mechanical properties acquired by the PLLA/d-HAp nanocomposites as key steps for the differentiation process.

Enhancing osteoconduction of PLLA-based nanocomposite scaffolds for bone regeneration using different biomimetic signals to MSCs.

In bone engineering, the adhesion, proliferation and differentiation of mesenchymal stromal cells rely on signaling from chemico-physical structure of the substrate, therefore prompting the design of mimetic "extracellular matrix"-like scaffolds. In this study, three-dimensional porous poly-L-lactic acid (PLLA)-based scaffolds have been mixed with different components, including single walled carbon nanotubes (CNT), micro-hydroxyapatite particles (HA), and BMP2, and treated with plasma (PT), to obtain four different nanocomposites: PLLA + CNT, PLLA + CNTHA, PLLA + CNT + HA + BMP2 and PLLA + CNT + HA + PT. Adult bone marrow mesenchymal stromal cells (MSCs) were derived from the femur of orthopaedic patients, seeded on the scaffolds and cultured under osteogenic induction up to differentiation and mineralization. The release of specific metabolites and temporal gene expression profiles of marrow-derived osteoprogenitors were analyzed at definite time points, relevant to in vitro culture as well as in vivo differentiation. As a result, the role of the different biomimetic components added to the PLLA matrix was deciphered, with BMP2-added scaffolds showing the highest biomimetic activity on cells differentiating to mature osteoblasts. The modification of a polymeric scaffold with reinforcing components which also work as biomimetic cues for cells can effectively direct osteoprogenitor cells differentiation, so as to shorten the time required for mineralization.

Design and Analysis of a Novel Ultraviolet-C Device for Surgical Face Mask Disinfection.

This paper deals with the design of a compact sanitization device and the definition of a specific protocol for UV-C disinfection of a surgical face mask. The system was designed considering the material properties, face mask shape, and UV-C light distribution. DIALux software was used to evaluate the irradiance distribution provided by the lamps emitting in the UV-C range. The irradiance needed for UV-C-decontaminated bacteria and virus, and other contaminating pathogens, without compromising their integrity and guaranteeing inactivation of the bacteria, was evaluated. The face mask's material properties were analyzed with respect to UV-C exposure in terms of physicochemical properties, breathability, and bacterial filtration performance. Information on the effect of time-dependent passive decontamination at room temperature storage was provided. Single and multiple cycles of UV-C sanitization did not adversely affect respirator breathability and bacterial filtration efficiency. This multidisciplinal approach may provide important information on how it is possible to correctly sanitize a face mask and, in case of shortage, safely reuse the face mask.

Thermal treatment of magnesium particles in polylactic acid polymer films elicits the expression of osteogenic differentiation markers and lipidome profile remodeling in human adipose stem cells.

The efficacy of polylactic acid (PLA)/Magnesium (Mg)-based materials for driving stem cells toward bone tissue engineering applications requires specific Mg surface properties to modulate the interface of stem cells with the film. Here, we have developed novel PLA/Mg-based composites and explored their osteogenic differentiation potential on human adipose stem cells (hASCs). Mg-particles/polymer interface was improved by two treatments: heating in oxidative atmosphere (TT) and surface modification with a compatibilizer (PEI). Different contents of Mg particles were dispersed in PLA and composite surface and bulk properties, protein adsorption, stem cell-PLA/Mg interactions, osteogenic markers expressions, and lipids composition profile were evaluated. Mg particles were uniformly distributed on the surface and in the bulk PLA polymer. Improved and modulated particle-polymer adhesion was observed in Mg particle-treated composites. After 21 days in canonical growth culture conditions, hASCs on PLA/MgTT displayed the highest expression of the general osteogenic markers, RUNX2, SSP1, and BGLAP genes, Alkaline Phosphatase, type I Collagen, Osteopontin, and Calcium deposits. Moreover, by LC/MS QTOF mass-spectrophotometry lipidomic analysis, we found in PLA/MgTT-cells, for the first time, a remodeling of the lipid classes composition associated with the osteogenic differentiation. We ascribed these results to MgTT characteristics, which improve Mg availability and composite osteoinductive performance.

Dielectric Spectroscopy of PP/MWCNT Nanocomposites: Relationship with Crystalline Structure and Injection Molding Condition.

In this paper, we study the correlation between the dielectric behavior of polypropylene/multi-walled carbon nanotube (PP/MWCNT) nanocomposites and the morphology with regard to the crystalline structure, nanofiller dispersion and injection molding conditions. As a result, in the range of the percolation threshold the dielectric behavior shifts to a more frequency-independent behavior, as the mold temperature increases. Moreover, the position further from the gate appears as the most conductive. This effect has been associated to a modification of the morphology of the MWCNT clusters induced by both the flow of the molten polymer during the processing phase and the variation of the crystalline structure, which is increasingly constituted by γ-phase as the mold temperature increases. The obtained results allow one to understand the effect of tuning the processing condition in the frequency-dependent electrical behavior of PP/MWCNT injection-molded nanocomposites, which can be successfully exploited for an advanced process/product design.

Microstructure and cytocompatibility of electrospun nanocomposites based on poly(epsilon-caprolactone) and carbon nanostructures.

Carbon nanostructures (CNSs) are attractive and promising nanomaterials for the next generation of tissue engineering scaffolds, especially in neural prosthesis. Optimizing scaffold vascularization may be an important strategy to promote the repair of damaged brain tissue. In this context, the idea was to evaluate the cell response of electrospun nanohybrid scaffolds loaded with CNSs. Fibrous composites based on poly(epsilon-caprolactone) (PCL) and CNSs were fabricated by means of electrospinning technique. High-purity carbon nanofibers (CNFs) and single-wall carbon nanotubes (SWNTs) were studied. A detailed microstructural characterization was performed to evaluate the most favorable experimental conditions for the realization of fibrous PCL/CNS fabrics. Electrospun mats comprised of rather uniform and homogeneous submicrometric fibers were obtained starting from 1:1 v/v mixture of tetrahydrofuran (THF) and N,N dimethylformamide (DMF). In vitro cytocompatibility tests were performed using rat cerebro-microvascular endothelial cells (CECs). Acquired results showed an increased cell viability for PCL/CNS nanocomposites, suggesting these materials as a suitable environment for endothelial cells. These results are indicative of the promising potential of CNS-based nanocomposites in biomedical devices for tissue engineering applications where endothelial functional properties are required.

Effect of Injection Molding Conditions on Crystalline Structure and Electrical Resistivity of PP/MWCNT Nanocomposites.

Polypropylene (PP) / multi-walled carbon nanotube (MWCNT) nanocomposites were prepared by melt-mixing and used to manufacture samples by injection molding. The effect of processing conditions on the crystallinity and electrical resistivity was studied. Accordingly, samples were produced varying the mold temperature and injection rate, and the DC electrical resistivity was measured. The morphology of MWCNT clusters was studied by optical and electron microscopy, while X-ray diffraction was used to study the role of the crystalline structure of PP. As a result, an anisotropic electrical behavior induced by the process was observed, which is further influenced by the injection molding processing condition. It was demonstrated that a reduction of electrical resistivity can be obtained by increasing mold temperature and injection rate, which was associated to the formation of the γ-phase and the related inter-cluster morphology of the MWCNT conductive network.

Improving the flexibility and compostability of starch/poly(butylene cyclohexanedicarboxylate)-based blends.

Fully biobased blends of thermoplastic starch and a poly(butylene cyclohexanedicarboxylate)-based random copolyester containing 25 % of adipic acid co-units (PBCEA) are prepared by melt blending and direct extrusion film casting. The obtained films are characterized from the physicochemical and mechanical point of view and their fragmentation under composting conditions is evaluated. The results demonstrate that the introduction of adipic acid co-units in the PBCE macromolecular chains permits to decrease the blending temperature, thus avoiding unwanted starch degradation reactions. Moreover, the presence of small amounts of citric acid as compatibilizer further improves the interfacial adhesion between the two components and promotes the formation of micro-porosities within the films. The synergistic combination of these factors leads to the development of materials showing an elastomeric behavior, i.e. no evident yield and elongation at break higher than 450 %, good moisture resistance and fast fragmentation in compost.

Synergic Effect of Nanolignin and Metal Oxide Nanoparticles into Poly(l-lactide) Bionanocomposites: Material Properties, Antioxidant Activity, and Antibacterial Performance.

Binary and ternary poly(l-lactide) (PLLA)-based nanocomposites, containing nanolignin (1 wt %) and different metal oxide nanoparticles (0.5 wt %, Ag2O, TiO2, WO3, Fe2O3, and ZnFe2O4), were realized by solvent casting, and their morphological, thermal, surface, optical, antioxidant, and antimicrobial characterizations were performed. The presence of metal oxide nanoparticles at the selected weight concentration affects the surface microstructure of the PLLA polymer, and this outcome is particle-type dependent, according to the shape, morphology, and chemical properties of the selected nanoparticles (NPs). Analogously, wettability of PLLA-based nanocomposites was slightly modified by the presence of hydrophobic lignin nanoparticles and different shaped metal oxides. Results of differential scanning calorimetry (DSC) and X-ray powder diffraction (XRD) tests confirmed that nanoparticle addition confined the mobility of the amorphous phase, increasing at the same time the formation of more numerous but less perfect PLLA crystals. Interestingly, antioxidant activity was also obtained in ternary-based nanocomposites, where a synergic effect of lignin and metal oxide nanoparticles was obtained. Antibacterial tests showed manifest activity of TiO2 and Ag2O nanoparticles containing PLLA films, and the time dependence was more evident for Staphylococcus aureus than for Escherichia coli. Lignin nanoparticles are able to provide protection against UV light while still allowing visible light to pass and even surpass the UV-protection capacity provided by many inorganic nanoparticles. This makes them an attractive renewable additive for the realization of PLLA/metal oxide nanocomposites in the fields of food, drug packaging, and biomedical industry, where antibacterial and antioxidant properties are required.

Multifunctional ternary composite films based on PLA and Ag/alginate microbeads: Physical characterization and silver release kinetics.

Novel multifunctional composite poly(lactic acid) (PLA) films with alginate microbeads containing silver nanoparticles (AgNPs) were developed for potential antimicrobial food packaging applications. AgNPs, 10-20 nm in size, were synthesized in a Na-alginate solution by a hydrothermal method yielding a sterile, pH neutral colloid solution of low viscosity that was electrostatically extruded to produce Ag/alginate microbeads (190 µm in size) with retained AgNPs. Dried microbeads were uniformly dispersed in PLA films with retained AgNPs as confirmed by UV-Vis spectroscopy and scanning electron microscopy. The films were characterized regarding thermal and mechanical properties as well as silver release in different food simulants. Results show that PLA matrix served as a diffusion barrier so that the released silver concentration in water after 10 days was within the prescribed limit of 0.05 mg kg-1 while the films induced inhibitory effects against Staphylococcus aureus in the agar diffusion test.

Combined effect of cellulose nanocrystals, carvacrol and oligomeric lactic acid in PLA_PHB polymeric films.

Biodegradable multicomponent films based on poly(lactic acid) (PLA) and poly(3-hydroxybutyrate) (PHB) plasticized with oligomeric lactic acid (OLA), reinforced with synthetized cellulose nanocrystals (CNC) and modified by a natural additive with antimicrobial activity (carvacrol) were formulated and processed by extrusion. Morphological, mechanical, thermal, migration and barrier properties were tested to determine the effect of different components in comparison with neat poly(lactic acid). Results showed the positive effect of CNC in the five components based films, with the increase of the Young's modulus of the PLA_PHB_10Carv_15OLA, associated with an increase in the elongation at break (from 150% to 410%), by showing an OTR reduction of 67%. Disintegrability in compost conditions and enzymatic degradation were tested to evaluate the post-use of these films. All formulations disintegrated in less than 17 days, while proteinase K preferentially degraded the amorphous regions, and crystallinity degree of the nanocomposite films increased as a consequence of enzyme action.

Nanocomposites Based on Biodegradable Polymers.

In the present review paper, our main results on nanocomposites based on biodegradable polymers (on a time scale from 2010 to 2018) are reported. We mainly focused our attention on commercial biodegradable polymers, which we mixed with different nanofillers and/or additives with the final aim of developing new materials with tunable specific properties. A wide list of nanofillers have been considered according to their shape, properties, and functionalization routes, and the results have been discussed looking at their roles on the basis of different adopted processing routes (solvent-based or melt-mixing processes). Two main application fields of nanocomposite based on biodegradable polymers have been considered: the specific interaction with stem cells in the regenerative medicine applications or as antimicrobial materials and the active role of selected nanofillers in food packaging applications have been critically revised, with the main aim of providing an overview of the authors' contribution to the state of the art in the field of biodegradable polymeric nanocomposites.

Surface Hydrophilicity of Poly(l-Lactide) Acid Polymer Film Changes the Human Adult Adipose Stem Cell Architecture.

Current knowledge indicates that the molecular cross-talk between stem cells and biomaterials guides the stem cells' fate within a tissue engineering system. In this work, we have explored the effects of the interaction between the poly(l-lactide) acid (PLLA) polymer film and human adult adipose stem cells (hASCs), focusing on the events correlating the materials' surface characteristics and the cells' plasma membrane. hASCs were seeded on films of pristine PLLA polymer and on a PLLA surface modified by the radiofrequency plasma method under oxygen flow (PLLA+O2). Comparative experiments were performed using human bone-marrow mesenchymal stem cells (hBM-MSCs) and human umbilical matrix stem cells (hUCMSCs). After treatment with oxygen-plasma, the surface of PLLA films became hydrophilic, whereas the bulk properties were not affected. hASCs cultured on pristine PLLA polymer films acquired a spheroid conformation. On the contrary, hASCs seeded on PLLA+O2 film surface maintained the fibroblast-like morphology typically observed on tissue culture polystyrene. This suggests that the surface hydrophilicity is involved in the acquisition of the spheroid conformation. Noteworthy, the oxygen treatment had no effects on hBM-MSC and hUCMSC cultures and both stem cells maintained the same shape observed on PLLA films. This different behavior suggests that the biomaterial-interaction is stem cell specific.

Processing and characterization of nanocomposite based on poly(butylene/triethylene succinate) copolymers and cellulose nanocrystals.

A new class of biodegradable materials developed by a combination of random eco-friendly copolyesters containing butylene succinate (BS) and triethylene succinate (TES) sequences with cellulose nanocrystals (CNC), is proposed and studied. Polymers and nanocomposite films were prepared by an optimized extrusion process to improve the processability and mechanical response for flexible film manufacturing. Poly(butylene succinate) (PBS) homopolymer and two random copolyesters containing different amounts of TES co-units, P(BS85TES15) and P(BS70TES30), were synthesized by melt polycondensation. The effect of TES and CNC presence and content on the microstructure, tensile properties, thermal characteristics and disintegration under composting conditions, as well as on the toughening mechanism of the blends was investigated. Material properties were modulated by varying the chemical composition. CNC were used as reinforcement additive and their effect is modulated by the interaction with the three polymeric matrices. The extruded films displayed tunable degradation rates, mechanical properties and wettability, and showed promising results for different industrial applications.