Chitosan-Collagen Electrospun Nanofibers Loaded with Curcumin as Wound-Healing Patches.

Composite chitosan-collagen nanofibrous mats embedded with curcumin were prepared via a single-step electrospinning procedure and explored as wound-healing patches with superior biological activity. A mild crosslinking protocol consisting of a short exposure to ammonia vapor and UV radiation was developed to ensure proper stability in physiological-like conditions without affecting the intrinsic biocompatibility of chitosan and collagen. The fabricated composite patches displayed a highly porous, homogeneous nanostructure consisting of fibers with an average diameter of 200 nm, thermal stability up to 200 °C, mechanical features able to ensure protection and support to the new tissues, and water-related properties in the ideal range to allow exudate removal and gas exchange. The release kinetic studies carried out in a simulated physiological environment demonstrated that curcumin release was sustained for 72 h when the mats are crosslinked hence providing prolonged bioactivity reflected by the displayed antioxidant properties. Remarkably, combining chitosan and collagen not only ensures prolonged stability and optimal physical-chemical properties but also allows for better-promoting cell adhesion and proliferation and enhanced anti-bacteriostatic capabilities with the addition of curcumin, owing to its beneficial anti-inflammatory effect, ameliorating the attachment and survival/proliferation rates of keratinocytes and fibroblasts to the fabricated patches.

Electrospun Materials Based on Polymer and Biopolymer Blends-A Review.

This review covers recent developments and progress in polymer and biopolymer blending and material preparation by electrospinning. Electrospinning is a technique that is used to produce nanofibers to improve the quality of membranes. Electrospun nanofibers are highly applicable in biomedical sciences, supercapacitors, and in water treatment following metal ion adsorption. The key affecting factors of electrospinning have been checked in the literature to obtain optimal conditions of the electrospinning process. Future research directions and outlooks have been suggested to think about innovative ideas for research in this field.

Seasonal Molecular Difference in Fibrillar Collagen Extracts Derived from the Marine Sponge Chondrosia reniformis (Nardo, 1847) and Their Impact on Its Derived Biomaterials.

Chondrosia reniformis (Nardo, 1847) is a marine sponge of high biotechnological interest both for its natural compound content and for its peculiar collagen, which is suitable for the production of innovative biomaterials in the form, for instance, of 2D membranes and hydrogels, exploitable in the fields of tissue engineering and regenerative medicine. In this study, the molecular and chemical-physical properties of fibrillar collagen extracted from specimens collected in different seasons are studied to evaluate the possible impact of sea temperature on them. Collagen fibrils were extracted from sponges harvested by the Sdot Yam coast (Israel) during winter (sea temperature: 17 °C) and during summer (sea temperature: 27 °C). The total AA composition of the two different collagens was evaluated, together with their thermal stability and glycosylation level. The results showed a lower lysyl-hydroxylation level, lower thermal stability, and lower protein glycosylation level in fibrils extracted from 17 °C animals compared to those from 27 °C animals, while no differences were noticed in the GAGs content. Membranes obtained with fibrils deriving from 17 °C samples showed a higher stiffness if compared to the 27 °C ones. The lower mechanical properties shown by 27 °C fibrils are suggestive of some unknown molecular changes in collagen fibrils, perhaps related to the creeping behavior of C. reniformis during summer. Overall, the differences in collagen properties gain relevance as they can guide the intended use of the biomaterial.

Electrospun alginate mats embedding silver nanoparticles with bioactive properties.

Polysaccharide-based composites embedding silver nanoparticles (AgNPs) represent a promising alternative to common antimicrobial materials because of the effective, broad-spectrum biocidal properties of AgNPs combined with the biocompatibility and environmental safety of the naturally occurring polymeric component. In this work, AgNPs stabilized with alginate chains (Alg@AgNPs) were successfully synthesized in situ within the polysaccharide solution through a wet chemical approach carried out at different concentrations of the silver salt precursor. Once obtained, the aqueous suspensions were electrospun to prepare non-woven membranes, showing a homogeneous nanostructured texture (with fiber diameter between 100 and 150 nm), which was found to be influenced by the size (between 20 and 35 nm) of the embedded metal nanoparticles. The biocidal potential of the nanocomposite mats was preliminarily tested against Gram-negative E. coli. The results showed that the antimicrobial response of the investigated samples occurred within a day of incubation and can be observed for AgNPs content in the polysaccharide fibers far below the nanomolar regime.

Potential Biomedical Applications of Collagen Filaments derived from the Marine Demosponges Ircinia oros (Schmidt, 1864) and Sarcotragus foetidus (Schmidt, 1862).

Collagen filaments derived from the two marine demosponges Ircinia oros and Sarcotragus foetidus were for the first time isolated, biochemically characterised and tested for their potential use in regenerative medicine. SDS-PAGE of isolated filaments revealed a main collagen subunit band of 130 kDa in both of the samples under study. DSC analysis on 2D membranes produced with collagenous sponge filaments showed higher thermal stability than commercial mammalian-derived collagen membranes. Dynamic mechanical and thermal analysis attested that the membranes obtained from filaments of S. foetidus were more resistant and stable at the rising temperature, compared to the ones derived from filaments of I. oros. Moreover, the former has higher stability in saline and in collagenase solutions and evident antioxidant activity. Conversely, their water binding capacity results were lower than that of membranes obtained from I. oros. Adhesion and proliferation tests using L929 fibroblasts and HaCaT keratinocytes resulted in a remarkable biocompatibility of both developed membrane models, and gene expression analysis showed an evident up-regulation of ECM-related genes. Finally, membranes from I. oros significantly increased type I collagen gene expression and its release in the culture medium. The findings here reported strongly suggest the biotechnological potential of these collagenous structures of poriferan origin as scaffolds for wound healing.

Composite Poly(vinyl alcohol)-Based Nanofibers Embedding Differently-Shaped Gold Nanoparticles: Preparation and Characterization.

Poly(vinyl alcohol) nanofibrous mats containing ad hoc synthesized gold nanostructures were prepared via a single-step electrospinning procedure and investigated as a novel composite platform with several potential applications. Specifically, the effect of differently shaped and sized gold nanostructures on the resulting mat physical-chemical properties was investigated. In detail, nearly spherical nanoparticles and nanorods were first synthesized through a chemical reduction of gold precursors in water by using (hexadecyl)trimethylammonium bromide as the stabilizing agent. These nanostructures were then dispersed in poly(vinyl alcohol) aqueous solutions to prepare nanofibrous mats, which were then stabilized via a humble thermal treatment able to enhance their thermal stability and water resistance. Remarkably, the nanostructure type was proven to influence the mesh morphology, with the small spherical nanoparticles and the large nanorods leading to thinner well defined or bigger defect-rich nanofibers, respectively. Finally, the good mechanical properties shown by the prepared composite mats suggest their ease of handleability thereby opening new perspective applications.

Effect of sodium alginate molecular structure on electrospun membrane cell adhesion.

Alginate-based electrospun nanofibers prepared via electrospinning technique represent a class of materials with promising applications in the biomedical and pharmaceutical industries. However, to date, the effect of alginate molecular mass and block composition on the biological response of such systems remains to some extent unclear. As such, in the present work, three alginates (i.e., M.pyr, L.hyp, A.nod) with different molecular features are employed to prepare nanofibers whose ability to promote cell adhesion is explored by using both skin and bone cell lines. Initially, a preliminary investigation of the raw materials is carried out via rheological and zeta-potential measurements to determine the different grade of polyelectrolyte behaviour of the alginate samples. Specifically, both the molecular mass and block composition are found to be important factors affecting the alginate response, with long chains and a predominance of guluronic moieties leading to a marked polyelectrolyte nature (i.e., lower dependence of the solution viscosity upon the polymer concentration). Subsequently, physically crosslinked alginate nanofibrous mats are first morphologically characterized via both scanning electron and atomic force microscopy, which show a homogenous and defect-free structure, and their biological response is then evaluated. Noticeably, fibroblast and keratinocyte cell lines do not show significant differences in terms of cell adhesion on the three mats (i.e., 30-40% and 10-20% with respect to the seeded cells, respectively), with the formers presenting a greater affinity toward the alginate-based nanofibers. Conversely, both the investigated osteoblast cells are characterized by a distinct behaviour depending on the alginate type. Specifically, polysaccharide samples with an evident polyelectrolyte nature are found to better promote cell viability (i.e., cell adhesion in the range 15-36% with respect to seeded cells) compared to the ones displaying a nearly neutral behaviour (i.e., cell adhesion in the range 5-25% with respect to seeded cells). Therefore, the obtained results, despite being preliminary, suggest that the alginate type (i.e., molecular structure properties) may play a topical role in conditioning the efficiency of healing patches for bone reparation, but it has a negligible effect in the case of skin regeneration.

Effect of Crosslinking Type on the Physical-Chemical Properties and Biocompatibility of Chitosan-Based Electrospun Membranes.

Chitosan nanofibrous membranes are prepared via an electrospinning technique and explored as potential wound healing patches. In particular, the effect of a physical or chemical crosslinking treatment on the mat morphological, mechanical, water-related, and biological properties is deeply evaluated. The use of phosphate ions (i.e., physical crosslinking) allows us to obtain smooth and highly homogenous nanofibers with an average size of 190 nm, whereas the use of ethylene glycol diglycidyl ether (i.e., chemical crosslinking) leads to rougher, partially coalesced, and bigger nanofibers with an average dimension of 270 nm. Additionally, the physically crosslinked mats show enhanced mechanical performances, as well as greater water vapour permeability and hydrophilicity, with respect to the chemically crosslinked ones. Above all, cell adhesion and cytotoxicity experiments demonstrate that the use of phosphate ions as crosslinkers significantly improves the capability of chitosan mats to promote cell viability owing to their higher biocompatibility. Moreover, tuneable drug delivery properties are achieved for the physically crosslinked mats by a simple post-processing impregnation methodology, thereby indicating the possibility to enrich the prepared membranes with unique features. The results prove that the proposed approach may lead to the preparation of cheap, biocompatible, and efficient chitosan-based nanofibers for biomedical and pharmaceutical applications.

Polymer-free cyclodextrin and natural polymer-cyclodextrin electrospun nanofibers: A comprehensive review on current applications and future perspectives.

The present review discusses the use of cyclodextrins and their derivatives to prepare electrospun nanofibers with specific features. Cyclodextrins, owing to their unique capability to form inclusion complexes with hydrophobic and volatile molecules, can indeed facilitate the encapsulation of bioactive compounds in electrospun nanofibers allowing fast-dissolving products for food, biomedical, and pharmaceutical purposes, filtering materials for wastewater and air purification, as well as a variety of other technological applications. Additionally, cyclodextrins can improve the processability of naturally occurring biopolymers helping the fabrication of "green" materials with a strong industrial relevance. Hence, this review provides a comprehensive state-of-the-art of different cyclodextrins-based nanofibers including those made of pure cyclodextrins, of polycyclodextrins, and those made of natural biopolymer functionalized with cyclodextrins. To this end, the advantages and disadvantages of such approaches and their possible applications are investigated along with the current limitations in the exploitation of electrospinning at the industrial level.

Experimental assessment of the performance of vitreous cutters with fluids with different rheological properties.

PURPOSE: To assess the influence of rheological properties of an artificial vitreous (AV) on the performance of double-blade (DB) and single-blade (SB) guillotine vitreous cutters, with 23-, 25-, and 27-gauge (G) probes. METHODS: We evaluate the aspiration flow rate, using an optical method, based on image processing. Experiments are conducted using ten viscoelastic vitreous phantoms, with different properties that are measured with rheological tests. RESULTS: Aspiration rate strongly varies with fluid properties. Regardless of cutter geometry and operational conditions, the flow rate significantly decreases as vitreous viscosity and elasticity increase. CONCLUSIONS: All tested vitreous probes are very sensitive to changes in fluid rheology. SB cutters produce smaller flow rates compared with DB ones of the same caliber; however, they are less sensitive to fluid properties at low aspiration pressures. The use of vitreous substitutes for test performance guarantees comparability between flow rate results achieved with different vitrectomy systems operating in different media. This outcome is further confirmed by the low values of estimated flow rate relative errors.

Nanocomposite alginate-based electrospun membranes as novel adsorbent systems.

Alginate-based membranes embedding zinc oxide nanoparticles are prepared via electrospinning and exploited as biosorbent materials. The mats exhibit a uniform texture characterized by the presence of nanofibers with an average diameter of 100 nm and interconnected voids of 140 nm average size. UV-vis spectrophotometric tests were performed to evaluate the membrane uptake/release performances by employing aqueous solutions of Methylene Blue (MB) and Congo Red (CR), chosen as model probes of basic and acidic type, respectively. Isotherm kinetics and equilibrium data are fitted with theoretical models to acquire information on the process mechanisms and rates. At low dosage, the mats show comparable adsorption capacity toward both dyes with limited selectivity for the cationic one suggesting that the process is conditioned by the macroporous structure of the membranes. Moreover, a good reusability for achieved for MB after simple washing steps in deionized water. Remarkably, the desorption efficacy under physiological-like conditions turn out to be very high for MB but reduced for CR indicating that the release process is affected by ionic interactions. Based on the results, the electrospun membranes reveal potential as innovative, low-cost, and versatile absorbent platforms to be used in drug delivery applications as well as in purification processes.

Investigation of the Mechanical and Dynamic-Mechanical Properties of Electrospun Polyvinylpyrrolidone Membranes: A Design of Experiment Approach.

Polyvinylpyrrolidone electrospun membranes characterized by randomly, partially, or almost completely oriented nanofibers are prepared using a drum collector in static (i.e., 0 rpm) or rotating (i.e., 250 rpm or 500 rpm) configuration. Besides a progressive alignment alongside the tangential speed direction, the nanofibers show a dimension increasing with the collector rotating speed in the range 410-570 nm. A novel design of experiment approach based on a face-centred central composite design is employed to describe membrane mechanical properties using the computation of mathematical models and their visualization via response surface methodology. The results demonstrate the anisotropic nature of the fibre-oriented membranes with Young's modulus values of 165 MPa and 71 MPa parallelly and perpendicularly to the alignment direction, respectively. Above all, the proposed approach is proved to be a promising tool from an industrial point of view to prepare electrospun membranes with a tailored mechanical response by simply controlling the collector speed.

Rheological, Mechanical and Morphological Characterization of Fillers in the Nautical Field: The Role of Dispersing Agents on Composite Materials.

Coatings have a fundamental role in covering the external surface of yachts by acting both as protective and aesthetic layers. In particular, fillers represent the essential layer from the point of view of mechanical properties and consist of a polymeric matrix, different extenders and additives, and dispersing agents, with the latter having the role to provide good extender-matrix compatibility. In the present work, the effects of dispersing agents with an ionic or steric action on the interactions between hollow glass microspheres and an epoxy-polyamide resin are evaluated. Un-crosslinked filler materials are studied via rheological tests, whereas the mechanical and morphological properties of the crosslinked samples are assessed. The results clearly indicate that steric dispersing agents provide a much greater compatibility effect compared to ionic ones, owing to their steric hindrance capability, thus leading to better-performing filler materials with a less-marked Payne effect, which is here proved to be an efficient tool to provide information concerning the extent of component interactions in nautical fillers. To the best of our knowledge, this work represents the first attempt to deeply understand the role of dispersing agents, which are until now empirically used in the preparation of fillers.

Multilayer Alginate-Polycaprolactone Electrospun Membranes as Skin Wound Patches with Drug Delivery Abilities.

A multilayer nanofibrous membrane consisting of a layer of polycaprolactone and one of physically cross-linked alginate-embedding ZnO nanoparticles is prepared via electrospinning technique as potential wound healing patches with drug delivery capabilities. A washing-cross-linking protocol is developed to obtain stable materials at the same time removing poly(ethylene oxide), which was used here as a cospinning agent for alginate, without interfering with the membrane's peculiar nanofibrous structure. The mechanical behavior of the samples is assessed via a uniaxial tensile test showing appropriate resistance and manageability together with a good thermal stability as proved via thermogravimetric analysis. The polycaprolactone external layer enriches the samples with good liquid-repellent properties, whereas the alginate layer is able to promote tissue regeneration owing to its capability to promote cell viability and allow exudate removal and gas exchanges. Moreover, using methylene blue and methyl orange as model molecules, promising drug delivery abilities are observed for the mats. Indeed, depending on the nature and on the dye-loading concentration, the release kinetic can be easily tuned to obtain a slow controlled or a fast burst release. Consequently, the proposed alginate-polycaprolactone membrane represents a promising class of innovative, simple, and cost-effective wound healing patches appropriate for large-scale production.

Chitosan-based electrospun membranes: Effects of solution viscosity, coagulant and crosslinker.

Chitosan-based membranes were prepared via electrospinning technique using a low concentrated acetic acid solution as solvent and poly(ethylene oxide) as co-spinning agent. Different solutions were rheologically characterized and increasing the solution viscosity was found to correspond to a better-defined morphology. The membranes were first subjected to a coagulation process with different baths in order to stabilize chitosan and the mats were found not able to withstand a strongly basic environment. Subsequently, a physical and a chemical crosslinking approach were separately optimized to obtain stable mats whose composition was assessed via thermogravimetric and spectroscopic techniques, proving in both cases the elimination of the co-spinning agent. Above all, the ionically crosslinked mats represent a class of extremely promising biomedical products being probably highly biocompatible and characterized by thin and homogenous nanofibers with a diameter of 200 nm, thus showing the ideal structure to foster cell viability.

Preparation of composite alginate-based electrospun membranes loaded with ZnO nanoparticles.

In the present work alginate-based nanofibrous membranes embedding zinc oxide nanoparticles (ZnO-NPs) were prepared via electrospinning technique. ZnO-NPs were synthesized by means of a "green" sol-gel method by using alginate itself as stabilizing agent and characterized through UV-vis spectroscopy, thermogravimetric and morphological analysis. Formulations containing sodium alginate, poly(ethylene oxide) and ZnO-NPs were rheologically studied to identify the most suitable ones to be electrospun; alginate molecular structure played an important role on the solution spinnability due to the polysaccharide capability to establish electrostatic interactions and hydrogen bonds with ZnO-NPs. An innovative washing-crosslinking protocol was developed to obtain stable products which composition was assessed using Fourier Transform InfraRed spectroscopy and thermogravimetric analysis. Morphological investigation combined with EDX spectroscopy proved the obtained mats were highly porous and composed by thin homogenous nanofibers with a good distribution of the used nanofillers, thus representing potential products for several purposes (e.g. biomedical, pharmaceutical and environmental applications).

Alginate-Based Electrospun Membranes Containing ZnO Nanoparticles as Potential Wound Healing Patches: Biological, Mechanical, and Physicochemical Characterization.

In the present work, alginate-based mats with and without ZnO nanoparticles were prepared via an electrospinning technique and subjected to a washing-cross-linking process to obtain highly stable products characterized by thin and homogeneous nanofibers with a diameter of 100 ± 30 nm. Using a commercial collagen product as control, the biological response of the prepared mats was carefully evaluated with particular attention paid to the influence of the used cross-linking agent (Ca2+, Sr2+, or Ba2+ ions) and to the presence of nanofillers. Fibroblast and keratinocyte cultures successfully proved the safety of the prepared alginate-based mats, whereas ZnO nanoparticles were found to provide strong antibacteriostatic and antibacterial properties; above all, the strontium- and barium-cross-linked samples showed performances in terms of cell adhesion and growth very similar to those of the commercial collagen membrane despite them showing a significantly lower protein adsorption. Moreover, the mechanical and water-related properties of the strontium-cross-linked mats embedding ZnO nanoparticles were proven to be similar to those of human skin (i.e., Young modulus of 470 MPa and water vapor permeability of 3.8 × 10-12 g/m Pa s), thus proving the ability of the prepared mats to be able to endure considerable stress, maintaining at the same time the fundamental ability to remove exudates. Taking into account the obtained results, the proposed alginate-based products could lead to harmless and affordable surgical patches and wound dressing membranes with a simpler and safer production procedure than the commonly employed animal collagen-derived systems.

A micro-rheological and rheological study of biopolymers solutions: Hyaluronic acid.

Hyaluronic acid (HA) solutions represent an important class of biomedical products, mostly used as viscosupplements in orthopaedics and as fillers in the cosmetic industry. The focus of the present work is the hydrodynamic, micro-rheological and rheological characterization of HA in physiological saline. Standard viscoelastic characterization techniques were coupled with micro-rheological measurements, i.e. by measuring the passive motions of particles embedded in the samples via Dynamic Light Scattering (DLS), effectively extending the accessible frequency range typical of standard rheometers. The influence of molecular weight and polymer concentration on the storage modulus (G'), loss modulus (G") and complex viscosity (η*) of HA saline solutions was investigated. A brief comparison with theoretical models was made showing such concentrated solutions to be of a semi-flexible nature. In addition, the entanglement concentration ν, the critical molecular weight M¯c, and the mesh size ξ of the physical network created by the entangled polymer chains were calculated.

The effects of morpholine pre-treated and carboxymethylated cellulose nanofibrils on the properties of alginate-based hydrogels.

The effects of varying percentage loadings of morpholine pre-treated cellulose nanofibrils (MCNF) and carboxymethylated cellulose nanofibrils (CMCNF) on the aqueous swelling, compressive modulus and viscoelastic properties of calcium-ion-crosslinked alginate hydrogels were investigated. In addition, the pore structures of hydrogels with the highest compressive modulus were studied. The incorporation of 5 wt. % MCNF resulted in a slightly reduced aqueous swelling, a 36% increase in compressive modulus and a layered pore structure when compared with the neat alginate hydrogel. On the other hand, the addition of CMCNF at the same loading led to a slightly improved aqueous swelling, an increase in compressive modulus (17%) and high porosity. Further increases in CNF loadings did not result in significant increase in material properties. The alginate/CNF composite materials have potentials to be used in applications where good swelling and mechanical robustness are required.

Production, Characterization and Biocompatibility Evaluation of Collagen Membranes Derived from Marine Sponge Chondrosia reniformis Nardo, 1847.

Collagen is involved in the formation of complex fibrillar networks, providing the structural integrity of tissues. Its low immunogenicity and mechanical properties make this molecule a biomaterial that is extremely suitable for tissue engineering and regenerative medicine (TERM) strategies in human health issues. Here, for the first time, we performed a thorough screening of four different methods to obtain sponge collagenous fibrillar suspensions (FSs) from C. reniformis demosponge, which were then chemically, physically, and biologically characterized, in terms of protein, collagen, and glycosaminoglycans content, viscous properties, biocompatibility, and antioxidant activity. These four FSs were then tested for their capability to generate crosslinked or not thin sponge collagenous membranes (SCMs) that are suitable for TERM purposes. Two types of FSs, of the four tested, were able to generate SCMs, either from crosslinking or not, and showed good mechanical properties, enzymatic degradation resistance, water binding capacity, antioxidant activity, and biocompatibility on both fibroblast and keratinocyte cell cultures. Finally, our results demonstrate that it is possible to adapt the extraction procedure in order to alternatively improve the mechanical properties or the antioxidant performances of the derived biomaterial, depending on the application requirements, thanks to the versatility of C. reniformis extracellular matrix extracts.