Engineered Highly Porous Polyvinyl Alcohol Hydrogels with Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and Graphene Nanosheets for Musculoskeletal Tissue Engineering: Morphology, Water Sorption, Thermal, Mechanical, Electrical Properties, and Biocompatibility.

Electroactive composite materials are very promising for musculoskeletal tissue engineering because they can be applied in combination with electrostimulation. In this context, novel graphene-based poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/polyvinyl alcohol (PHBV/PVA) semi-interpenetrated networks (semi-IPN) hydrogels were engineered with low amounts of graphene (G) nanosheets dispersed within the polymer matrix to endow them with electroactive properties. The nanohybrid hydrogels, obtained by applying a hybrid solvent casting-freeze-drying method, show an interconnected porous structure and a high water-absorption capacity (swelling degree > 1200%). The thermal characterization indicates that the structure presents microphase separation, with PHBV microdomains located between the PVA network. The PHBV chains located in the microdomains are able to crystallize; even more after the addition of G nanosheets, which act as a nucleating agent. Thermogravimetric analysis indicates that the degradation profile of the semi-IPN is located between those of the neat components, with an improved thermal stability at high temperatures (>450 °C) after the addition of G nanosheets. The mechanical (complex modulus) and electrical properties (surface conductivity) significantly increase in the nanohybrid hydrogels with 0.2% of G nanosheets. Nevertheless, when the amount of G nanoparticles increases fourfold (0.8%), the mechanical properties diminish and the electrical conductivity does not increase proportionally, suggesting the presence of G aggregates. The biological assessment (C2C12 murine myoblasts) indicates a good biocompatibility and proliferative behavior. These results reveal a new conductive and biocompatible semi-IPN with remarkable values of electrical conductivity and ability to induce myoblast proliferation, indicating its great potential for musculoskeletal tissue engineering.

Pro-Myogenic Environment Promoted by the Synergistic Effect of Conductive Polymer Nanocomposites Combined with Extracellular Zinc Ions.

A new strategy based on the combination of electrically conductive polymer nanocomposites and extracellular Zn2+ ions as a myogenic factor was developed to assess its ability to synergically stimulate myogenic cell response. The conductive nanocomposite was prepared with a polymeric matrix and a small amount of graphene (G) nanosheets (0.7% wt/wt) as conductive filler to produce an electrically conductive surface. The nanocomposites' surface electrical conductivity presented values in the range of human skeletal muscle tissue. The biological evaluation of the cell environment created by the combination of the conductive surface and extracellular Zn2+ ions showed no cytotoxicity and good cell adhesion (murine C2C12 myoblasts). Amazingly, the combined strategy, cell-material interface with conductive properties and Zn bioactive ions, was found to have a pronounced synergistic effect on myoblast proliferation and the early stages of differentiation. The ratio of differentiated myoblasts cultured on the conductive nanocomposites with extracellular Zn2+ ions added in the differentiation medium (serum-deprived medium) was enhanced by more than 170% over that of non-conductive surfaces (only the polymeric matrix), and more than 120% over both conductive substrates (without extracellular Zn2+ ions) and non-conductive substrates with extracellular Zn2+. This synergistic effect was also found to increase myotube density, myotube area and diameter, and multinucleated myotube formation. MyoD-1 gene expression was also enhanced, indicating the positive effect in the early stages of myogenic differentiation. These results demonstrate the great potential of this combined strategy, which stands outs for its simplicity and robustness, for skeletal muscle tissue engineering applications.

Engineering alginate hydrogel films with poly(3-hydroxybutyrate-co-3-valerate) and graphene nanoplatelets: Enhancement of antiviral activity, cell adhesion and electroactive properties.

A new biodegradable semi-interpenetrated polymer network (semi-IPN) of two US Food and Drug Administration approved materials, poly(3-hydroxybutyrate-co-3-valerate) (PHBV) and calcium alginate (CA) was engineered to provide an alternative strategy to enhance the poor adhesion properties of CA. The synthesis procedure allows the additional incorporation of 10 % w/w of graphene nanoplatelets (GNPs), which have no cytotoxic effect on human keratinocytes. This quantity of multilayer graphene provides superior antiviral activity to the novel semi-IPN against a surrogate virus of SARS-CoV-2. Adding GNPs hardly affects the water absorption or electrical conductivity of the pure components of CA and PHBV. However, the semi-IPN's electrical conductivity increases dramatically after adding GNP due to molecular rearrangements of the intertwined polymer chains that continuously distribute the GNP nanosheets, This new hydrophilic composite biomaterial film shows great promise for skin biomedical applications, especially those that require antiviral and/or biodegradable electroconductive materials.

Graphene Oxide versus Carbon Nanofibers in Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Films: Degradation in Simulated Intestinal Environments.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a microbial biodegradable polymer with a broad range of promising industrial applications. The effect of incorporation of low amounts (1% w/w) of carbon nanomaterials (CBNs) such as 1D carbon nanofibers (CNFs) or 2D graphene oxide (GO) nanosheets into the PHBV polymer matrix affects its degradation properties, as it is reported here for the first time. The study was performed in simulated gut conditions using two different media: an acidic aqueous medium (pH 6) and Gifu anaerobic medium. The results of this study showed that the incorporation of low amounts of filamentous 1D hydrophobic CNFs significantly increased the degradability of the hydrophobic PHBV after 3 months in simulated intestinal conditions as confirmed by weight loss (~20.5% w/w in acidic medium) and electron microscopy. We can attribute these results to the fact that the long hydrophobic carbon nanochannels created in the PHBV matrix with the incorporation of the CNFs allowed the degradation medium to penetrate at ultrafast diffusion speed increasing the area exposed to degradation. However, the hydrogen bonds formed between the 2D hydrophilic GO nanosheets and the hydrophobic PHBV polymer chains produced a homogeneous composite structure that exhibits lower degradation (weight loss of ~4.5% w/w after three months in acidic aqueous medium). Moreover, the water molecules present in both degradation media can be linked to the hydroxyl (-OH) and carboxyl (-COOH) groups present on the basal planes and at the edges of the GO nanosheets, reducing their degradation potential.

Non-Woven Infection Prevention Fabrics Coated with Biobased Cranberry Extracts Inactivate Enveloped Viruses Such as SARS-CoV-2 and Multidrug-Resistant Bacteria.

The Coronavirus Disease (COVID-19) pandemic is demanding the rapid action of the authorities and scientific community in order to find new antimicrobial solutions that could inactivate the pathogen SARS-CoV-2 that causes this disease. Gram-positive bacteria contribute to severe pneumonia associated with COVID-19, and their resistance to antibiotics is exponentially increasing. In this regard, non-woven fabrics are currently used for the fabrication of infection prevention clothing such as face masks, caps, scrubs, shirts, trousers, disposable gowns, overalls, hoods, aprons and shoe covers as protective tools against viral and bacterial infections. However, these non-woven fabrics are made of materials that do not exhibit intrinsic antimicrobial activity. Thus, we have here developed non-woven fabrics with antimicrobial coatings of cranberry extracts capable of inactivating enveloped viruses such as SARS-CoV-2 and the bacteriophage phi 6 (about 99% of viral inactivation in 1 min of viral contact), and two multidrug-resistant bacteria: the methicillin-resistant Staphylococcus aureus and the methicillin-resistant Staphylococcus epidermidis. The morphology, thermal and mechanical properties of the produced filters were characterized by optical and electron microscopy, differential scanning calorimetry, thermogravimetry and dynamic mechanical thermal analysis. The non-toxicity of these advanced technologies was ensured using a Caenorhabditis elegans in vivo model. These results open up a new prevention path using natural and biodegradable compounds for the fabrication of infection prevention clothing in the current COVID-19 pandemic and microbial resistant era.

Novel Semi-Interpenetrated Polymer Networks of Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate)/Poly (Vinyl Alcohol) with Incorporated Conductive Polypyrrole Nanoparticles.

This paper reports the preparation and characterization of semi-interpenetrating polymer networks (semi-IPN) of poly(3-hydroxybutirate-co-3-hydroxyvalerate), PHBV, and poly (vinyl alcohol), PVA, with conductive polypirrole (PPy) nanoparticles. Stable hybrid semi-IPN (PHBV/PVA 30/70 ratio) hydrogels were produced by solvent casting, dissolving each polymer in chloroform and 1-methyl-2-pyrrolidone respectively, and subsequent glutaraldehyde crosslinking of the PVA chains. The microstructure and physical properties of this novel polymeric system were analysed, including thermal behaviour and degradation, water sorption, wettability and electrical conductivity. The conductivity of these advanced networks rose significantly at higher PPy nanoparticles content. Fourier transform infrared spectroscopy (FTIR) and calorimetry characterization indicated good miscibility and compatibility between all the constituents, with no phase separation and strong interactions between phases. A single glass transition was observed between those of pure PHBV and PVA, although PVA was dominant in its contribution to the glass transition process. Incorporating PPy nanoparticles significantly reduced the hydrogel swelling, even at low concentrations, indicating molecular interactions between the PPy nanoparticles and the hydrogel matrix. The PHBV/PVA semi-IPN showed higher thermal stability than the neat polymers and PHBV/PVA blend, which also remained in the tertiary systems.