Further Step in the Transition from Conventional Plasticizers to Versatile Bioplasticizers Obtained by the Valorization of Levulinic Acid and Glycerol.

In the last two decades, the use of phthalates has been restricted worldwide due to their well-known toxicity. Nonetheless, phthalates are still widely used for their versatility, high plasticization effect, low cost, and lack of valuable alternatives. This study presents the fully bio-based and versatile glycerol trilevulinate plasticizer (GT) that was obtained by the valorization of glycerol and levulinic acid. The mild-conditions and solvent-free esterification used to synthesize GT was optimized by investigating the product by Fourier transform infrared and NMR spectroscopy. An increasing content of GT, from 10 to 40 parts by weight per hundred parts of resin (phr), was tested with poly(vinyl chloride), poly(3-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(lactic acid), and poly(caprolactone), which typically present complicated processability and/or mechanical properties. GT produced a significant plasticization effect on both amorphous and semicrystalline polymers, reducing their glass-transition temperature and stiffness, as observed by differential scanning calorimetry measurements and tensile tests. Remarkably, GT also decreased both the melting temperature and crystallinity degree of semicrystalline polymers. Furthermore, GT underwent enzyme-mediated hydrolysis to its initial constituents, envisioning a promising prospective for environmental safety and upcycling. Furthermore, 50% inhibitory concentration (IC50) tests, using mouse embryo fibroblasts, proved that GT is an unharmful alternative plasticizer, which makes it potentially applicable in the biomedical field.

Fully Biobased Polyhydroxyalkanoate/Tannin Films as Multifunctional Materials for Smart Food Packaging Applications.

Fully biobased and biodegradable materials have attracted a growing interest in the food packaging sector as they can help to reduce the negative impact of fossil-based plastics on the environment. Moreover, the addition of functionalities to these materials by introducing active molecules has become an essential requirement to create modern packaging able to extend food's shelf-life while informing the consumer about food quality and freshness. In this study, we present an innovative bioplastic formulation for food packaging based on poly(hydroxybutyrate-co-valerate) (PHBV) and tannins as multifunctional additives. As a proof of concept, PHBV/tannin films were prepared by solvent casting, increasing the tannin content from 1 to 10 per hundred of resin (phr). Formic acid was used to reach a homogeneous distribution of the hydrophilic tannins into hydrophobic PHBV, which is remarkably challenging by using other solvents. Thanks to their well-known properties, the effect of tannins on the antioxidant, UV protection, and gas barrier properties of PHBV was evaluated. Samples containing 5 phr bioadditive revealed the best combination of these properties, also maintaining good transparency. Differential scanning calorimetry (DSC) investigations revealed that films are suitable for application from the fridge to potentially high temperatures for food heating (up to 200 °C). Tensile tests have also shown that Young's modulus (900-1030 MPa) and tensile strength (20 MPa) are comparable with those of the common polymers and biopolymers for packaging. Besides the improvement of the PHBV properties for extending food's shelf-life, it was also observed that PHBV/tannin could colorimetrically detect ammonia vapors, thus making this material potentially applicable as a smart indicator for food spoilage (e.g., detection of fish degradation). The presented outcomes suggest that tannins can add multifunctional properties to a polymeric material, opening up a new strategy to obtain an attractive alternative to petroleum-based plastics for smart food packaging applications.

Fine Tuning of the Mechanical Properties of Bio-Based PHB/Nanofibrillated Cellulose Biocomposites to Prevent Implant Failure Due to the Bone/Implant Stress Shielding Effect.

A significant mechanical properties mismatch between natural bone and the material forming the orthopedic implant device can lead to its failure due to the inhomogeneous loads distribution, resulting in less dense and more fragile bone tissue (known as the stress shielding effect). The addition of nanofibrillated cellulose (NFC) to biocompatible and bioresorbable poly(3-hydroxybutyrate) (PHB) is proposed in order to tailor the PHB mechanical properties to different bone types. Specifically, the proposed approach offers an effective strategy to develop a supporting material, suitable for bone tissue regeneration, where stiffness, mechanical strength, hardness, and impact resistance can be tuned. The desired homogeneous blend formation and fine-tuning of PHB mechanical properties have been achieved thanks to the specific design and synthesis of a PHB/PEG diblock copolymer that is able to compatibilize the two compounds. Moreover, the typical high hydrophobicity of PHB is significantly reduced when NFC is added in presence of the developed diblock copolymer, thus creating a potential cue for supporting bone tissue growth. Hence, the presented outcomes contribute to the medical community development by translating the research results into clinical practice for designing bio-based materials for prosthetic devices.

Effect of polyhydroxyalkanoates on the microbial reductive dechlorination of polychlorinated biphenyls and competing anaerobic respirations in a marine microbial culture.

The effect of polyhydroxyalkanoates (PHAs) with different composition on the reductive dechlorination activity of a polychlorinated biphenyls (PCBs) dechlorinating marine microbial community and on the activity of sulfate-reducing (SRB) and methanogenic bacteria (MB), were investigated in marine sediment microcosms and compared with the main monomer, 3-hydroxybutyric acid (3HB). Despite PHAs were fermented more slowly than 3HB, all electron donors stimulated constantly sulfate-reduction, methanogenesis and, only transiently, PCB reductive dechlorination. No relevant differences were observed with different compositions of PHAs. According to electron balances, the majority of the supplied electrons (50 %) were consumed by SRB and to less extent by MB (9-31 %), while a small percentage (0.01 %) was delivered to OHRB. In the studied conditions PHAs were confirmed as potential slow­hydrogen releasing compounds in marine environment but their fermentation rate was sufficiently high to mainly stimulate the competitors of organohalide respring bacteria for electron donors.

Mixed Matrix Membranes Based on Torlon® and ZIF-8 for High-Temperature, Size-Selective Gas Separations.

Torlon® is a thermally and plasticization-resistant polyamide imide characterized by low gas permeability at room temperature. In this work, we aimed at improving the polymer performance in the thermally-enhanced He/CO2 and H2/CO2 separations, by compounding Torlon® with a highly permeable filler, ZIF-8, to fabricate Mixed Matrix Membranes (MMMs). The effect of filler loading, gas size, and temperature on the MMMs permeability, diffusivity, and selectivity was investigated. The He permeability increased by a factor of 3, while the He/CO2 selectivity decreased by a factor of 2, when adding 25 wt % of ZIF-8 at 65 °C to Torlon®; similar trends were observed for the case of H2. The MMMs permeability and size-selectivity were both enhanced by temperature. The behavior of MMMs is intermediate between the pure polymer and pure filler ones, and can be described with models for composites, indicating that such materials have a good polymer/filler adhesion and their performance could be tailored by acting on the formulation. The behavior observed is in line with previous investigations on MMMs based on glassy polymers and ZIF-8, in similar conditions, and indicates that ZIF-8 can be used as a polymer additive when the permeability is a controlling aspect, with a proper choice of loading and operative temperature.

Carbon Nanotubes/Regenerated Silk Composite as a Three-Dimensional Printable Bio-Adhesive Ink with Self-Powering Properties.

In this study, regenerated silk (RS) obtained from Bombyx Mori cocoons is compounded with carboxyl-functionalized carbon nanotubes (f-CNTs) in an aqueous environment for the fabrication of functional bio-adhesives. Molecular interactions between RS and carboxyl groups of CNTs result in structural increase of the ß-sheet formation, obtaining a resistant adhesive suitable for a wet biological substrate. Moreover, the functionalization of CNTs promotes their dispersion in RS, thus enabling the production of films with controlled electrical conductivity. The practical utility of such a property is demonstrated through the fabrication of a piezoelectric device implanted in a rat to monitor the breathing in vivo and to be used as a self-powered system. Finally, RS/f-CNTs were used as a printable biomaterial ink to three dimensionally print bilayer hollow tubular structures composed of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and RS. Initial tests carried out by seeding and growing human skin fibroblasts demonstrated that the 3D printed bilayer hollow cylindrical structures offer a suitable surface for the seeded cells to attach and proliferate. In general, the herein proposed RS/f-CNT composite serves as a versatile material for solvent-free dispersion processing and 3D printing, thus paving a new approach to prepare multifunctional materials with potential applications of great interest in sealing biological substrates and implantable devices for regenerative medicine.

The role of biotechnology in the transition from plastics to bioplastics: an opportunity to reconnect global growth with sustainability.

Building new value chains, through the valorization of biomass components for the development of innovative bio-based products (BBPs) aimed at specific market sectors, will accelerate the transition from traditional production technologies to the concept of biorefineries. Recent studies aimed at mapping the most relevant innovations undergoing in the field of BBPs (Fabbri et al. 2019, Final Report of the Task 3 BIOSPRI Tender Study on Support to R&I Policy in the Area of Bio-based Products and Services, delivered to the European Commission (DG RTD)), clearly showed the dominant position played by the plastics sector, in which new materials and innovative technical solutions based on renewable resources, concretely contribute to the achievement of relevant global sustainability goals. New sustainable solutions for the plastic sector, either bio-based or bio-based and biodegradable, have been intensely investigated in recent years. The global bioplastics and biopolymers market size is expected to grow from USD 10.5 billion in 2020 to USD 27.9 billion by 2025 (Markets and Markets, 2020, Bioplastics & Biopolymers Market by Type (Non-Biodegradable/Bio-Based, Biodegradable), End-Use Industry (Packaging, Consumer Goods, Automotive & Transportation, Textiles, Agriculture & Horticulture), Region - Global Forecast to 2025), and this high growth is driven primarily by the growth of the global packaging end-use industry. Such relevant opportunities are the outcomes of intensive scientific and technological research devoted to the development of new materials with selected technical features, which can represent feasible substitutes for the fossil-based plastic materials currently used in the packaging sectors and other main fields. This article offers a map of the latest developments connected to the plastic sector, achieved through the application of biotechnological routes for the preparation of completely new polymeric structures, or drop-in substitutes derived from renewable resources, and it describes the specific role played by biotechnology in promoting and making this transition faster.

Two-Step Solvent-Free Synthesis of Poly(hydroxybutyrate)-Based Photocurable Resin with Potential Application in Stereolithography.

A bio-based polymeric ink for stereolithography developed through a two-step solvent-free process is herein proposed. Specifically, low-molecular-weight poly(hydroxybutyrate) (PHB)-diol oligomers are prepared via molten transesterification of bacterial PHB with 1,4-butanediol. Transesterification conditions such as diol concentration, catalyst amount, and reaction time are studied for optimizing the final oligomers' molecular weight and structural features. In the second step, the oligomeric hydroxyl terminals are converted into methacrylate moieties through a solvent-free end-capping reaction and diluted in propylene carbonate in order to obtain a photo-polymerizable ink with suitable viscosity. The ink is UV-cured, and the obtained material properties are investigated by FT-IR and differential scanning calorimetry measurements. The proposed method provides a valuable and environmentally friendly alternative to currently available synthetic routes, overcoming their typical disadvantages related to the used solvents and harsh conditions. Moreover, it opens up a sustainable route for converting polyesters into functionalized oligomeric derivatives, which can potentially find application in 3D printing of customized biomedical devices.

Vascular Wall-Mesenchymal Stem Cells Differentiation on 3D Biodegradable Highly Porous CaSi-DCPD Doped Poly (α-hydroxy) Acids Scaffolds for Bone Regeneration.

Vascularization is a crucial factor when approaching any engineered tissue. Vascular wall-mesenchymal stem cells are an excellent in vitro model to study vascular remodeling due to their strong angiogenic attitude. This study aimed to demonstrate the angiogenic potential of experimental highly porous scaffolds based on polylactic acid (PLA) or poly-e-caprolactone (PCL) doped with calcium silicates (CaSi) and dicalcium phosphate dihydrate (DCPD), namely PLA-10CaSi-10DCPD and PCL-10CaSi-10DCPD, designed for the regeneration of bone defects. Vascular wall-mesenchymal stem cells (VW-MSCs) derived from pig thoracic aorta were seeded on the scaffolds and the expression of angiogenic markers, i.e. CD90 (mesenchymal stem/stromal cell surface marker), pericyte genes α-SMA (alpha smooth muscle actin), PDGFR-ß (platelet-derived growth factor receptor-ß), and NG2 (neuron-glial antigen 2) was evaluated. Pure PLA and pure PCL scaffolds and cell culture plastic were used as controls (3D in vitro model vs. 2D in vitro model). The results clearly demonstrated that the vascular wall mesenchymal cells colonized the scaffolds and were metabolically active. Cells, grown in these 3D systems, showed the typical gene expression profile they have in control 2D culture, although with some main quantitative differences. DNA staining and immunofluorescence assay for alpha-tubulin confirmed a cellular presence on both scaffolds. However, VW-MSCs cultured on PLA-10CaSi-10DCPD showed an individual cells growth, whilst on PCL-10CaSi-10DCPD scaffolds VW-MSCs grew in spherical clusters. In conclusion, vascular wall mesenchymal stem cells demonstrated the ability to colonize PLA and PCL scaffolds doped with CaSi-DCPD for new vessels formation and a potential for tissue regeneration.

Highly porous polycaprolactone scaffolds doped with calcium silicate and dicalcium phosphate dihydrate designed for bone regeneration.

Polycaprolactone (PCL), dicalcium phosphate dihydrate (DCPD) and/or calcium silicates (CaSi) have been used to prepare highly porous scaffolds by thermally induced phase separation technique (TIPS). Three experimental mineral-doped formulations were prepared (PCL-10CaSi, PCL-5CaSi-5DCPD, PCL-10CaSi-10DCPD); pure PCL scaffolds constituted the control group. Scaffolds were tested for their chemical-physical and biological properties, namely thermal properties by differential scanning calorimetry (DSC), mechanical properties by quasi-static parallel-plates compression testing, porosity by a standard water-absorption method calcium release, alkalinizing activity, surface microchemistry and micromorphology by Environmental Scanning electronic Microscopy (ESEM), apatite-forming ability in Hank Balanced Saline Solution (HBSS) by Energy Dispersive X-ray Spectroscopy (EDX) and micro-Raman, and direct contact cytotoxicity. All mineral-doped scaffolds released calcium and alkalinized the soaking medium, which may favor a good biological (osteogenic) response. ESEM surface micromorphology analyses after soaking in HBSS revealed: pure PCL, PCL-10CaSi and PCL-10CaSi-10DCPD kept similar surface porosity percentages but different pore shape modifications. PCL-5CaSi-5DCPD revealed a significant surface porosity increase despite calcium phosphates nucleation (p < 0.05). Micro-Raman spectroscopy detected the formation of a B-type carbonated apatite (Ap) layer on the surface of PCL-10CaSi-10DCPD aged for 28 days in HBSS; a similar phase (but of lower thickness) formed also on PCL-5CaSi-5DCPD and PCL; the deposit formed on PCL-10CaSi was mainly composed of calcite. All PCL showed bulk open porosity higher than 94%; however, no relevant brittleness was observed in the materials, which retained the possibility to be handled without collapsing. The thermo-mechanical properties showed that the reinforcing and nucleating action of the inorganic fillers CaSi and DCPD improved viscoelastic properties of the scaffolds, as confirmed by the increased value of storage modulus and the slight increase in the crystallization temperature for all the biomaterials. A detrimental effect on the mechanical properties was observed in samples with the highest amount of inorganic particles (PCL-10CaSi-10DCPD). All the scaffolds showed absence of toxicity, in particular PCL-10CaSi-10DCPD. The designed scaffolds are biointeractive (release biologically relevant ions), nucleate apatite, possess high surface and internal open porosity and can be colonized by cells, creating a bone forming osteoblastic microenvironment and appearing interesting materials for bone regeneration purposes.

Highly porous PHB-based bioactive scaffolds for bone tissue engineering by in situ synthesis of hydroxyapatite.

In this study bioactive and bioresorbable porous scaffolds for bone tissue regeneration, based on poly(3-hydroxybutyrate) (PHB), are presented. The porous structure is obtained by thermally induced phase separation (TIPS) technique, whereas the osteoinductivity and osteoconductivity are enhanced through the incorporation of hydroxyapatite (HA). The HA particles are generated in PHB using an innovative filler in situ synthesis, and the properties of the composite scaffolds are then compared to scaffolds obtained by conventional mechanical dispersion of ex situ synthesized HA particles. The in situ synthesis leads to composite materials with improved porosity, even at high filler content, without any degradation of the polymeric matrix as confirmed by GPC and DSC measurements. On the contrary, the samples prepared by ex situ method show a suppressed porosity by increasing the inorganic filler content, therefore limiting the amount of HA that can be loaded in PHB and the resulting bioactivity. The possibility to use PHB/HA porous composites as scaffolds for bone tissue regeneration, is assessed by preliminary cell viability in vitro studies. In particular, it is observed that the composites are fully cytocompatible and able to sustain MC3T3-E1 mouse pre-osteoblast cells adhesion and proliferation. Investigations on cell morphology reveal, for all PHB/HA scaffolds, the presence of differentiated cells with a predominance of osteocyte-like morphology, which are not observed for neat PHB scaffolds. Moreover, the MC3T3-E1 cells differentiation towards osteoblastic phenotype is further supported by the evaluation of the early osteogenic markers. In particular, samples loaded with HA in situ synthesized showed the highest ALP production and typical morphology of the terminal differentiation stages of osteoblasts.

Facile method based on 19F-NMR for the determination of hydroxyl value and molecular weight of hydroxyl terminated polymers.

A new, facile and low-sample consuming technique for the determination of hydroxyl values (OHVs) in poly(hydroxyalkanoate)s (PHAs) is herein presented. After a fast, uncatalyzed and non-destructive trifluoroacetylation of all PHA hydroxyl groups, OHVs are calculated through 19F-NMR measurements, comparing integral values of the only two signals in the spectra: the fluorinated hydroxyl groups and the internal standard (trifluorotoluene) signals. Furthermore, the combined ex situ functionalization with a simple workup of the fluorinated polymer allowed to obtain spectra with an improved signal/noise ratio and short acquisition time if compared to 1H-NMR-based methods. The results obtained by 19F-NMR measurements are then validated by the established standard method based on acid/base titration and the 1H-NMR-based method. The proposed approach shows to be valuable not only for low molecular weight PHAs, but also for high molecular weight samples, which typically cannot be analysed by conventional 1H-NMR-based methods due to the very low concentration of hydroxyl terminal groups. Thanks to the observed high reliability, the method was also successfully applied to evaluate the OHVs of various commercial hydroxyl terminated polymers with different molecular weights in order to further show its wide applicability.

Effectiveness of polymeric coated films containing bacteriocin-producer living bacteria for Listeria monocytogenes control under simulated cold chain break.

Nisin, enterocin 416K1 and living bacteriocin-producer Enterococcus casseliflavus IM 416K1 have been entrapped in polyvinyl alcohol (PVOH) based coatings applied to poly (ethylene terephthalate) (PET) films, and their effectiveness in the control of the growth of Listeria monocytogenes ATCC 19117 has been tested. The anti-listerial activity of the doped coated films was evaluated by both a modified agar diffusion assay and a direct contact with artificially contaminated precooked chicken fillets stored at 4 °C, 22 °C and under simulated cold chain break conditions (1 day at 30 °C). The live-Enterococcus-doped film showed a more remarkable activity than nisin- and enterocin-doped films over long times both at 4 °C and 22 °C. The use of this film at 22 °C resulted in full inactivation of L. monocytogenes from the seventh day of the test. Live-Enterococcus-doped film displayed a much better antilisterial activity in comparison to nisin- and enterocin-doped films also in samples incubated at 4 °C, and submitted at one day (3rd or 7th day) of storage at 30 °C, to simulate cold chain break conditions. All results suggest that the live-Enterococcus-doped film can behave as a smart active food packaging, very effective in cold chain break conditions when the Listeria growth is fast.

Polylactic acid-based porous scaffolds doped with calcium silicate and dicalcium phosphate dihydrate designed for biomedical application.

Polylactic acid (PLA), dicalcium phosphate dihydrate (DCPD) and/or hydraulic calcium silicate (CaSi) have been used to prepare highly-porous scaffolds by thermally induced phase separation technique (TIPS). Three experimental mineral-doped formulations were prepared (PLA-10CaSi, PLA-5CaSi-5DCPD, PLA-10CaSi-10DCPD). Pure PLA scaffolds constituted the control group. Scaffolds were tested for their chemical-physical and biological properties, namely calcium release, alkalinizing activity, surface microchemistry and micromorphology by ESEM, apatite-forming ability by EDX, micro-Raman and IR spectroscopy, thermal properties by differential scanning calorimetry, mechanical properties by quasi-static parallel-plates compression testing, porosity by a standard water-absorption method and direct-contact cytotoxicity. All mineral-doped scaffolds released biologically relevant ions (biointeractive). A B-type carbonated apatite layer (thickness decreasing along the series PLA-10CaSi-10DCPD>PLA-10CaSi>PLA-5CaSi-5DCPD>PLA) was detected on the surface of all the 28d-aged scaffolds. Surface pores of fresh scaffolds ranged from 10 to 20µm in pure PLA to 10-100µm in PLA-10CaSi. An increase in porosity was detected in 28d-aged pure PLA scaffolds (approx. 30% of material loss with decrease of the PLA chain length); differently, in mineral-doped scaffolds, the PLA degradation was balanced by deposition/nucleation of apatite. All scaffolds showed absence of toxicity, in particular PLA-10CaSi-10DCPD. The designed scaffolds are biointeractive (release biologically relevant ions), nucleate apatite, possess high surface and internal open porosity and can be colonized by cells, appearing interesting materials for bone regeneration.