Development and Characterization of Electrospun Biopapers of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Derived from Cheese Whey with Varying 3-Hydroxyvalerate Contents

[EN] In the present study, three different newly developed copolymers of poly(3-hydroxybutyrate-co-3-hydroxyval-erate) (PHBV) with 20, 40, and 60 mol % contents in 3-hydroxyvalerate (3HV) were produced by the biotechnological process of mixed microbial cultures (MMCs) using cheese whey (CW), a by-product from the dairy industry, as feedstock. The CW-derived PHBV copolyesters were first purified and then processed by solution electrospinning, yielding fibers of approximately 2 mu m in cross-section in all cases. The resultant electrospun PHBV mats were, thereafter, post-processed by annealing at different temperatures, below their maximum of melting, selected according to their 3HV content in order to obtain continuous films based on coalesced fibers, so-called biopapers. The resultant PHBV films were characterized in terms of their morphology, crystallinity, and mechanical and barrier properties to assess their potential application in food packaging. The CW-derived PHBV biopapers showed high contact transparency but a slightly yellow color. The fibers of the 20 mol % 3HV copolymer were seen to contain mostly poly(3-hydroxybutyrate) (PHB) crystals, the fibers of the 40 mol % 3HV copolymer a mixture of PHB and poly(3-hydroxyvalerate) (PHV) crystals and lowest crystallinity, and the fibers of the 60 mol % 3HV sample were mostly made of PHV crystals. To understand the interfiber coalesce process undergone by the materials during annealing, the crystalline morphology was also assessed by variable-temperature both combined small-angle and wide-angle X-ray scattering synchrotron and Fourier transform infrared experiments. From these experiments and, different from previously reported biopapers with lower 3HV contents, all samples were inferred to have a surface energy reduction mechanism for interfiber coalescence during annealing, which is thought to be activated by a temperature-induced decrease in molecular order. Due to their reduced crystallinity and molecular order, the CW-derived PHBV biopapers, especially the 40 mol % 3HV sample, were found to be more ductile and tougher. In terms of barrier properties, the three copolymers performed similarly to water and limonene, but to oxygen, the 40 mol % sample showed the highest relative permeability. Overall, the materials developed, which are compatible with the Circular Bioeconomy organic recycling strategy, can have an excellent potential as barrier interlayers or coatings of application interest in food packaging.This research work was funded by the H2020 EU project YPACK (reference number 773872) and by the Spanish Ministry of Science and Innovation (MICI) project RTI2018-097249-B-C21. B.M.-R. would like to acknowledge the MICI for her FPI fellowship (BES-2016-077972) and S.T.-G. for his MICI Juan de la Cierva-Incorporacion contract (IJCI-2016-29675). The ALBA Synchrotron is also acknowledged for the funding received through the project "Time-resolved Combined Wide-and Small-angle X-ray Scattering Characterization as a Function of Temperature of Electrospun Polyhydroxyalkanoates Derived from Biowaste" (2018022619). The authors would also like to thank the Unidad Asociada IATA(CSIC)-UJI in "Plastics Technology".Meléndez-Rodríguez, B.; Reis, MAM.; Carvalheira, M.; Sammon, C.; Cabedo, L.; Torres-Giner, S.; Lagaron, JM. (2021). Development and Characterization of Electrospun Biopapers of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Derived from Cheese Whey with Varying 3-Hydroxyvalerate Contents. Biomacromolecules. 22(7):2935-2953. https://doi.org/10.1021/acs.biomac.1c00353S2935295322

Barrier biopaper multilayers obtained by impregnation of electrospun poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with protein and polysaccharide hydrocolloids

[EN] Multilayer biopapers composed of two electrospun layers of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) were impregnated, at the inner side of one of the layers, with cellulose nanocrystals (CNCs) and their composites with hydrocolloids, to develop high-barrier fully biobased structures. The study aimed for the first time at comparing the impregnation of electrospun fibers with several biopolymer solutions. Thus, neat CNCs, and CNCs mixed as a minor fraction, that is, 2 wt%, with gelatin (GE), agar (AG), xanthan gum (XG), and gum arabic (GA) were assessed in their potential to improve the barrier properties of PHBV. Glycerol plasticizer was added to the composite formulations. The impregnated electrospun multilayer mats were subsequently annealed, below the PHBV melting point, to yield continuous films by an interfiber coalescence process, so-called biopapers, and thereafter characterized to evaluate their potential for high barrier food packaging applications. The morphological characterization revealed good interlayer adhesion, more noticeably for those containing CNCs and their nanocomposites with AG and XG. From their mechanical response, it was inferred that the material behavior was governed mainly by the rigidity of the PHBV substrates, and this could not be significantly improved by impregnation with any of the various hydrocolloids. Whereas the water vapor barrier was not seen to improve in any of the samples, the barrier to the organic vapor limonene, used as a standard for aroma barrier, was however improved in the samples impregnated with AG and XG composites. Interestingly, the oxygen barrier properties were significantly improved but only by impregnation with pure CNCs. This study reports for the first time a scalable impregnation technology approach to produce fully biobased barrier multilayers.This research work was funded by the H2020 EU project USABLE PAKAGING (reference number 836884) and by the Spanish Ministry of Science and Innovation (MICI) project RTI2018-097249-B-C21. Ms. Beatriz Melendez-Rodriguez would like to acknowledge the MICI for her FPI fellowship (BES-2016-077972) and Dr. Torres-Giner for his MICI Juan de la Cierva¿Incorporacion contract (IJCI-2016-29675). The authors would also like to acknowledge the Unidad Asociada IATA (CSIC)-UJI in Polymer Technology.Melendez-Rodriguez, B.; M'bengue, M.; Torres-Giner, S.; Cabedo, L.; Prieto, C.; Lagaron, JM. (2021). Barrier biopaper multilayers obtained by impregnation of electrospun poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with protein and polysaccharide hydrocolloids. Carbohydrate Polymer Technologies and Applications. 2:1-9. https://doi.org/10.1016/j.carpta.2021.100150S19

High-oxygen-barrier multilayer films based on polyhydroxyalkanoates and cellulose nanocrystals

This study reports on the development and characterization of organic recyclable high-oxygen-barrier multilayer films based on different commercial polyhydroxyalkanoate (PHA) materials, including a blend with commercial poly(butylene adipate-co-terephthalate) (PBAT), which contained an inner layer of cellulose nanocrystals (CNCs) and an electrospun hot-tack adhesive layer of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) derived from cheese whey (CW). As a result, the full multilayer structures were made from bio-based and/or compostable materials. A characterization of the produced films was carried out in terms of morphological, optical, mechanical, and barrier properties with respect to water vapor, limonene, and oxygen. Results indicate that the multilayer films exhibited a good interlayer adhesion and contact transparency. The stiffness of the multilayers was generally improved upon incorporation of the CNC interlayer, whereas the enhanced elasticity of the blend was reduced to some extent in the multilayer with CNCs, but this was still much higher than for the neat PHAs. In terms of barrier properties, it was found that 1 µm of the CNC interlayer was able to reduce the oxygen permeance between 71% and 86%, while retaining the moisture and aroma barrier of the control materials.This research work was funded by the H2020 EU project YPACK (reference number 773872) and by the Spanish Ministry of Science and Innovation (MICI) project RTI2018-097249-B-C21.Beatriz Melendez-Rodriguez would like to acknowledge the MICI for her FPI fellowship (BES-2016-077972) and Sergio Torres-Giner for his MICI Juan de la Cierva–Incorporación contract (IJCI-2016-29675). The authors would also like to acknowledge the Unidad Asociada in Polymer Technology, joint unit IATA(CSIC)-UJI

Valorization of Municipal Biowaste into Electrospun Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopapers for Food Packaging Applications

American Chemical Society[EN] The present study reports on the production and characterization of a new biopackaging material made of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) derived from municipal biowaste (MBW) and produced by the mixed bacterial culture technology. After purification and extraction, the MBW-derived PHBV was processed by electrospinning to yield defect-free ultrathin fibers, which were thermally post-treated. Annealing at 130 degrees C, well below the biopolymer's melting temperature (T-m), successfully yielded a continuous film resulting from coalescence of the electrospun fibrillar morphology, the so-called biopaper, exhibiting enhanced optical and color properties compared to traditional melt compounding routes. The crystallinity and crystalline morphology were comprehensively studied as a function of temperature by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy and combined time-resolved synchrotron small- and wide-angle X-ray scattering (SAXS and WAXS) experiments, which clearly indicated that the molecular order within the copolyester was improved up to a maximum at 130 degrees C, and then it decreased at the biopolymer's T-m. It was hypothesized that by annealing at the temperature at which the thermally induced molecular order is maximized, the fibers generated sufficient mobility to align alongside, hence reducing surface energy and porosity. The data suggest that this material shows a good balance between enhanced mechanical and improved barrier properties to vapors and gases in comparison to traditional paper and other currently used petroleum-derived polymers, thus presenting significant potential to be part of innovative food biopackaging designs for the protection and preservation of foods in a circular bioeconomy scenario.The Spanish Ministry of Science and Innovation (MICI) project RTI2018-097249-B-C21 and EU projects H2020 YPACK (reference number 773872) and H2020 USABLE (reference number 836884) are acknowledged for funding support. B.M.-R. and S.T.-G. would also like to thank MICI for the FPI fellowship (BES-2016-077972) and the Juan de la Cierva IncorporaciOn contract (IJCI-2016-29675), respectively. The ALBA Synchrotron, Spain, is also acknowledged for the funding received through the project proposal 2018022619. 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Development of Active Barrier Multilayer Films Based on Electrospun Antimicrobial Hot-Tack Food Waste Derived Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and Cellulose Nanocrystal Interlayers

[EN] Active multilayer films based on polyhydroxyalkanoates (PHAs) with and without high barrier coatings of cellulose nanocrystals (CNCs) were herein successfully developed. To this end, an electrospun antimicrobial hot-tack layer made of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) derived from cheese whey, a by-product from the dairy industry, was deposited on a previously manufactured blown film of commercial food contact PHA-based resin. A hybrid combination of oregano essential oil (OEO) and zinc oxide nanoparticles (ZnONPs) were incorporated during the electrospinning process into the PHBV nanofibers at 2.5 and 2.25 wt%, respectively, in order to provide antimicrobial properties. A barrier CNC coating was also applied by casting from an aqueous solution of nanocellulose at 2 wt% using a rod at 1m/min. The whole multilayer structure was thereafter assembled in a pilot roll-to-roll laminating system, where the blown PHA-based film was located as the outer layers while the electrospun antimicrobial hot-tack PHBV layer and the barrier CNC coating were placed as interlayers. The resultant multilayer films, having a final thickness in the 130-150 mu m range, were characterized to ascertain their potential in biodegradable food packaging. The multilayers showed contact transparency, interlayer adhesion, improved barrier to water and limonene vapors, and intermediate mechanical performance. Moreover, the films presented high antimicrobial and antioxidant activities in both open and closed systems for up to 15 days. Finally, the food safety of the multilayers was assessed by migration and cytotoxicity tests, demonstrating that the films are safe to use in both alcoholic and acid food simulants and they are also not cytotoxic for Caco-2 cells.The Spanish Ministry of Science and Innovation (MICI) through the RTI2018-097249-B-C21 program number and the EU H2020 YPACK (reference number 773872) projects funded this research.Figueroa-Lopez, KJ.; Torres-Giner, S.; Angulo, I.; Pardo-Figuerez, M.; Escuin, JM.; Bourbon, AI.; Cabedo, L.... (2020). Development of Active Barrier Multilayer Films Based on Electrospun Antimicrobial Hot-Tack Food Waste Derived Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and Cellulose Nanocrystal Interlayers. Nanomaterials. 10(12):1-24. https://doi.org/10.3390/nano10122356S124101

Development of active bio-based multilayer systems: encapsulation of cinnamaldehyde and their physicochemical characterization

[Excerpt] In this work, different multilayer structures, using a polyhydroxybutyrate-co-valerate film with a valerate content of 8% (PHBV8) as support, were developed aiming the development of active bio-based multilayer systems. An interlayer based on zein nanofibers with and without cinnamaldehyde were electrospun in the PHBV8 film and three multilayer systems were developed: 1) without an outer layer; 2) using a PHBV8 film as outer layer; and 3) using an alginate-based film as outer layer. Their physico-chemical properties were evaluated through: water vapour and oxygen permeabilities and colour measurements, Fourier Transform Infrared Spectroscopy (FTIR) and thermal analyses. Results showed that the presence of different outer layers affected the water vapour permeability and transparency of the multilayer films. (...

Layer-by-layer technique to developing functional nanolaminate films with antifungal activity

The layer-by-layer (LbL) deposition method was used to build up alternating layers (five) of different polyelectrolyte solutions (alginate, zein-carvacrol nanocapsules, chitosan and chitosan-carvacrol emulsions) on an aminolysed/charged polyethylene terephthalate (A/C PET) film. These nanolaminated films were characterised by contact angle measurements and through the determination of water vapour (WVTR) and oxygen (O2TR) transmission rates. The effect of active nanolaminated films against the Alternaria sp. and Rhizopus stolonifer was also evaluated. This procedure allowed developing optically transparent nanolaminated films with tuneable water vapour and gas properties and antifungal activity. The water and oxygen transmission rate values for the multilayer films were lower than those previously reported for the neat alginate or chitosan films. The presence of carvacrol and zein nanocapsules significantly decreased the water transmission rate (up to 40 %) of the nanolaminated films. However, the O2TR behaved differently and was only improved (up to 45 %) when carvacrol was encapsulated, i.e. nanolaminated films prepared by alternating alginate with nanocapsules of zein-carvacrol layers showed better oxygen barrier properties than those prepared as an emulsion of chitosan and carvacrol. These films containing zein-carvacrol nanocapsules also showed the highest antifungal activity (30 %), which did not significantly differ from those obtained with the highest amount of carvacrol, probably due to the controlled release of the active agent (carvacrol) from the zein-carvacrol nanocapsules. Thus, this work shows that nanolaminated films prepared with alternating layers of alginate and zein-carvacrol nanocapsules can be considered to improve the shelf-life of foodstuffs.The authors acknowledge financial support from FP7 IP project BECOBIOCAP^. M. J. Fabra is recipients of a Juan de la Cierva contract from the Spanish Ministry of Economy and Competitivity. Maria L. Flores-López thanks Mexican Science and Technology Council (CONACyT, Mexico) for PhD fellowship support (CONACyT Grant Number 215499/310847). The author Miguel A. Cerqueira is a recipient of a fellowship (SFRH/BPD/72753/2010) supported by Fundação para a Ciência e Tecnologia, POPH-QREN and FSE (FCT, Portugal). The authors also thank the FCT Strategic Project of UID/ BIO/04469/2013 unit, the project RECI/BBB-EBI/0179/2012 (FCOMP- 01-0124-FEDER-027462) and the project BBioInd - Biotechnology and Bioengineering for improved Industrial and Agro-Food processes,^ REF. NORTE-07-0124-FEDER-000028 Co-funded by the Programa Operacional Regional do Norte (ON.2–O Novo Norte), QREN, FEDER. The support of EU Cost Action FA0904 is gratefully acknowledged

Optimization of electrospun polylactide-based ultrathin fibers for osteoconductive bone scaffolds

This is the peer reviewed version of the following article: Torres-Giner, S., Gimeno-Alcañiz, J. V., Ocio, M. J., & Lagaron, J. M. (2011). Optimization of electrospun polylactide-based ultrathin fibers for osteoconductive bone scaffolds. Journal of Applied Polymer Science, 122(2), 914-925, which has been published in final form at 10.1002/app.34208. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving."[EN] Bone tissue interfacial scaffolds, which encourage cell growth, are critical determinants for clinical success after implant surgery. Over the years, a number of resorbable configurations have emerged for bone cell support and growth, but only a few have demonstrated clinical efficacy. Polymer coatings produced by electrospinning are regarded as very promising bone interfaces because of the ultrathin-scaled dimensions of its physical structure. In this study, the morphology, composition, thermal properties, and cell growth viability of a number of polylactide-based systems containing different binary and ternary formulations of this biomaterial with collagen and commercial hydroxyapatite nanoparticles were characterized. The best performance in terms of biocompatibility was obtained for the tricomponent system in which the submicron fibers were further subjected to uniaxial orientation process during formation. The in vitro proliferation of the cells, which harbored on these ultrathin-structured mats, was examined by means of a metabolic activity indicator and ensured by means of scanning electron microscopy, and cell anchorage was checked by fluorescent optical microscopy. Finally, the optimum tricomponent material was successfully sterilized for the first time by gamma radiation without noticeable losses in cell-seeding capacity.Torres-Giner, S.; Gimeno-Alcañiz, JV.; Ocio, MJ.; Lagaron, JM. (2011). Optimization of electrospun polylactide-based ultrathin fibers for osteoconductive bone scaffolds. Journal of Applied Polymer Science. 122(2):914-925. https://doi.org/10.1002/app.34208914925122

Comparative Performance of Electrospun Collagen Nanofibers Cross-linked by Means of Different Methods

[EN] Collagen, as the major structural protein of the extracellular matrix in animals, is a versatile biomaterial of great interest in various engineering applications. Electrospun nanofibers of collagen are regarded as very promising materials for tissue engineering applications because they can reproduce the morphology of the natural bone but have as a drawback a poor structural consistency in wet conditions. In this paper, a comparative study between the performance of different cross-linking methods such as a milder enzymatic treatment procedure using transglutaminase, the use of N-[3-(dimethylamino)propyl]-N¿-ethylcarbodiimide hydrochloride/ N-hydroxysuccinimide, and genipin, and the use of a physical method based on exposure to ultraviolet light was carried out. The chemical and enzymatic treatments provided, in this order, excellent consistency, morphology, cross-linking degree, and osteoblast viability for the collagen nanofibers. Interestingly, the enzymatically cross-linked collagen mats, which are considered to be a more biological treatment, promoted adequate cell adhesion, making the biomaterial biocompatible and with an adequate degree of porosity for cell seeding and in-growth.The authors acknowledge the EU integrated project NEWBONE, the company Nanobiomatters Ltd. (Paterna, Spain), and the Spanish MEC (Project MAT2006- 10261-C03) for financial support and Impex Química Corp. for supplying TGTorres-Giner, S.; Gimeno-Alcañiz, JV.; Ocio, MJ.; Lagaron, JM. (2009). Comparative Performance of Electrospun Collagen Nanofibers Cross-linked by Means of Different Methods. ACS Applied Materials & Interfaces. 1(1):218-223. https://doi.org/10.1021/am800063x2182231

Development of an Electrospun Patch Platform Technology for the Delivery of Carvedilol in the Oral Mucosa

The work herein presented aims to develop and characterize carvedilol (CVD) releasable non-water-soluble monolayers and a multilayer patch made of ultrathin micron and submicron fibers for drug delivery into the sublingual mucosa. Firstly, the developed formulations containing CVD within different biopolymers (PDLA, PCL, and PHB) were characterized by scanning electron microscopy (SEM), attenuated total reflectance Fourier transformed infrared spectroscopy (ATR-FTIR), differential scanning calorimetry (DSC), wide-angle X-ray scattering (WAXS), and for their in vitro drug release. SEM micrographs assessed the fiber morphology attained by adding carvedilol. ATR-FTIR spectra revealed good chemical compatibility between CVD and the tested biopolymers, whereas DSC and WAXS confirmed that CVD was in an amorphous state within the biopolymeric fibers. In vitro release studies showed enhanced CVD release kinetics from the electrospun biopolymer monolayers compared to the dissolution rate of the commercial form of the pure drug, except for the slow-releasing PDLA fibers. Finally, the selected CVD-loaded layer, i.e., electrospun PHB, was built into a three-layer patch to tackle mucosa adhesion and unidirectional release, while retaining the enhanced release kinetics. The patch design proposed here further demonstrates the potential of the electro-hydrodynamic processing technology to render unique mucoadhesive controlled delivery platforms for poorly water-soluble drugs