Because of the physiological and cardiac changes associated with cardiovascular disease, tissue engineering can potentially restore the biological functions of cardiac tissue through the fabrication of scaffolds. In the present study, hybrid nanofiber scaffolds of poly (vinyl alcohol) (PVA) and bioglass type 58S (58SiO2-33CaO-9P2O5, Bg) were fabricated, and their effect on the spontaneous activity of chick embryonic cardiomyocytes in vitro was determined. PVA/Bg nanofibers were produced by electrospinning and stabilized by chemical crosslinking with glutaraldehyde. The electrospun scaffolds were analyzed to determine their chemical structure, morphology, and thermal transitions. The crosslinked scaffolds were more stable to degradation in water. A Bg concentration of 25% in the hybrid scaffolds improved thermal stability and decreased degradation in water after PVA crosslinking. Cardiomyocytes showed increased adhesion and contractility in cells seeded on hybrid scaffolds with higher Bg concentrations. In addition, the effect of Ca2+ ions released from the bioglass on the contraction patterns of cultured cardiomyocytes was investigated. The results suggest that the scaffolds with 25% Bg led to a uniform beating frequency that resulted in synchronous contraction patterns.

Electrospun fibers have emerged as promising materials in the field of biomedicine, due to their superior physical and cell supportive properties. In particular, electrospun mats are being developed for advanced wound dressing applications. Such applications require the firers to possess excellent antimicrobial properties in order to inhibit potential microbial colonization from resident and non-resident bacteria. In this study, we have developed Poly-ε-Caprolactone /gelatin hybrid composite mats loaded with natural herbal extract (Gymnema sylvestre) to prevent bacterial colonization. As-spun scaffolds exhibited good wettability and desirable mechanical properties retaining their fibrous structure after immersing them in phosphate buffered saline (pH 7.2) for up to 30 days. The initial burst release of Gymnema sylvestre prevented the colonization of bacteria as confirmed by the radial disc diffusion assay. Furthermore, the electrospun mats promoted cellular attachment, spreading and proliferation of human primary dermal fibroblasts and cultured keratinocytes, which are crucial parenchymal cell-types involved in the skin recovery process. Overall these results demonstrated the utility of Gymnema sylvestre impregnated electrospun PCL/Gelatin nanofibrous mats as an effective antimicrobial wound dressing.

The development of resistance against photodamage triggered by photodynamic therapy (PDT) is ascribed mainly to the cellular redox defenses and repair. If the tumor tissue is not promptly eliminated by the first few PDT sessions, PDT-resistance can be favored, challenging the efficacy of the treatment. Although the mechanism of PDT resistance is still unclear, in vitro assays have evidenced that it can be developed through the PARP damage-repair signaling pathway. Therefore, inhibition of poly(adenosine diphosphate (ADP)-ribose) polymerase (PARP) has the potential to increase PDT efficacy. This work reports on the synthesis of a controlled release system of a photosensitizer, methylene blue (MB) and a PARP-inhibitor, the veliparib. MB and veliparib were co-encapsulated in poly(lactic-co-glycolic acid) (PLGA) nanoparticles (VMB-NPs). A colloidal stable aqueous suspension of nanoparticles was obtained. The average hydrodynamic diameter was 90 nm and a narrow size distribution was obtained, with a polydispersity index (PDI) of 0.08. The release kinetics of MB and veliparib from VMB-NPs showed an initial burst of 8.7% and 58.3% release of the total amounts of MB and veliparib respectively, in the first 6 h, and a delayed release of up to 11.3% and 70%, in 19 days, for MB and veliparib, respectively. The VMB-NPs showed no cytotoxicity in the dark but the viability of B16F10-Nex2 cells decreased by 36% when the cells were irradiated (102 J/cm2, 660 nm) and treated with VMB-NPs containing 1.0 µM of MB and 8.3 µM of veliparib. Considering the increased photoactivity even at low MB and veliparib concentrations and the absence of cytotoxicity in dark, the co-encapsulation of MB and veliparib was shown to be a promising strategy to improve the PDT efficacy.

In this study, magnetite nanoparticles were prepared and coated with poly(ethylene glycol) terminated by alendronate to ensure firm binding to the iron oxide surface. Magnetic nanoparticles, designated as magnetite coated with poly(ethylene glycol)-alendronate (Fe3O4@PEG-Ale), were characterized in terms of number-average (Dn) and hydrodynamic (Dh) size, ζ-potential, saturation magnetization, and composition. The effect of particles on blood pressure, vascular functions, nitric oxide (NO), and superoxide production in the tissues of spontaneously hypertensive rats, as well as the effect on red blood cell (RBC) parameters, was investigated after intravenous administration (1 mg Fe3O4/kg of body weight). Results showed that Fe3O4@PEG-Ale particles did negatively affect blood pressure, heart rate and RBC deformability, osmotic resistance and NO production. In addition, Fe3O4@PEG-Ale did not alter functions of the femoral arteries. Fe3O4@PEG-Ale induced increase in superoxide production in the kidney and spleen, but not in the left heart ventricle, aorta and liver. NO production was reduced only in the kidney. In conclusion, the results suggest that acute intravenous administration of Fe3O4@PEG-Ale did not produce negative effects on blood pressure regulation, vascular function, and RBCs in hypertensive rats.

Galectin-3 is considered a cancer biomarker and bioindicator of fibrosis and cardiac remodeling and, therefore, it is desirable to develop convenient methods for its detection. Herein, an approach based on the development of multivalent electrochemical probes with high galectin-3 sensing abilities is reported. The probes consist of multivalent presentations of lactose-ferrocene conjugates scaffolded on poly (amido amine) (PAMAM) dendrimers and gold nanoparticles. Such multivalent lactose-ferrocene conjugates are synthesized by coupling of azidomethyl ferrocene-lactose building blocks on alkyne-functionalized PAMAM, for the case of the glycodendrimers, and to disulfide-functionalized linkers that are then used for the surface modification of citrate-stabilized gold nanoparticles. The binding and sensing abilities toward galectin-3 of both ferrocene-containing lactose dendrimers and gold nanoparticles have been evaluated by means of isothermal titration calorimetry, UV-vis spectroscopy, and differential pulse voltammetry. The highest sensitivity by electrochemical methods to galectin-3 was shown by lactosylferrocenylated gold nanoparticles, which are able to detect the lectin in nanomolar concentrations.

Electrodes from carbonized Zn₄O(1,4-benzodicarboxylic acid) (MOF-5) structures were prepared successfully via evaporating the solvent with a poly(vinylidene fluoride) (PVDF) binder. The solvent used for a nanocomposite cast was easily removed. Such an elegant method for preparing electrodes provides a facile, cost-effective, and void/cracking-free nanocomposite distribution on the current collector. The highly porous nanoparticles containing pure carbon attach well to the PVDF membrane which results in an increased active surface area of the electrode to 847 m²/g. The electrochemical analysis shows that the best weight ratio of CMOF-5 to PVDF equals 85:15, 80:20, and 75:25, respectively. The specific capacitance of these samples is 218 F/g, 210 F/g, and 180 F/g, correspondingly. An additional advantage of the electrode prepared from the carbonized MOF-5 is the possibility to synthesis MOF structures from recovered substrates used in its synthesis (distilled N,N-Dimethylformamide DMF and terephthalic acid recovered from polyethylene terephthalate waste). We will demonstrate this in this contribution as well. Furthermore, the carbonized MOF-5 can be recovered from the spent electrode and reused again in the electrochemical device.

Novel three-dimensional (3D) nanohydroxyapatite-PLGA scaffolds with high porosity was developed to better mimic mineral component and microstructure of natural bone. To perform a final assessment of this nanomaterial as a potential bone substitute, its toxicological profile was particularly investigated. Therefore, we performed a comet assay on human monocytes for in vitro genotoxicity investigation, and the systemic subchronic toxicity investigation on rats being per oral feed with exactly administrated extract quantities of the nano calcium hydroxyapatite covered with tiny layers of PLGA (ALBO-OS) for 120 days. Histological and stereological parameters of the liver, kidney, and spleen tissue were analyzed. Comet assay revealed low genotoxic potential, while histological analysis and stereological investigation revealed no significant changes in exposed animals when compared to controls, although the volume density of blood sinusoids and connective tissue, as well as numerical density and number of mitosis were slightly increased. Additionally, despite the significantly increased average number of the Ki67 and slightly increased number of CD68 positive cells in the presence of ALBO-OS, immunoreactive cells proliferation was almost neglected. Blood analyses showed that all of the blood parameters in rats fed with extract nanomaterial are comparable with corresponding parameters of no feed rats, taken as blind probe. This study contributes to the toxicological profiling of ALBO-OS scaffold for potential future application in bone tissue engineering.

Herein, we present a new heterogeneous catalyst active toward glucose to formic acid methyl ester oxidation. The catalyst was fabricated via electrostatic immobilization of the inorganic polyoxometalate HPA-5 catalyst H8[PMo7V5O40] onto the pore surface of amphiphilic block copolymer membranes prepared via non-solvent-induced phase separation (NIPS). The catalyst immobilization was achieved via wet impregnation due to strong coulombic interactions between protonated tertiary amino groups of the polar poly(2-(dimethylamino)ethyl methacrylate) block and the anionic catalyst. Overall, three sets of five consecutive catalytic cycles were performed in an autoclave under 90 °С and 11.5 bar air pressure in methanol, and the corresponding yields of formic acid methyl ester were quantified via head-space gas chromatography. The obtained results demonstrate that the membrane maintains its catalytic activity over multiple cycles, resulting in high to moderate yields in comparison to a homogeneous catalytic system. Nevertheless, presumably due to leaching, the catalytic activity declines over five catalytic cycles. The morphological and chemical changes of the membrane during the prolonged catalysis under harsh conditions were examined in detail using different analytic tools, and it seems that the underlying block copolymer is not affected by the catalytic process.

In this work the nanocomposite of vanadium oxide with conducting polymer poly(3,4-ethylenedioxythiophene) (VO@PEDOT) was obtained by microwave-assisted hydrothermal synthesis. The detailed study of its structural and electrochemical properties as cathode of aqueous zinc-ion battery was performed by scanning electron microscopy, energy dispersive X-ray analysis, X-ray diffraction analysis, X-ray photoelectron spectroscopy, thermogravimetric analysis, cyclic voltammetry, galvanostatic charge-discharge, and electrochemical impedance spectroscopy. The initial VO@PEDOT composite has layered nanosheets structure with thickness of about 30-80 nm, which are assembled into wavy agglomerated thicker layers of up to 0.3-0.6 μm. The phase composition of the samples was determined by XRD analysis which confirmed lamellar structure of vanadium oxide V10O24∙12H2O with interlayer distance of about 13.6 Å. The VO@PEDOT composite demonstrates excellent electrochemical performance, reaching specific capacities of up to 390 mA∙h∙g-1 at 0.3 A∙g-1. Moreover, the electrodes retain specific capacity of 100 mA∙h∙g-1 at a high current density of 20 A∙g-1. The phase transformations of VO@PEDOT electrodes during the cycling were studied at different degrees of charge/discharge by using ex situ XRD measurements. The results of ex situ XRD allow us to conclude that the reversible zinc ion intercalation occurs in stable zinc pyrovanadate structures formed during discharge.

Polymeric nanoparticles made of the copolymer Poly(L-lactide-co-caprolactone-co-glycolide) were prepared using the solvent evaporation method. Two different surfactants, polyvinyl alcohol and dextran, and a mixture of the two were employed. The three types of nanoparticles were used as hosting carriers of two chemotherapeutic drugs, the hydrophilic doxorubicin and the hydrophobic SN-38. The morphostructural characterization showed similar features for the three types of nanoparticles, while the drug encapsulation efficiency indicated that the dextran-based systems are the most effective with both drugs. Cellular studies with breast cancer cells were performed to compare the delivery capability and the cytotoxicity profile of the three nanosystems. The results show that the unloaded nanoparticles are highly biocompatible at the administered concentrations and confirmed that dextran-coated nanoparticles are the most efficient vectors to release the two drugs, exerting cytotoxic activity. PVA, on the other hand, shows limited drug release in vitro, probably due to strong interactions with both drugs. Data also show the release is more efficient for doxorubicin than for SN-38; indeed, the doxorubicin IC50 value for the dextran-coated nanoparticles was about 35% lower than the free drug. This indicates that these nanocarriers are suitable candidates to deliver hydrophilic drugs while needing further modification to host hydrophobic molecules.

This research reports about the development by electrospinning of fiber-based films made of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) derived from fermented fruit waste, so-called bio-papers, with enhanced antimicrobial performance. To this end, different combinations of oregano essential oil (OEO) and zinc oxide nanoparticles (ZnONPs) were added to PHBV solutions and electrospun into mats that were, thereafter, converted into homogeneous and continuous films of ~130 μm. The morphology, optical, thermal, mechanical properties, crystallinity, and migration into food simulants of the resultant PHBV-based bio-papers were evaluated and their antimicrobial properties were assessed against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) in both open and closed systems. It was observed that the antimicrobial activity decreased after 15 days due to the release of the volatile compounds, whereas the bio-papers filled with ZnONPs showed high antimicrobial activity for up to 48 days. The electrospun PHBV biopapers containing 2.5 wt% OEO + 2.25 wt% ZnONPs successfully provided the most optimal activity for short and long periods against both bacteria.

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.

A systematic study for the preparation and structural analysis of poly(styrene-co-acrylic acid) composite nanospheres (PSA) and silver nanoparticles loaded poly(styrene-co-acrylic acid) composite nanospheres (nAg@PSA) is reported. Poly(styrene-co-acrylic acid) nanospheres were synthesized by soap-free emulsion polymerization of styrene (St) and acrylic acid (AA) in water. Ag nanoparticles (Ag-NPs) were well-dispersed on the surfaces of poly(styrene-co-acrylic acid) composite nanospheres by in situ chemical reduction of AgNO₃ using NaBH₄ as a reducing agent in water. The particle size of PSA nanospheres was uniform. The surfaces of PSA nanospheres were distributed by highly uniform half-sphere arrays. Those half-sphere protruded more with the increase of the feeding amount of AA or the feed ratios of AA and St. The carboxyl groups content of nanospheres was directly proportional to the nanosphere surface area. This relationship and X-ray photoelectron spectroscopy and transmission electron microscopy images of the PSA nanospheres indicate that the acrylic acid was mainly distributed on the surface of the polystyrene spheres with unnegligible thickness. The number of Ag-NPs depends on immobilized carboxyl groups on the surface of PSA, according to thermogravimetry, ultraviolet-visible, X-ray diffraction and transmission electron microscopy results.

Food quality is mainly affected by oxygen through oxidative reactions and the proliferation of microorganisms, generating changes in its taste, odor, and color. The work presented here describes the generation and further characterization of films with active oxygen scavenging properties made of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) loaded with cerium oxide nanoparticles (CeO2NPs) obtained by electrospinning coupled to a subsequent annealing process, which could be used as coating or interlayer in a multilayer concept for food packaging applications. The aim of this work is to explore the capacities of these novel biopolymeric composites in terms of O2 scavenging capacity, as well as antioxidant, antimicrobial, barrier, thermal, and mechanical properties. To obtain such biopapers, different ratios of CeO2NPs were incorporated into a PHBV solution with hexadecyltrimethylammonium bromide (CTAB) as a surfactant. The produced films were analyzed in terms of antioxidant, thermal, antioxidant, antimicrobial, optical, morphological and barrier properties, and oxygen scavenging activity. According to the results, the nanofiller showed some reduction of the thermal stability of the biopolyester but exhibited antimicrobial and antioxidant properties. In terms of passive barrier properties, the CeO2NPs decreased the permeability to water vapor but increased the limonene and oxygen permeability of the biopolymer matrix slightly. Nevertheless, the oxygen scavenging activity of the nanocomposites showed significant results and improved further by incorporating the surfactant CTAB. The PHBV nanocomposite biopapers developed in this study appear as very interesting constituents for the potential design of new active organic recyclable packaging materials.

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-β (platelet-derived growth factor receptor-β), 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.

Centella asiatica, propolis, and hinokitiol, as natural antibacterial reagents, were integrated into the poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] (PHBH) polymer to produce antibacterial wound dressings, using electrospinning process. The results showed that the fiber diameters and surface morphology of PHBH composite nanofibers were influenced by the addition of ethanol-centella (EC), methanol-centella (MC), ethanol-propolis (EP), and ethanol-hinokitiol (EH) at various ratios compared to pristine PHBH nanofibers. From FT-IR, the nanofibrous samples with higher contents of natural antibacterial substances showed the peaks of carboxylic acid, aromatic ring, and tropolone carbon ring from centella, propolis, and hinokitiol, respectively. Furthermore, the tensile strength of neat PHBH nanofibers was increased from 8.00 ± 0.71 MPa up to 16.35 ± 1.78 MPa by loading of propolis (EP) 7% into PHBH. X-ray analysis explained that the loading of propolis (EP) was also able to increase the crystallinity in PHBH composite nanofibers from 47.0% to 54.5%. The antibacterial results demonstrated that PHBH composite nanofibers containing natural antibacterial products were potent inhibitors against the growth of Escherichia coli and Staphylococcus aureus, amongst them hinokitiol and propolis proved to be the most effective. Additionally, the release studies displayed that centella and hinokitiol had faster release from PHBH composite nanofibers in comparison to propolis.

Uncalcined/unsintered hydroxyapatite and poly-l-lactide-co-glycolide (u-HA/PLLA/PGA) is a new bioresorbable nanomaterial with superior characteristics compared with current bioresorbable materials, including appropriate mechanical properties, outstanding bioactive/osteoconductive features, and remarkably shorter resorption time. Nevertheless, the bone regeneration characteristics of this nanomaterial have not been evaluated in maxillofacial reconstructive surgery. In this study, we used a rat mandible model to assess the bone regeneration ability of u-HA/PLLA/PGA material, compared with uncalcined/unsintered hydroxyapatite and poly-l-lactide acid (u-HA/PLLA) material, which has demonstrated excellent bone regenerative ability. A 4-mm-diameter defect was created at the mandibular angle area in 28 Sprague Dawley male rats. The rats were divided into three groups: u-HA/PLLA/PGA (u-HA/PLLA/PGA graft + defect), u-HA/PLLA (u-HA/PLLA graft + defect), and sham control (defect alone). At 1, 3, 8, and 16 weeks after surgeries, the rats were sacrificed and assessed by micro-computed tomography, histological analysis with hematoxylin and eosin staining, and immunohistochemical analyses. The results confirmed that the accelerated bone bioactive/regenerative osteoconduction of u-HA/PLLA/PGA was comparable with that of u-HA/PLLA in the rat mandible model. Furthermore, this new regenerative nanomaterial was able to more rapidly induce bone formation in the early stage and had great potential for further clinical applications in maxillofacial reconstructive surgery.

We demonstrate an accessible and effective technique for exfoliating graphite foil and graphite powder into graphene in a water solution of inorganic salt. In our research, we report an electrochemical cathodic exfoliation in an aqueous solution of Na2SO4. After electro-exfoliation, the resulting graphene was premixed with furfuryl alcohol (FA) and an inorganic template (CaCO3 and Na2CO3). Once FA was polymerized to poly(furfuryl alcohol) (PFA), the mixture was carbonized. Carbon bridges originating in thermally-decomposed PFA joined exfoliated graphene flakes and stabilized the whole sponge-type structure after the nano-template was removed. Gases evolved at the graphite electrode (cathode) played an important role in the process of graphene-flake splitting and accelerated the change of graphite into graphene flakes. Starting graphite materials and graphene sponges were characterized using Raman spectroscopy, SEM, high-resolution transmission electron microscopy (HRTEM), elemental analysis, and low-temperature adsorption of nitrogen to determine their structure, morphology, and chemical composition. The discovered manufacturing protocol had a positive influence on the specific surface area and porosity of the sponges. The SEM and HRTEM studies confirmed a high separation degree of graphite and different agglomeration pathways. Raman spectra were analyzed with particular focus on the intensities of ID and IG peaks; the graphene-type nature of the sponges was confirmed.

Nanocomposite scaffolds combining carbon nanomaterials (CNMs) with a biocompatible matrix are able to favor the neuronal differentiation and growth of a number of cell types, because they mimic neural-tissue nanotopography and/or conductivity. We performed comparative analysis of biomimetic scaffolds with poly-L-lactic acid (PLLA) matrix and three different p-methoxyphenyl functionalized carbon nanofillers, namely, carbon nanotubes (CNTs), carbon nanohorns (CNHs), and reduced graphene oxide (RGO), dispersed at varying concentrations. qRT-PCR analysis of the modulation of neuronal markers in human circulating multipotent cells cultured on nanocomposite scaffolds showed high variability in their expression patterns depending on the scaffolds' inhomogeneities. Local stimuli variation could result in a multi- to oligopotency shift and commitment towards multiple cell lineages, which was assessed by the qRT-PCR profiling of markers for neural, adipogenic, and myogenic cell lineages. Less conductive scaffolds, i.e., bare poly-L-lactic acid (PLLA)-, CNH-, and RGO-based nanocomposites, appeared to boost the expression of myogenic-lineage marker genes. Moreover, scaffolds are much more effective on early commitment than in subsequent differentiation. This work suggests that biomimetic PLLA carbon-nanomaterial (PLLA-CNM) scaffolds combined with multipotent autologous cells can represent a powerful tool in the regenerative medicine of multiple tissue types, opening the route to next analyses with specific and standardized scaffold features.

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 µm 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.