Search

Approximately 740 million symptomatic patients are affected by otitis media every year. Being an inflammatory disease affecting the middle ear, it is one of the primary causes of tympanic membrane (TM) perforations, often resulting in impaired hearing abilities. Antibiotic therapy using broad-spectrum fluoroquinolones, such as ciprofloxacin (CIP), is frequently employed and considered the optimal route to treat otitis media. However, patients often get exposed to high dosages to compensate for the low drug concentration reaching the affected site. Therefore, this study aims to integrate tissue engineering with drug delivery strategies to create biomimetic scaffolds promoting TM regeneration while facilitating a localized release of CIP. Distinct electrospinning (ES) modalities were designed in this regard either by blending CIP into the polymer ES solution or by incorporating nanoparticles-based co-ES/electrospraying. The combination of these modalities was investigated as well. A broad range of release kinetic profiles was achieved from the fabricated scaffolds, thereby offering a wide spectrum of antibiotic concentrations that could serve patients with diverse therapeutic needs. Furthermore, the incorporation of CIP into the TM patches demonstrated a favorable influence on their resultant mechanical properties. Biological studies performed with human mesenchymal stromal cells confirmed the absence of any cytotoxic or anti-proliferative effects from the released antibiotic. Finally, antibacterial assays validated the efficacy of CIP-loaded scaffolds in suppressing bacterial infections, highlighting their promising relevance for TM applications.

Abstract Recently, in vitro models emerge as valuable tools in biomedical research by enabling the investigation of complex physiological processes in a controlled environment, replicating some traits of interest of the biological tissues. This study focuses on the development of tubular polymeric scaffolds, made of electrospun fibers, aimed to generate three‐dimensional (3D) in vitro intestinal models resembling the lumen of the gut. Polycaprolactone (PCL) and polyacrylonitrile (PAN) are used to produce tightly arranged ultrafine fiber meshes via electrospinning in the form of continuous tubular structures, mimicking the basement membrane on which the epithelial barrier is formed. Morphological, physical, mechanical, and piezoelectric properties of the PCL and PAN tubular scaffolds are investigated. They are cultured with Caco‐2 cells using different biological coatings (i.e., collagen, gelatin, and fibrin) and their capability of promoting a compact epithelial layer is assessed. PCL and PAN scaffolds show 42% and 50% porosity, respectively, with pore diameters and size suitable to impede cell penetration, thus promoting an intestinal epithelial barrier formation. Even if both polymeric structures allow Caco‐2 cell adhesion, PAN fiber meshes best suit many requirements needed by this model, including highest mechanical strength upon expansion, porosity and piezoelectric properties, along with the lowest pore size.

Many tissues and cells are influenced by mechano-electric stimulation, thus the application of piezoelectric materials has recently received considerable attention in tissue engineering. This report investigated electrospun fiber meshes based on poly(vinylidenefluoride-co-trifluoroethylene) [P(VDF-TrFE)] as instructive biomaterials for osteogenic differentiation of human mesenchymal stromal cells (hMSCs). The influence played by methyl ethyl ketone (MEK), used as a solvent in place of dimethylformamide (DMF)/acetone mixture, and the effect of rotating velocity of the electrospinning collector on fiber morphology, mechanical and piezoelectric properties were studied. The solvent had noticeable effects on morphology and piezoelectric properties of electrospun fibers, with MEK outperforming DMF/acetone. By increasing the collector velocity up to 4000 rpm, the fiber diameter reduced and the mutual alignment of the fibers increased, corresponding to enhanced mechanical properties and piezo-active β-phase content. However, as a consequence of the diverse mechanical properties of random and aligned fibrous architectures, which ultimately affected the piezoelectric properties, randomly-oriented fibers exhibited higher remnant piezoelectric properties (Vout and d31 piezoelectric coefficient) than aligned ones. On these scaffolds, hMSCs showed an excellent capability of early osteogenic differentiation, leading to high calcium production. Fiber surface topology, fiber mesh morphology and remnant piezoelectric properties played a determinant role on hMSC osteogenic commitment.

Sensorineural hearing loss (SNHL) occurs when the sound transduction mechanism in the inner ear is compromised, because of impairments affecting the sensory hair cells—the actual biological transducers (90% of cases)—or the neurons. SNHL results in a broad spectrum of developmental, cognitive and psycho-social damages. To date, only cochlear implants (CIs) can offer a therapeutic solution to patients. They are multi-component electronic devices, surgically implanted, which capture, elaborate and convert the sound into electric stimuli delivered to the cochlea. Due to inherent limitations of the current electronic-based CIs, a new class of devices has been envisioned, which is based on piezoelectric materials. However, using piezoelectric membranes, the obtained sensitivity was not enough. The new frontiers for piezoelectric material-based CI aim at synergizing micro/nanofabrication aided by multiscale materials modeling with an in vivo tissue engineering approach to provide an implantable biomaterial-based system for SNHL, acting as a next-generation CI. Specifically, the envisioned device will move forward the primitive concept of bulk-structured piezoelectric CIs by designing a nanostructured material (e.g., based on nanofibers) to be precisely delivered and be intimately and efficiently integrated with the cochlear microenvironment. Piezoelectric material-based CIs are indeed hypothesized to have a much higher resolution of electrical stimulation with more than hundreds of channels, compared to maximum 22 stimulating elements present in electronic-based CIs. Moreover, the stimulation site will be closest to peripheral nerve fiber endings for maximal resolution. This would be the first sensory implant with a feedback mechanism on a micrometer scale.

Bone defects are a significant health problem worldwide. Novel treatment approaches in the tissue engineering field rely on the use of biomaterial scaffolds to stimulate and guide the regeneration of damaged tissue that cannot repair or regrow spontaneously. This work aimed at developing and characterizing new piezoelectric scaffolds to provide electric bio-signals naturally present in bone and vascular tissues. Mixing and extrusion were used to obtain nanocomposites made of polyhydroxybutyrate (PHB) as a matrix and barium titanate (BaTiO3) nanoparticles as a filler, at BaTiO3/PHB compositions of 5/95, 10/90, 15/85 and 20/80 (w/w%). The morphological, thermal, mechanical and piezoelectric properties of the nanocomposites were studied. Scanning electron microscopy analysis showed good nanoparticle dispersion within the polymer matrix. Considerable increases in the Young’s modulus, compressive strength and the piezoelectric coefficient d31 were observed with increasing BaTiO3 content, with d31 = 37 pm/V in 20/80 (w/w%) BaTiO3/PHB. 3D printing was used to produce porous cubic-shaped scaffolds using a 90° lay-down pattern, with pore size ranging in 0.60–0.77 mm and good mechanical stability. Biodegradation tests conducted for 8 weeks in saline solution at 37 °C showed low mass loss (∼4%) for 3D printed scaffolds. The results obtained in terms of piezoelectric, mechanical and chemical properties of the nanocomposite provide a new promising strategy for vascularized bone tissue engineering.

Bacteriotherapy is emerging as a strategic and effective approach to treat infections by providing putatively harmless bacteria (i.e., probiotics) as antagonists to pathogens. Proper delivery of probiotics or their metabolites (i.e., post-biotics) can facilitate their availing of biomaterial encapsulation via innovative manufacturing technologies. This review paper aims to provide the most recent biomaterial-assisted strategies proposed to treat infections or dysbiosis using bacteriotherapy. We revised the encapsulation processes across multiscale biomaterial approaches, which could be ideal for targeting different tissues and suit diverse therapeutic opportunities. Hydrogels, and specifically polysaccharides, are the focus of this review, as they have been reported to better sustain the vitality of the live cells incorporated. Specifically, the approaches used for fabricating hydrogel-based devices with increasing dimensionality (D)—namely, 0D (i.e., particles), 1D (i.e., fibers), 2D (i.e., fiber meshes), and 3D (i.e., scaffolds)—endowed with probiotics, were detailed by describing their advantages and challenges, along with a future overlook in the field. Electrospinning, electrospray, and 3D bioprinting were investigated as new biofabrication methods for probiotic encapsulation within multidimensional matrices. Finally, examples of biomaterial-based systems for cell and possibly post-biotic release were reported.

Antimicrobial resistance poses a growing challenge in respiratory tract diseases like otitis media, often necessitating surgical interventions due to pharmacological treatment limitations. Bacteriotherapy, involving probiotics and/or their bioproducts, emerges as a promising alternative in such a scenario. This study aims to pave the way to middle ear bacteriotherapy by developing an innovative sodium alginate (SA)-based probiotic delivery system using electrospinning and electrospray techniques. Electrospray enabled the precise production of probiotic-laden SA microparticles, demonstrating potential for targeted bacterial delivery. By overcoming challenges due to the SA molecular structure, we successfully electrospun SA-based fiber meshes with poly(ethylene oxide) (PEO) as a support polymer. The rheologic behavior of the probiotic/SA solutions and the morphology of the obtained microparticles and fibers was evaluated, along with the diameter variation over time. The cytocompatibility of the produced microparticles and fibers was assessed using human dermal keratinocytes and their antimicrobial activity was tested against E. coli. The incorporation of probiotic-laden SA microparticles within electrospun SA/PEO fiber meshes finally offered a patch-like structure to be applied on the tympanic membrane or on the outer auditory canal, which could be a versatile and ideally safe treatment strategy in chronic otitis media. This innovative approach holds promise for clinical applications dealing with inflammatory processes, infections and dysbiosis, thus possibly addressing the complex healing process of chronic upper respiratory diseases while mitigating antimicrobial resistance.

High-performance and low-cost indicators are important in food and cosmetics industry but market uptake is low due to several challenges such as toxicity, cost and unclear reading. Here, the authors report on optically-programmed, non-colorimetric indicators based on nanotextured organic non-wovens, encoded by controlling their cross-linking degree.

Type-1 diabetes is one of the most prevalent metabolic disorders worldwide. It results in a significant lack of insulin production by the pancreas and the ensuing hyperglycemia, which needs to be regulated through a tailored administration of insulin throughout the day. Recent studies have shown great advancements in developing an implantable artificial pancreas. However, some improvements are still required, including the optimal biomaterials and technologies to produce the implantable insulin reservoir. Here, we discuss the employment of two types of cyclic olefin copolymers (Topas 5013L-10 and Topas 8007S-04) for an insulin reservoir fabrication. After a preliminary thermomechanical analysis, Topas 8007S-04 was selected as the best material to fabricate a 3D-printed insulin reservoir due to its higher strength and lower glass transition temperature (Tg). Fiber deposition modeling was used to manufacture a reservoir-like structure, which was employed to assess the ability of the material to prevent insulin aggregation. Although the surface texture presents a localized roughness, the ultraviolet analysis did not detect any significant insulin aggregation over a timeframe of 14 days. These interesting results make Topas 8007S-04 cyclic olefin copolymer a potential candidate biomaterial for fabricating structural components in an implantable artificial pancreas.

The aim of this study was to investigate the effect of different types of nanocellulose, i.e., cellulose nanocrystal (CNC), cellulose nanofiber (CNF) and bacterial nanocellulose (BNC), and also different drying methods (oven-drying and freeze-drying) on the properties of acrylic acid (AA)/sodium alginate (SA) super absorbent polymers (SAPs). In addition, the presence of ammonium per sulfate as an initiator and N-N methylene-bis-acrylamide as a cross-linker were considered. Synthesized SAPs were characterized by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). The absorption and rheological properties (i.e., storage modulus and loss modulus) were also investigated. The results of FTIR spectroscopy demonstrated several types of interactions, such as hydrogen and esterification, between SA, AA and nanocellulose. SEM analysis revealed a microporous structure in the SAPs. All SAPs had a centrifuge retention capacity (CRC)/free swelling capacity (FSC) ≥ 69%. The absorption behavior showed that the oven-dried SAPs had superior (about 2×) CRC and FRC in different aqueous media compared to the freeze-dried counterparts. The freeze-dried SAPs showed increased rheological properties in comparison to the oven-dried ones, with SAPs containing BNC and CNC having the highest rheological properties, respectively. Overall, it can be concluded that oven-dried SAPs containing CNC had better absorption properties than the other ones tested in this study.

Polyhydroxyalkanoates (PHAs) are a family of biopolyesters synthesized by various microorganisms. Due to their biocompatibility and biodegradation, PHAs have been proposed for biomedical applications, including tissue engineering scaffolds. Olive leaf extract (OLE) can be obtained from agri-food biowaste and is a source of polyphenols with remarkable antioxidant properties. This study aimed at incorporating OLE inside poly(hydroxybutyrate-co-hydroxyvalerate) (PHBHV) fibers via electrospinning to obtain bioactive bio-based blends that are useful in wound healing. PHBHV/OLE electrospun fibers with a size of 1.29 ± 0.34 µm were obtained. Fourier transform infrared chemical analysis showed a uniform surface distribution of hydrophilic -OH groups, confirming the presence of OLE in the electrospun fibers. The main OLE phenols were released from the fibers within 6 days. The biodegradation of the scaffolds in phosphate buffered saline was investigated, demonstrating an adequate stability in the presence of metalloproteinase 9 (MMP-9), an enzyme produced in chronic wounds. The scaffolds were preliminarily tested in vitro with HFFF2 fibroblasts and HaCaT keratinocytes, suggesting adequate cytocompatibility. PHBHV/OLE fiber meshes hold promising features for wound healing, including the treatment of ulcers, due to the long period of durability in an inflamed tissue environment and adequate cytocompatibility.

In this study, for the first time, a composite fluff pulp was produced based on the combination of softwood (i.e., long-length fiber), hardwood (i.e., short-length fiber), non-wooden pulps (i.e., bagasse) and bentonite, with specific amounts to be used in hygienic pads (e.g., baby diapers and sanitary napkins). After the defibration process, the manufactured fluff pulp was placed as an absorbent mass in diapers and sanitary napkins. Therefore, tests related to the fluff pulp, such as grammage, thickness, density, ash content, humidity percentage, pH and brightness, tests related to the manufactured baby diapers, such as absorption capacity, retention rate, retention capacity, absorption time and rewet, and tests related to the sanitary napkin, such as absorption capacity and rewet, were performed according to the related standards. The results demonstrated that increasing the amount of bagasse pulp led to increasing the ash content, pH and density of fluff pulp and decreasing the brightness. The addition of bentonite as a filler also increased ash content and pH of fluff pulp. The results also demonstrated that increasing of bagasse pulp up to 30% in combination with softwood pulp led to increasing absorption capacity, retention rate, retention capacity, absorption time and rewet of baby diapers and of sanitary napkins.

Perforation is the most common illness of the tympanic membrane (TM), which is commonly treated with surgical procedures. The success rate of the treatment could be improved by novel bioengineering approaches. In fact, a successful restoration of a damaged TM needs a supporting biomaterial or scaffold able to meet mechano-acoustic properties similar to those of the native TM, along with optimal biocompatibility. Traditionally, a large number of biological-based materials, including paper, silk, Gelfoam®, hyaluronic acid, collagen, and chitosan, have been used for TM repair. A novel biopolymer with promising features for tissue engineering applications is cellulose. It is a highly biocompatible, mechanically and chemically strong polysaccharide, abundant in the environment, with the ability to promote cellular growth and differentiation. Bacterial cellulose (BC), in particular, is produced by microorganisms as a nanofibrous three-dimensional structure of highly pure cellulose, which has thus become a popular graft material for wound healing due to a number of remarkable properties, such as water retention, elasticity, mechanical strength, thermal stability, and transparency. This review paper provides a comprehensive overview of the current experimental studies of BC, focusing on the application of BC patches in the treatment of TM perforations. In addition, computational approaches to model cellulose and TM are summarized, with the aim to synergize the available tools toward the best design and exploitation of BC patches and scaffolds for TM repair and regeneration.

Chitin nanofibrils (CN) and nanolignin (NL) were used to embed active molecules, such as vitamin E, sodium ascorbyl phosphate, lutein, nicotinamide and glycyrrhetinic acid (derived from licorice), in the design of antimicrobial, anti-inflammatory and antioxidant nanostructured chitin nanofibrils–nanolignin (CN-NL) complexes for skin contact products, thus forming CN-NL/M complexes, where M indicates the embedded functional molecule. Nano-silver was also embedded in CN-NL complexes or on chitin nanofibrils to exploit its well-known antimicrobial activity. A powdery product suitable for application was finally obtained by spray-drying the complexes co-formulated with poly(ethylene glycol). The structure and morphology of the complexes was studied using infrared spectroscopy and field emission scanning electron microscopy, while their thermal stability was investigated via thermo-gravimetry. The latter provided criteria for evaluating the suitability of the obtained complexes for subsequent demanding industrial processing, such as, for instance, incorporation into bio-based thermoplastic polymers through conventional melt extrusion. In vitro tests were carried out at different concentrations to assess skin compatibility. The obtained results provided a physical–chemical, morphological and cytocompatibility knowledge platform for the correct selection and further development of such nanomaterials, allowing them to be applied in different products. In particular, chitin nanofibrils and the CN-NL complex containing glycyrrhetinic acid can combine excellent thermal stability and skin compatibility to provide a nanostructured system potentially suitable for industrial applications.

Poly(lactic acid)(PLA) is an aliphatic polyester that can be derived from natural and renewable resources. Owing to favorable features, such as biocompatibility, biodegradability, good thermal and mechanical performance, and processability, PLA has been considered as one of the most promising biopolymers for biomedical applications. Particularly, electrospun PLA nanofibers with distinguishing characteristics, such as similarity to the extracellular matrix, large specific surface area and high porosity with small pore size and tunable mechanical properties for diverse applications, have recently given rise to advanced spillovers in the medical area. A variety of PLA-based nanofibrous structures have been explored for biomedical purposes, such as wound dressing, drug delivery systems, and tissue engineering scaffolds. This review highlights the recent advances in electrospinning of PLA-based structures for biomedical applications. It also gives a comprehensive discussion about the promising approaches suggested for optimizing the electrospun PLA nanofibrous structures towards the design of specific medical devices with appropriate physical, mechanical and biological functions.

The COVID-19 outbreak has increased the incidence of tracheal lesions in patients who underwent invasive mechanical ventilation. We measured the pressure exerted by the cuff on the walls of a test bench mimicking the laryngotracheal tract. The test bench was designed to acquire the pressure exerted by endotracheal tube cuffs inflated inside an artificial model of a human trachea. The experimental protocol consisted of measuring pressure values before and after applying a maneuver on two types of endotracheal tubes placed in two mock-ups resembling two different sized tracheal tracts. Increasing pressure values were used to inflate the cuff and the pressures were recorded in two different body positions. The recorded pressure increased proportionally to the input pressure. Moreover, the pressure values measured when using the non-armored (NA) tube were usually higher than those recorded when using the armored (A) tube. A periodic check of the cuff pressure upon changing the body position and/or when performing maneuvers on the tube appears to be necessary to prevent a pressure increase on the tracheal wall. In addition, in our model, the cuff of the A tube gave a more stable output pressure on the tracheal wall than that of the NA tube.

The demand for bone substitutes is increasing in Western countries. Bone graft substitutes aim to provide reconstructive surgeons with off-the-shelf alternatives to the natural bone taken from humans or animal species. Under the tissue engineering paradigm, biomaterial scaffolds can be designed by incorporating bone stem cells to decrease the disadvantages of traditional tissue grafts. However, the effective clinical application of tissue-engineered bone is limited by insufficient neovascularization. As bone is a highly vascularized tissue, new strategies to promote both osteogenesis and vasculogenesis within the scaffolds need to be considered for a successful regeneration. It has been demonstrated that bone and blood vases are piezoelectric, namely, electric signals are locally produced upon mechanical stimulation of these tissues. The specific effects of electric charge generation on different cells are not fully understood, but a substantial amount of evidence has suggested their functional and physiological roles. This review summarizes the special contribution of piezoelectricity as a stimulatory signal for bone and vascular tissue regeneration, including osteogenesis, angiogenesis, vascular repair, and tissue engineering, by considering different stem cell sources entailed with osteogenic and angiogenic potential, aimed at collecting the key findings that may enable the development of successful vascularized bone replacements useful in orthopedic and otologic surgery.

Chitin nanofibrils (CNs) are an emerging bio-based nanomaterial. Due to nanometric size and high crystallinity, CNs lose the allergenic features of chitin and interestingly acquire anti-inflammatory activity. Here we investigate the possible advantageous use of CNs in tympanic membrane (TM) scaffolds, as they are usually implanted inside highly inflamed tissue environment due to underlying infectious pathologies. In this study, the applications of CNs in TM scaffolds were twofold. A nanocomposite was used, consisting of poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT) copolymer loaded with CN/polyethylene glycol (PEG) pre-composite at 50/50 (w/w %) weight ratio, and electrospun into fiber scaffolds, which were coated by CNs from crustacean or fungal sources via electrospray. The degradation behavior of the scaffolds was investigated during 4 months at 37 °C in an otitis-simulating fluid. In vitro tests were performed using cell types to mimic the eardrum, i.e., human mesenchymal stem cells (hMSCs) for connective, and human dermal keratinocytes (HaCaT cells) for epithelial tissues. HMSCs were able to colonize the scaffolds and produce collagen type I. The inflammatory response of HaCaT cells in contact with the CN-coated scaffolds was investigated, revealing a marked downregulation of the pro-inflammatory cytokines. CN-coated PEOT/PBT/(CN/PEG 50:50) scaffolds showed a significant indirect antimicrobial activity.

Cosmetics has recently focused on biobased skin-compatible materials. Materials from natural sources can be used to produce more sustainable skin contact products with enhanced bioactivity. Surface functionalization using natural-based nano/microparticles is thus a subject of study, aimed at better understanding the skin compatibility of many biopolymers also deriving from biowaste. This research investigated electrospray as a method for surface modification of cellulose tissues with chitin nanofibrils (CNs) using two different sources—namely, vegetable (i.e., from fungi), and animal (from crustaceans)—and different solvent systems to obtain a biobased and skin-compatible product. The surface of cellulose tissues was uniformly decorated with electrosprayed CNs. Biological analysis revealed that all treated samples were suitable for skin applications since human dermal keratinocytes (i.e., HaCaT cells) successfully adhered to the processed tissues and were viable after being in contact with released substances in culture media. These results indicate that the use of solvents did not affect the final cytocompatibility due to their effective evaporation during the electrospray process. Such treatments did not also affect the characteristics of cellulose; in addition, they showed promising anti-inflammatory and indirect antimicrobial activity toward dermal keratinocytes in vitro. Specifically, cellulosic substrates decorated with nanochitins from shrimp showed strong immunomodulatory activity by first upregulating then downregulating the pro-inflammatory cytokines, whereas nanochitins from mushrooms displayed an overall anti-inflammatory activity via a slight decrement of the pro-inflammatory cytokines and increment of the anti-inflammatory marker. Electrospray could represent a green method for surface modification of sustainable and biofunctional skincare products.

Structurally and mechanically similar to the extracellular matrix (ECM), biomimetic hydrogels offer a number of opportunities in medical applications. However, the generation of synthetic microenvironments that simulate the effects of natural tissue niches on cell growth and differentiation requires new methods to control hydrogel feature resolution, biofunctionalization and mechanical properties. Here we show how these goals can be achieved by using a pullulan-based hydrogel, engineered in composition and server as cell-adhesive hydrogel, 3D photo-printable in dimension, ranging from the macro- to the micro-scale dimensions, and of tunable mechanical properties. For this, we used absorbers that limit light penetration, achieving 3D patterning through stereolithography with feature vertical resolution of 200 μm and with overall dimension up to several millimeters. Furthermore, we report the fabrication of 3D pullulan-modified hydrogels by two-photon lithography, with sub-millimetric dimensions and minimum feature sizes down to some microns. These materials open the possibility to produce multiscale printed scaffolds that here we demonstrate to be inert for cell adhesion, but biologically compatible and easily functionalizable with cell adhesive proteins. Under these conditions, successful cell cultures were established in 2D and 3D. Keywords: Hydrogel, Biomaterials, Polysaccharide, Pullulan, 3D printing, Two photon laser lithography, Mesenchymal stromal cells

Need for regeneration and repair of nasal tissues occurs as a consequence of several pathologies affecting the nose, including, but not limited to infective diseases, traumas and tumor resections. A platform for nasal tissue regeneration was set up using poly(vinyl alcohol)/gelatin sponges with 20%–30% (w/w) gelatin content to be used as scaffolds, for their intrinsic hydrophilic, cell adhesive and shape recovery properties. We propose mesodermal progenitor cells (MPCs) isolated from the bone marrow as a unique stem cell source for obtaining different connective tissues of the nose, including vascular tissue. Finally, epithelial cell immune response to these scaffolds was assessed in vitro in an environment containing inflammatory molecules. The results showed that mesenchymal stromal cells (MSCs) deriving from MPCs could be used to differentiate into cartilage and fibrous tissue; whereas, in combination with endothelial cells still deriving from MPCs, into pre-vascularized bone. Finally, the scaffold did not significantly alter the epithelial cell response to inflammatory insults derived from interaction with bacterial molecules.

Olive tree is a well-known source of polyphenols. We prepared an olive leaf extract (OLE) and characterized it via high performance liquid chromatography (HPLC) analysis. OLE was blended with different polyhydroxyalkanoates (PHAs), namely, poly(hydroxybutyrate-co-hydroxyvalerate) (PHBHV) and polyhydroxybutyrate/poly(hydroxyoctanoate-co-hydroxydecanoate) (PHB/PHOHD), to produce fiber meshes via electrospinning: OLE/PHBV and OLE/ (PHB/PHOHD), respectively. An 80–90% (w/w%) release of the main polyphenols from the OLE/PHA fibers occurred in 24 h, with a burst release in the first 30 min. OLE and the produced fiber meshes were assayed using human dermal keratinocytes (HaCaT cells) to evaluate the expression of a panel of cytokines involved in the inflammatory process and innate immune response, such as the antimicrobial peptide human beta defensin 2 (HBD-2). Fibers containing OLE were able to decrease the expression of the pro-inflammatory cytokines at 6 h up to 24 h. All the PHA fibers allowed an early downregulation of the pro-inflammatory cytokines in 6 h, which is suggestive of a strong anti-inflammatory activity exerted by PHA fibers. Differently from pure OLE, PHB/PHOHD fibers (both with and without OLE) upregulated the expression of HBD-2. Our results showed that PHA fiber meshes are suitable in decreasing pro-inflammatory cytokines and the incorporation of OLE may enable indirect antibacterial properties, which is essential in wound healing and tissue regeneration.

A novel strategy involving Olive Leaf Extract (OLE) and Cold Atmospheric Plasma (CAP) was developed as a green antimicrobial treatment. Specifically, we reported a preliminary investigation on the combined use of OLE + CAP against three pathogens, chosen to represent medical and food industries (i.e., E. coli, S. aureus and L. innocua). The results indicated that a concentration of 100 mg/mL (total polyphenols) in OLE can exert an antimicrobial activity, but still insufficient for a total bacterial inactivation. By using plain OLE, we significantly reduced the growth of Gram positive S. aureus and L. innocua, but not Gram-negative E. coli. Instead, we demonstrated a remarkable decontamination effect of OLE + CAP in E. coli, S. aureus and L. innocua samples after 6 h. This effect was optimally maintained up to 24 h in S. aureus strain. E. coli and L. innocua grew again in 24 h. In the latter strain, OLE alone was most effective to significantly reduce bacterial growth. By further adjusting the parameters of OLE + CAP technology, e.g., OLE amount and CAP exposure, it could be possible to prolong the initial powerful decontamination over a longer time. Since OLE derives from a bio-waste and CAP is a non-thermal technology based on ionized air, we propose OLE + CAP as a potential green platform for bacterial decontamination. As a combination, OLE and CAP can lead to better antimicrobial activity than individually and may replace or complement conventional thermal procedures in food and biomedical industries.

The world of cosmetics is now aiming at biobased materials which are skin-compatible and can be used to generate more sustainable beauty masks with enhanced bioactivity. This work presents, in this line of interest, the combination of two innovative technologies, namely electrospinning and dry powder impregnation, to generate biobased skin soluble electrospun pullulan carriers dry impregnated with chitin nanofibrils-nanolignin-glycyrrethinic acid (CLA) complexes, as effective biobased and skin compatible beauty masks. The scalability of the pullulan electrospun carrier and bioactive complexes impregnation were optimized and the morphology evaluated. Subsequently, skin compatibility and mask effectiveness were investigated in vitro and in vivo. The results showed that cell viability was optimal for both impregnated and neat pullulan fibers. Additionally, the CLA impregnated pullulan fibers were able to upregulate the endogenous antimicrobial molecule HBD-2. Preliminary studies in vivo indicated that the beauty mask containing the CLA complexes significantly decreased area, length and depth of forehead and crow’s feet wrinkles, and significantly increased moisturizing levels in the skin. The developed beauty mask was also seen to increase skin firmness, while it did not show skin irritation after the test. The work demonstrates that the combination of these two technologies may open new alternatives to more sustainable bioactive cosmetic products for the skin.

The applications of tissue engineered constructs have witnessed great advances in the last few years, as advanced fabrication techniques have enabled promising approaches to develop structures and devices for biomedical uses. (Bio-)printing, including both plain material and cell/material printing, offers remarkable advantages and versatility to produce multilateral and cell-laden tissue constructs; however, it has often revealed to be insufficient to fulfill clinical needs. Indeed, three-dimensional (3D) (bio-)printing does not provide one critical element, fundamental to mimic native live tissues, i.e., the ability to change shape/properties with time to respond to microenvironmental stimuli in a personalized manner. This capability is in charge of the so-called “smart materials”; thus, 3D (bio-)printing these biomaterials is a possible way to reach four-dimensional (4D) (bio-)printing. We present a comprehensive review on stimuli-responsive materials to produce scaffolds and constructs via additive manufacturing techniques, aiming to obtain constructs that closely mimic the dynamics of native tissues. Our work deploys the advantages and drawbacks of the mechanisms used to produce stimuli-responsive constructs, using a classification based on the target stimulus: humidity, temperature, electricity, magnetism, light, pH, among others. A deep understanding of biomaterial properties, the scaffolding technologies, and the implant site microenvironment would help the design of innovative devices suitable and valuable for many biomedical applications.

The skin is the body outermost tissue and acts as a barrier and defense line to protect our organs [...]

Polyhydroxyalkanoates (PHAs) are a family of bio-based polyesters that have found different biomedical applications. Chitin and lignin, byproducts of fishery and plant biomass, show antimicrobial and anti-inflammatory activity on the nanoscale. Due to their polarities, chitin nanofibril (CN) and nanolignin (NL) can be assembled into micro-complexes, which can be loaded with bioactive factors, such as the glycyrrhetinic acid (GA) and CN-NL/GA (CLA) complexes, and can be used to decorate polymer surfaces. This study aims to develop completely bio-based and bioactive meshes intended for wound healing. Poly(3-hydroxybutyrate)/Poly(3-hydroxyoctanoate-co-3-hydroxydecanoate), P(3HB)/P(3HO-co-3HD) was used to produce films and fiber meshes, to be surface-modified via electrospraying of CN or CLA to reach a uniform distribution. P(3HB)/P(3HO-co-3HD) fibers with desirable size and morphology were successfully prepared and functionalized with CN and CLA using electrospinning and tested in vitro with human keratinocytes. The presence of CN and CLA improved the indirect antimicrobial and anti-inflammatory activity of the electrospun fiber meshes by downregulating the expression of the most important pro-inflammatory cytokines and upregulating human defensin 2 expression. This natural and eco-sustainable mesh is promising in wound healing applications.

Being designated to protect other tissues, skin is the first and largest human body organ to be injured and for this reason, it is accredited with a high capacity for self-repairing. However, in the case of profound lesions or large surface loss, the natural wound healing process may be ineffective or insufficient, leading to detrimental and painful conditions that require repair adjuvants and tissue substitutes. In addition to the conventional wound care options, biodegradable polymers, both synthetic and biologic origin, are gaining increased importance for their high biocompatibility, biodegradation, and bioactive properties, such as antimicrobial, immunomodulatory, cell proliferative, and angiogenic. To create a microenvironment suitable for the healing process, a key property is the ability of a polymer to be spun into submicrometric fibers (e.g., via electrospinning), since they mimic the fibrous extracellular matrix and can support neo- tissue growth. A number of biodegradable polymers used in the biomedical sector comply with the definition of bio-based polymers (known also as biopolymers), which are recently being used in other industrial sectors for reducing the material and energy impact on the environment, as they are derived from renewable biological resources. In this review, after a description of the fundamental concepts of wound healing, with emphasis on advanced wound dressings, the recent developments of bio-based natural and synthetic electrospun structures for efficient wound healing applications are highlighted and discussed. This review aims to improve awareness on the use of bio-based polymers in medical devices.

Bacterial colonization of implanted biomedical devices is the main cause of healthcare-associated infections, estimated to be 8.8 million per year in Europe. Many infections originate from damaged skin, which lets microorganisms exploit injuries and surgical accesses as passageways to reach the implant site and inner organs. Therefore, an effective treatment of skin damage is highly desirable for the success of many biomaterial-related surgical procedures. Due to gained resistance to antibiotics, new antibacterial treatments are becoming vital to control nosocomial infections arising as surgical and post-surgical complications. Surface coatings can avoid biofouling and bacterial colonization thanks to biomaterial inherent properties (e.g., super hydrophobicity), specifically without using drugs, which may cause bacterial resistance. The focus of this review is to highlight the emerging role of degradable polymeric micro- and nano-structures that show intrinsic antifouling and antimicrobial properties, with a special outlook towards biomedical applications dealing with skin and skin damage. The intrinsic properties owned by the biomaterials encompass three main categories: (1) physical–mechanical, (2) chemical, and (3) electrostatic. Clinical relevance in ear prostheses and breast implants is reported. Collecting and discussing the updated outcomes in this field would help the development of better performing biomaterial-based antimicrobial strategies, which are useful to prevent infections.

The interaction of immune cells with drugs and/or with other cell types should be mechanistically investigated in order to reduce attrition of new drug development. However, they are currently only limited technologies that address this need. In our work, we developed initial but significant building blocks that enable such immune-drug studies. We developed a novel microfluidic platform replicating the Lymph Node (LN) microenvironment called LN-on-a-chip, starting from design all the way to microfabrication, characterization and validation in terms of architectural features, fluidics, cytocompatibility, and usability. To prove the biomimetics of this microenvironment, we inserted different immune cell types in a microfluidic device, which showed an in-vivo-like spatial distribution. We demonstrated that the developed LN-on-a-chip incorporates key features of the native human LN, namely, (i) similarity in extracellular matrix composition, morphology, porosity, stiffness, and permeability, (ii) compartmentalization of immune cells within distinct structural domains, (iii) replication of the lymphatic fluid flow pattern, (iv) viability of encapsulated cells in collagen over the typical timeframe of immunotoxicity experiments, and (v) interaction among different cell types across chamber boundaries. Further studies with this platform may assess the immune cell function as a step forward to disclose the effects of pharmaceutics to downstream immunology in more physiologically relevant microenvironments.

The tympanic membrane (TM) primes the sound transmission mechanism due to special fibrous layers mainly of collagens II, III, and IV as a product of TM fibroblasts, while type I is less represented. In this study, human mesenchymal stromal cells (hMSCs) were cultured on star-branched poly(ε-caprolactone) (*PCL)-based nonwovens using a TM bioreactor and proper differentiating factors to induce the expression of the TM collagen types. The cell cultures were carried out for one week under static and dynamic conditions. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) were used to assess collagen expression. A Finite Element Model was applied to calculate the stress distribution on the scaffolds under dynamic culture. Nanohydroxyapatite (HA) was used as a filler to change density and tensile strength of *PCL scaffolds. In dynamically cultured *PCL constructs, fibroblast surface marker was overexpressed, and collagen type II was revealed via IHC. Collagen types I, III and IV were also detected. Von Mises stress maps showed that during the bioreactor motion, the maximum stress in *PCL was double that in HA/*PCL scaffolds. By using a *PCL nonwoven scaffold, with suitable physico-mechanical properties, an oscillatory culture, and proper differentiative factors, hMSCs were committed into fibroblast lineage-producing TM-like collagens.

Nanobiocomposites suitable for preparing skin compatible films by flat die extrusion were prepared by using plasticized poly(lactic acid) (PLA), poly(butylene succinate-co-adipate) (PBSA), and Chitin nanofibrils as functional filler. Chitin nanofibrils (CNs) were dispersed in the blends thanks to the preparation of pre-nanocomposites containing poly(ethylene glycol). Thanks to the use of a melt strength enhancer (Plastistrength) and calcium carbonate, the processability and thermal properties of bionanocomposites films containing CNs could be tuned in a wide range. Moreover, the resultant films were flexible and highly resistant. The addition of CNs in the presence of starch proved not advantageous because of an extensive chain scission resulting in low values of melt viscosity. The films containing CNs or CNs and calcium carbonate resulted biocompatible and enabled the production of cells defensins, acting as indirect anti-microbial. Nevertheless, tests made with Staphylococcus aureus and Enterobacter spp. (Gram positive and negative respectively) by the qualitative agar diffusion test did not show any direct anti-microbial activity of the films. The results are explained considering the morphology of the film and the different mechanisms of direct and indirect anti-microbial action generated by the nanobiocomposite based films.

In the cosmetic sector, natural and sustainable products with a high compatibility with skin, thus conjugating wellness with a green-oriented consumerism, are required by the market. Poly(hydroxyalkanoate) (PHA)/starch blends represent a promising alternative to prepare flexible films as support for innovative beauty masks, wearable after wetting and releasing starch and other selected molecules. Nevertheless, preparing these films by extrusion is difficult due to the high viscosity of the polymer melt at the temperature suitable for processing starch. The preparation of blends including poly(butylene succinate-co-adipate) (PBSA) or poly(butylene adipate-co-terephthalate) (PBAT) was investigated as a strategy to better modulate melt viscosity in view of a possible industrial production of beauty mask films. The release properties of films in water, connected to their morphology, was also investigated by extraction trials, infrared spectroscopy and stereo and electron microscopy. Then, the biocompatibility with cells was assessed by considering both mesenchymal stromal cells and keratinocytes. All the results were discussed considering the morphology of the films. This study evidenced the possibility of modulating thanks to the selection of composition and the materials processing of the properties necessary for producing films with tailored properties and processability for beauty masks.

The present review had the aim of describing the methodologies of synthesis and properties of biobased pullulan, a microbial polysaccharide investigated in the last decade because of its interesting potentialities in several applications. After describing the implications of pullulan in nano-technology, biodegradation, compatibility with body and skin, and sustainability, the current applications of pullulan are described, with the aim of assessing the potentialities of this biopolymer in the biomedical, personal care, and cosmetic sector, especially in applications in contact with skin.

It has been demonstrated that three-dimensional (3D) cell culture models represent fundamental tools for the comprehension of cellular phenomena both for normal and cancerous tissues. Indeed, the microenvironment affects the cellular behavior as well as the response to drugs. In this study, we performed a morphological analysis on a hepatocarcinoma cell line, HepG2, grown for 24 days inside a bioartificial hydrogel composed of poly(vinyl alcohol) (PVA) and gelatin (G) to model a hepatocellular carcinoma (HCC) in 3D. Morphological features of PVA/G hydrogels were investigated, resulting to mimic the trabecular structure of liver parenchyma. A histologic analysis comparing the 3D models with HepG2 cell monolayers and tumor specimens was performed. In the 3D setting, HepG2 cells were viable and formed large cellular aggregates showing different morphotypes with zonal distribution. Furthermore, β-actin and α5β1 integrin revealed a morphotype-related expression; in particular, the frontline cells were characterized by a strong immunopositivity on a side border of their membrane, thus suggesting the formation of lamellipodia-like structures apt for migration. Based on these results, we propose PVA/G hydrogels as valuable substrates to develop a long term 3D HCC model that can be used to investigate important aspects of tumor biology related to migration phenomena.

Dysphagia refers to difficulty in swallowing often associated with syndromic disorders. In dysphagic patients’ rehabilitation, tongue motility is usually treated and monitored via simple exercises, in which the tongue is pushed against a depressor held by the speech therapist in different directions. In this study, we developed and tested a simple pressure/force sensor device, named “Tonic Tongue (ToTo)”, intended to support training and monitoring tasks for the rehabilitation of tongue musculature. It consists of a metallic frame holding a ball bearing support equipped with a sterile disposable depressor, whose angular displacements are counterbalanced by extensional springs. The conversion from angular displacement to force is managed using a simple mechanical model of ToTo operation. Since the force exerted by the tongue in various directions can be estimated, quantitative assessment of the outcome of a given training program is possible. A first prototype of ToTo was tested on 26 healthy adults, who were trained for one month. After the treatment, we observed a statistically significant improvement with a force up to 2.2 N (median value) in all tested directions of pushing, except in the downward direction, in which the improvement was slightly higher than 5 N (median value). ToTo promises to be an innovative and reliable device that can be used for the rehabilitation of dysphagic patients. Moreover, since it is a self-standing device, it could be used as a point-of-care solution for in-home rehabilitation management of dysphasia.

The preparation and selection of biobased materials compatible with skin is essential for producing innovative and highly eco-friendly beauty masks. The use of a commercial elastomeric poly(hydroxyalkanoate) and starch was fundamental to select materials for bioplastic films with the necessary resistance in wet conditions, skin compatibility and capacity for a fast release of polysaccharides and similar active and functional molecules. Micrometric calcium carbonate was also used to control the stickiness of film during moulding. Starch release in water was investigated by gravimetric and infrared analyses. The compatibility with skin was investigated via two different in vitro tests based on human keratinocytes and human mesenchymal stromal cells. The materials were highly cytocompatible with skin, enabled immune modulation by keratinocytes and starch release in water up to 49% by weight in 30 min. These outcomes are a good starting point for boosting the production of biobased and biodegradable beauty masks, thus decreasing the impact onto environment of cosmetic products that are currently still mainly produced using petrol-based substrates.

BackgroundTissue engineering appears to be an attractive alternative to the traditional approach in the treatment of fracture non-unions. Mesenchymal stromal cells (MSCs) are considered an appealing cell source for clinical intervention. However, ex vivo cell expansion and differentiation towards the osteogenic lineage, together with the design of a suitable scaffold have yet to be optimized. Major concerns exist about the safety of MSC-based therapies, including possible abnormal overgrowth and potential cancer evolution.AimsWe examined the long-term efficacy and safety of ex vivo expanded bone marrow MSCs, embedded in autologous fibrin clots, for the healing of atrophic pseudarthrosis of the upper limb. Our research work relied on three main issues: use of an entirely autologous context (cells, serum for ex vivo cell culture, scaffold components), reduced ex vivo cell expansion, and short-term MSC osteoinduction before implantation.Methods and findingsBone marrow MSCs isolated from 8 patients were expanded ex vivo until passage 1 and short-term osteo-differentiated in autologous-based culture conditions. Tissue-engineered constructs designed to embed MSCs in autologous fibrin clots were locally implanted with bone grafts, calibrating their number on the extension of bone damage. Radiographic healing was evaluated with short- and long-term follow-ups (range averages: 6.7 and 76.0 months, respectively). All patients recovered limb function, with no evidence of tissue overgrowth or tumor formation.ConclusionsOur study indicates that highly autologous treatment can be effective and safe in the long-term healing of bone non-unions. This tissue engineering approach resulted in successful clinical and functional outcomes for all patients.