Improvement of Osteogenic Differentiation of Mouse Pre-Osteoblastic MC3T3-E1 Cells on Core-Shell Polylactic Acid/Chitosan Electrospun Scaffolds for Bone Defect Repair.

Electrospun hybrid scaffolds composed of synthetic and natural polymers have gained increasing interest in tissue engineering applications over the last decade. In this work, scaffolds composed of polylactic acid electrospun fibers, either treated (P-PLA) or non-treated (PLA) with air-plasma, were coated with high molecular weight chitosan to create a core-shell microfibrous structure. The effective thickness control of the chitosan layer was confirmed by gravimetric, spectroscopic (FTIR-ATR) and morphological (SEM) investigations. The chitosan coating increased the fiber diameter of the microfibrous scaffolds while the tensile mechanical tests, conducted in dry and wet environments, showed a reinforcing action of the coating layer on the scaffolds, in particular when deposited on P-PLA samples. The stability of the Chi coating on both PLA and P-PLA substrates was confirmed by gravimetric analysis, while their mineralization capacity was evaluated though scanning electron microscopy (SEM) and energy-dispersive spectroscopy (EDS) after immersing the scaffolds in simulated body fluids (SBF) at 37 °C for 1 week. Sample biocompatibility was investigated through cell viability assay and SEM analysis on mouse pre-osteoblastic MC3T3-E1 cells grown on scaffolds at different times (1, 7, 14 and 21 days). Finally, Alizarin Red assay and qPCR analysis suggested that the combination of plasma treatment and chitosan coating on PLA electrospun scaffolds influences the osteoblastic differentiation of MC3T3-E1 cells, thus demonstrating the great potential of P-PLA/chitosan hybrid scaffolds for bone tissue engineering applications.

Green and Integrated Wearable Electrochemical Sensor for Chloride Detection in Sweat.

Wearable sensors for sweat biomarkers can provide facile analyte capability and monitoring for several diseases. In this work, a green wearable sensor for sweat absorption and chloride sensing is presented. In order to produce a sustainable device, polylactic acid (PLA) was used for both the substrate and the sweat absorption pad fabrication. The sensor material for chloride detection consisted of silver-based reference, working, and counter electrodes obtained from upcycled compact discs. The PLA substrates were prepared by thermal bonding of PLA sheets obtained via a flat die extruder, prototyped in single functional layers via CO2 laser cutting, and bonded via hot-press. The effect of cold plasma treatment on the transparency and bonding strength of PLA sheets was investigated. The PLA membrane, to act as a sweat absorption pad, was directly deposited onto the membrane holder layer by means of an electrolyte-assisted electrospinning technique. The membrane adhesion capacity was investigated by indentation tests in both dry and wet modes. The integrated device made of PLA and silver-based electrodes was used to quantify chloride ions. The calibration tests revealed that the proposed sensor platform could quantify chloride ions in a sensitive and reproducible way. The chloride ions were also quantified in a real sweat sample collected from a healthy volunteer. Therefore, we demonstrated the feasibility of a green and integrated sweat sensor that can be applied directly on human skin to quantify chloride ions.

Efficacy of poly(lactic acid)/carvacrol electrospun membranes against Staphylococcus aureus and Candida albicans in single and mixed cultures.

Carvacrol (CAR) is one of the most promising essential oil components with antimicrobial activity. New technologies aimed to incorporate this active molecule into carrier matrix to improve the stability and prolong the biological activity. The goal of this study was to investigate the feasibility of incorporating CAR into electrospun membranes of poly(lactic acid) (PLA) for potential applications as active antimicrobial system. To this end, PLA membranes containing homogeneously dispersed CAR were successfully prepared and a series of systematic tests including morpho-mechanical properties, in vitro release rate, and antimicrobial/antibiofilm activities against Staphylococcus aureus and Candida albicans were carried out. The results revealed that CAR has a good compatibility with PLA and acts as a plasticizer, improving flexibility and extensibility of the matrix. The gradual release of CAR from PLA membranes warranted a significant antimicrobial activity up to 144 h and reduced the biofilm production by 92-96 and 88-95% of S. aureus and C. albicans in single and mixed cultures. A strong decrease of cell count, biomass, metabolic activity, and vitality of established 24- and 48-h biofilms were also demonstrated. In conclusion, this work highlights the potential of electrospun nanofibrous membranes as efficient stabilizers-carriers of CAR and opens up interesting perspectives on the use of this system as new tool for skin and wound bacterial-fungal infections.

Antimicrobial additives for poly(lactic acid) materials and their applications: current state and perspectives.

Poly(lactic acid)-based antimicrobial materials received considerable attention as promising systems to control microbial growth. The remarkable physicochemical properties of PLA such as renewability, biodegradability, and US Food and Drug Administration (FDA) approval for clinical use open up interesting perspectives for application in food packaging and biomedical materials. Nowadays, there is an increasing consumer demands for fresh, high-quality, and natural foods packaged with environmentally friendly materials that prolong the shelf life. The incorporation of antimicrobial agents into PLA-based polymers is likely to lead to the next generation of packaging materials. The development of antimicrobial PLA materials as a delivery system or coating for biomedical devices is also advantageous in order to reduce possible dose-dependent side effects and limit the phenomena of antibiotic resistance. This mini-review summarizes the most recent advances made in antimicrobial PLA-based polymers including their preparation, biocidal action, and applications. It also highlights the potential of PLA systems as efficient stabilizers-carriers of various kinds of antimicrobial additives including essential oils and other natural compounds, active particles and nanoparticles, and conventional and synthetic molecules.

Correction to: Antimicrobial additives for poly (lactic acid) materials and their applications: current state and perspectives.

The published online version contains mistake in Tables 1 and 3. instead of using "g/mol" it was presented as "g/mol mol-1" and "g/mol-1".

Poly-l-Lactic Acid (PLLA)-Based Biomaterials for Regenerative Medicine: A Review on Processing and Applications.

Synthetic biopolymers are effective cues to replace damaged tissue in the tissue engineering (TE) field, both for in vitro and in vivo application. Among them, poly-l-lactic acid (PLLA) has been highlighted as a biomaterial with tunable mechanical properties and biodegradability that allows for the fabrication of porous scaffolds with different micro/nanostructures via various approaches. In this review, we discuss the structure of PLLA, its main properties, and the most recent advances in overcoming its hydrophobic, synthetic nature, which limits biological signaling and protein absorption. With this aim, PLLA-based scaffolds can be exposed to surface modification or combined with other biomaterials, such as natural or synthetic polymers and bioceramics. Further, various fabrication technologies, such as phase separation, electrospinning, and 3D printing, of PLLA-based scaffolds are scrutinized along with the in vitro and in vivo applications employed in various tissue repair strategies. Overall, this review focuses on the properties and applications of PLLA in the TE field, finally affording an insight into future directions and challenges to address an effective improvement of scaffold properties.

Behavior of Calcium Phosphate-Chitosan-Collagen Composite Coating on AISI 304 for Orthopedic Applications.

Calcium phosphate/chitosan/collagen composite coating on AISI 304 stainless steel was investigated. Coatings were realized by galvanic coupling that occurs without an external power supply because it begins with the coupling between two metals with different standard electrochemical potentials. The process consists of the co-deposition of the three components with the calcium phosphate crystals incorporated into the polymeric composite of chitosan and collagen. Physical-chemical characterizations of the samples were executed to evaluate morphology and chemical composition. Morphological analyses have shown that the surface of the stainless steel is covered by the deposit, which has a very rough surface. XRD, Raman, and FTIR characterizations highlighted the presence of both calcium phosphate compounds and polymers. The coatings undergo a profound variation after aging in simulated body fluid, both in terms of composition and structure. The tests, carried out in simulated body fluid to scrutinize the corrosion resistance, have shown the protective behavior of the coating. In particular, the corrosion potential moved toward higher values with respect to uncoated steel, while the corrosion current density decreased. This good behavior was further confirmed by the very low quantification of the metal ions (practically absent) released in simulated body fluid during aging. Cytotoxicity tests using a pre-osteoblasts MC3T3-E1 cell line were also performed that attest the biocompatibility of the coating.

Occurrence of Microplastics in Waste Sludge of Wastewater Treatment Plants: Comparison between Membrane Bioreactor (MBR) and Conventional Activated Sludge (CAS) Technologies.

In this study, the presence of microplastics in the sludge of three wastewater treatment plants (WWTPs) was examined. The investigated WWTPs operated based on a conventional activated sludge (CAS) process, with (W1) or without (W2) primary clarification, and a membrane bioreactor process (MBR) (W3). The microplastics (MPs) concentration in the samples of W3 was approximately 81.1 ± 4.2 × 103 particles/kg dry sludge, whereas MPs concentrations in W1 and W2 were 46.0 ± 14.8 × 103 particles/kg dry sludge and 36.0 ± 5.2 × 103 particles/kg dry sludge, respectively. Moreover, MPs mainly consisted of fragments (66-68%) in the CAS plants, whereas the fractions of MPs shapes in the MBR sludge were more evenly distributed, although fiber (47%) was the most abundant fraction. Furthermore, samples from the MBR showed a greater diversity in MPs composition. Indeed, all the main polyesters (i.e., textile fibers and polyethylene terephthalate), polyolefins (i.e., polyethylene and polypropylene) and rubber (i.e., polybutadiene) were observed, whereas only polybutadiene, cellulose acetate and polyester were detected in the CAS plants. These findings confirmed that MPs from wastewater are transferred and concentrated in the waste sludge. This is a critical finding since sludge disposal could become a new pathway for microplastic release into the environment and because MPs might affect the fouling behavior of the membrane.

Effect of Polyhydroxyalkanoate (PHA) Concentration on Polymeric Scaffolds Based on Blends of Poly-L-Lactic Acid (PLLA) and PHA Prepared via Thermally Induced Phase Separation (TIPS).

Hybrid porous scaffolds composed of both natural and synthetic biopolymers have demonstrated significant improvements in the tissue engineering field. This study investigates for the first time the fabrication route and characterization of poly-L-lactic acid scaffolds blended with polyhydroxyalkanoate up to 30 wt%. The hybrid scaffolds were prepared by a thermally induced phase separation method starting from ternary solutions. The microstructure of the hybrid porous structures was analyzed by scanning electron microscopy and related to the blend composition. The porosity and the wettability of the scaffolds were evaluated through gravimetric and water contact angle measurements, respectively. The scaffolds were also characterized in terms of the surface chemical properties via Fourier transform infrared spectroscopy in attenuated total reflectance. The mechanical properties were analyzed through tensile tests, while the crystallinity of the PLLA/PHA scaffolds was investigated by differential scanning calorimetry and X-ray diffraction.

Biocomposite PBAT/lignin blown films with enhanced photo-stability.

Lignin can be obtained as a byproduct during cellulose-rich pulp fibers production and it is habitually treated as waste or intended for low-value destinations. However, due to UV absorption and mechanical properties, lignin can contribute to the fabrication of biodegradable blown films with superior performances. In this study, it was established the suitability of lignin for manufacturing biocomposite PBAT blown films with higher stiffness and photo-oxidation resistance. The effect of the filler concentration on the melt rheological behavior in non-isothermal elongational flow was investigated. The results allowed us to choose the correct filler concentration for producing films through a film blowing operation. The PBAT/lignin blown film composites displayed an increase of the elastic modulus if compared to neat PBAT films without affecting their elongation at break. Furthermore, the filler delayed the photo-oxidative degradation of PBAT hence potentially allowing open-air applications.

Solution-Based Processing for Scaffold Fabrication in Tissue Engineering Applications: A Brief Review.

The fabrication of 3D scaffolds is under wide investigation in tissue engineering (TE) because of its incessant development of new advanced technologies and the improvement of traditional processes. Currently, scientific and clinical research focuses on scaffold characterization to restore the function of missing or damaged tissues. A key for suitable scaffold production is the guarantee of an interconnected porous structure that allows the cells to grow as in native tissue. The fabrication techniques should meet the appropriate requirements, including feasible reproducibility and time- and cost-effective assets. This is necessary for easy processability, which is associated with the large range of biomaterials supporting the use of fabrication technologies. This paper presents a review of scaffold fabrication methods starting from polymer solutions that provide highly porous structures under controlled process parameters. In this review, general information of solution-based technologies, including freeze-drying, thermally or diffusion induced phase separation (TIPS or DIPS), and electrospinning, are presented, along with an overview of their technological strategies and applications. Furthermore, the differences in the fabricated constructs in terms of pore size and distribution, porosity, morphology, and mechanical and biological properties, are clarified and critically reviewed. Then, the combination of these techniques for obtaining scaffolds is described, offering the advantages of mimicking the unique architecture of tissues and organs that are intrinsically difficult to design.

Use of Biochar as Filler for Biocomposite Blown Films: Structure-Processing-Properties Relationships.

In this work, biocomposite blown films based on poly(butylene adipate-co-terephthalate) (PBAT) as biopolymeric matrix and biochar (BC) as filler were successfully fabricated. The materials were subjected to a film-blowing process after being compounded in a twin-screw extruder. The preliminary investigations conducted on melt-mixed PBAT/BC composites allowed PBAT/BC 5% and PBAT/BC 10% to be identified as the most appropriate formulations to be processed via film blowing. The blown films exhibited mechanical performances adequate for possible application as film for packaging, agricultural, and compost bags. The addition of BC led to an improvement of the elastic modulus, still maintaining high values of deformation. Water contact angle measurements revealed an increase in the hydrophobic behavior of the biocomposite films compared to PBAT. Additionally, accelerated degradative tests monitored by tensile tests and spectroscopic analysis revealed that the filler induced a photo-oxidative resistance on PBAT by delaying the degradation phenomena.

Combining carvacrol and nisin in biodegradable films for antibacterial packaging applications.

In this work, the feasibility of antibacterial biopolymeric films containing carvacrol (CRV) and a nisin commercial formulation (Nis) for potential food packaging applications was investigated. As polymer matrix, a commercial biodegradable polymer formulation of Mater-Bi (MB) was chosen due to its significant food packaging applications. CRV and Nis were chosen due to their well-established antibacterial properties and their potential synergistic effect. MB/CRV, MB/Nis, and MB/CRV/Nis systems were produced by melt mixing and compression molding. The mechanical properties of the films were evaluated by tensile tests. Differential scanning calorimetry was assessed aiming at investigating the effect of the two compounds and their mixture on the thermal properties of MB. The release profile of CRV and Nis from the MB-based films was evaluated in water at 4 °C by UV-Vis measurements and it was fitted with a power-law model. The antibacterial activity of MB-based films was tested in vitro against Listeria monocytogenes, Salmonella enteritidis, Escherichia coli, and Staphylococcus aureus. The combination of CRV and Nis strongly affected the properties of the MB-based films and ensured higher antibacterial activity if compared to MB/CRV and MB/Nis systems.

Physical and biological properties of electrospun poly(d,l-lactide)/nanoclay and poly(d,l-lactide)/nanosilica nanofibrous scaffold for bone tissue engineering.

Electrospun scaffolds exhibiting high physical performances with the ability to support cell attachment and proliferation are attracting more and more scientific interest for tissue engineering applications. The inclusion of inorganic nanoparticles such as nanosilica and nanoclay into electrospun biopolymeric matrices can meet these challenging requirements. The silica and clay incorporation into polymeric nanofibers has been reported to enhance and improve the mechanical properties as well as the osteogenic properties of the scaffolds. In this work, for the first time, the physical and biological properties of polylactic acid (PLA) electrospun mats filled with different concentrations of nanosilica and nanoclay were evaluated and compared. The inclusion of the particles was evaluated through morphological investigations and Fourier transform infrared spectroscopy. The morphology of nanofibers was differently affected by the amount and kind of fillers and it was correlated to the viscosity of the polymeric suspensions. The wettability of the scaffolds, evaluated through wet contact angle measurements, slightly increased for both the nanocomposites. The crystallinity of the systems was investigated by differential scanning calorimetry highlighting the nucleating action of both nanosilica and nanoclay on PLA. Scaffolds were mechanically characterized with tensile tests to evaluate the reinforcing action of the fillers. Finally, cell culture assays with pre-osteoblastic cells were conducted on a selected composite scaffold in order to compare the cell proliferation and morphology with that of neat PLA scaffolds. Based on the results, we can convince that nanosilica and nanoclay can be both considered great potential fillers for electrospun systems engineered for bone tissue regeneration.

Core-shell PLA/Kef hybrid scaffolds for skin tissue engineering applications prepared by direct kefiran coating on PLA electrospun fibers optimized via air-plasma treatment.

Over the recent years, there is a growing interest in electrospun hybrid scaffolds composed of synthetic and natural polymers that can support cell attachment and proliferation. In this work, the physical and biological properties of polylactic acid (PLA) electrospun mats coated with kefiran (Kef) were evaluated. Gravimetric, spectroscopic (FTIR-ATR) and morphological investigations via scanning electron microscopy confirmed the effective formation of a thin kefiran layer wrapped on the PLA fibers with an easy-tunable thickness. Air plasma pre-treatment carried out on PLA (P-PLA) affected both the morphology and the crystallinity of Kef coating as confirmed by differential scanning calorimetry and X-ray diffraction analyses. Scaffolds were mechanically characterized with tensile tests to evaluate the reinforcing action of the Kef coating. The water resistance of Kefiran coating in distilled water at 37 °C evaluated on both PLA/Kef and P-PLA/Kef was carried out by gravimetric and morphological analyses. Finally, cell culture assays with embryonic fibroblast cells were conducted on selected hybrid scaffolds to compare the cell proliferation, morphology, and collagen production with PLA and P-PLA electrospun scaffolds. Based on the results, we can demonstrate that direct coating of PLA from Kef/water solutions is an effective approach to prepare hybrid scaffolds with tunable properties and that the plasma pre-treatment enhances the affinity between PLA and Kefiran. In vitro tests demonstrated the great potential of PLA/Kef hybrid scaffolds for skin tissue engineering.

PBAT Based Composites Reinforced with Microcrystalline Cellulose Obtained from Softwood Almond Shells.

This study explores the processability, mechanical, and thermal properties of biocompostable composites based on poly (butylene adipate-co-terephthalate) (PBAT) as polymer matrix and microcrystalline cellulose (MCC) derived from softwood almond (Prunus dulcis) shells (as-MCC) as filler at two different weight concentration, i.e., 10 wt% and 20 wt%. The materials were processed by melt mixing and a commercial MCC (c-MCC) was used as filler comparison. The fibrillar shape of as-MCC particles was found to change the rheological behavior of PBAT, particularly at the highest concentration. The melt mixing processing allowed obtaining a uniform dispersion of both kinds of fillers, slightly reducing the L/D ratio of as-MCC fibers. The as-MCC particles led to a higher increase of the elastic modulus of PBAT if compared to the c-MCC counterparts. Both the MCC fillers caused a drastic reduction of the elongation at break, although it was higher than 120% also at the highest filler concentrations. DSC analysis revealed that both MCC fillers poorly affected the matrix crystallinity, although as-MCC induced a slight PBAT crystallinity increase from 8.8% up to 10.9% for PBAT/as-MCC 20%. Therefore, this work demonstrates the great potential of MCC particles derived from almond shells as filler for biocompostable composites fabrication.

Engineered Membranes for Residual Cell Trapping on Microfluidic Blood Plasma Separation Systems: A Comparison between Porous and Nanofibrous Membranes.

Blood-based clinical diagnostics require challenging limit-of-detection for low abundance, circulating molecules in plasma. Micro-scale blood plasma separation (BPS) has achieved remarkable results in terms of plasma yield or purity, but rarely achieving both at the same time. Here, we proposed the first use of electrospun polylactic-acid (PLA) membranes as filters to remove residual cell population from continuous hydrodynamic-BPS devices. The membranes hydrophilicity was improved by adopting a wet chemistry approach via surface aminolysis as demonstrated through Fourier Transform Infrared Spectroscopy and Water Contact Angle analysis. The usability of PLA-membranes was assessed through degradation measurements at extreme pH values. Plasma purity and hemolysis were evaluated on plasma samples with residual red blood cell content (1, 3, 5% hematocrit) corresponding to output from existing hydrodynamic BPS systems. Commercially available membranes for BPS were used as benchmark. Results highlighted that the electrospun membranes are suitable for downstream residual cell removal from blood, permitting the collection of up to 2 mL of pure and low-hemolyzed plasma. Fluorometric DNA quantification revealed that electrospun membranes did not significantly affect the concentration of circulating DNA. PLA-based electrospun membranes can be combined with hydrodynamic BPS in order to achieve high volume plasma separation at over 99% plasma purity.

Innovative, ecofriendly biosorbent-biodegrading biofilms for bioremediation of oil- contaminated water.

Immobilization of microorganisms capable of degrading specific contaminants significantly promotes bioremediation processes. In this study, innovative and ecofriendly biosorbent-biodegrading biofilms have been developed in order to remediate oil-contaminated water. This was achieved by immobilizing hydrocarbon-degrading gammaproteobacteria and actinobacteria on biodegradable oil-adsorbing carriers, based on polylactic acid and polycaprolactone electrospun membranes. High capacities for adhesion and proliferation of bacterial cells were observed by scanning electron microscopy. The bioremediation efficiency of the systems, tested on crude oil and quantified by gas chromatography, showed that immobilization increased hydrocarbon biodegradation by up to 23 % compared with free living bacteria. The resulting biosorbent biodegrading biofilms simultaneously adsorbed 100 % of spilled oil and biodegraded more than 66 % over 10 days, with limited environmental dispersion of cells. Biofilm-mediated bioremediation, using eco-friendly supports, is a low-cost, low-impact, versatile tool for bioremediation of aquatic systems.

Effect of hydroxyapatite concentration and size on morpho-mechanical properties of PLA-based randomly oriented and aligned electrospun nanofibrous mats.

The growing demand for nanofibrous biocomposites able to provide peculiar properties requires systematic investigations of processing-structure-property relationships. Understanding the morpho-mechanical properties of an electrospun scaffold as a function of the filler features and mat microstructure can aid in designing these systems. In this work, the reinforcing effect of micrometric and nanometric hydroxyapatite particles in polylactic acid-based electrospun scaffold presenting random and aligned fibers orientation, was evaluated. The particles incorporation was investigated trough Fourier transform infrared spectroscopy in attenuated total reflectance. The morphology of the nanofibers was analyzed through scanning electron microscopy and it was correlated with the viscosity of polymeric solutions studied by rheological measurements. Scaffolds were mechanical characterized with tensile tests in order to find a correlation between the preparation method and the strength of the mats. The influence of hydroxyapatite particles on the crystallinity of the composites was investigated by differential scanning calorimetry. Finally, cell culture assays with pre-osteoblatic cells were conducted on a selected composite scaffold in order to compare the cell proliferation and morphology with that of polylactic acid scaffolds. Based on the results, we can prove that polylactic acid/hydroxyapatite composites can be one of the biomaterials with the greatest potential for bone tissue regeneration.

Flexible mats as promising antimicrobial systems via integration of Thymus capitatus (L.) essential oil into PLA.

Aim: To develop electrospun mats loaded with Thymus capitatus (L.) essential oil (ThymEO) and to study their morpho-mechanical and antimicrobial properties. Materials & methods: Poly(lactic acid) (PLA) mats containing ThymEO were prepared by electrospinning. The effect of ThymEO on the morpho-mechanical properties of fibers was assayed by scanning electron microscopy and dynamometer measurements. The antimicrobial activity of ThymEO delivered either in liquid or vapor phase was assessed through killing curves and invert Petri dishes method. The cytotoxicity was also investigated. Results: The mechanical properties were enhanced by integrating ThymEO into PLA. Both liquid and vapors of ThymEO released from mats caused reductions of microbial viable cells. Negligible cytotoxicity was demonstrated. Conclusion: PLA/ThymEO delivery systems could be suitable for treating microbial infections.