Development of highly bacterial filtration efficient and antibacterial cellulose acetate/gum rosin composite nanofibers for facemask application.

Cellulose acetate (CA) is a non-toxic, renewable, and biodegradable polymeric material that can be effectively electrospuned into bacterial filtration efficient nanofiber membrane for face mask application. However, its fragile and non-antibacterial nature influenced its scalability. In this context, natural antibacterial gum rosin (GR) additive can be explored. Therefore, the present study aimed to produce a CA/GR composite nanofibers membrane for the finest bacterial filtration, excellent antibacterial moiety, and improved tensile properties for facemask application. Hence, in this work, we have studied the effect of GR concentrations (0-15 g) on the needleless electrospinning behavior and fibers' morphology through rheology, electrical conductivity, and SEM analysis. These analyses revealed that GR significantly affects the fibers' spinning behavior, morphology, and diameter of the produced fibers. Later, ATR-FTIR spectroscopy mapped the functional changes in the produced nanofibers that affirmed the integration of GR with CA polymer. This modification resulted in a 3-fold rise in tensile strength and an 11-fold decline in elongation% in 15 g CA/GR composite nanofibers membrane than the control sample. Furthermore, it has shown 98.79 ± 0.10% bacterial filtration efficiency and âˆ¼ 93 % reduction in Staphylococcus Aureus and Klebsiella Pneumoniae bacterial growth, elucidating a high-efficiency level for potential facemask application.

Bacterial Biofilms on Polyamide Nanofibers: Factors Influencing Biofilm Formation and Evaluation.

Electrospun polyamide (PA) nanofibers have great potential for medical applications (in dermatology as antimicrobial compound carriers or surgical sutures). However, little is known about microbial colonization on these materials. Suitable methods need to be chosen and optimized for the analysis of biofilms formed on nanofibers and the influence of their morphology on biofilm formation. We analyzed 11 PA nanomaterials, both nonfunctionalized and functionalized with AgNO3, and tested the formation of a biofilm by clinically relevant bacteria (Escherichia coli CCM 4517, Staphylococcus aureus CCM 3953, and Staphylococcus epidermidis CCM 4418). By four different methods, it was confirmed that all of these bacteria attached to the PAs and formed biofilms; however, it was found that the selected method can influence the outcomes. For studying biofilms formed by the selected bacteria, scanning electron microscopy, resazurin staining, and colony-forming unit enumeration provided appropriate and comparable results. The values obtained by crystal violet (CV) staining were misleading due to the binding of the CV dye to the PA structure. In addition, the effect of nanofiber morphology parameters (fiber diameter and air permeability) and AgNO3 functionalization significantly influenced biofilm maturation. Furthermore, the correlations between air permeability and surface density and fiber diameter were revealed. Based on the statistical analysis, fiber diameter was confirmed as a crucial factor influencing biofilm formation (p ≤ 0.01). The functionalization of PAs with AgNO3 (from 0.1 wt %) effectively suppressed biofilm formation. The PA functionalized with a concentration of 0.1 wt % AgNO3 influenced the biofilm equally as nonfunctionalized PA 8% 2 g/m2. Therefore, biofilm formation could be affected by the above-mentioned morphology parameters, and ultimately, the risk of infections from contaminated medical devices could be reduced.

Antibacterial properties and cytocompatibility of biobased nanofibers of fish scale gelatine, modified polylactide, and freshwater clam shell.

We report herein new nanofibers prepared from fish scale gelatine (FSG), modified polylactide (MPLA), and a natural antibacterial agent of freshwater clam (Corbicula fluminea Estefanía) shell powder (FCSP). A preparation of FSG from Mullet scales is also described. To improve the biocompatibility and antibacterial activity of the non-woven nanofibers, MPLA/FCSP was added to enhance their antibacterial properties. FSG was then combined with MPLA/FCSP using an electrospinning technique to improve the biocompatibility of the as-fabricated 100-500-nm-diameter non-woven MPLA/FCSP/FSG nanofibers. The resulting tensile properties and morphological characteristics indicated enhanced adhesion among FSG, FCSP, and MPLA in the MPLA/FCSP/FSG nanofibers, as well as improved water resistance and tensile strength, compared with the PLA/FSG nanofibers. MTT assay, cell-cycle, and apoptosis analyses showed that both PLA/FSG and MPLA/FCSP/FSG nanofibers had good biocompatibility. Increasing the FSG content in PLA/FSG and MPLA/FCSP/FSG nanofibers enhanced cell proliferation and free-radical scavenging ability, but did not affect cell viability. Quantitative analysis of bacteria inhibition revealed that FCSP imparts antibacterial activity.

Viability improvement of Bifidobacterium animalis Bb12 by encapsulation in chitosan/poly(vinyl alcohol) hybrid electrospun fiber mats.

For the improvement of the probiotics' viability, a novel nanofiber mats consisting of chitosan (CS)/poly(vinyl alcohol) (PVA) loaded with the probiotic Bifidobacterium animalis subsp. lactis Bb12 and inulin (INU) as a prebiotic were fabricated via electrospinning approach. Scanning electron microscopy revealed that cells were well encapsulated in nanofibers with a diameter ranging from 117.5 ± 70.6 to 217.6 ± 62.7 nm and fluorescence microscopy images also revealed the viability of bacteria. Thermal analysis of fibrous mats proposed potential use of them in heat-treated foods owing to high melting temperatures (>200 °C). Moreover, in comparison to free cells, the survivability of cells especially enclosed in CS/PVA/INU electrospun fibers were remarkably increased under simulated gastric (SGF) and intestinal fluids (SIF). This study shows that the electrospun fiber mats are acceptable platforms for the protection of living probiotic cells and provide a substitute way for the expansion of functional food.

Novel electrospun fibers with incorporated commensal bacteria for potential preventive treatment of the diabetic foot.

AIM: A novel electrospun biocompatible nanofibrous material loaded with commensal bacteria for potential preventive treatment of the diabetic foot was developed. MATERIALS & METHODS: Two biocompatible polymers (carboxymethylcellulose and polyethylene oxide) were combined with a bacterium isolate from the skin located between the toes of a healthy adult (identified using a matrix-assisted laser desorption/ionization mass spectrometry-based method as a strain of Staphylococcus epidermidis). Higher bacteria loads in the material were assured through their encapsulation in polyethylenimine. The nanofibrous material was characterized using scanning electron microscopy, zeta-potential measurements and through evaluation of cell growth and viability. RESULTS & DISCUSSION: nanometer formation was confirmed using scanning electron microscopy, while the zeta-potential measurements revealed successful bacteria encapsulation. Viable and sufficiently growing cells were confirmed prior and after their incorporation. CONCLUSION: The prepared materials were proven suitable to deliver viable commensal bacteria in a comparable share to the Staphylococcaceae in the foot microbiome making this approach promising for preventive diabetic foot treatment.

Encapsulation of living bacteria in electrospun cyclodextrin ultrathin fibers for bioremediation of heavy metals and reactive dye from wastewater.

Cyclodextrins (CD) are cyclic oligosaccharides produced from the enzymatic degradation of starch as a white powder form; on the other hand, they can be transformed into ultrathin electrospun fiber form by electrospinning technique. The electrospun cyclodextrin fibers (CD-F) can be quite attractive materials to encapsulate bacteria for bioremediation purposes. For instance, CD-F not only serve as a carrier matrix but also it serves as a feeding source for the encapsulated bacteria. In the present study, we demonstrate a facile approach by encapsulation of bacteria into CD-F matrix for wastewater treatment application. The natural and non-toxic properties of CD-F render a better bacterial viability for fibrous biocomposite. The encapsulated bacteria in CD-F exhibit cell viability for more than 7days at 4°C storage condition. Furthermore, we have tested the bioremediation capability of bacteria/CD-F biocomposite for the treatment of heavy metals (Nickel(II) and Chromium(VI)) and textile dye (Reactive Black 5, RB5). The bacteria/CD-F biocomposite has shown removal efficiency of Ni(II), Cr(VI) and RB5 as 70±0.2%, 58±1.4% and 82±0.8, respectively. As anticipated, the pollutants removal capabilities of the bacteria/CD-F was higher compare to free bacteria since bacteria can use CD as an extra carbon source which promotes their growth rate. This study demonstrates that CD-F are suitable platforms for the encapsulation of bacterial cells to develop novel biocomposites that have bioremediation capabilities for wastewater treatment.

Bacteria encapsulated electrospun nanofibrous webs for remediation of methylene blue dye in water.

In this study, preparation and application of novel biocomposite materials that were produced by encapsulation of bacterial cells within electrospun nanofibrous webs are described. A commercial strain of Pseudomonas aeruginosa which has methylene blue (MB) dye remediation capability was selected for encapsulation, and polyvinyl alcohol (PVA) and polyethylene oxide (PEO) were selected as the polymer matrices for the electrospinning of bacteria encapsulated nanofibrous webs. Encapsulation of bacterial cells was monitored by scanning electron microscopy (SEM) and fluorescence microscopy, and the viability of encapsulated bacteria was checked by live/dead staining and viable cell counting assay. Both bacteria/PVA and bacteria/PEO webs have shown a great potential for remediation of MB, yet bacteria/PEO web has shown higher removal performances than bacteria/PVA web, which was probably due to the differences in the initial viable bacterial cells for those two samples. The bacteria encapsulated electrospun nanofibrous webs were stored at 4°C for three months and they were found as potentially storable for keeping encapsulated bacterial cells alive. Overall, the results suggest that electrospun nanofibrous webs are suitable platforms for preservation of living bacterial cells and they can be used directly as a starting inoculum for bioremediation of water systems.

Pectin-non-starch nanofibers biocomposites as novel gastrointestinal-resistant prebiotics.

Incorporation of nanofibers of chitin (NC), lignocellulose (NLC) and bacterial cellulose (BNC) in pectin was studied to improve prebiotic activity and gastrointestinal resistance of the pectin-nanofibers biocomposites for protection of probiotics under simulated gastrointestinal conditions. The biocomposites were prepared using various compositions of pectin and nanofibers, which were designed using D-optimal mixture method. The incorporation of the nanofibers in pectin led to a slow degradation of the pectin-nanofibers biocomposites in contrast to their rapid swelling. AFM analysis indicated the homogenous distribution of interconnected nanofibers network structure in the pectin-nanofibers biocomposite. FTIR spectra demonstrated fabrication of the biocomposites based on the inter- and intra-molecular hydrogen bonding and ionic interaction of pectin-Ca2+. XRD patterns revealed the amorphous structures of the biocomposites as compared to the crystalline structures of the nanofibers. Among the compositions, the optimal compositions were as follows: 60% pectin+40% NC, 50% pectin+50% NLC and 60% pectin+40% BNC, where the prebiotic score, probiotic survival under simulated gastric and intestinal conditions were optimum. The optimal biocomposite pectin-NC exhibited the highest survival of the entrapped probiotic bacteria under simulated gastric (97.7%) and intestinal (95.8%) conditions when compared with the corresponding to free cells (76.2 and 73.4%).

Ozone Gas as a Benign Sterilization Treatment for PLGA Nanofiber Scaffolds.

The use of electrospun nanofibers for tissue engineering and regenerative medicine applications is a growing trend as they provide improved support for cell proliferation and survival due, in part, to their morphology mimicking that of the extracellular matrix. Sterilization is a critical step in the fabrication process of implantable biomaterial scaffolds for clinical use, but many of the existing methods used to date can negatively affect scaffold properties and performance. Poly(lactic-co-glycolic acid) (PLGA) has been widely used as a biodegradable polymer for 3D scaffolds and can be significantly affected by current sterilization techniques. The aim of this study was to investigate pulsed ozone gas as an alternative method for sterilizing PLGA nanofibers. The morphology, mechanical properties, physicochemical properties, and response of cells to PLGA nanofiber scaffolds were assessed following different degrees of ozone gas sterilization. This treatment killed Geobacillus stearothermophilus spores, the most common biological indicator used for validation of sterilization processes. In addition, the method preserved all of the characteristics of nonsterilized PLGA nanofibers at all degrees of sterilization tested. These findings suggest that ozone gas can be applied as an alternative method for sterilizing electrospun PLGA nanofiber scaffolds without detrimental effects.

Effects of the anaerobic respiration of Shewanella oneidensis MR-1 on the stability of extracellular U(VI) nanofibers.

Uranium (VI) is considered to be one of the most widely dispersed and problematic environmental contaminants, due in large part to its high solubility and great mobility in natural aquatic systems. We previously reported that under anaerobic conditions, Shewanella oneidensis MR-1 grown in medium containing uranyl acetate rapidly accumulated long, extracellular, ultrafine U(VI) nanofibers composed of polycrystalline chains of discrete meta-schoepite (UO(3)·2H2O) nanocrystallites. Wild-type MR-1 finally transformed the uranium (VI) nanofibers to uranium (IV) nanoparticles via further reduction. In order to investigate the influence of the respiratory chain in the uranium transformation process, a series of mutant strains lacking a periplasmic cytochrome MtrA, outer membrane (OM) cytochrome MtrC and OmcA, a tetraheme cytochrome CymA anchored to the cytoplasmic membrane, and a trans-OM protein MtrB, were tested in this study. Although all the mutants produced U(VI) nanofibers like the wild type, the transformation rates from U(VI) nanofibers to U(IV) nanoparticles varied; in particular, the mutant with deletion in tetraheme cytochrome CymA stably maintained the uranium (VI) nanofibers, suggesting that the respiratory chain of S. oneidensis MR-1 is probably involved in the stability of extracellular U(VI) nanofibers, which might be easily treated via the physical processes of filtration or flocculation for the remediation of uranium contamination in sediments and aquifers, as well as the recovery of uranium in manufacturing processes.

Poly(vinyl alcohol)/sodium alginate/layered silicate based nanofibrous mats for bacterial inhibition.

Poly(vinyl alcohol) (PVA)/sodium alginate (ALG)/organic rectorite (OREC) composite nanofibrous mats are fabricated by electrospinning aqueous solutions with different mixing ratios. Both good fiber shape and three-dimensional structure of nanofibrous mats can be observed by Field Emission Scanning Electron Microscopy. Energy-dispersive X-ray spectroscopy shows the existence of OREC in the as-spun composite mats. In addition, small-angle X-ray diffraction confirms that the interlayer of OREC is intercalated by ALG/PVA chains, and the distance between OREC interlayers is increased from 4.50 to 4.74 nm. Wide angle X-ray diffraction and Fourier transform infrared spectra further verify the intercalation is between polymers and layered silicate. Moreover, the thermal gravimetric analysis shows that the addition of OREC has only a small effect on the thermal stability of composites. Furthermore, the antibacterial experiments illustrate that OREC can enhance the bacterial inhibition ability of nanofibrous mats against Escherichia coli and Staphylococcus aureus.

Regenerated silk fibroin nanofibrous matrices treated with 75% ethanol vapor for tissue-engineering applications.

As an excellent biocompatible and biodegradable protein polymer, silk fibroin (SF) has found wide applications, particularly serving as therapeutic agent for tissue-engineering applications, on which both post-spin treatment and sterilization processing are crucial to drug-loaded matrices. To find a safe, effective and appropriate post-spin treatment and sterilization approach for drug-loaded biomaterial matrices is one of the major problems in the field of tissue engineering at present. In this work, a simple, safe and effective approach skillfully integrating post-spin treatment with sterilization processing was developed to drug-loaded SF nanofibrous matrices. Electrospun SF nanofibrous matrices from its aqueous solution were post-treated with 75% ethanol vapor. (13)C-NMR and WAXD analysis demonstrated that such post-spin treatment rendered the structure of SF nanofibrous matrices transform from the silk I form to the silk II form. Furthermore, biological assays suggested that as-treated SF nanofibrous matrices significantly promoted the development of murine connective tissue fibroblasts. Skillfully integrated with novel sterilization processing, 75% ethanol vapor treatment could be a potential approach to designing and fabricating diverse drug-loaded SF nanofibrous matrices serving as therapeutic agents for tissue-engineering applications in that it can effectively protect the drug from losing compared with traditional post-spin treatment and sterilization processing.

Bacterial cellulose-based materials and medical devices: current state and perspectives.

Bacterial cellulose (BC) is a unique and promising material for use as implants and scaffolds in tissue engineering. It is composed of a pure cellulose nanofiber mesh spun by bacteria. It is remarkable for its strength and its ability to be engineered structurally and chemically at nano-, micro-, and macroscales. Its high water content and purity make the material biocompatible for multiple medical applications. Its biocompatibility, mechanical strength, chemical and morphologic controllability make it a natural choice for use in the body in biomedical devices with broader application than has yet been utilized. This paper reviews the current state of understanding of bacterial cellulose, known methods for controlling its physical and chemical structure (e.g., porosity, fiber alignment, etc.), biomedical applications for which it is currently being used, or investigated for use, challenges yet to be overcome, and future possibilities for BC.

Production of peritrichate bacterionanofibers and their proteinaceous components by Acinetobacter sp. Tol 5 cells affected by growth substrates.

The toluene-degrading bacterium Acinetobacter sp. Tol 5 is highly adhesive through cell-surface nanofibers. Previously, we identified two morphologically distinct nanofibers on Tol 5 cells, namely, nonperitrichate anchor-like and peritrichate pilus-like nanofibers. In the present study, the application of improved electron microscopy techniques enabled discrimination of three distinct types of peritrichate nanofibers on Tol 5 cells. Interestingly, production of these nanofibers was affected by the available growth substrate. Thick, long, straight nanofibers a, which were present on cells grown on toluene, lactate, and ethanol, were not observed on cells grown on triacylglycerol (TAG). In contrast, cells grown on TAG were covered with long, curved nanofibers c, which only existed sparsely on cells grown on toluene, lactate, and ethanol. Thin, short, straight nanofibers b were found densely covering the margin of cells grown on all four growth substrates. SDS-PAGE of Tol 5 cell-surface proteins detected a protein of 17.5 kDa that was expressed at a high level on ethanol, but was undetectable on TAG. Conversely, a 26kDa protein was identified that was exclusively expressed on TAG, but was only faintly expressed by cells grown on the other substrates. Based on N-terminal amino acid sequences, the 17.5 and 26 kDa proteins were identified as the major subunits of type 1 and Fil pili, respectively, which are typical bacterionanofibers. From these results, we deduced that nanofibers a and c are type 1 and Fil pili, respectively. The adhesiveness of Tol 5 cells was low only when they were grown on TAG.

Potential of biocellulose nanofibers production from agricultural renewable resources: preliminary study.

In the present preliminary study, we report results for the biocellulose nanofibres production by Gluconacetobacter xylinus. Production was examined by utilizing different feedstocks of single sugars and sugar mixtures with compositions similar to the acid hydrolyzates of different agriculture residues. Profiles for cell proliferation, sugar consumption, and the subsequent pH changes were thoroughly analyzed. Highest biocellulose production of 5.65 g/L was achieved in fructose medium with total sugar consumption of 95.57%. Moreover, the highest production using sugar mixtures was 5.2 g/L, which was achieved in feedstock with composition identical to the acid hydrolyzate of wheat straws. This represented the highest biocellulose yield of 17.72 g/g sugars compared with 14.77 g/g fructose. The lowest production of 1.1 and 1.75 g/L were obtained in xylose and glucose media, respectively, while sucrose and arabinose media achieved relatively higher production of 4.7 and 4.1 g/L, respectively. Deviation in pH of the fermentation broths from the optimum value of 4-5 generally had marked effect on biocellulose production with single sugars in feedstock. However, the final pH values recorded in the different sugar mixtures were approximately 3.3-3.4, which had lower effect on production hindrance. Analyzing profiles for sugars' concentrations and cell growth showed that large amount of the metabolized sugars were mainly utilized for bacterial cell growth and maintenance, rather than biocellulose production. This was clearly observed with single sugars of low production, while sugar consumption was rather utilized for biocellulose production with sugar mixtures. Results reported in this study demonstrate that agriculture residues might be used as potential feedstocks for the biocellulose nanofibres production. Not only this represents a renewable source of feedstock, but also might lead to major improvements in production if proper supplements and control were utilized in the fermentation process.

Preparation of nanofibers containing the microalga Spirulina (Arthrospira).

Spirulina is a microalga which offers biological functions highly favorable for tissue engineering. Highly porous scaffolds can be produced by electrospinning containing biomass of Spirulina. The goal of this contribution was therefore to establish spinning conditions allowing to produce well defined nanofibers with diameters down to about 100 nm and to produce nanofibers with various concentration of the biomass for subsequent studies in tissue engineering applications. The experimental results reveal that the blend system PEO/biomass is behaved surprisingly well in electrospinning. Very thin bead-free nanofibers with diameters of about 110 nm can be produced for different biomass contents of up to 67 wt.% of the nanofibers and for PEO concentrations in the spinning solution well below 4 wt.%. These results suggest to us the use of the biomass containing nanofibers as extracellular matrices for stem cell culture and future treatment of spinal chord injury.