Detailed Structural Analyses of Nanofibrillated Bacterial Cellulose and Its Application as Binder Material for a Display Device.

Nanofibrillated bacterial cellulose (NFBC) is produced by culturing a cellulose-producing bacterium under agitated aerobic conditions in a carboxymethylcellulose (CMC)-supplemented medium. Detailed structural analyses revealed that NFBC fiber widths varied with the degree of substitution of the CMC used, and zeta potential values decreased with the increment of CMC concentration in the medium. Transmission electron microscopy observation after immunostaining demonstrated that CMC molecules were present on the NFBC microfibril surfaces. We tested NFBC for utility as a binder for a display device that uses electrochromic (EC) material. Introduction of a quaternary ammonium group into the EC molecules enhanced their interactions with the negatively charged NFBC microfibrils. A casting process homogeneously adsorbed the EC molecules onto the surface of a transparent electrode with NFBC. A homogeneous color change was successfully observed upon applying an electric field, suggesting that NFBC could be used as a binder material for uniform surface adsorption.

One-Step Production of Amphiphilic Nanofibrillated Cellulose Using a Cellulose-Producing Bacterium.

Nanofibrillated bacterial cellulose (NFBC) is produced by culturing a cellulose-producing bacterium (Gluconacetobacter intermedius NEDO-01) with rotation or agitation in medium supplemented with carboxymethylcellulose (CMC). Despite a high yield and dispersibility in water, the product immediately aggregates in organic solvents. To broaden its applicability, we prepared amphiphilic NFBC by culturing strain NEDO-01 in medium supplemented with hydroxyethylcellulose or hydroxypropylcellulose instead of CMC. Transmission electron microscopy analysis revealed that the resultant materials (HE-NFBC and HP-NFBC, respectively) comprised relatively uniform fibers with diameters of 33 ± 7 and 42 ± 8 nm, respectively. HP-NFBC was dispersible in polar organic solvents such as methanol, acetone, isopropyl alcohol, acetonitrile, tetrahydrofuran (THF), and dimethylformamide, and was also dispersible in poly(methyl methacrylate) (PMMA) by solvent mixing using THF. HP-NFBC/PMMA composite films were highly transparent and had a higher tensile strength than neat PMMA film. Thus, HP-NFBC has a broad range of applications, including as a filler material.

Low Molecular Weight Branched PEI Binding to Linear DNA.

Polyethylenimine (PEI) is one of the most versatile non-viral vectors used in gene therapy, especially for delivering plasmid DNA to human cells. However, a good understanding of PEI binding to DNA, the fundamental basis for the functioning of PEI as a vector, has been missing in the literature. In this study, PEI (branched, 600 Da) binding to DNA was examined by isothermal titration calorimetry (ITC), quartz crystal microbalance (QCM) and a complementary set of analysis tools. We demonstrated that a separation between the binding heat and the condensation heat is needed and that the excluded site model should be used for PEI binding stage in the ITC analysis. The equilibrium constant for PEI binding to DNA was determined to be 2.5×105 M-1 from the ITC analysis, and as 2.3×105 M-1 from the QCM analysis. Additionally, we suggested that the 600 Da branched PEI binds to the major groove of DNA and the rearrangement of PEI on DNA may be difficult to occur because of the small dissociation rate. The binding analysis presented here can be employed to improve our understanding of the functioning of PEI and PEI-like non-viral vectors.

Cellulose/glucomannan blends prepared from aqueous sodium hydroxide solution and their practical use as food materials.

Aqueous sodium hydroxide solution is one of the simplest and most environmentally friendly solvents of cellulose. Regenerated cellulose/glucomannan blends were prepared from an aqueous sodium hydroxide solution, and the mechanical properties and structure of the blends were investigated. In addition, the noodle-shaped blends were sensory evaluated as food materials. The blends exhibited porous structures, which corresponded with high water and oil absorption. The strength and modulus of the blends were markedly low, probably due to the highly porous structure. The viscoelastic measurement indicated that cellulose and glucomannan existed in a rubbery state under wet conditions. The blends had a good oral sensation, probably derived from the remarkable porous structure, high water content, low strength and modulus, and rubbery state in wet conditions. The sensory evaluation suggested that the noodle-shaped blends can be used as a food material with a texture similar to capellini pasta.

Substituent distribution of propyl cellulose studied by nuclear magnetic resonance.

A series of propyl cellulose (PC) samples with different degrees of substitution (DS) ranging from 0.34 to 2.02 were prepared by a slurry method using propyl bromide as the etherification reagent. Two-dimensional nuclear magnetic resonance (NMR) studies were performed to identify the 1H and 13C chemical shifts of eight anhydroglucose units (AGUs) in PC chains including un-, 2-mono-, 3-mono-, 6-mono-, 2,3-di-, 2,6-di-, 3,6-di-, and 2,3,6-tri-substituted ones. In addition, the mole fractions (χ) of these AGUs in the studied PC samples and their changes with DS were determined from the quantitative 13C NMR spectra. The obtained χ-DS profiles were different from those of methyl and ethyl celluloses prepared by a similar slurry method, indicating that the molecular sizes of the substituent reagents utilized for cellulose ethers strongly affected their substituent distributions.

Determination of mole fractions of ethyl-cellulose-containing monomers by NMR.

Three samples of ethyl cellulose (EC) with different degrees of substitution (DS)-0.51, 1.41, and 2.28-were prepared by a slurry method using ethyl bromide as the etherification reagent. 1H-13C HSQC and HSQC-TOCSY NMR spectral analysis allowed for complete assignment of the 1H and 13C chemical shifts, respectively, of eight anhydroglucose units (AGUs) comprising EC chains-un-, 2-mono-, 3-mono-, 6-mono-, 2,3-di-, 2,6-di-, 3,6-di-, and 2,3,6-tri-substituted AGUs. In addition, the lineshape of the quantitative 13C NMR spectra of the three EC samples provided change in the mole fractions of these AGUs against DS, making it possible to estimate the reaction mechanism for the production of EC, elucidating reactivities of the hydroxyl groups at the 2, 3, and 6 positions of cellulose and interactions between the substituent groups within the same AGU and vicinal AGUs.

NMR characterization of cellulose acetate: Mole fraction of monomers in cellulose acetate determined from carbonyl carbon resonances.

Cellulose acetate (CA) samples with varying degrees of substitution were prepared via homogeneous acetylation in 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) and by the acid-hydrolysis of cellulose triacetate in acetic acid. Quantitative analysis of the 13C NMR spectra facilitated the assignment of the carbonyl carbon shifts of the 2-mono-, 3-mono-, 6-mono-, 2,3-di-, 2,6-di-, 3,6-di-, and 2,3,6-tri-substituted anhydroglucose units (AGUs), and the determination of the mole fraction of 7 AGUs and unsubstituted AGU in the CA chains. This shed some light on the mechanism of CA production in homogeneous reaction systems. In addition, comparison of the mole fractions of the 8 AGUs suggested that the acetone solubility of CA strongly related to the AGU composition.

Structural and mechanical characterization of bacterial cellulose-polyethylene glycol diacrylate composite gels.

This study explores the structural and mechanical properties of bacterial cellulose-polyethylene glycol diacrylate (BC-PEGDA) composite gels. The molecular dynamics results obtained by solid-state 13C nuclear magnetic resonance analyses suggested that BC and PEGDA molecules were incompatible as composite gels, though BC fibers and PEGDA interact with each other. The mechanical strength of the gels depended on the amount of PEGDA, becoming softer and more stretchable when a tensile force was applied, but for a large amount of PEGDA, they became brittle. The BC-3% and 5% PEGDA gels had similar viscoelastic behaviors as a BC gel, and these composite gels could stick to human skin. Since BC-PEGDA composite gels are composed of BC and PEGDA-both of which are biocompatible, it is thought that these composite gels also have excellent biocompatibility. Taken together, we concluded that the BC-3% and 5% PEGDA gels have great potential for use in medical and cosmetic fields.

Cationic cellulose hydrogels cross-linked by poly(ethylene glycol): Preparation, molecular dynamics, and adsorption of anionic dyes.

Cationic cellulose hydrogels (CCGs) were prepared from quaternized celluloses with degrees of substitution (DS) of 0.56, 0.84, and 1.33, by the cross-linking reaction with poly(ethylene glycol) diglycidyl ether as a cross-linker. The CCGs exhibited swelling behavior in aqueous solutions, which was not affected by pH and temperature of the solution because of the presence of quaternary ammonium groups in their structures. The CCGs showed adsorption ability toward anionic dyes in aqueous solution, which increased with increasing DS. The dye adsorption was found to follow the pseudo-second order kinetic model and the equilibrium isotherm data can be described by the Langmuir adsorption model. In addition, the CCGs could be regenerated and proved to be recyclable adsorbents for wastewater treatment.

NMR characterization of sodium carboxymethyl cellulose: Substituent distribution and mole fraction of monomers in the polymer chains.

A series of sodium carboxymethyl cellulose (CMC) samples with degrees of substitution (DS) from 0.68 to 2.84 were prepared from cellulose by multistep carboxymethylation. Two-dimensional NMR spectroscopic analyses allowed complete assignment of the (1)H and (13)C chemical shifts of the eight anhydroglucose units (AGUs) found in CMC: 2,3,6-tri-, 2,3-di-, 2,6-di-, 3,6-di-, 2-mono-, 3-mono-, 6-mono-, and unsubstituted AGUs. A comparative analysis of the chemical shift data revealed the substituent effects of the carboxymethyl groups at the 2-, 3-, and 6-positions on the (1)H and (13)C chemical shifts of the AGUs and that additivity could be applied to the (1)H and (13)C chemical shifts of CMC. Quantitative (13)C NMR spectra of the CMC samples allowed determination of the AGU composition, as well as the substituent distribution at the 2-, 3-, and 6-positions of cellulose, which allowed estimation of the reactivity of the hydroxyl groups toward the carboxymethylation.

NMR characterization of cellulose acetate: chemical shift assignments, substituent effects, and chemical shift additivity.

A series of cellulose acetates (CA) with degrees of substitution (DS) ranging from 2.92-0.92 dissolved in dimethylsulfoxide (DMSO)-d6 and cellulose dissolved in tetrabutylammonium fluoride (TBAF)/DMSO-d6 were investigated by two-dimensional NMR spectroscopy. The NMR spectroscopic analysis allowed the determination of the (1)H and (13)C NMR chemical shifts of the eight anhydroglucose units (AGUs) that contain CA: 2,3,6-tri-, 2,3-di-, 2,6-di-, 3,6-di-, 2-mono-, 3-mono-, 6-mono-, and unacetylated AGUs. A comparative analysis of the chemical shift data revealed the substituent effect of acetyl groups at the 2-, 3-, and 6-positions on the (1)H and (13)C nuclei in the same AGU. In addition, chemical shift additivity could be applied to the (1)H and (13)C chemical shifts of CA because the chemical shifts of the diacetylated and triacetylated AGUs could be almost completely explained by the acetyl substituent effects at the 2-, 3-, and 6-positions.

Characterization, molecular dynamics, and encapsulation ability of ß-cyclodextrin polymers crosslinked by polyethylene glycol.

A series of water-insoluble cyclodextrin polymers (CDP) was prepared by crosslinking ß-cyclodextrin (CD) with polyethylene glycol diglycidyl ether (PEGDE). Similarly, a reference CDP was prepared using ethylene glycol diglycidyl ether (EGDE). Increasing the feed ratio of PEGDE to CD in the reaction mixture led to high degrees of crosslinking. Relaxation measurements revealed structural homogeneity among the CDPs, which exhibited mobilities that strongly depended on the chain lengths of the crosslinking agents. In addition, all the CDPs displayed high encapsulation abilities toward bisphenol A (BPA) in aqueous media. In particular, the CDP sample with a low degree of crosslinking by PEGDE showed the highest encapsulation ability toward BPA. In contrast, the CDP crosslinked by EGDE exhibited low encapsulation ability because its highly dense structure, which results from the short chain lengths of the crosslinking agents, hinders the penetration of BPA molecules.

Characterization and properties of carboxymethyl cellulose hydrogels crosslinked by polyethylene glycol.

Novel hydrogels were prepared from carboxymethyl cellulose (CMC) sodium salt by crosslinking with polyethylene glycol diglycidyl ether (PEGDE). The detailed structures of the hydrogels were determined via FTIR and solid-state NMR spectroscopic analyses. Increasing the feed ratio of PEGDE to CMC in the reaction mixture led to an increase in the crosslinking degree, which enhanced the physical strength of the hydrogels. The hydrogels exhibited enzyme degradability, and after 3 days of incubation with cellulase, 62-28 wt% of the CMC in the hydrogel was degraded under the conditions employed in this study. In addition, the hydrogels exhibited protein adsorption and release abilities, and the amounts of proteins adsorbed on the hydrogels and the release profile of the proteins depended on the protein sizes and crosslinking degree of the hydrogels. These unique properties might enable the use of CMC-based hydrogels as drug delivery system carriers for protein-based drugs if the biological safety of the hydrogel can be verified.

Chemical shift assignment of the complicated monomers comprising cellulose acetate by two-dimensional NMR spectroscopy.

(1)H and (13)C chemical shift assignment of the anhydroglucose units (AGUs) that comprise cellulose acetate with the total degree of substitution of 2.33 was performed by a combination of two-dimensional NMR spectra obtained from COSY, TOCSY, HSQC-TOCSY, and HSQC experiments. In the TOCSY spectra, there are 13 kinds of correlation networks for the H1 to H6' atoms of AGU; each (1)H resonance of the 13 AGUs was assigned from the changes in the intensities of the (1)H resonances with TOCSY spin-lock time during the TOCSY experiments. The ring (13)C resonances were assigned via analysis of HSQC spectra. Comparison of the shift data enabled the assignment of the 13 AGUs as three types of 2,3,6-tri-acetylated, one type of 2-mono-, 3-mono-, 6-mono-, 2,3-di-, 2,6-di-, and 3,6-di-acetylated AGU, and unsubstituted AGU, and three types of AGUs located in end units. These shift data will be useful for characterizing the molecular structures of cellulose derivatives as well as cellulose acetate (CA).

Characterization and bisphenol A adsorption capacity of ß-cyclodextrin-carboxymethylcellulose-based hydrogels.

Novel hydrogel beads having molecular adsorption abilities were prepared from carboxymethylcellulose sodium salt (CMC) and ß-cyclodextrin (ß-CD) by suspension crosslinking, using ethylene glycol diglycidyl ether (EGDE) in basic medium as a crosslinking agent. FTIR and solid-state NMR spectroscopic analysis revealed that the amount of incorporated ß-CD and crosslinking densities within the hydrogel bead structures are strongly dependent on the molar feed ratio of ß-CD to CMC during preparation. The hydrogel beads showed water-swelling capacities of 70-200 mL/g-polymer, with decreases in capacity associated with increased amounts of ß-CD incorporated in the gel structure. The hydrogel beads also showed a high adsorption capacity toward bisphenol A (BPA) in water. Batch BPA-adsorption experiments were analyzed employing Langmuir isotherm models; hydrogel bead adsorption isotherms for BPA could be fitted to the Langmuir model. The maximum BPA-adsorption among the prepared series of hydrogel beads amounted to 167 µmol g(-1).

Regulation of endoglucanase gene (cmcax) expression in Acetobacter xylinum.

Although cellulose is the most abundant biopolymer in nature, the detailed mechanisms of cellulose biosynthesis remain unknown. Acetobacter xylinum is one of the best-studied model organisms for cellulose biosynthesis. Interestingly, the over-expression of the cmcax gene cause enhancement of cellulose production in A. xylinum, while its product (CMCax) has cellulose degradation activity. The addition of CMCax into medium also promotes cellulose production, suggesting that CMCax is involved in cellulose synthetic pathway. In the present study, we reveal the regulation mechanism of cmcax expression in A. xylinum. First, we treated cells with four kinds of beta-glucodisaccharide. Using an enzyme assay and real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), we observed an increase in CMCax activity and an induction of cmcax expression by gentiobiose treatment. Therefore, we concluded that gentiobiose induced cmcax expression. Although gentiobiose does not originally exist in the cultivation medium, we have revealed that membrane and intra-cellular proteins extracted from A. xylinum produce gentiobiose from glucose, which is one of the components in the cultivation medium. Furthermore, we confirmed that cmcax expression in a wild-type strain increased gradually after 5 d cultivation using real-time qRT-PCR. These results have led us to conclude that the increase in cmcax expression after 5 d cultivation is caused by the increase in gentiobiose, which could be synthesized by a condensation reaction in A. xylinum. Since CMCax plays a pivotal role in the cellulose production system, our results will contribute to the elucidation of mechanisms of cellulose biosynthesis.

Shear bond strength of resin composite to magnetic Fe-Pt alloy.

STATEMENT OF PROBLEM: It is important to obtain strong adhesion between resin composite material and magnetic Fe-Pt alloy for fabrication of an indirect veneered crown. However, little is known about the bond strength between these materials. PURPOSE: This study evaluated the bond strength of resin composite to magnetic Fe-Pt alloy, using 4 adhesive metal primers. MATERIAL AND METHODS: Seventy disk-shaped patterns (7.5 mm in diameter, 2.0 mm thick) were cast from a custom-made Fe-Pt ingot using a high-frequency centrifugal casting machine. After the bonding surfaces were airborne-particle abraded with 50 microm Al2O3, specimens were divided into groups (n=7) and primed with either Metal Primer II (MP), Meta Fast Bonding Liner (BL), New Meta Color INFIS Opaque Primer (IN), or Epricord Opaque Primer (EP). Nonprimed specimens served as controls (n=7). A resin composite (New Meta Color INFIS) was then applied to all specimens and polymerized. After the specimens were immersed in 37 degrees C water for 24 hours or thermal cycled at 4 degrees C to 60 degrees C with a 1-minute dwell time of up to 10,000 cycles, the shear bond strengths (MPa) were measured using a universal testing machine at a crosshead speed of 0.5 mm/min. The data were statistically analyzed by a 2-way analysis of variance and the Scheffe test (alpha=.05). RESULTS: Although IN had the lowest strength, there were no significant differences in bond strengths of the other 3 primers (MP, BL, and EP) before thermal cycling. The bond strengths of MP and EP were slightly altered by thermal cycling (P >.05); however, BL and IN underwent a significant decrease (P <.05). CONCLUSION: The results of this in vitro study indicate that MP and EP provide superior long-term shear strength when bonding the resin composite evaluated to Fe-Pt alloy.

CP/MAS (13)C NMR study of cellulose and cellulose derivatives. 2. Complete assignment of the (13)C resonance for the ring carbons of cellulose triacetate polymorphs.

Complex ring (13)C resonance lines of the cross-polarization/magic angle spinning (CP/MAS) (13)C NMR spectra of cellulose triacetate (CTA) I and CTA II were completely assigned, for the first time, by (13)C-enriched CTA allomorphs. The (13)C-enriched CTA I was prepared by heterogeneous acetylation of bacterial cellulose which was biosynthesized by Acetobacter xylinum (A. xylinum) ATCC10245 from culture medium containing D-(2-(13)C)-, D-(3-(13)C)-, or D-(5-(13)C)glucose as a carbon source, while CTA II samples were obtained by solution acetylation of the (13)C-enriched bacterial celluloses. From comparison of the spectra of normal CTA prepared from ramie with those of the enriched CTA samples, it was revealed that all carbons composed of CTA I appeared as a singlet, while those of CTA II except for C1 were shown as equal-intensity doublets in the CP/MAS (13)C NMR spectrum. This finding suggests that CTA I is made up of one kind of glucopyranose residue while there are two magnetically inequivalent sites in the unit cell of CTA II in the same population.

CP/MAS (13)C NMR study of cellulose and cellulose derivatives. 1. Complete assignment of the CP/MAS (13)C NMR spectrum of the native cellulose.

The precise assignments of cross polarization/magic angle spinning (CP/MAS) (13)C NMR spectra of cellulose I(alpha) and I(beta) were performed by using (13)C labeled cellulose biosynthesized by Acetobacter xylinum (A. xylinum) ATCC10245 strain from culture medium containing D-[1,3-(13)C]glycerol or D-[2-(13)C]glucose as a carbon source. On the CP/MAS (13)C NMR spectrum of cellulose from D-[1,3-(13)C]glycerol, the introduced (13)C labeling were observed at C1, C3, C4, and C6 of the biosynthesized cellulose. In the case of cellulose biosynthesized from D-[2-(13)C]glucose, the transitions of (13)C labeling to C1, C3, and C5 from C2 were observed. With the quantitative analysis of the (13)C transition ratio and comparing the CP/MAS (13)C NMR spectrum of the Cladophora cellulose with those of the (13)C labeled celluloses, the assignments of the cluster of resonances which belong to C2, C3, and C5 of cellulose, which have not been assigned before, were performed. As a result, all carbons of cellulose I(alpha) and I(beta) except for C1 and C6 of cellulose I(alpha) and C2 of cellulose I(beta) were shown in equal intensity of doublet in the CP/MAS spectrum of the native cellulose, which suggests that two inequivalent glucopyranose residues were contained in the unit cells of both cellulose I(alpha) and I(beta) allomorphs.