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We revealed that the encapsulation of enzyme-immobilized silica particles in hollow-type spherical bacterial cellulose (HSBC) gels enables the use of the inside of HSBC gels as a reaction field. The encapsulation of horseradish peroxidase (HRP)-immobilized silica particles (Si-HRPs, particle size: 40-50 µm) within HSBC gels was performed by using a BC gelatinous membrane produced at the interface between Komagataeibacter xylinus suspension attached onto an alginate gel containing Si-HRPs and silicone oil. After the biosynthesis of the BC gelatinous membrane, formed from cellulose nanofiber networks, the alginate gel was removed via immersion in a phosphate-buffered solution. Si-HRP encapsulated HSBC gels were reproducibly produced using our method with a yield of over 90%. The pore size of the network structure of the BC gelatinous membrane was less than 1 µm, which is significantly smaller than the encapsulated Si-HRPs. Consequently, the encapsulated Si-HRPs could neither pass through the BC gelatinous membrane nor leak from the interior cavity of the HSBC gel. The activity of the encapsulated HRPs was detected using the 3,3',5,5'-tetramethylbenzidine (TMB)-H2O2 system, demonstrating that this method can encapsulate the enzyme without inactivation. Since HSBC gels are composed of a network structure of biocompatible cellulose nanofibers, immune cells cannot enter the hollow interior, thus, the enzyme-immobilized particles encapsulated inside the HSBC gel are protected from immune-cell attacks. The encapsulation technique demonstrated in this study is expected to facilitate the delivery of enzymes and catalysts that are not originally present in the in vivo environment.
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To reduce the risk of the adsorption of granular activated carbon in the gastrointestinal tract, we successfully produced a hollow-type spherical bacterial cellulose (HSBC) gel containing activated carbon with a particle size of 6 µm. In this study, the aim of which was to develop an effective formulation, we evaluated the stability of activated-carbon-encapsulating HSBC gels under various pH conditions. Activated-carbon-encapsulating HSBC gels (ACEGs) retained the activated carbon without leaking when subjected to agitation in acidic or basic environments. The saturated adsorption amount, calculated using the Langmuir adsorption isotherm, was affected by the target adsorbate and pH conditions. These results indicate that ACEGs can adsorb uremic toxins and their precursors similarly to conventional uremic toxin adsorbents while preventing direct contact between the encapsulated activated carbon and the gastrointestinal tract. Compared to powdered activated carbon, the ACEG is less likely to be adsorbed in the gastrointestinal tract. Therefore, the proposed ACEG is a promising new formulation that will contribute to the treatment of renal failure and improve patients' compliance with medication.
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We encountered a pediatric case of cubitus varus deformity with a sheared olecranon fracture in an 8-year-old boy who underwent corrective osteotomy without relevant medical history. The patient fell, resulting in a sheared olecranon fracture. He underwent a closed reduction and casting. As the displacement slightly increased within a week, we followed him without secondary reduction to expect remodeling. No remodeling occurred; corrective osteotomy was performed one-year post-injury for a marked cubitus varus deformity. At 2.5 years after corrective osteotomy, little difference existed in the carrying angle (CA) and varus angulation (VA) of the proximal ulna than that of the contralateral side, without pain or limited range of motion. The acceptable displacement range for pediatric forearm fractures is <1 cm shortening and 15° angular deformation in patients under 10 years old, and 10° angular deformation in older children. Here, the deformity of the ulna in the coronal plane did not remodel. Proximal forearm deformity can be accurately evaluated in flexion contracture elbows by measuring VA. Ulnar osteotomies are commonly performed on Monteggia fractures to reduce the radial head, and the osteotomy site is at the center of the deformity of the diaphysis. Corrective osteotomy for cubitus varus deformity after supracondylar humerus fracture improves function and cosmetic appearance, with good clinical results. In addition, it could prevent cubitus varus deformity from causing posterolateral rotatory instability. The coronal-plane deformity of the proximal ulnar was not expected to remodel. We recommended early accurate reduction and consideration of additional internal fixation for preventing re-displacement. Corrective osteotomy for cubitus varus deformity of the proximal ulna was an effective treatment.
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In this study, the introduction of a positive charge on the surface of a shape memory material was investigated to enhance cell affinity. To achieve this, the direct chemical modification of a material surface was proposed. Sheet-type, crosslinked poly(caprolactone-co-α-bromo-ɤ-butyrolactone) (poly(CL-co-BrBL)) were prepared, and the direct reaction of amino compounds with bromo groups was conducted on the material surface with a positive charge. Branched poly(CL-co-BrBL) was prepared, followed by the introduction of methacryloyl groups to each chain end. Using the branched macromonomers, stable and sheet-type materials were derived through UV-light irradiation. Then, the materials were soaked in an amino compound solution to react with the bromo groups under various conditions. Differential scanning calorimetry and surface analysis of the modified materials indicated that 10 vol% of N, N-dimethylethylenediamine in n-hexane and 1 h soaking time were optimal to maintain the inherent thermal properties. The achievement of increased luminance and a positive zeta potential proved that the direct modification method effectively introduced the positive charge only on the surface, thereby enhancing cell affinity.
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For reducing side effects and improvement of swallowing, we studied the encapsulation of activated carbon formulations with a hollow-type spherical bacterial cellulose (HSBC) gel using two kinds of encapsulating methods: Methods A and B. In Method A, the BC gelatinous membrane was biosynthesized using Komagataeibacter xylinus (K. xylinus) at the interface between the silicone oil and cell suspension containing activated carbon. In Method B, the bacterial cellulose (BC) gelatinous membrane was formed at the interface between the cell suspension attached to the alginate gel containing activated carbon and the silicone oil. After the BC gelatinous membrane was biosynthesized by K. xylnus, alginate gel was removed by soaking in a phosphate buffer. The activated carbon encapsulated these methods could neither pass through the BC gelatinous membrane of the HSBC gel nor leak from the interior cavity of the HSBC gel. The adsorption ability was evaluated using indole, which is a precursor of the uremic causative agent. From curve-fitting, the adsorption process followed the pseudo-first-order and intra-particle diffusion models, and the diffusion of the indole molecules at the surface of the encapsulated activated carbon within the HSBC gel was dominant at the initial stage of adsorption. It was observed that the adsorption of the encapsulated activated carbon by the intraparticle diffusion process became dominant with longer adsorption times.
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A hollow-type spherical bacterial cellulose (HSBC) gel prepared using conventional methods cannot load particles larger than the pore size of the cellulose nanofiber network of bacterial cellulose (BC) gelatinous membranes. In this study, we prepared a HSBC gel encapsulating target substances larger than the pore size of the BC gelatinous membranes using two encapsulating methods. The first method involved producing the BC gelatinous membrane on the surface of the core that was a spherical alginate gel with a diameter of 2 to 3 mm containing the target substances. With this method, the BC gelatinous membrane was biosynthesized using Gluconacetobacter xylinus at the interface between the cell suspension attached onto the alginate gel and the silicone oil. The second method involved producing the BC gel membrane on the interface between the silicone oil and cell suspension, as well as the spherical alginate gel with a diameter of about 1 mm containing target substances. After the BC gelatinous membrane was biosynthesized, an alginate gel was dissolved in a phosphate buffer to prepare an HSBC gel with the target substances. These encapsulated substances could neither pass through the BC gelatinous membrane of the HSBC gel nor leak from the interior space of the HSBC gel. These results suggest that the HSBC gel had a molecular sieving function. The HSBC gel walls prepared using these methods were observed to be uniform and would be useful for encapsulating bioactive molecules, such as immobilized enzymes in HSBC gel, which is expected to be used as a drug carrier.
Assuntos
Cápsulas/química , Celulose/análogos & derivados , Gluconacetobacter xylinus/química , Microgéis/química , Alginatos/química , Membranas Artificiais , Silicones/químicaRESUMO
We developed a novel cultivating system for hollow-type spherical bacterial cellulose (HSBC) gel production without any molds or template. It consisted of floating aqueous medium droplet containing Gluconacetobacter xylinus (G. xylinus) at the boundary of two non-mixed silicone oil layers. The fibrils of bacterial cellulose (BC) were produced at the interface of water and oil phases. Fibril layers effectively thickened layer-by-layer and eventually formed a shell structure. The size of the HSBC gel can be controlled by the volume of dropped cell suspension. For cell suspensions of 50 µL and 10 µL, HSBC gels of approximately 4.0 mm and 2.5 mm were obtained, respectively. The shell of the HSBC gel is the gelatinous membrane formed by well-organized fibril networks; they comprised type-I crystal structure of cellulose. Additionally, we studied release profile of the fluorescein isothiocyanate-dextran (FITC-Dex) and observed that it released rapidly from the HSBC gels compared to from the BC gels obtained by the static culture method. The release behavior from HSBC gel agreed satisfactorily with Higuchi model. Therefore, the shell of HSBC gel is surely a thin gelatinous membrane of BC, and would be useful as a drug release device.
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2-Methacryloyloxyethyl phosphorylcholine (MPC) has a PC group and is one of the most well-known bioinert polymers. In this study, we evaluated the interaction between MPC and DNA, which specifically interacts with the phospholipid head group via Ca2+ ions. A MPC monolayer and poly(MPC) brush were fabricated to observe the effect of the structure on the interaction between MPC and DNA via Ca2+ ions. The poly(MPC) brush, which shows higher MPC unit density, more efficiently interacted with DNA via Ca2+ ions. Also, serum protein could interact with the poly(MPC) brush via DNA, although the brush itself hardly interacted with serum proteins. Cell adhesion was significantly provoked on poly(MPC)/DNA compared with poly(MPC) because serum protein adsorption was induced on poly(MPC)/DNA.
Assuntos
Cálcio/química , DNA/química , Fosforilcolina/química , Espermatozoides/química , Animais , Adesão Celular , Íons/química , Masculino , Estrutura Molecular , Salmão , Espermatozoides/citologia , Propriedades de SuperfícieRESUMO
We developed a label-free and reagent-less potentiometric biosensor with improved affinity for thrombin. Two different oligomeric DNA aptamers that can recognize different epitopes in thrombin were introduced in parallel or serial manners on the sensing surface to capture the target via multiple contacts as found in many biological systems. The spacer and linker in the aptamer probes were optimized for exerting the best performance in molecular recognition. To gain the specificity of the sensor to the target, an antifouling molecule, sulfobeaine-3-undecanethiol (SB), was introduced on the sensor to form a self-assembled monolayer (SAM). Surface characterization revealed that the aptamer probe density was comparable to the distance of the two epitopes in thrombin, while the backfilling SB SAM was tightly aligned on the surface to resist nonspecific adsorption. The apparent binding parameters were obtained by thrombin sensing in potentiometry using the 1:1 Langmuir adsorption model, showing the improved dissociation constants (Kd) with the limit of detection of 5.5 nM on the dual aptamer-immobilized surfaces compared with single aptamer-immobilized ones. A fine control of spacer and linker length in the aptamer ligand was essential to realize the multivalent binding of thrombin on the sensor surface. The findings reported herein are effective for improving the sensitivity of potentiometric biosensor in an affordable way towards detection of tiny amount of biomolecules.