1.Immobilization of Lactobacillus bulgaricus with gellan gum and its application in continuous fermentation of D-lactic acid from corn straw.
Yongxin GUO ; Gang WANG ; Kexin LI ; Jiaqi HAN ; Huan CHEN ; Sitong ZHANG ; Yanli LI ; Guang CHEN
Chinese Journal of Biotechnology 2023;39(3):1083-1095
Biorefinery of chemicals from straw is an effective approach to alleviate the environmental pollution caused by straw burning. In this paper, we prepared gellan gum immobilized Lactobacillus bulgaricus T15 gel beads (LA-GAGR-T15 gel beads), characterized their properties, and established a continuous cell recycle fermentation process for D-lactate (D-LA) production using the LA-GAGR-T15 gel beads. The fracture stress of LA-GAGR-T15 gel beads was (91.68±0.11) kPa, which was 125.12% higher than that of the calcium alginate immobilized T15 gel beads (calcium alginate-T15 gel beads). This indicated that the strength of LA-GAGR-T15 gel beads was stronger, and the strain was less likely to leak out. The average D-LA production was (72.90±2.79) g/L after fermentation for ten recycles (720 h) using LA-GAGR-T15 gel beads as the starting strain and glucose as the substrate, which was 33.85% higher than that of calcium alginate-T15 gel beads and 37.70% higher than that of free T15. Subsequently, glucose was replaced by enzymatically hydrolyzed corn straw and fermented for ten recycles (240 h) using LA-GAGR-T15 gel beads. The yield of D-LA reached (1.74±0.79) g/(L·h), which was much higher than that of using free bacteria. The wear rate of gel beads was less than 5% after ten recycles, which indicated that LA-GAGR is a good carrier for cell immobilization and can be widely used in industrial fermentation. This study provides basic data for the industrial production of D-LA using cell-recycled fermentation, and provides a new way for the biorefinery of D-LA from corn straw.
Fermentation
;
Lactobacillus delbrueckii
;
Zea mays
;
Lactic Acid
;
Alginates/chemistry*
;
Glucose
2.Synthesis of a novel injectable alginate impression material and impression accuracy evaluation.
Xingzi LIU ; Xinhui WANG ; Jingya WU ; Jingjing LUO ; Yun WANG ; Quanli LI
West China Journal of Stomatology 2022;40(6):662-667
OBJECTIVES:
This work aimed to synthesize a novel injectable alginate impression material and evaluate its accuracy.
METHODS:
Certain proportions of sodium alginate, trisodium phosphate dodecahydrate, potassium fluorotitanate, diatomaceous earth, and other ingredients were dissolved in water and mixed evenly with a planetary centrifugal mixer to obtain a certain viscosity base paste. Certain proportions of calcium sulfate hemihydrate, magnesium oxide, glycerin, and polyethylene glycol (PEG) 400 were mixed evenly with a planetary centrifugal mixer to obtain the reactor paste with the same viscosity as the base paste. The base and reactor pastes were poured into a two-cylinder cartridge at a 2∶1 volume ratio. A gun device was used to accomplish mixing by compressing materials into a mixing tip. The samples were divided into three groups: injectable alginate impression materials (IA group) as the experimental group, and Jeltrate alginate impression materials (JA group) and Silagum-putty/light addition silicone rubber impression materials (SI group) as the two control groups.
RESULTS:
Scanning electron microscopy (SEM) showed that the injectable alginate impression materials had a denser structure and fewer bubbles than the commercial alginate impression material. The accuracy of the three kinds of impression materials was evaluated by 3D image superposition. The deviations between the three test group models and the standard model (trueness) were 49.58 μm±1.453 μm (IA group), 54.75 μm±7.264 μm (JA group), and 30.92 μm±1.013 μm (SI group). The deviations of the models within each test group (precision) were 85.79 μm±8.191 μm (IA group), 97.65 μm±11.060 μm (JA group), and 56.51 μm±4.995 μm (SI group). Significant differences in trueness and precision were found among the three kinds of impression materials (P<0.05).
CONCLUSIONS
The accuracy of the new injectable alginate impression material was better than that of the traditional powder-type alginate impression material but worse than that of the addition silicone rubber impression materials. The novel injec-table alginate impression material demonstrated good operation performance and impression accuracy, showing broad application prospect.
Alginates/chemistry*
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Silicone Elastomers/chemistry*
;
Dental Impression Materials/chemistry*
;
Powders
3.A Noval Method for Producing Antibacterial Wound Dressing by Using Fused Deposition Molding with Post-3D-printed Process.
Chinese Journal of Medical Instrumentation 2019;43(4):275-278
Using three-dimensional printing to produce antibacterial wound dressing is a new topic that will change the production style of wound dressing industry. Combining with post-3D-printed process, a desktop fused deposition molding equipment can be used to produce wound dressing containing polyvinyl alcohol, alginate and chitosan. The wound dressing produced by FDM has good aspects of absorbency, moisture vapour transmission rate and mechanical property. After loaded with antibacterial agent iodine and silver nano particle, the antibacterial activity rate increases to 99% and it is suitable to use as antibacterial wound dressing. This method affects the production of wound dressing to a more cost-effective way, and provides a possible individualized treatment for patient in the future.
Alginates
;
chemistry
;
Anti-Bacterial Agents
;
administration & dosage
;
Bacteria
;
drug effects
;
Bandages
;
economics
;
standards
;
Chitosan
;
chemistry
;
Humans
;
Iodine
;
administration & dosage
;
pharmacology
;
Nanoparticles
;
administration & dosage
;
Polyvinyl Alcohol
;
chemistry
;
Printing, Three-Dimensional
;
Silver
;
administration & dosage
;
pharmacology
;
Wound Healing
4.Formulation development and evaluation of gastroretentive floating beads with Brucea javanica oil using ionotropic gelation technology.
Yue ZHANG ; Xi-Tong ZHANG ; Qi ZHANG ; Bing WANG ; Tong ZHANG
Chinese Journal of Natural Medicines (English Ed.) 2018;16(4):293-301
In the present study, a gastric retention floating system for Brucea javanica oil, composed of alginate and carrageenan, was prepared using ionotropic gelation. Parameters for floatability, drug load, encapsulation efficiency, bead morphology, in vitro release, and in vivo gastric retention were evaluated. The optimized formulation via Box-Behnken design consisted of 1.7% alginate (W/V), 1.02% carrageenan (W/V), 1.4% CaCO (W/V), and a gelling bath of pH 0.8. The alginate-carrageenan-Brucea javanica oil beads had a porous structure and exhibited up to 24 h of in vitro floatability with a load capacity of 45%-55% and an encapsulation efficiency of 70%-80%. A 6-h sustained release was observed in vitro. The beads had a prolonged gastric retention (> 60% at 6 h) in fasted rats, compared to non-floating beads (15% at 6 h), as measured by gamma scintigraphy with single-photon emission tomography/computed tomography (SPET/CT). In conclusion, the alginate-carrageenan-Brucea javanica oil system showed enhanced oil encapsulation efficiency, excellent floating and gastric retention abilities, and a favorable release behavior.
Alginates
;
chemistry
;
Animals
;
Biological Availability
;
Brucea
;
chemistry
;
Carrageenan
;
chemistry
;
Delayed-Action Preparations
;
administration & dosage
;
chemistry
;
pharmacokinetics
;
Drug Carriers
;
chemistry
;
Drug Delivery Systems
;
methods
;
Drug Evaluation, Preclinical
;
Gastric Mucosa
;
metabolism
;
Glucuronic Acid
;
chemistry
;
Hexuronic Acids
;
chemistry
;
Microspheres
;
Plant Oils
;
administration & dosage
;
chemistry
;
pharmacokinetics
;
Rats
;
Rats, Sprague-Dawley
5.Three-dimensional Culture of Chondrocyte Using Methacrylic Alginate Gel Beads Cross-linked with Mixed Metal-cation.
Yang WANG ; Yuxia FENG ; Xing FAN ; Liling REN
Journal of Biomedical Engineering 2015;32(3):599-604
This study was to explore a better three-dimensional (3-D) culture method of chondrocyte. The interpenetrating network (IPN) gel beads were developed through a photo-cross linking reaction with mixed barium ions and calcium ions at the ratio of 5:5 with the methacrylic alginate (MA), which was a chemically conjugated alginate with methacrylic groups. The second generation of primary cartilage cells was encapsulated in the MA gel beads for three weeks. In the designated timing, HE stain, Alamar blue method and Scanning electron microscopic were used to determine the cartilage cells growth, proliferation and the cell distribution in the scaffolds, respectively. The expression of type II collagen was investigated by an immunohistochemistry assay and the glycosaminoglycan content was quantitatively evaluated with the spectrophotometry of 1, 9 dimethylene blue assay. Compared to the alginate control group, the deposition of glycosaminoglycan was significantly upregulated in IPN-MA gel beads with higher cell proliferation. The secretion of extracellular matrix and proliferation of chondrocyte in methacrylic alginate gel beads were higher than that in Alginate beads. Cells were able to attach, to grow well on the scaffolds under scanning electron microscopy. The result of immunohistochemistry staining of collagen type II was positive, confirming the maintenance of chondrocyte phenotype in methacrylic alginate gel beads. This study shows a great potential for three-dimensional culture of cartilage.
Alginates
;
chemistry
;
Barium
;
chemistry
;
Calcium
;
chemistry
;
Cartilage
;
cytology
;
Cations
;
Cell Culture Techniques
;
instrumentation
;
Cells, Cultured
;
Chondrocytes
;
cytology
;
Collagen Type II
;
chemistry
;
Glucuronic Acid
;
chemistry
;
Glycosaminoglycans
;
chemistry
;
Hexuronic Acids
;
chemistry
;
Metals
;
chemistry
;
Microscopy, Electron, Scanning
6.Production of L(+)-tartaric acid by immobilized Rhizobium strain BK-20.
Xiang LAN ; Wenna BAO ; Haifeng PAN ; Zhipeng XIE ; Jianguo ZHANG
Chinese Journal of Biotechnology 2014;30(2):315-319
The cis-epoxysuccinate hydrolase (CESH) from Rhizobium strain BK-20 is the key enzyme for L(+)-tartaric acid production. To establish a highly efficient and stable production process, we first optimized the enzyme production from Rhizobium strain BK-20, and then developed an immobilized cell-culture process for sustained production of L(+)-tartaric acid. The enzyme activity of free cells reached (3 498.0 +/- 142.6) U/g, and increased by 643% after optimization. The enzyme activity of immobilized cells reached (2 817.2 +/- 226.7) U/g, under the optimal condition with sodium alginate as carrier, cell concentration at 10% (W/V) and gel concentration at 1.5% (W/V). The immobilized cells preserved high enzyme activity and normal structure after 10 repeated batches. The conversion rate of the substrate was more than 98%, indicating its excellent production stability.
Alginates
;
chemistry
;
Cells, Immobilized
;
Glucuronic Acid
;
chemistry
;
Hexuronic Acids
;
chemistry
;
Hydrolases
;
metabolism
;
Rhizobium
;
enzymology
;
metabolism
;
Tartrates
;
metabolism
7.Differentiation of adipose-derived stem cells toward nucleus pulposus-like cells induced by hypoxia and a three-dimensional chitosan-alginate gel scaffold in vitro.
Zhicheng ZHANG ; Fang LI ; Haiquan TIAN ; Kai GUAN ; Guangmin ZHAO ; Jianlin SHAN ; Dajiang REN
Chinese Medical Journal 2014;127(2):314-321
BACKGROUNDInjectable three-dimensional (3D) scaffolds have the advantages of fluidity and moldability to fill irregular-shaped defects, simple incorporation of bioactive factors, and limited surgical invasiveness. Adipose-derived stem cells (ADSCs) are multipotent and can be differentiated toward nucleus pulposus (NP)-like cells. A hypoxic environment may be important for differentiation to NP-like cells because the intervertebral disc is an avascular tissue. Hence, we investigated the induction effects of hypoxia and an injectable 3D chitosan-alginate (C/A) gel scaffold on ADSCs.
METHODSThe C/A gel scaffold consisted of medical-grade chitosan and alginate. Gel porosity was calculated by liquid displacement method. Pore microstructure was analyzed by light and scanning electron microscopy. ADSCs were isolated and cultured by conventional methods. Passage 2 BrdU-labeled ADSCs were co-cultured with the C/A gel. ADSCs were divided into three groups (control, normoxia-induced, and hypoxia-induced groups). In the control group, cells were cultured in 10% FBS/DMEM. Hypoxia-induced and normoxia-induced groups were induced by adding transforming growth factor-β1, dexamethasone, vitamin C, sodium pyruvate, proline, bone morphogenetic protein-7, and 1% ITS-plus to the culture medium and maintaining in 2% and 20% O2, respectively. Histological and morphological changes were observed by light and electron microscopy. ADSCs were characterized by flow cytometry. Cell viability was investigated by BrdU incorporation. Proteoglycan and type II collagen were measured by safranin O staining and the Sircol method, respectively. mRNA expression of hypoxia-inducing factor-1α (HIF-1α), aggrecan, and Type II collagen was determined by reverse transcription-polymerase chain reaction.
RESULTSC/A gels had porous exterior surfaces with 80.57% porosity and 50-200 üm pore size. Flow cytometric analysis of passage 2 rabbit ADSCs showed high CD90 expression, while CD45 expression was very low. The morphology of induced ADSCs resembled that of NP cells. BrdU immunofluorescence showed that most ADSCs survived and proliferated in the C/A gel scaffold. Scanning electron microscopy showed that ADSCs grew well in the C/A gel scaffold. ADSCs in the C/A gel scaffold were positive for safranin O staining. Hypoxia-induced and normoxia-induced groups produced more proteoglycan and Type II collagen than the control group (P < 0.05). Proteoglycan and Type II collagen levels in the hypoxia-induced group were higher than those in the normoxia-induced group (P < 0.05). Compared with the control group, higher mRNA expression of HIF-1α, aggrecan, and Type II collagen was detected in hypoxia-induced and normoxiainduced groups (P < 0.05). Expression of these genes in the hypoxia-induced group was significantly higher than that in the normoxia-induced group (P < 0.05).
CONCLUSIONADSCs grow well in C/A gel scaffolds and differentiate toward NP-like cells that produce the same extracellular matrix as that of NP cells under certain induction conditions, which is promoted in a hypoxic state.
Adipose Tissue ; cytology ; Alginates ; chemistry ; Animals ; Cell Differentiation ; physiology ; Cells, Cultured ; Chitosan ; chemistry ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Rabbits ; Stem Cells ; cytology ; physiology ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry
8.Preparation of cinnamomi cortex oil microspheres based on porous silicon dioxide and its property characterizations.
Chun-Xia ZHU ; Yan-Rong JIANG ; Zhen-Hai ZHANG ; Dong-Mei DING ; Xiao-Bin JIA
China Journal of Chinese Materia Medica 2013;38(20):3479-3483
To determine the optimum process for preparing Cinnamomi Cortex oil microspheres based on porous silicon dioxide. After porous silica dioxide adsorbed Cinnamomi Cortex oil, Cinnamomi Cortex oil microspheres were prepared by the dropping method, with sodium alginate as the skeleton materials. The preparation process was optimized through the L(9) (3(4)) orthogonal test design, with microspheres diameter, distribution, drug loading capacity and entrapment efficiency as the indexes. The cinnamon volatile oil microspheres were characterized by scanning election microscope (SEM), thermogravimetric analysis (TGA), and infrared (IR) spectroscopy. An in vitro drug release experiment was conducted. The results showed that the microspheres prepared with the optimal process parameters were in good shape, even in size and good in dispersibility, with an average diameter of 1.61 mm, an average drug loading capacity of 32.85%, an entrapment efficiency of 94.79%. The maximum drug release capacity reached 72.6%, 95.0%, 97.4%, respectively, under pH 4.0, 6.8, 7.4 in 6 hours. Meanwhile, microsphere generation was tested by IR, TGA and other methods. The established optimum process for preparing Cinnamomi Cortex oil microspheres was proved to be stable and practical.
Alginates
;
chemistry
;
Chemistry, Pharmaceutical
;
Cinnamomum
;
chemistry
;
Drug Carriers
;
chemistry
;
Drugs, Chinese Herbal
;
chemistry
;
Glucuronic Acid
;
chemistry
;
Hexuronic Acids
;
chemistry
;
Microspheres
;
Particle Size
;
Porosity
;
Silicon Dioxide
;
chemistry
;
Solubility
9.Preparation of microspheres of superoxide dismutase and their activities.
Journal of Zhejiang University. Medical sciences 2013;42(6):666-670
OBJECTIVETo prepare the poly(lactic-co-glycolic acid) (PLGA) microspheres and composite alginate-chitosan-PLGA microspheres containing superoxide dismutase (SOD) and to evaluate their SOD activities.
METHODSThe SOD-PLGA microspheres were prepared by W/O/W emulsification method, and the composite microspheres were prepared by two steps:alginate-chitosan microcapsules were first prepared by a modified emulsification and ion crosslinking method, and then they were further dispersed in PLGA to form the composite microspheres. The SOD concentration was determined by Coomassie method, its activity was measured by xanthine oxidase system.
RESULTSThe SOD activity was less sensitive to temperature and sensitive to pH, organic solvents, ultrasound and vigorous stir without iced bath. The entrapment efficiencies of SOD in PLGA (50:50) microspheres, PLGA (70:30) microspheres, alginate-chitosan microcapsules, the composite PLGA (50:50) microspheres and the composite PLGA (70:30) microspheres were 36.42%±1.81%, 66.18%±0.05%, 91.08%±1.28%, 87.30%±3.89% and 83.19%±3.48%, respectively. In vitro release tests demonstrated that the SOD activities in 50:50 composite microspheres were higher than that in the PLGA ones at 1 h, 8 h and 1 w.
CONCLUSIONThe composite alginate-chitosan-PLGA microspheres for SOD sustained release can significantly improve the protein entrapment efficiency and maintain its protein activity.
Alginates ; chemistry ; Chitosan ; chemistry ; Delayed-Action Preparations ; chemistry ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Lactic Acid ; chemistry ; Microspheres ; Polyglycolic Acid ; chemistry ; Superoxide Dismutase ; chemistry
10.Preparation and properties evaluation of methacrylic alginate gell beads cross-linked by mixed metal-cation.
Xing FAN ; Shengnan LEI ; Liling REN
Journal of Biomedical Engineering 2013;30(6):1272-1275
The aim of this study was to obtain the alginate gels which could have proper compressive strength and excellent permeability for cell proliferation and could have more promising potentials in the application of tissue engineering. Through the reaction of the carboxyl of the alginate and the amino of methacrylic acid, methylacrylic was generated into alginate long chain which could be enhanced by the polymerization of double bond under thermal reaction condition. And then alginate gel beads were prepared using the mixture of calcium chloride and barium chloride solution as cationic crosslinker, and the compressive modulus and permeability of the prepared alginate gel beads were investigated. When the ratio of barium ions to calcium ions was 5:5, the compression modulus was 189.7 kPa, and it showed the best permeability for trypsin with molecular weight of 24 kDa and entrapment effect for bovine serum albumin with molecular weight of 67 kDa. Compared to compositions of other ratios, the alginate gel beads made in 5:5 mixture indicated excellent compressive modulus and permeability. These results indicated that the alginate hydrogel beads with the ratio of barium ions to calcium ions being 5:5 have a potential application in tissue engineering as a support material and encapsulating materials in cell culture.
Alginates
;
chemistry
;
Barium Compounds
;
chemistry
;
Calcium Chloride
;
chemistry
;
Cations
;
Cell Culture Techniques
;
Cell Proliferation
;
Chlorides
;
chemistry
;
Cross-Linking Reagents
;
chemistry
;
Gels
;
chemistry
;
Metals
;
Polymethacrylic Acids
;
chemistry
;
Tissue Engineering

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