• Title/Summary/Keyword: scaffold complex

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Enchancement of Gamma-Aminobutyric Acid Production by Co-Localization of Neurospora crassa OR74A Glutamate Decarboxylase with Escherichia coli GABA Transporter Via Synthetic Scaffold Complex

  • Somasundaram, Sivachandiran;Maruthamuthu, Murali Kannan;Ganesh, Irisappan;Eom, Gyeong Tae;Hong, Soon Ho
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1664-1669
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    • 2017
  • Gamma-aminobutyric acid is a precursor of nylon-4, which is a promising heat-resistant biopolymer. GABA can be produced from the decarboxylation of glutamate by glutamate decarboxylase. In this study, a synthetic scaffold complex strategy was employed involving the Neurospora crassa glutamate decarboxylase (GadB) and Escherichia coli GABA antiporter (GadC) to improve GABA production. To construct the complex, the SH3 domain was attached to the N. crassa GadB, and the SH3 ligand was attached to the N-terminus, middle, and C-terminus of E. coli GadC. In the C-terminus model, 5.8 g/l of GABA concentration was obtained from 10 g/l glutamate. When a competing pathway engineered strain was used, the final GABA concentration was further increased to 5.94 g/l, which corresponds to 97.5% of GABA yield. With the introduction of the scaffold complex, the GABA productivity increased by 2.9 folds during the initial culture period.

BMP-2 Immoblized in BCP-Chitosan-Hyaluronic Acid Hybrid Scaffold for Bone Tissue Engineering

  • Nath, Subrata Deb;Abueva, Celine;Sarkar, Swapan Kumar;Lee, Byong Taek
    • Korean Journal of Materials Research
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    • v.24 no.12
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    • pp.704-709
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    • 2014
  • In this study, we fabricated a novel micro porous hybrid scaffold of biphasic calcium phosphate (BCP) and a polylectrolyte complex (PEC) of chitosan (CS) and hyaluronic acid (HA). The fabrication process included loading of CS-HA PEC in a bare BCP scaffold followed by lypophilization. SEM observation and porosimetry revealed that the scaffold was full of micro and macro pores with total porosity of more than 60 % and pore size in the range of $20{\sim}200{\mu}m$. The composite scaffold was mechanically stronger than the bare BCP scaffold and was significantly stronger than the CS-HA PEC polymer scaffold. Bone morphogenetic growth factor (BMP-2) was immobilized in CS-HA PEC in order to integrate the osteoinductive potentiality required for osteogenesis. The BCP frame, prepared by sponge replica, worked as a physical barrier that prolonged the BMP-2 release significantly. The preliminary biocompatibility data show improved biological performance of the BMP-2 immobilized hybrid scaffold in the presence of rabbit bone marrow stem cells (rBMSC).

Experimental Research of ZrO2/BCP/PCL Scaffold with Complex Pore Pattern for Bone Tissue Regeneration (골 조직 재생을 위한 복합 공극 패턴을 가진 ZrO2/BCP/PCL 인공지지체의 실험적 평가)

  • Sa, Min-Woo;Shim, Hae-Ri;Kim, Jong Young
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.39 no.11
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    • pp.1153-1159
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    • 2015
  • Recently, synthetic biopolymers and bioceramics such as poly (${\varepsilon}$-caprolactone)(PCL), hydroxyapatite, tricalcium phosphate, biphasic calcium phosphate(BCP), and zirconia have been used as substrates to generate various tissues or organs in tissue engineering. Thus, the purpose of this study was the characterization of $ZrO_2$/BCP/PCL(ZBP) scaffold for bone tissue regeneration. Based on the result of single-line test, blended 3D ZBP scaffolds with fully interconnected pores and new complex pore pattern of $45^{\circ}+135^{\circ}$-type and staggered-type were successfully fabricated using a polymer deposition system. Furthermore, the effect of ZBP scaffold on mechanical property was analyzed. In addition, in vitro cell interaction of ZBP scaffold on MG63 cells was evaluated using a cell counting kit-8(CCK-8) assay.

Development of bone scaffold using HA(Hydroxyapatite) nano powder (HA(Hydroxyapatite) 나노 입자를 이용한 bone scaffold의 개발)

  • Kim J.Y.;Lee S.J.;Lee J.W.;Kim Shin-Yoon;Cho D.W.
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2006.05a
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    • pp.159-160
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    • 2006
  • A novel approach to the manufacture of biocompatible ceramic scaffold for tissue engineering using micro-stereolithography system is introduced. Micro-stereolithography is a newly proposed technology that enables to make a 3D micro structure. The 3D micro structures made by this technology can have accurate and complex shape within a few micron error. Therefore, the application based on this technology can vary greatly in nano-bio fields. Recently, tissue-engineering techniques have been regarded as alternative candidate to treat patients with serious bone defects. So many techniques to design and fabricate 3D scaffolds have been developed. But the imperfection of scaffold such as random pore size and porosity causes a limitation in developing optimum scaffold. So scaffold development with controllable pore size and fully interconnected shape have been needed for a more progress in tissue engineering. In this paper, bone scaffold was developed by applying the micro-stereolithography to the mold technology. The scaffold material used was HA(Hydroxyapatite) nano powder. HA is a type of calcium phosphate ceramic with similar characteristic to human inorganic bone component. The bone scaffold made by HA is expected, in the near future, to be an efficient therapy for bone defect.

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Co-Localization of GABA Shunt Enzymes for the Efficient Production of Gamma-Aminobutyric Acid via GABA Shunt Pathway in Escherichia coli

  • Pham, Van Dung;Somasundaram, Sivachandiran;Park, Si Jae;Lee, Seung Hwan;Hong, Soon Ho
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.710-716
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    • 2016
  • Gamma-aminobutyric acid (GABA) is a non-protein amino acid, which is an important inhibitor of neurotransmission in the human brain. GABA is also used as the precursor of biopolymer Nylon-4 production. In this study, the carbon flux from the tricarboxylic acid cycle was directed to the GABA shunt pathway for the production of GABA from glucose. The GABA shunt enzymes succinate-semialdehyde dehydrogenase (GabD) and GABA aminotransferase (GabT) were co-localized along with the GABA transporter (GadC) by using a synthetic scaffold complex. The co-localized enzyme scaffold complex produced 0.71 g/l of GABA from 10 g/l of glucose. Inactivation of competing metabolic pathways in mutant E. coli strains XBM1 and XBM6 increased GABA production 13% to reach 0.80 g/l GABA by the enzymes co-localized and expressed in the mutant strains. The recombinant E. coli system developed in this study demonstrated the possibility of the pathway of the GABA shunt as a novel GABA production pathway.

Performance Improvement of Polymer Deposition System by Nozzle Guide and Its Application to Washer Scaffold Fabrication (노즐 가이드를 적용한 폴리머 적층 시스템의 Washer Scaffold 제작을 위한 성능 개선)

  • Sa, Min-Woo;Kim, Jong Young
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.37 no.3
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    • pp.249-257
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    • 2013
  • Rapid prototyping was used to design and develop a three-dimensional (3D) scaffold for tissue engineering application. In this study, the nozzle guide (TB-CP-HN, MUSASHI ENGINEERING, INC., JAPAN) used with the syringe of the polymer deposition system (PDS) was evaluated by measuring the scaffold line width and height. 3D scaffolds were fabricated using a biodegradable polymer called poly-caprolactone (PCL). The PCL polymer can be deposited from the needle of a syringe using a 200-${\mu}m$ precision nozzle, at a pressure of 600 kPa and temperature of $125^{\circ}C$. The advantages and improvements in this nozzle guide were addressed through washer scaffold fabrication. Overall, this research indicated that the fabrication of a complex-shaped scaffold using an enhanced polymer deposition system may have potential for tissue engineering.

Three-Dimensional Porous Collagen/Chitosan Complex Sponge for Tissue Engineering

  • Kim, Sung Eun;Cho, Yong Woo;Kang, Eun Jung;Kwon, Ick Chan;Lee, Eunhee Bae;Kim, Jung Hyun;Chung, Hesson;Jeong, Seo Young
    • Fibers and Polymers
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    • v.2 no.2
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    • pp.64-70
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    • 2001
  • A three-dimensional, porous collagen/chitosan complex sponge was prepared to closely simulate basic extracellular matrix (ECM) constitutes, collagen and glycosaminoglycan. The complex sponge was prepared by a lyophilization method and had the regular network with highly porous structure, suitable for cell adhesion and growth. The pores were well interconnected, and their distribution was fairly homogeneous. The complex sponge was crosslinked using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to increase its boilogical stability and enhance its mechanical properties. The crosslinking medium has a great effect on the inner structure of the sponge. The homogeneous, porous structure of the sponge was remarkably collapsed in an aqueous crosslinking medium. However, the morphology of the sponge remained almost intact in a water/ethanol mixture crosslinking milieu. Mechanical properties of the collagen/chitosan sponge were significantly enhanced by EDC-mediated crosslinking. The potential of the sponge as a scaffold for tissue engineering was investigated using a Chinese hamster ovary cell (CHO-K1) line.

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Fabrication of Biodegradable Microstructures using Projection Microstereolithography Technology (프로젝션 마이크로광조형 기술을 이용한 생분해성 마이크로구조물 제작)

  • Choi, Jae-Won;Ha, Young-Myoung;Park, In-Baek;Ha, Chang-Sik;Lee, Seok-Hee
    • Proceedings of the KSME Conference
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    • 2007.05a
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    • pp.1259-1264
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    • 2007
  • Microstereolithography technology has potential capability for fabrication of 3D microstructures. It evolved from conventional SLA which is one of the RP processes. In a microstereolithography process, 3D microstructures can be easily fabricated by continuously stacking 2D layer which is photopolymerized using a liquid prepolymer. Combination between biocompatible/biodegradable photocurable prepolymer and 3D complex fabrication in microstereolithography makes broad application areas such as medical, pharmaceutic, and bio devices. In particular, a 3D microneedle for transdermal drug delivery and a scaffold for tissue engineering are fabricated using this technology. In this paper, the authors address development of microstereolithography system adapted to large surface and fabrication of various microstructures. In addition, to apply human body we suggest a biodegradable 3D microneedle and a scaffold using biodegradable photocurable prepolymer.

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An Overview of Laser-assisted Bioprinting (LAB) in Tissue Engineering Applications

  • Ventura, Reiza Dolendo
    • Medical Lasers
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    • v.10 no.2
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    • pp.76-81
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    • 2021
  • Biological tissues and organs are composed of different arrays of cells, biochemical cues, and extracellular matrices arranged in a complex microarchitecture. Laser-Assisted Bioprinting (LAB) is an emerging and promising technology that is reproducible with high accuracy that can be used for fabricating complex bioengineered scaffolds that mimic tissues and organs. The LAB process allows researchers to print intricate structural scaffolds using cells and different biomaterials essential for facilitating cell-scaffold interaction and to induce tissue and organ regeneration which cannot be achieved in a traditional scaffold fabrication. This process can fabricate artificial cell niches or architecture without affecting cellular viability and material integrity. This review tackles the basic principles and key aspects of Laser-Assisted Bioprinting. Recent advances, limitations, and future perspectives are also discussed.

Development of Three-dimensional Scaffold for Cartilage Regeneration using Microstereolithography (마이크로 광 조형 기술을 이용한 연골조직 재생용 3 차원 인공지지체 개발)

  • Lee, Seung-Jae;Kang, Tae-Yun;Park, Jung- Kyu;Rhie, Jong-Won;Hahn, Sei-Kwang;Cho, Dong-Woo
    • Proceedings of the KSME Conference
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    • 2007.05a
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    • pp.1265-1270
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    • 2007
  • Conventional methods for fabricating three-dimensional (3-D) scaffolds have substantial limitations. In this paper, we present 3-D scaffolds that can be made repeatedly with the same dimensions using a microstereolithography system. This system allows the fabrication of a pre-designed internal structure, such as pore size and porosity, by stacking photopolymerized materials. The scaffolds must be manufactured in a material that is biocompatible and biodegradable. In this regard, we synthesized liquid photocurable biodegradable TMC/TMP, followed by acrylation at terminal ends. And also, solidification properties of TMC/TMP polymer are to be obtained through experiments. Cell adhesion to scaffolds significantly affects tissue regeneration. As a typical example, we seeded chondrocytes on two types of 3-D scaffold and compared the adhesion results. Based on these results, the scaffold geometry is one of the most important factors in chondrocyte adhesion. These 3-D scaffolds could be key factors for studying cell behavior in complex environments and eventually lead to the optimum design of scaffolds for the regeneration of various tissues, such as cartilage and bone.

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