• Title/Summary/Keyword: cell scaffolds

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In vitro Cartilage Regeneration using Primary Chondrocytes Cultured within Porous Poly(lactide-co-glycolide) Scaffolds

  • Yun, Jun-Jin;Go, Ye-Jeong;Baek, Jeong-Hwan;Park, Tae-Gwan
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.421-424
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    • 2000
  • Cartilage injuries are frequent nowadays. The previous surgical treatment of cartilage defect was limited. Another approach in the treatment of cartilage injuries is the use of reconstitute cartilage consisting of chondrocytes cultured in suitable biodegradable scaffolds. Current studies have demonstrated the compatibility of chondrocytes with different biomaterials and the chondrogenesis in various types of porous scaffolds. The cell ingrowth into the porous scaffolds is modulated by initial cell loading efficiency. Therefore, well-interconnected pore structure and even pore distribution of the scaffolds are essential for efficient cell seeding. According to our previous work, well-interconnected macroporous scaffolds can be prepared by gas-foaming/salt-leaching method using ammonium bicarbonate salt as porogen additives. In this work, primary chondrocytes were cultured in PLGA 65/35 scaffolds fabricated by using our method. Cells seeded in the scaffolds showed well distribution by agitated seeding method. Histochemical staining of proteoglycans present in the scaffolds was used to visualize the chondrocyte ingrowth in the scaffolds. At 3 weeks, the population of chondrocytes was increased for the most part of the scaffolds, and extra cellular matrix (ECM) secretion was increased as culture periods progressed.

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Continuos-Flow culture of Hepatocytes in Sugar-derivatized poly (lactide-co-glycolide) Scaffolds Prepared by Gas-foaming/salt-leaching Method

  • Yun, Jun-Jin;Park, Tae-Gwan
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.141-144
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    • 2000
  • Highly open porous polymer matrices are required for high density cell seeding, efficient nutrient, and oxygen supply to the cells cultured in the three dimensional matrices. However, there are severe problems of mass transfer limitations within the cell/scaffolds culture system. Thus we hypothesize that continuos-flow culture conditioning of cells with the scaffolds may improve the cell viability and the differentiated function. In this study, we fabricated porous PLGA scaffolds by using gas-foaming/salt-leaching method as previous described. Viscous PLGA gel paste contains ammonium bicarbonate particulates, acting as a gas-foaming agent as well as a salt-leaching porogen, were cast into Teflon mold and dried. Ammonium bicarbonate salt upon contact to an acidic aqueous solution evloves gaseous ammonia and carbon dioxide by itself. And we conjugated galactose moiety [AGA; $N-(aminobuty1)-O-{\beta}-D-galactopyranosyl-(1{\rightarrow}4)-D-glucoamide]$ to the terminal end group of a PLGA to increase the cell adhesion and matain the differentiated function of hepatocytes. Cell-seeded scaffolds were secured in a flow bioreactor chamber and exposed to continuous flow at 5 ml/min. As a result of our study, the high yield of hepatocytes attachment was accomplished by increasing the concentration of PLGA-AGA conjugate in polymer scaffolds and cells in the scaffolds under continuos flow condition maintained a high level of viability and albumin secretion rate of cultured hepatocytes showed a higher level that of control groups.

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Effects of Three-dimensional Scaffolds on Cell Organization and Tissue Development

  • Yan Li;Yang, Shang-Tian
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.5
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    • pp.311-325
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    • 2001
  • Tissue engineering scaffolds play a critical role in regulating the reconstructed human tissue development. Various types of scaffolds have been developed in recent years, including fibrous matrix and foam-like scaffolds. The design of scaffold materials has been investigated extensively. However, the design of physical structure of the scaffold, especially fibrous matrices, has not received much attention. This paper compares the different characteristics of fibrous and foam-like scaffolds, and reviews regulatory roles of important scaffold properties, including surface geometry, scaffold configuration, pore structure, mechanical property and bioactivity. Tissue regeneration, cell organization, proliferation and differentiation under different microstructures were evaluated. The importance of proper scaffold selection and design is further discussed with the examples of bone tissue engineering and stem cell tissue engineering. This review addresses the importance of scaffold microstructure and provides insights in designing appropriate scaffold structure for different applications of tissue engineering.

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The Need for Research on the Comparison of Sensory Characteristics between Cultured Meat Produced Using Scaffolds and Meat

  • Sol-Hee Lee;Jungseok Choi
    • Food Science of Animal Resources
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    • v.44 no.2
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    • pp.269-283
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    • 2024
  • Cultured meat is one of the research areas currently in the spotlight in the agricultural and livestock industry, and refers to cells obtained from livestock that are proliferated and differentiated and processed into edible meat. These cell-cultured meats are mainly studied at the lab-scale by culturing them in flasks, and for commercial use, they are produced using scaffolds that mimic cell supports. Scaffolds are broadly divided into fiber scaffolds, hydrogels, and micro-carrier beads, and these are classified according to processing methods and materials. In particular, a scaffold is essential for mass production, which allows it to have appearance, texture, and flavor characteristics similar to meat. Because cultured meat is cultured in a state where oxygen is blocked, it may be lighter in color or produce less flavor substances than edible meat, but these can be compensated for by adding natural substances to the scaffolds or improving fat adhesion. In addition, it has the advantage of being able to express the texture characteristics of the scaffolds that make up the meat in various ways depending on the materials and manufacturing methods of the scaffolds. As a result, to increase consumers' preference for cultured meat and its similarity to edible meat, it is believed that manufacturing scaffolds taking into account the characteristics of edible meat will serve as an important factor. Therefore, continued research and interest in scaffolds is believed to be necessary.

Polymer brush: a promising grafting approach to scaffolds for tissue engineering

  • Kim, Woonjung;Jung, Jongjin
    • BMB Reports
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    • v.49 no.12
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    • pp.655-661
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    • 2016
  • Polymer brush is a soft material unit tethered covalently on the surface of scaffolds. It can induce functional and structural modification of a substrate's properties. Such surface coating approach has attracted special attentions in the fields of stem cell biology, tissue engineering, and regenerative medicine due to facile fabrication, usability of various polymers, extracellular matrix (ECM)-like structural features, and in vivo stability. Here, we summarized polymer brush-based grafting approaches comparing self-assembled monolayer (SAM)-based coating method, in addition to physico-chemical characterization techniques for surfaces such as wettability, stiffness/elasticity, roughness, and chemical composition that can affect cell adhesion, differentiation, and proliferation. We also reviewed recent advancements in cell biological applications of polymer brushes by focusing on stem cell differentiation and 3D supports/implants for tissue formation. Understanding cell behaviors on polymer brushes in the scale of nanometer length can contribute to systematic understandings of cellular responses at the interface of polymers and scaffolds and their simultaneous effects on cell behaviors for promising platform designs.

Preparation and Biocompatibility of Composite Bone Scaffolds Using Gnotobiotic Pig Bones (무균돼지뼈를 이용한 복합 골지지체의 제조와 생체적합성 평가)

  • Im, Ae-Lee;Chung, Jong-Hoon;Lim, Ki-Taek;Choung, Pill-Hoon;Hong, Ji-Hyang
    • Journal of Biosystems Engineering
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    • v.32 no.1 s.120
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    • pp.50-56
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    • 2007
  • Highly porous composite bioceramic bone scaffolds were developed using sintered gnotobiotic pig bones. These scaffolds consisted of poly-D,L-lactic acid (P(D,L)LA) and bioceramic materials of pig bone powder. The bone scaffolds were able to promote biocompatibility and possess interconnected pores that would support cell adhesion and proliferation adequately. The composite scaffolds were tested with dental pulp stem cells for cytotoxicity test. Cells seeded on the composite scaffolds were readily attached, well proliferated, as confirmed by cytotoxicity test, and cell adhesion assessment. The composite bone scaffold had no toxicity in cytotoxicity test on the extract of 0.013 g scaffold to 2 ml culture medium. The cells on the composite bone scaffold proliferated better than cells on the P(D,L)LA scaffolds.

Fabrication of Nanofiber-Combined 3D Scaffolds using Dual-Head Deposition Technology (듀얼헤드 적층 기술을 이용한 나노섬유로 결합된 3D 인공지지체 제작)

  • Sa, Min-Woo;Lee, Chang-Hee;Kim, Jong Young
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.17 no.1
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    • pp.108-115
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    • 2018
  • In bone tissue engineering, polycaprolactone (PCL) is one of the most widely used biomaterials to manufacture scaffolds as a synthetic polymer with biodegradability and biocompatibility. The polymer deposition system (PDS) with four axis heads, which can dispense bio-polymers, has been used in scaffold fabrication for tissue engineering applications. A dual-head deposition technology of PDS is an effective technique to fabricate 3D scaffolds. The electrospinning technology has been widely used to fabricate porous and highly interconnected polymer fibers. Thus, PDS can fabricate nanofiber-combined hybrid scaffolds using fused deposition modeling (FDM) and electrospinning methods. This study aims to fabricate nanofiber-combined scaffolds with uniform nanofibers using PDS. The PCL nanofibers were fabricated and evaluated according to the fabrication process parameters. PCL nanofibers were successfully fabricated when the applied voltage, tip-to-collector distance, flow rate, and solution concentration were 5 kV, 1 cm, 0.1 ml/h, and 8 wt%, respectively. The cell proliferation was evaluated according to the electrospinning time. Scanning electron microscopy was used to acquire images of the cross-sectioned hybrid scaffolds. The cell proliferation test of the PCL and nanofiber-combined hybrid scaffolds was performed using a CCK-8 assay according to the electrospinning time. The result of in-vitro cell proliferation using osteosarcoma MG-63 cells shows that the hybrid scaffold has good potential for bone regeneration.

Evaluation of polyglycolic acid as an animal-free biomaterial for three-dimensional culture of human endometrial cells

  • Sadegh Amiri;Zohreh Bagher;Azadeh Akbari Sene;Reza Aflatoonian;Mehdi Mehdizadeh;Peiman Broki Milan;Leila Ghazizadeh;Mahnaz Ashrafi;FatemehSadat Amjadi
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.4
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    • pp.259-269
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    • 2022
  • Objective: Animal-free scaffolds have emerged as a potential foundation for consistent, chemically defined, and low-cost materials. Because of its good potential for high biocompatibility with reproductive tissues and well-characterized scaffold design, we investigated whether polyglycolic acid (PGA) could be used as an animal-free scaffold instead of natural fibrin-agarose, which has been used successfully for three-dimensional human endometrial cell culture. Methods: Isolated primary endometrial cells was cultured on fibrin-agarose and PGA polymers and evaluated various design parameters, such as scaffold porosity and mean fiber diameter. Cytotoxicity, scanning electron microscopy (SEM), and immunostaining experiments were conducted to examine cell activity on fabricated scaffolds. Results: The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and SEM results showed that endometrial cells grew and proliferated on both scaffolds. Immunostaining showed cytokeratin and vimentin expression in seeded cells after 7 days of culture. On both scaffolds, an epithelial arrangement of cultured cells was found on the top layer and stromal arrangement matrix on the bottom layer of the scaffolds. Therefore, fibrin-agarose and PGA scaffolds successfully mimicked the human endometrium in a way suitable for in vitro analysis. Conclusion: Both fibrin-agarose and PGA scaffolds could be used to simulate endometrial structures. However, because of environmental and ethical concerns and the low cost of synthetic polymers, we recommend using PGA as a synthetic polymer for scaffolding in research instead of natural biomaterials.

Latent Transforming Growth Factor-beta1 Functionalised Electrospun Scaffolds Promote Human Cartilage Differentiation: Towards an Engineered Cartilage Construct

  • Lim, Erh-Hsuin;Sardinha, Jose Paulo;Myers, Simon;Stevens, Molly
    • Archives of Plastic Surgery
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    • v.40 no.6
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    • pp.676-686
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    • 2013
  • Background To overcome the potential drawbacks of a short half-life and dose-related adverse effects of using active transforming growth factor-beta 1 for cartilage engineering, a cell-mediated latent growth factor activation strategy was developed incorporating latent transforming growth factor-${\beta}$1 (LTGF) into an electrospun poly(L-lactide) scaffold. Methods The electrospun scaffold was surface modified with NH3 plasma and biofunctionalised with LTGF to produce both random and orientated biofunctionalised electrospun scaffolds. Scaffold surface chemical analysis and growth factor bioavailability assays were performed. In vitro biocompatibility and human nasal chondrocyte gene expression with these biofunctionalised electrospun scaffold templates were assessed. In vivo chondrogenic activity and chondrocyte gene expression were evaluated in athymic rats. Results Chemical analysis demonstrated that LTGF anchored to the scaffolds was available for enzymatic, chemical and cell activation. The biofunctionalised scaffolds were non-toxic. Gene expression suggested chondrocyte re-differentiation after 14 days in culture. By 6 weeks, the implanted biofunctionalised scaffolds had induced highly passaged chondrocytes to re-express Col2A1 and produce type II collagen. Conclusions We have demonstrated a proof of concept for cell-mediated activation of anchored growth factors using a novel biofunctionalised scaffold in cartilage engineering. This presents a platform for development of protein delivery systems and for tissue engineering.

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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