• Title/Summary/Keyword: 지지세포

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Effects of SIS/PLGA Porous Scaffolds and Muscle-Derived Stem Cell on the Formation of Tissue Engineered Bone (SIS/PLGA 담체와 근육유래 줄기세포를 이용한 생체조직공학적 골재생)

  • Kim Soon Hee;Yun Sun Jung;Jang Ji Wook;Kim Moon Suk;Khang Gilson;Lee Hai Bang
    • Polymer(Korea)
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    • v.30 no.1
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    • pp.14-21
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    • 2006
  • Tissue engineering techniques require the use of a porous biodegradable/bioresorbable scaffold, which server as a three-dimensional template for initial cell attachment and subsequent tissue formation in both in vitro and in vivo. Small intestinal submucosa (SIS) has been investigated as a source of collagenous tissue with the potential to be used as biomaterials because of its inherent strength and biocompatibility. SIS-loaded poly(L-lactide-co-glicolide)(PLGA) scaffolds were prepared by solvent casting/particle leaching. Characterizations of SIS/PLGA scaffold were carried out by SEM, mercury porosimeter, and so on. Muscle-derived stem cells can be differentiated in culture into osteoblasts, chondrocytes, and even myoblasts by the controlling the culture environment. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide(MTT) test. Osteogenic differential cells were analyzed by alkaline phosphatase(ALP) activity. SIS/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of SIS on the osteoinduction compared with controlled PLGA scaffolds. Thin sections were cut from paraffin embedded tissues and histological sections were conducted hematoxylin and eosin (H&E), Trichrome, and von Kossa. We observed that bone formatioin of SIS/PLGA hybrid scaffold as natural/synthetic scaffold was better thean that of only PLGA scaffold. It canb be explained that SIS contains various kinds of bioactive molecules for osteoinduction.

Tissue Engineered Bone Formation Using Porous Chitosan and Chitosan/Tricalcium Phosphate Matrices (Chitosan 및 Chitosan/Tricalcium Phosphate 다공성 기질을 이용한 조직공학적 골형성에 관한 연구)

  • Lee, Yong-Moo;Choi, Sang-Mook;Park, Yoon-Jeong;Lee, Seung-Jin;Ku, Young;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.28 no.4
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    • pp.577-604
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    • 1998
  • chitosan은 골치유증진 및 골세포의 분화를 촉진하는 것으로 알려진 천연의 생분해성 고분자이다. 이연구에서는 chitosan 및 chitosan/tricalcium phosphate(TCP) 다공성 기질을 제조하여 골이식재 및 조직공학적 골형성을 위한 3차원적 세포배양 지지체로서의 가능성을 평가하고자 하였다. chitosan 용액 및 TCP가 포함된 chitosan 용액을 동결건조함으로써 소공의 크기가 $100-200{\mu}m$인 스폰지형태의 chitosan 및 chitosan/TCP 다공성 기질을 제작하였다. 골이식재로서의 효과를 평가하기 위하여 백서의 두개골 결손부에 제작된 chiosan 및 chitosan/TCP 다공성 기질을 각각 이식하고 2주 및 4주 후에 동물을 희생하여 조직학적으로 치유양상을 관찰하였다. 조직공학적 골형성을 위한 세포배양 지지체로서의 가능성을 평가하기 위하여 백서 태자의 두개골에서 분리된 골아세포를 chitosan 및 chitosan/TCP 다공성 기질에 각각 접종하고 56일간 배양하면서 각 기간 별로 세포수, 염기성 인산효소 활성, 축적된 calcium의 양을 측정하였고 배양된 세포-기질 혼합체를 광학현미경 및 주사전자현 미경하에서 조직학적 관찰을 시행하였다. 백서 두개골결손부에 이식된 chitosan 및 chiosan/TCP 다공성 기질은 별다른 이물반응 없이 자연 분해되면서 신생골조직 내에 매립되었으며 이식하지 않은 대조군에 비해 유의하게 높은 신생골형성 효과를 나타내어 우수한 골전도성이 있음이 확인되었다. 신생골형성 양상이나 형성된 양에 있어서 두 가지 기질간의 유의한 차이는 없었다. 골아세포-기질 혼합체의 배양결과, 접종후 배양 28일 경과 시까지 골아세포수는 지속적으로 증가하다가 이후에는 5 8일까지 성장정도가 둔화되었다. 염기성 인산효소의 활성 및 calcium 축적량은 접종후 배양시간경과에 따라 56일까지 지속적으로 증가하였다. 세포수 및 염기성 인산효소의 활성에서 두 기질간의 유의한 차이는 없었고, calcium 축적량에 있어서는 chitosan/TCP 기질에서 유의하게 높았고 증가속도도 컸다. 배양된 골아세포가 접종된 다공성 기질의 조직학적 관찰결과, 골아세포는 다공성 기질에 잘 부착하여 중층의 형태로 성장하면서 광화된 골기질을 형성함이 관찰되었다. 배양 14일부터 작은 골편형태의 골형성이 기질 표면에 부착되어 관찰되었고, 배양기간이 길어짐에 따라 성장하여 배양 56일째에는 상당한 양의 광화된 골질이 형성됨이 관찰되었다. 배양 56일 경과후의 광화된 골질의 양은 chitosan/TCP 기질에서 더 많았다. 이 연구의 결과, chitosan 및 chitosan/TCP 다공성 기질이 골이식재로서 뿐만 아니라, 조직공학적 골형성에 적용되는 골아세포의 배양을 위한 3차원구조의 세포지지체로 이용되어 골재생술식에 유용한 생체재료로 활용될 수 있음이 확인되었다.

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Effects of Demineralized Bone Particle Loaded Poly(lactic-co-glycolic acid) Scaffolds on the Attachment and Proliferation of Costal Cartilage Cells (탈미네랄화된 골분/PLGA 지지체에서 늑연골 세포의 부착과 성장에 미치는 영향)

  • Cho, Sun Ah;Song, Jeong Eun;Kim, Kyoung Hee;Ko, Hyun Ah;Lee, Dongwon;Kwon, Soon Yong;Chung, Jin Wha;Khang, Gilson
    • Polymer(Korea)
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    • v.37 no.5
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    • pp.632-637
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    • 2013
  • It has been widely accepted that costal cartilage cells (CCs) have more excellent initial proliferation capacity than articular cartilage cells as well as the easiness for isolation and collection. This study demonstrated that CCs might be one of the substitutes for articular cartilage cells by tissue engineered cartilage. Poly(lactic-co-glycolic acid) (PLGA) has been extensively tested and used as scaffold material but it was limited by the low attachment of cells and the induction of inflammatory cells. Base on previous our studies, we confirmed demineralized bone particle (DBP) had the power of the reduction of inflammatory reaction and the stimulation proliferation of cells. We fabricated PLGA scaffold loaded with 10, 20, 40 and 80 wt% DBP and then tested the possibility of the regeneration of cartilage using CCs. Assays of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and scanning electron microscope (SEM) carried out to evaluate the attachment and proliferation of CCs in DBP/PLGA scaffolds. Glycosaminoglycan (sGAG) and collagen contents assay were conducted to confirm the effects of DBP on formation of extracellular matrix. This study demonstrated that DBP/PLGA scaffolds showed significant positive effects on cell growth and proliferation due to the vitality of DBP as well as the possibility of the application of CCs for tissue engineered cartilage.

Characteristic Analysis and Fabrication of Bioceramic Scaffold using Mixing Ratios of TCP/HA by Fused Deposition Modeling (압출 적층 조형 기술을 이용한 TCP/HA 의 혼합비율에 따른 바이오 세라믹 인공지지체의 제작 및 특성 연구)

  • Sa, Min-Woo;Kim, Jong Young
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.38 no.11
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    • pp.1273-1281
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    • 2014
  • Tissue engineering is an emerging research field that has the potential to restore, regenerate and repair damaged bone tissue and organs. Tricalcium phosphate and hydroxyapatite biomaterials-based calcium phosphate are excellent materials that have both osteoconduction and biocompatibility for bone tissue regeneration. In this study, solution structures were successfully fabricated using a fused deposition modeling system based on deposition and heating devices. The morphology characteristics of the bioceramic scaffolds sintered at a temperature of $1,300^{\circ}C$ were analyzed by scanning electron microscopy. The effects of various blended TCP/HA ratio on the microstructure and shrinkage were studied. The mechanical properties of the scaffolds were measured using a compression testing machine from stress-strain curves on the crosshead velocity of 1 mm/min. The fabricated scaffolds were evaluated by cell proliferation tests of MG-63 cells. The results of this study suggest that the blended TCP(75 wt%)/HA(25 wt%) scaffold is an appropriate scaffold for bone tissue regeneration.

Controlling Pore Size of Electrospun Silk Fibroin Scaffold for Tissue Engineering (전기방사를 이용한 조직공학용 실크 피브로인 나노 섬유 지지체의 기공 크기 조절)

  • Cho, Se-Youn;Park, Hyun-Ho;Jin, Hyoung-Joon
    • Polymer(Korea)
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    • v.36 no.5
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    • pp.651-655
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    • 2012
  • Considerable effort has been directed toward the use of silk fibroin as a biotechnological material in biomedical applications on account of its excellent biodegradability, biocompatibility, and unique mechanical properties. For use in tissue engineering, it is very important to design and control the pore architecture of polymeric scaffolds, which provide the vital framework for seeded cells to organize into functioning tissue. In the present study, a silk fibroin scaffold with controlled interconnectivity and pore size was prepared using an electrospinning method with poly(ethylene oxide).

Effect of the Mechanical Properties of Cell-Interactive Hydrogels on a Control of Cell Phenotype (세포친화적 하이드로젤의 기계적 물성이 세포 표현형 제어에 미치는 영향)

  • Kim, Do Yun;Park, Honghyun;Lee, Kuen Yong
    • Polymer(Korea)
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    • v.39 no.3
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    • pp.412-417
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    • 2015
  • A critical element in tissue engineering approaches is a control of the mechanical properties of polymer scaffolds to regulate cell phenotype, which may lead to clinically successful tissue regeneration. In this study, we hypothesized that gel stiffness could be a key factor to manipulate adhesion and proliferation of different types of cells. RGD-modified alginate gels with various mechanical properties were prepared and used as a substrate for MC3T3-E1 and H9C2 cells. Adhesion and growth rate of MC3T3-E1 cells in vitro were increased in parallel with an increase of gel stiffness. In contrast, those of H9C2 cells were decreased. This approach to control the mechanical properties of polymer scaffolds depending on the cell types may find useful applications in the tissue engineering.

Preparation and Characterization of Ipriflavone-Loaded Poly(L-lactide-co-glycolide) Scaffold for Tissue Engineered Bone (조직공학적 골을 위한 애프리플라본을 함유한 다공성 지지체의 제조 및 그 특성)

  • Jang, Ji-Wook;Lee, Bong;Han, Chang-Whan;Lee, Il-Woo;Lee, Hai-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.27 no.3
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    • pp.226-234
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    • 2003
  • Ipriflavone (IP), a non-hormonal isoflavone derivative, has been shown to interfere with bone remodeling by inhibiting bone resorption and stimulating bone formation. IP consistently increased the amount of Ca incorporated into the cell layer by mesenchymal stem cells (MSCs). In this study, we developed the novel IP loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds for the possibility of the application of the tissue engineered bone. IP/PLGA scaffo1ds were prepared by solvent casting/salt leaching method and were characterized by porosimeter, scanning electron microscopy, determination of residual salt amount, differential scanning calorimetry, and X-ray diffractometer, respectively. IP/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of IP on the osteoinduction compared with control PLGA scaffo1ds. Thin sections were cut from paraffin embedded tissues and histological sections were stained H&E, von Kossa, and immunohistochemical staining for Type I collagen and osteocalcin. It can be observed that the porosity was above 91.7% and the pore size was above 101 $\mu\textrm{m}$. Control scaffo1d and IP/PLGA scaffo1ds of 50% IP were implanted on the back of athymic nude mouse to observe the effect of IP on the induction of cells proliferation for 9 weeks. The evidence of calcification, osteoblast, and osteoid from the undifferentiated stem cells in the subcutaneous sites and other soft connective tissue sites having a preponderance of stem cells has been observed. From these results, it seems that IP plays an important role for bone induction in IP/PLCA scaffolds.

Preparation and Characterization of Hyaluronic Acid Loaded PLGA Scaffold by Emulsion Freeze-Drying Method (히알루론산을 함유한 PLGA 지지체의 제조 및 특성결정)

  • Ko, Youn-Kyung;Kim, Soon-Hee;Jeong, Jae-Soo;Park, Jung-Soo;Lim, Ji-Ye;Kim, Moon-Suk;Lee, Hae-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.31 no.6
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    • pp.505-511
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    • 2007
  • Poly(lactide-co-glycolide)(PLGA) and hyaluronic acid (HA) has been widely used as biocompatible scaffold materials to regenerate tissue. In this present study, we fabricated microporous PLGA and HA loaded PLGA scaffolds by a emusion freeze-drying method. In order to confirm that the release profile of cytokine or water-soluble drugs, we manufactured the granulocyte macrophage colony stimulating factor(GM-CSF) loaded PLGA and HA-PLGA scaffold. All scaffolds were characterized using scanning electron microscope(SEM), mercury porosimeter and wettability measurement. Cell proliferation and viability were assessed by a 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) test. The porosity of HA-PLGA scaffold was greater than 95% with the total pore area of $261\;m^2/g$. The HA-FLGA scaffold exhibited well interconnected pores to allow greater cell adhesion and prolixferation. It was proven by higher cell viability in the HA-PLGA scaffold than PLGA alone. This may be due to the enhanced natural properties and higher water retention capacity of HA.

Mechanical Property and Cell Compatibility of Silk/PLGA Hybrid Scaffold; In Vitro Study (실크/PLGA 하이브리드 지지체의 기계적 물성과 세포친화력; in vitro 연구)

  • Song, Yi-Seul;Yoo, Han-Na;Eum, Shin;Kim, On-You;Yoo, Suk-Chul;Kim, Hyung-Eun;Lee, Dong-Won;Khang, Gil-Son
    • Polymer(Korea)
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    • v.35 no.3
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    • pp.189-195
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    • 2011
  • The design of new bioactive scaffolds offering physiologic environment for tissue formation is an important frontier in biomaterials research. In this study, we performed compressive strength, water-uptake ability, and SEM analysis for physical property assessment of 3-D silk/PLGA scaffold, and investigated the adhesion, proliferation, phenotype maintenance, and inflammatory responses of RAW 264.7 and NIH/3T3 for cell compatibility. Scaffolds were prepared by the solvent casting/salt leaching method and their compressive strength and water-uptake ability were excellent at 20 wt% silk content. Result of cell compatibility assay showed that inflammatory responses distinctly decreased, and initial adhesion and proliferation were maximized at 20 wt% silk content. In conclusion, we suggest that silk/PLGA scaffolds may be useful to tissue engineering applications.

Preparation and Characterization of PLGA Scaffold Impregnated Keratin for Tissue Engineering Application (케라틴이 함유된 조직공학적 PLGA 지지체의 제조 및 특성 분석)

  • Oh, A-Young;Kim, Soon-Hee;Lee, Sang-Jin;Yoo, James J.;Dyke, Mark van;Rhee, John M.;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.5
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    • pp.403-408
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    • 2008
  • Keratin is the major structural fibrous protein providing outer covering such as wool, hair, and nail. Keratin is useful as natural protein. We developed the keratin loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds (keratin/PLGA) for the possibility of the application of the tissue engineering using bone marrow mesenchymal (BMSCs). Keratin/PLGA (contents 0%, 10%, 20% and 50% of PLGA weight) scaffolds were prepared by solvent casting/salt leaching method. We characterized porosity, wettability, and water uptake ability, DSC of keratin/PLGA scaffold. We seeded BMSCs isolated from the femurs of rat into the inner core of the hybrid scaffold. Celluar viability were assayed by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) test. We confirmed that keratin/PLGA scaffold is hydrophilic by wettability, and water uptake ability measurement results. In MTT assay results, cell viability in scaffolds impregnated 10 and 20 wt% of keratin were higher than other scaffolds. In conclusion, we suggest that keratin/PLGA scaffold may be useful to tissue engineering using BMSCs.