• 제목/요약/키워드: tissue-engineering

검색결과 1,851건 처리시간 0.028초

온도감응성 수화젤을 이용한 골수간엽줄기세포의 골분화 유도 (Osteogenic Differentiation of Bone Marrow Stem Cells Using Thermo-Sensitive Hydrogels)

  • 김선경;현훈;김순희;윤선중;김문석;이종문;강길선;이해방
    • 폴리머
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    • 제30권3호
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    • pp.196-201
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    • 2006
  • 온도감응성 고분자인 폴리 (에틸렌 글리콜)을 기본으로 하는 다이블록 및 트리블록 폴리에스테르 공중합체들은 비독성과 생체적합성 그리고 생분해성 특징 때문에 조직공학 분야 및 주사제형의 약물전달체에서 많은 응용이 이루어지고 있다. 본 연구에서는 다이블록 공중합체를 이용한 솔-젤 전이 현상을 갖는 고분자를 평균분자량 750 g/mole의 메톡시 폴리 (에틸렌 글리콜)과 카프로락톤을 실온에서 HCl $Et_2O$ 존재 하에서 개환중합을 통하여 합성하였다. 이 공중합체를 이용하여 in vivo상에서 골수간엽줄기세포와 골유도물질인 덱사메타손의 존재 여부에 따라 골분화의 가능성을 조사하였다. 조직을 파라핀으로 고정시켜 슬라이드를 제조한 후 hematoxylin & eosin, 본쿠사 및 오스테오칼신 염색을 통하여 골형성 정도를 확인하였다. 결론적으로 덱사메타손이 함유된 젤이 골형성에 도움을 주지만 젤만으로는 강한 골형성을 유도할 수 없으므로 줄기세포나 다른 골형성 유도재료를 사용하면 우수하게 골형성 효과를 가져올 것이라고 판단되었다.

유산균(Lactobacillus fermentum) 함유 미생물제제의 크리핑 벤트그래스 밀도개선 효과 (Microbial Fertilizer Containing Lactobacillus fermentum Improved Creeping Bentgrass Density)

  • 조기웅;김영선;함선규;배은지;이재필;김두환;김우성;이긍주
    • Weed & Turfgrass Science
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    • 제6권4호
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    • pp.322-332
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    • 2017
  • 골프장 잔디 관리에서 미생물제제는 잔디의 생육 및 품질을 개선하고, 병을 예방하기 위해 사용되어왔다. 본 연구는 유산균(Lactobacillus fermentum)을 함유하는 미생물제제의 처리에 따른 크리핑 벤트그래스의 생육과 품질에 미치는 영향을 조사하였다. 처리구는 무처리구(NF), 대조구(CF), 미생물제제 처리구(MFcL: microbial fertilizer containing L. fermentum; MFL: CF+MFcL $1.0g\;m^{-2}$; 2MFL: CF+ MFcL $2.0g\;m^{-2}$) 및 미생물제제 단독 처리구(OMF; MFcL $1.0g\;m^2$)로 설정하였다. 시험 종료 후 토양 화학성은 처리구별 차이를 나타내지 않아 MFcL의 처리가 토양 이화학성에 미치는 영향은 미미하였다. MFcL 처리 시 잔디의 시각적 품질, 잔디 예지물, 잔디 중 무기 성분 함량 및 흡수량은 대조구와 비슷하였다. 잔디 밀도는 MFcL처리구에서 대조구에 비해 약 20% 정도 증가하였다. 이들 결과는 유산균을 함유하는 미생물제제의 처리는 크리핑 벤트그래스의 밀도를 향상에 기여하는 것을 확인하였다.

Changes of the Structural and Biomechanical Properties of the Bovine Pericardium after the Removal of ${\alpha}$-Gal Epitopes by Decellularization and ${\alpha}$-Galactosidase Treatment

  • Nam, Jinhae;Choi, Sun-Young;Sung, Si-Chan;Lim, Hong-Gook;Park, Seong-Sik;Kim, Soo-Hwan;Kim, Yong Jin
    • Journal of Chest Surgery
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    • 제45권6호
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    • pp.380-389
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    • 2012
  • Background: Bovine pericardium is one of the most widely used materials in bioprosthetic heart valves. Immunologic responses have been implicated as potential causes of limited durability of xenogenic valves. This study aimed to determine the effectiveness of decellularization and ${\alpha}$-galactosidase (${\alpha}$-gal) to remove major xenoreactive antigens from xenogenic tissues. Materials and Methods: Recombinant Bacteroides thetaiotaomicron (B. thetaiotaomicron) ${\alpha}$-gal or decellularization, or both were used to remove ${\alpha}$-gal from bovine pericardium. It was confirmed by ${\alpha}$-gal-bovine serum albumin-based enzyme-linked immunosorbent assay (ELISA), high-performance anion exchange chromatography, flow cytometry, 3,3'-diaminobenzidine-staining, and lectin-based ELISA. The mechanical properties of bovine pericardium after decellularization or ${\alpha}$-gal treatment were investigated by tests of tensile-strength, permeability, and compliance. Collagen fiber rearrangement was also evaluated by a 20,000${\times}$ transmission electron microscope (TEM). Results: Recombinant B. thetaiotaomicron ${\alpha}$-gal could effectively remove ${\alpha}$-gal from bovine pericardium B. thetaiotaomicron (0.1 U/mL, pH 7.2) while recombinant human ${\alpha}$-gal removed it recombinant human ${\alpha}$-gal (10 U/mL, pH 5.0). There was no difference in the mechanical properties of fresh and recombinant ${\alpha}$-gal-treated bovine pericardium. Furthermore, the TEM findings demonstrated that recombinant ${\alpha}$-gal made no difference in the arrangement of collagen fiber bundles with decellularization. Conclusion: Recombinant B. thetaiotaomicron ${\alpha}$-gal effectively removed ${\alpha}$-gal from bovine pericardium with a small amount under physiological conditions compared to human recombinant ${\alpha}$-gal, which may alleviate the harmful xenoreactive immunologic responses of ${\alpha}$-gal. Recombinant ${\alpha}$-gal treatment had no adverse effects on the mechanical properties of bovine pericardium.

키토산이 배양중인 치주인대세포에 미치는 영향 (Effects of Chitosan on Human Periodontal Ligament Cells in Vitro)

  • 김옥수;정현주
    • Journal of Periodontal and Implant Science
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    • 제31권1호
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    • pp.163-180
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    • 2001
  • The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.

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수종의 생약추출물이 MC3T3-E1 세포의 염기성 인산분해 효소 활성에 미치는 영향 (Effects of Extracts of Natural Products on Alkaline Phosphatase Activity of MC3T3 - E1 Cells)

  • 박상기;김대겸;유승한;김현아;김명동;유형근;신형식
    • Journal of Periodontal and Implant Science
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    • 제31권1호
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    • pp.123-135
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    • 2001
  • Several growth factors and polypeptides were studied for the regeneration of periodontal supporting tissues which had been lost due to periodontal disease. But these are not commonly used for regenerators of bone tissue or alveolar bone, because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many natural products, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential or periodontal tissues. Cnidii Rhizoma, Rhinocerotis Cornu and Drynariae Rhizoma have been traditionally used as a drug for treatment of bone disease in oriental medicine. The purpose of this study was to examine the ability of alkaline phosphatase synthesis of MC3T3-E1 cells when above medicines were supplimented. MC3T3-E1 cells were cultured with ${\alpha}-MEM(negative control)$, dexamethasone(positive control), and each natural products for 3 and 5 days. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry. Except Cnidii Rhizoma, all of the natural products of this study induced higher activity of ALP synthesis than controls. Among them Drynariae Rhizoma induced the highest activity. In the aspects of culturing time, all medicines did not showed the difference between 3 and 5 days, but $10^{-7}g/ml$ group of Rhinocerotis Corun showed significant increase at 3 days than at 5 days. These results indicate that several natural products have a inducing ability of ALP synthesis on osteoblasts.

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GLCM알고리즘을 이용한 경직장 초음파 영상의 정량적 평가 (Quantitative assessment of Endorectal Ultrasonography by using GLCM Algorithm)

  • 노다정;강민지;김유경;서아름;이인호;정희성;조진영;고성진
    • 한국정보통신학회:학술대회논문집
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    • 한국정보통신학회 2015년도 춘계학술대회
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    • pp.383-387
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    • 2015
  • 현대인들의 불규칙한 생활과 서구화된 식습관에 의해 대장 및 직장 질환이 증가하고 있는 추세로, 특히 직장암은 전체 대장암의 50%를 차지하고 있다. 초기 직장암의 경우 표면으로 돌출되는 부분이 없으므로 초음파로 조직 내부를 보지 않으면 직장 농양으로 오진하는 오류를 범하기도 한다. 그러나 초음파 진단을 하더라도 직장암의 병기에 따라 농양과 육안으로 구별이 어려운 경우가 있기 때문에 보다 정확한 진단이 필요하다. 그러므로 본 연구에서는 직장암과 농양 영상에 대한 컴퓨터 알고리즘을 이용하여 정량적인 분석을 하였다. GLCM을 적용하여 정상 20증례와 농양, 직장암 각 20증례를 분석영역($50{\times}50$ 픽셀)으로 설정하고, 각 영상에서 Autocorrelation, Max probability, Sum average, Sum variance 4가지 파라미터를 비교하여 분석하였다. 결과적으로 Autocorrelation, Max probability, Sum variance 3개의 파라미터에서는 100%, Sum average 파라미터에서는 암 95%, 농양에서 90% 이상의 높은 병변 질감 검출 효율이 나타났다. 이러한 파라미터들이 직장에서의 정상조직, 농양조직, 암조직 간의 판별의 기준으로 가치가 있다고 사료된다. 임상에서의 활용정도에 따라 컴퓨터 보조진단으로서의 충분한 유용성을 볼 수 있을 것이다.

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대식세포가 β-tricalcium Phosphate 뼈이식제의 생분해에 미치는 영향 (Effects of Macrophage on Biodegradation of β-tricalcium Phosphate Bone Graft Substitute)

  • 김영희;;변인선;오익현;민영기;양훈모;이병택;송호연
    • 한국세라믹학회지
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    • 제45권10호
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    • pp.618-624
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    • 2008
  • Various calcium phosphate bioceramics are distinguished by their excellent biocompatibility and osteoconductivity. Especially, the exceptional biodegradability of $\beta$-TCP makes it a bone graft substitute of choice in many clinical applications. The activation of osteoclasts, differentiated from macrophage precursor cells, trigger a cell-mediated resorption mechanism that renders $\beta$-TCP biodegradable. Based on this evidence, we studied the biodegradation process of granular-type $\beta$-TCP bone graft substitute through in vitro and in vivo studies. Raw 264.7 cells treated with RANKL and M-CSF differentiated into osteoclasts with macrophage-like properties, as observed with TRAP stain. These osteoclasts were cultured with $\beta$-TCP nano powders synthesized by microwave-assisted process. We confirmed the phagocytosis of osteoclasts by observing $\beta$-TCP particles in their phagosomes via electron microscopy. No damage to the osteoclasts during phagocytosis was observed, nor did the $\beta$-TCP powders show any sign of cytotoxicity. We also observed the histological changes in subcutaneous tissues of rats implanted with granule-type $\beta$-TCP synthesized by fibrous monolithic process. The $\beta$-TCP bone graft substitute was well surrounded with fibrous tissue, and 4 months after implantation, 60% of its mass had been biodegraded. Also, histological findings via H&E stain showed a higher level of infiltration of lymphocytes as well as macrophages around the granule-type $\beta$-TCP. From the results, we have concluded that macrophages play an important role in the biodegradation process of $\beta$-TCP bone graft substitutes.

Optimization of growth inducing factors for colony forming and attachment of bone marrow-derived mesenchymal stem cells regarding bioengineering application

  • Quan, Hongxuan;Kim, Seong-Kyun;Heo, Seong-Joo;Koak, Jai-Young;Lee, Joo-Hee
    • The Journal of Advanced Prosthodontics
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    • 제6권5호
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    • pp.379-386
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    • 2014
  • PURPOSE. These days, mesenchymal stem cells (MSCs) have received worldwide attention because of their potentiality in tissue engineering for implant dentistry. The purpose of this study was to evaluate various growth inducing factors in media for improvement of acquisition of bone marrow mesenchymal stem cells (BMMSCs) and colony forming unit-fibroblast (CFU-F). MATERIALS AND METHODS. The mouse BMMSCs were freshly obtained from female C3H mouse femur and tibia. The cells seeded at the density of $10^6$/dish in media supplemented with different density of fetal bovine serum (FBS), $1{\alpha}$, 25-dihydroxyvitamin (VD3) and recombinant human epidermal growth factor (rhEGF). After 14 days, CFU-F assay was conducted to analyze the cell attachment and proliferation, and moreover for VD3, the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was additionally conducted. RESULTS. The cell proliferation was increased with the increase of FBS concentration (P<.05). The cell proliferation was highest at the density of 20 ng/mL rhEGF compared with 0 ng/mL and 200 ng/mL rhEGF (P<.05). For VD3, although the colony number was increased with the increase of its concentration, the difference was not statistically significant (P>.05). CONCLUTION. FBS played the main role in cell attachment and growth, and the growth factor like rhEGF played the additional effect. However, VD3 did not have much efficacy compare with the other two factors. Improvement of the conditions could be adopted to acquire more functional MSCs to apply into bony defect around implants easily.

사람 양수에서 호흡기세포의 분리 (Isolation and Identification of Respiratory Cells from Human Amniotic Fluid)

  • 김은정;박용원;김영한;김유선;오정탁
    • Advances in pediatric surgery
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    • 제15권1호
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    • pp.1-10
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    • 2009
  • 최근에 양수에 존재하는 세포들은 다양한 세포분화의 능력을 가지고 있으며 세포 치료와 조직공학의 세포원으로서의 가능성이 높다는 연구 결과가 많이 보고되고 있다. 그러나 양수 내에 어떤 종류의 세포가 정상적으로 존재하는 지에 대해서는 제한적으로 알려져 있다. 본 연구에서는 사람의 양수 내에 호흡기세포를 분리 배양하는 것을 목표로 하였다. 임신 17주에서 20주 사이의 산모 10명에서 각각 5 mL의 양수를 획득하여 small airway growth medium (SAGM) 배양액에서 계대 배양을 시행하였으며 type II alveolar cells이 존재하는 지에 대하여 면역형광염색 및 RT-PCR을 시행하였다. 배양된 세포는 광학현미경 상 상피세포와 유사한 다면체의 모양이었으며 계대 배양 시에 변화 없이 동일한 형체를 보였다. 배양된 세포는 면역형광염색 상 type II alveolar cell의 특이표지자인 surfactant protein C (SPC) 및 TTF-1 protein에 대해서는 양성이었으나 CD 31 및 vimentin에 대해서는 음성이었으며, RT-PCR 상 SPC mRNA 가 발현되었다. 이상의 결과로 사람의 양수를 특이 배양액에 배양하면 호흡기세포를 분리, 확인할 수 있음을 알았으며 이러한 결과가 향후 양수세포의 추가적인 연구에 활용될 수 있다고 생각된다.

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The Significance of SDF-1α-CXCR4 Axis in in vivo Angiogenic Ability of Human Periodontal Ligament Stem Cells

  • Bae, Yoon-Kyung;Kim, Gee-Hye;Lee, Jae Cheoun;Seo, Byoung-Moo;Joo, Kyeung-Min;Lee, Gene;Nam, Hyun
    • Molecules and Cells
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    • 제40권6호
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    • pp.386-392
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    • 2017
  • Periodontal ligament stem cells (PDLSCs) are multipotent stem cells derived from periodontium and have mesenchymal stem cell (MSC)-like characteristics. Recently, the perivascular region was recognized as the developmental origin of MSCs, which suggests the in vivo angiogenic potential of PDLSCs. In this study, we investigated whether PDLSCs could be a potential source of perivascular cells, which could contribute to in vivo angiogenesis. PDLSCs exhibited typical MSC-like characteristics such as the expression pattern of surface markers (CD29, CD44, CD73, and CD105) and differentiation potentials (osteogenic and adipogenic differentiation). Moreover, PDLSCs expressed perivascular cell markers such as NG2, ${\alpha}-smooth$ muscle actin, platelet-derived growth factor receptor ${\beta}$, and CD146. We conducted an in vivo Matrigel plug assay to confirm the in vivo angiogenic potential of PDLSCs. We could not observe significant vessel-like structures with PDLSCs alone or human umbilical vein endothelial cells (HUVECs) alone at day 7 after injection. However, when PDLSCs and HUVECs were co-injected, there were vessel-like structures containing red blood cells in the lumens, which suggested that anastomosis occurred between newly formed vessels and host circulatory system. To block the $SDF-1{\alpha}$ and CXCR4 axis between PDLSCs and HUVECs, AMD3100, a CXCR4 antagonist, was added into the Matrigel plug. After day 3 and day 7 after injection, there were no significant vessel-like structures. In conclusion, we demonstrated the perivascular characteristics of PDLSCs and their contribution to in vivo angiogenesis, which might imply potential application of PDLSCs into the neovascularization of tissue engineering and vascular diseases.