관절연골 재생 치료에서 줄기세포의 생착 및 분화능 향상을 위한 트랜스글루타미나제-4의 이용 검증

학위논문(박사) -- 서울대학교대학원 : 의과대학 의학과, 2021.8. 김종근.Purpose: Although mesenchymal stem cells (MSCs) transplantation with a hydrogel type scaffold is a promising strategy for repairing damaged articular cartilage, MSC-based cartilage tissue engineering has numerous limitations, including poor implanted cell engraftment, phenotypic alteration, soft mechanical properties, in vivo degradation and loss of strength. Transglutaminase (TG) has been used to crosslink proteins to generate biocompatible tissue scaffolds in cell engineering. However, there are no published reports regarding the use of TG in improving the engraftment and differentiation of stem cell. This study aimed to investigate the efficacy of transplantation of synovium-derived mesenchymal stem cells (SDSCs) encapsulated in a hyaluronic acid/ collagen/ fibrinogen (HA/COL/FG) composite gel by supplementing recombinant human transglutaminase-4 (rhTG-4) in treating osteochondral defects. Materials and Methods: Human SDSCs were prepared from the synovial tissues of knee from human donors who underwent total knee arthroplasty. HA/COL/FG composite gel was generated with a mixture ratio of HA: COL: FG = 0.5: 0.5: 2.0. RhTG-4 was prepared using baculovirus expression system. The adhesion assay was carried out by directly counting the number of attached cells to the fibronectin coating plate at different concentrations of rhTG-4. The distribution of adherent cells was analyzed by crystal violet staining. To visualize cell adhesion and cytoskeletal proteins, immunofluorescence assay was performed and cystamine was supplemented to inhibit the rhTG-4 activity. SDSCs cultured in the HA/COL/FG composite gel were assayed for cell proliferation and viability at different rhTG-4 concentrations. The proliferation assay was carried out by direct counting the number of cells. The Live/Dead Viability kit was used for the assessment of the viability. To evaluate the extra cellular matrix (ECM) hardness, the cells were incubated to generate micro mass, according to the concentrations of rhTG-4 and the hardness was evaluated using a nanoindentation instrument. RhTG-4 activity in the hydrogel was estimated by measuring incorporation of biotinylated pentylamine to N, N-dimethylcasein. In vitro chondrogenesis of SDSCs encapsulated HA/COL/FG composite gel supplementing rhTG-4 was verified. The expression of chondrogenesis-related genes (type I COL, type II COL type X COL, aggrecan, and SOX-9) was analyzed using reverse transcription- polymerase chain reaction (RT-PCR). To investigate the effect of integrin β1 on the TG-4-induced actin remodeling, the intracellular signal transduction pathway and the control of chondrogensis-related genes expression, integrin β1 siRNA was transfected to inhibit integrin β1 expression. To evaluate the in vivo cartilage regeneration, fifty-six knee joints of rabbit osteochondral defect models were made and divided into 3 groups: control group, in which the defect was left untreated; Gel/SDSC group, in which the defect was filled with the HA/COL/FG composite gel loaded with human SDSCs; Gel/SDSC+TG-4 composite gels group, in which the defect was filled with the supplement of rhTG-4 in HA/COL/FG composite gels loaded with human SDSCs. After 12 weeks, tissue specimens were assessed by macroscopic and histological evaluation, and DiI-labeled SDSCs distribution in the center of defect was evaluated. Results: RhTG-4 treatment improved the attachment of SDSCs with increased adhesion to fibronectin and in a concentration-dependent manner. The immunofluorescence assay revealed that pretreatment with cystamine markedly downregulated rhTG-4-induced integrin β1 activation and dynamic actin remodeling, demonstrating the possibility of modulating the phenotype. SDSCs proliferation was significantly increased in rhTG-4 group. Nanoindentation analysis revealed that rhTG-4 stimulation increased the hardness of 3D micro masses. Activity of rhTG-4 in hydrogel was maintained for 2 h. In the group supplemented with rhTG-4, the relative expression levels of type II COL and aggrecan mRNA were significantly increased as determined by RT-PCR. Immunofluorescence analysis revealed that pretreatment with integrin β1 siRNA markedly decreased rhTG4-induced actin remodeling. Intracellular signal transduction pathway and chondrogenesis-related genes including aggrecan and type II collagen were activated in an integrin β1 siRNA-dependent manner. In vivo study, Gel/SDSC+TG-4 group showed more repaired defect lesion compared to the other groups and yielded reconstructed tissue resembling native hyaline cartilage. The total macroscopic and histological scores were significantly higher and DiI-labeled SDSCs distribution was markedly increased at the center of defect in Gel/SDSC+TG-4 group. Conclusion: RhTG-4 supplementation in the HA/COL/FG composite gel mediates cartilage regeneration by enhancing the engraftment and differentiation of SDSCs.Introduction 1 Chapter I. Enhancement of the stem cell engraftment 3 I-1. Materials and Methods 4 I-2. Results 8 Chapter II. Enhancement of the stem cell differentiation 15 II-1. Materials and Methods 16 II-2. Results 21 Discussion 33 Conclusion 45 References 46 Abstract in Korean 53박

인공 슬관절 전치환술 시 안정성과 효과를 고려한 국소 트라넥삼산의 최적의 투여량: 무작위 대조 연구

학위논문(석사)--서울대학교 대학원 :의과대학 의학과,2019. 8. 이명철.Background: The optimal safe and effective dose of topical tranexamic acid (TXA) for controlling bleeding after total knee arthroplasty (TKA) is not known. Methods: A total of 325 patients who were scheduled to undergo TKA were recruited in a prospective randomized double-blinded and placebo-controlled comparative study. The patients were randomly assigned to the following five groups based on the TXA injection (n = 65 per group): control; group 1, 0.5 g TXA; group 2, 1.0 g TXA; group 3, 2.0 g TXA; and group 4, 3.0 g TXA. The primary outcome was decrease in postoperative hemoglobin level. The secondary outcomes were blood loss calculated using Good's method, drainage volume, frequency of transfusion, range of motion (ROM), and plasma TXA levels. Results: The mean hemoglobin decrease was 3.7±1.1 g/dl, 3.5±0.86 g/dl, 3.0±1.07 g/dl, 2.8±0.87 g/dl, and 2.8±0.40 g/dl in the control group and groups 1, 2, 3 and 4, respectively. There were significant differences in the decrease in hemoglobin levels between the control group and groups 2 (p=0.0027), 3 (p=0.005), and 4 (p=0.001). There were significance differences in total blood loss and frequency of transfusion between the control group and groups 2 (p=0.004, 0.002, respectively), 3 (p=0.007, 0.000, respectively), and 4 (p=0.001, 0.002, respectively). There were no significant differences in the drainage volume and ROM among the groups. Serum TXA levels increased proportionally with the dose of topical TXA immediately and at 3 and 6 h post-operatively. Symptomatic deep vein thrombosis did not occur in any group. There were no other significant complications. Conclusions: The topical use of 1.0 g or more of TXA has a meaningful effect without a dose-response relationship and the blood levels of TXA increase with the dose. To prevent overdosing and reduce potential complications, 1.0 g TXA is recommended for topical application.배 경: 인공 슬관절 전치환술 시 출혈 조절을 위해 국소적으로 배액관을 통해 트라넥삼산을 주입하여 사용하고 있다. 그러나 안정성과 효과를 고려한 국소 트라넥삼산의 최적의 투여량에 대해 아직까지 정확하게 밝혀진 바가 없어 이에 대해 알아 보고자 한다. 대상 및 방법: 인공 슬관절 전치환술을 받을 325 명의 환자를 모집하여 전향적 무작위 이중 눈가림 대조 연구를 시행하였다. 환자들은 각각 65명씩 생리 식염수만 주입하는 대조군, 0.5g 트라넥삼산을 주입하는 1군, 1.0g 트라넥삼산을 주입하는 2군, .2.0g 트라넥삼산을 주입하는 3군, 3.0g 트라넥삼산을 주입하는 4군으로 나뉘어 졌다. 시험약물은 상처를 봉합한 후 배액관을 통해 주입하였다. 일차 변수로 수술 후 헤모글로빈 감소 정도를 확인하였다. 이차 변수들로는 Good의 방법을 통한 혈액 손실량, 배액관을 통해 나온 배액량, 수혈 빈도, 슬관절의 굴곡 각도 및 혈중 트라넥삼산의 분포 정도를 확인하였다. 결 과: 헤모글로빈 감소 정도는 대조군부터 4군까지 각각 3.7±1.1 g/dl, 3.5±0.86 g/dl, 3.0±1.07 g/dl, 2.8±0.87 g/dl, 2.8±0.40 g/dl로 측정되었다. 이중 대조군과 2,3,4 군을 비교하였을 때 감소 정도가 유의한 차이를 보였다 (P <0.01). 또한, 대조군에 비해 2,3,4 군에서 유의하게 혈액 손실 및 수혈 빈도가 적었다. (P <0.01). 배액량 및 슬관절의 굴곡 각도는 군간의 유의한 차이가 없었다. 혈중 트라넥삼산의 분포 정도는 용량에 비례하여 증가하는 소견을 보였다. 트라넥삼산 사용과 관련된 합병증은 없었다. 결 론: 1.0g을 포함한 그 이상 용량의 트라넥삼산을 사용하였을 때 출혈 조절 효과를 보이는 것으로 확인되었으나, 용량에 비례하지는 않았다. 그러나 혈중 트라넥삼산 분포는 용량에 비례하여 증가하였다. 따라서 인공 슬관절 전치환술 시 출혈 조절을 위해 국소 트라넥삼산을 사용한다면, 약물 남용 및 잠재적 합병증을 예방하기 위하여 1.0g의 트라넥삼산을 주입하는 것이 가장 적합하다고 판단된다.Abstract i Contents iv List of figures and tables v List of abbreviations and symbols vii Introduction 1 Material and Methods 3 Results 13 Discussion 19 References 27 Abstract in Korean 33Maste

Todd A. Henry, 2014 Assimilating Seoul: Japanese Rule and the Politics of Public Space in Colonial Korea 1910-1945, University of California Press

일제강점기에 대한 한국 학계，나아가 한국 국민의 관심은 언제나 현재진행형이며，특히 일본의 우경화가 심해질수록 그에 대한 반작용으로 더 강도가 세어지고 있다. 1945년 광복이후 일제강점기를 연구하는 연구자들은 이와 같은 일본의 태도에 대해 어떻게 대처해 왔고 대처해 나아가야 할까? 1980~90년대까지는 이러한 일제강점기에 대해 일제가 우리의 근대화 기회를 강탈하고， 우리의 인적 물적 자원을 수탈해 왔다는 방식의 인식이 주류를 이루었다면， 20세기 말부터는 식민지 근대화론에서 부터 후기 구조주의를 포함한 포스트 모던적인 철학사조에 바탕을 둔 식민지 근대성론 등 다양한 목소리가 등장하고 있다. 아울러 관심영역의 경우도 점차 일상사, 생활사, 지역사 등으로 다양해지고 있다. 이러한 관심 분야의 확대와 새로운 분석은 비록 민족감정과 배치되는 부분이 일부 존재한다 하더라도, 학계의 건강함과 지속가능한 발전을 위해서는 바람직한 모습이라 할 수 있을 것이다

政策變動에 관한 硏究 : 서울가정도우미 在家老人福祉政策을 중심으로

학위논문(석사)--서울대학교 행정대학원 :행정학과 행정학전공,2000.Maste

비상용성 폴리에스테르 블렌드에서 에스테르 교환 반응이 상거동에 미치는 영향 : Monte Carlo 모사

Thesis (master`s)--서울대학교 대학원 :섬유고분자공학과,1998.Maste

GIS를 활용한 고등학교 지리수업 모형 개발에 관한 연구

학위논문(석사)--서울대학교 대학원 :사회교육과 지리전공,2000.Maste

Effect of Seeding Rates on the Forage Quality and Productivity of Direct-Seeded Whole Crop Rice

This experiment was conducted to evaluate the effect of seeding rates on the forage quality and productivity of direct-seeded whole crop rice at the experimental field of the Grassland and Forages Division, National Institute of Animal Science, RDA from2007 to 2008. Whole crop rice variety, Nokyang, developed by the National Institute of Crop Science, was directly seeded with four different seeding rates (30, 60, 90 and 120 kg/ha). There were no significant differences (p<0.05) found in emergence date, heading date, greenish and disease resistance based on the seeding rates. Plant height increased at higher seeding rates but the tiller number per plant decreased (p<0.05). The dry matter (DM) content showed no significant difference based on seeding rates (p<0.05) while the yield of fresh and dry matter increased with incremented seeding rates (p<0.05). Acid detergent fiber (ADF) content increased with higher seeding rates, but the neutral detergent fiber (NDF) content did not show a similar tendency. The total digestible nutrient (TDN) content was lowest with 120 kg/ha seeding rate. The grain ratio decreased at higher seeding rates. In economic analysis, the plot seeded at a rate of 60 kg/ha showed the highest income increment. Although high seeding rates increased the DM yield, a seeding rate of 60 kg/ha is recommended as the proper seeding rate for direct-seeded whole crop rice.본 시험은 사료용 총체 벼 직파 재배시 파종량이 수량 및 사료가치에 미치는 영향을 구명하기 위해 2007년부터 2008년까지 국립축산과학원에서 수행되었다. 총체 벼는 국립식량과학원에서 육종한 녹양벼를 이용하여 4개의 파종량을 두고(30, 60, 90 및 120 kg/ha) 수행하였다. 출현일, 출수기, 녹색도 및 내병성은 파종량에 따른 차이는 나타나지 않았다. 초장은 파종량이 늘어남에 따라 커졌으며(p<0.05) 분얼수는 줄어들었다. 건물함량은 유의적인 차이를 나타내지 않았으나 생초 및 건물수량은 파종량이 증가함에 따라 유의적으로 증가되었다(p<0.05). 조단백질 함량은 파종량이 늘어남에 따라 감소되었으며 ADF 함량은 파종량이 늘어남에 따라 높아졌고 NDF 함량은 일정한 경향이 없었다. TDN 함량은 120 kg/ha 처리구에서 가장 낮았고, 잎-줄기-정조 비율은 파종량이 늘어남에 따라 정조비율은 줄어들었으며 잎의 비율은 약간 높아졌고 줄기는 큰 차이가 없었다. 경제성 분석결과 60 kg/ha 파종구에서 가장 높은 소득 증가가 있는 것으로 나타났다. 따라서 이상의 결과를 종합하여 볼 때 총체 벼 직파재배를 위한 적정 파종량은 60kg/ha 내외가 적당한 것으로 추천된다.본 논문은 농촌진흥청 공동연구사업(과제번호 PJ010535)의 지원에 의해 이루어진 것임.OAIID:oai:osos.snu.ac.kr:snu2014-01/102/2014015967/1SEQ:1PERF_CD:SNU2014-01EVAL_ITEM_CD:102USER_ID:2014015967ADJUST_YN:YEMP_ID:A079570DEPT_CD:A101CITE_RATE:0DEPT_NM:국제농업기술학과SCOPUS_YN:NCONFIRM: