2026-09-21T19:50:07Zhttps://repository.ajou.ac.kr/oai/request

oai:repository.ajou.ac.kr:201003/13972024-10-11T02:17:36Zcom_201003_14369com_201003_14368col_201003_14370

민, 병현 김, 룡호 대학원 의학과 200724419 김, 룡호 2011-01-31T06:58:49Z 2011-01-31T06:58:49Z 2009 2009 http://repository.ajou.ac.kr/handle/201003/1397 http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000009741 000000009741 " Bone Marrow-Derived Buffy Coat Can Supplement the Bone Marrow Stimulation Technique for Articular Cartilage Repair Bone marrow stimulation (BMS) has been used as a popular operating technique to repair defect of knee articular cartilage. However, the regenerated tissue from BMS is known as a fibrocartilage rather than a hyaline cartilage and thinner than native cartilage. The objective of this study was to investigate the feasibility of autologous bone marrow-derived buffy coat transplantation in the regeneration of large full-thickness cartilage defect. Rabbits were divided into four groups: the defect was remained untreated as a negative control (Group 1), performed with BMS only using a drill (diameter: 5 mm) and a 17 gauge needle (Group 2), injected with concentrated autologous buffy coat isolated from the iliac crest after BMS (Group 3) and implanted with autologous osteochondral graft (AOG) as a positive control (Group 4). The cartilage defect was finally covered with a thin membrane made of cartilage matrix material using cross suture method in groups 1, 2 and 3. Bone marrow aspirated from iliac crest was diluted with phosphate-buffered saline. Buffy coat was isolated by centrifugation through a Ficoll density gradient at 2000 rpm for 30 min. The repaired cartilages in each experimental group were evaluated by macroscopic observation, histology with Safranin-O staining, immunohistochemistry for collagen type IIand ICRS scoring at 4 and 8 weeks post-operation. In the results of gross observation and Safanin-O staining, the fibrous/hyaline cartilage was regenerated in groups 1 and 2, while the hyaline cartilage tissues with mature matrix and columnar organization of chondrocytes were observed in groups 3 and 4. In the immunohistochemical staining, expression of type II collagen was gradually increased along with time at the pericellular region in the repaired tissues of groups 2, 3 and 4. In the total ICRS, histological score increased significantly along with time in all groups. The ICRS score was higher in groups 3 and 4 than in other groups. This study demonstrated that injection of autologous bone marrow buffy coat after BMS effectively regenerated cartilage defect in rabbit model, being more effective than the BMS alone and similar to AOG. It is speculated that the buffy coat could probably provide more mesenchymal stem cells to regenerate the articular cartilage defect. In addition the extracellular matrix membrane used to cover the defect efficiently prevented leakage of injected cells and blood clots from cartilage defect into the knee joint cavity. In conclusion, the injection of autologous bone marrow-derived buffy coat in case of BMS arthroplasty could be a useful clinical protocol for cartilage repair. " Approved for entry into archive by Hyun Chul Cho (hccho@ajou.ac.kr) on 2011-01-31T06:58:49Z (GMT) No. of bitstreams: 1 000000009741.pdf: 2915005 bytes, checksum: 0d02ef89b9f68d62956230c4408f6b8b (MD5) Made available in DSpace on 2011-01-31T06:58:49Z (GMT). No. of bitstreams: 1 000000009741.pdf: 2915005 bytes, checksum: 0d02ef89b9f68d62956230c4408f6b8b (MD5) Submitted by Hyun Chul Cho (hccho@ajou.ac.kr) on 2011-01-31T06:58:49Z No. of bitstreams: 1 000000009741.pdf: 2915005 bytes, checksum: 0d02ef89b9f68d62956230c4408f6b8b (MD5) "TABLE OF CONTENTS ABSTRACT ⅰ TABLE OF CONTENTS ⅲ LIST OF FIGURES ⅳ LIST OF TABLES ⅴ LIST OF ABBREVIATIONS ⅵ Ⅰ. INTRODUCTION 1 Ⅱ. MATERIALS AND METHODS 4 A. Experimental design and surgery 4 B. Macroscopic and histological evaluation 7 C. Histological scoring (ICRS score) 7 D. Chemical assay of repaired tissue 9 Ⅲ. RESULTS 10 1. Colony-forming unit-fibroblasts (CFU-Fs) assay 10 2. Gross finding of cartilage defects 13 3. Expression of type II collagens 17 4. Sirius red staining 19 5. Chemical assay of repaired tissue 21 Ⅳ. DISCUSSION 24 Ⅴ. CONCLUSION 28 REFERENCES 28 국문요약 31 LIST OF FIGURES Figure. 1. Procedure for the implantation of autologous buffy coat 6 Figure. 2. Number of MSCs colony 11 Figure. 3. Gross finding of cartilage defects 14 Figure. 4. Histological Safanin-O staining 15 Figure. 5. ICRS score at week 4 and week 8 16 Figure. 6. The immunohistochemistry for collagen type II expression 18 Figure. 7. Sirius red staining 20 Figure. 8. Changes and comparison of GAG Contents among the experimental groups along with time 22 Figure. 9. Changes and comparison of DNA Contents among the groups along with time 23 LIST OF TABLES Table 1. Histological scoring system 8 Table 2. Total MSC contents 12 " Master en Bone Marrow-Derived Buffy Coat Can Supplement the Bone Marrow Stimulation Technique for Articular Cartilage Repair 골수자극술의 관절연골 재생 증진을 위한 골수유래 Buffy coat 첨가 효과 Thesis Theses Cartilage Regeneration Bone marrow stimulation 2009 9999-12-31 9999-12-31