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Manufacture of cartilage gel with adhesive property using fetal cartilage derived progenitor cell

초록/요약

Adult cartilages have limitations in tissue repair because they do not have blood vessels and nerves. Damaged cartilage associated with diseases and injuries increases in pain as time passes. Tissue engineered cartilages are made as an alternative cure. However, the cost using tissue engineering is expensive and the knowledge of material safety is lack. Based on this, recently, several researchers have attempted to make engineered cartilages using cells only instead of biomaterials. Even though various studies have attempted to make matured cartilages using the scaffold-free system with growth factors and bioreactors, we intended to make engineered cartilages using cells and their own ECM on the scaffold-free system without any other stimuli. Primary fetal cartilage derived progenitor cells (FCPC) were isolated, proliferated, and differentiated on the scaffold-free system. And then scaffold-free engineered constructs were fabricated. These scaffold-free engineered constructs were cultured in vitro for 1, 2, and 3 weeks. Their histological, biochemical, and mechanical evaluations have been carried out. With culture time, volume, GAG and collagen contents of scaffold-free engineered constructs were increased, while their DNA amounts were rarely changed. The mechanical strength showed the highest point on week 3. In addition, their adhesive properties were observed. When they were transplanted to ex vivo model, defects were repaired and integrated with neighbor host tissues very well in all groups without any difference no matter their in vitro culture period. Also, the scaffold-free constructs used FCPC and cultured for two weeks in vitro that had similar ECM components with natural cartilage and the adhesive ability. Therefore, they were effective for repairing cartilage defect when they were transplanted to in vivo model. Generally, most efforts for tissue engineered cartilages have been focused on maturation of tissues. However, this study attempted to identify paste-type engineered cartilages had adhesive ability and show repairs of cartilage defects using them.

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목차

1. Introduction. 01
1.1. Tissue engineering 01
1.2. Advantages of Scaffold-free Tissue Engineering Technologies 02
1.3. Scaffold-free approaches. 03
1.3.1. Cell sheet engineering. 03
1.3.2. Aggregate tissue engineering. 04
1.3.3. Self-assembling process.. 04
1.4. Fetal cartilage derived progenitor cells (FCPCs) 04
2. Materials and Methods. 07
2.1. Cell isolation and culture. 07
2.2. Fabrication of scaffold-free engineered cartilage 07
2.3. Histological and immunohistochemical analysis of the scaffold-free engineered cartilage. 07
2.4. CT Analysis. 08
2.5. Biochemical analysis 08
2.6. Mechanical analysis. 08
2.7. Ex vivo organ culture chondral defect model 09
2.8. Adhesion ability in partial cartilage defect of scaffold-free engineered cartilage. 09
2.9. Implantation of scaffold-free engineered cartilage into partial cartilage defect 09
2.10. Implantation of scaffold-free engineered cartilage into full-thickness cartilage defect 10
2.11. Statistics 10
3. Results 11
3.1. Gross examination and volume of scaffold-free engineered cartilage constructs 11
3.2. Histological evaluation and immunohistochemistry. 13
3.3. Biochemical analysis 15
3.4. Mechanical analysis. 17
3.5. Macroscopic observation of the ex vivo model 19
3.6. Hematoxylin Eosin and Safranin O staining of the ex vivo model 21
3.7. Macroscopic and histological observation of partial thickness defect in vivo model. 23
3.8. Cell tracking with PKH26 label 25
3.9. Macroscopic and histological observation of full thickness in vivo model. 27
4. Discussion 29
CONCLUSION. 33
REFERENCE. 34
국문요약 40

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