Development of platelet-rich fibrin (PRF) as biodegradation scaffold for application in cartilage engineering

• Main Authors: Pai-Hung Ko, 柯百鴻
• Other Authors: 陳建和
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/88097638314545469537

碩士 === 臺北醫學大學 === 醫學科學研究所 === 96 === Because of articular cartilage has a limited ability for self- repair. Thus, treatment of cartilage lesions is a challenge. Osteoarthritis (OA), is known as degenerative joint disease and a type of arthritis that is caused by the breakdown and eventual loss of the cartilage of one or more joints. This thesis investigates to incorporate PRF that involves developmental signals into fibrin for use in cell biological studies and as a regeneration matrix employing tissue-engineering (TE). The goal is to explore the fibrin gel that adds PRF scaffold for in vitro culture and in vivo of chondrocytes proliferation and differentiation. Platelet-rich fibrin (PRF) is a new generation of platelet concentration, it’s abundant in platelet cytokines, like platelet-derived growth factors (PDGF-BB), insulin-like growth factor-1 (IGF-1), transforming growth factor (TGF-??1) and bone morphogenetic proteins (BMP-2). They have the ability of chondrocytes proliferation and differentiation. In the thesis, fibrin scaffolds have two types. One is produced from bovine fibrinogen and thrombin that mix to fabricate the fibrin gel, and the other is add PRF from human blood by centrifugation without anticoagulant into fibrin scaffold. And the fibrins are made two forms, the fibrin gel and it is to make a fibrin sponge by freeze-drying. We set up the primary human chondrocytes and SW-1353 . The primary chondrocytes and SW-1353 are embedded and developed in these scaffolds. Additionally, the agarose scaffold is to be the control. Three types of three-dimension (3D) and two-dimension (2D) culture are fibrin, fibrin with PRF and agarose scaffold are evaluated，which is the adaptive one to enable the chondrocytes proliferation and differentiation . We compare SW-1353 and primary human chondrocytes on 2D and in 3D of three kinds of scaffold. In order to carry out a comparative study, we undertake the inner microstructure of fibrin, PRF sponge that has an interconnected pore structure is observed by scanning electron microscope (SEM). And we observe cell morphology by using microscopy. To quantify the concentration of growth factors in the PRF exduates, PDGF-BB, IGF-1, TGF-??1, and BMP-2 are stimulators that correlated with proliferation and differentiation by ELISA. MTT assay and RT-PCR are used for estimate cell survivability and the mRNA expression of type II collagen and aggrecan. To detect the glycosaminoglycan (GAG) from chondrocytes are in different scaffold by PAS and Alician blue staining. The results showed that the chondrocytes on 2D and in 3D fibrin + PRF scaffold structure could provided the more available proliferation and differentiation than cells just on 2D and in 3D fibrin or agarose. It is concluded that the fibrin with PRF gel including abundant cytokines and growth factors is a promising three-dimension scaffold of cells for cartilage tissue engineering.