The Effect of Platelet-rich Fibrin (PRF) Releasate Combined with Bone Marrow Stem Cells on Osteoporosis with a Mouse Model

• Main Authors: Chen-Hao Liao, 廖政豪
• Other Authors: Tzong-Fu Kuo
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/39221159761768569916

碩士 === 臺灣大學 === 獸醫學研究所 === 103 === Stem cells are a kind of undifferentiated cells which may differentiate into other cell types. And they can also divide indefinitely into stem cell line, but may become cancer cells at about the tenth generation. We collect the bone marrow stem cells (BMSCs) and differentiate BMSCs into osteoblast in vitro after cell sorting by flow cytometer. Platelet-rich fibrin (PRF) is a natural source of growth factors, which contains a large amount of growth factors that can facilitate bone regeneration, Such as: TGFb-1 (Transforming growth factor) , PDGF (platelet-derived growth factor) , VEGF (vascular endothelial growth factor) and Matrix proteins (thrombospondin-1, fibronectin, vitronectin). The growth factors secreted from PRF will cause cell transformation by promoting cell proliferation、matrix formation、bone formation and synthesis of collagen. We co-cultured mouse bone marrow stem cells with platelet-rich fibrin releasate (PRFr), to investigate whether the release would improve stem cell proliferation and promote cell differentiation or not. The results show that proliferation rate of the bone marrow stem cells which co-cultured with releasate were more rapid, and cell viability was much higher. Therefore, we deduced that platelet-rich fiber release solution have significant assistance on the growth and differentiation of bone marrow stem cells. To evaluate therapeutic efficacy of cell-based therapy in osteoporosis: a bone marrow stem cells (BMSCs) and platelet-rich fibrin releasate (PRFr) were used for osteoporosis treatment. An osteoporosis was established with a mouse model by ovariectomy (OVX). Transplantation of BMSCs, PRFr and BMSCs + PRFr into OVX mice for investigating the therapeutic potential for bone regeneration and recovered bone mass loss. OVX or sham operations were performed on virgin ICR mice at 16-weeks old, which were randomLy divided into three parts: Non-surgical, SHAM, and OVX. 6 mice in Non-surgical group was no surgery, 6 mice in SHAM group was subjected to sham surgery, and 30 mice in OVX group mice will accept four different treatments (1)Control group,non injection (2)BMSC-I group, injected BMSCs 3x105 cells/0.6mL PBS once a week for one weeks ; (3)PRF-I group, injected PRFr 0.6mL once a week for one weeks ; (4)BMSCs + PRF-I group, injected BMSCs 3x105 cells combine 0.6 mL PRFr) once a week for one weeks. (5)BMSC-IV group, injected BMSCs 1.2x106 BMSCs/0.6mL PBS once a week for four weeks; (6)PRF-IV group, injected 2.4mL PRFr once a week for four weeks; (7)BMSCs + PRF-IV group, injected 1.2x106 BMSCs / 2.4mL PRFr once a week for four weeks; (8)BMSCs + PRF-IV-i group, intra bone injected 1.2x106 BMSCs/ 2.4 mL PRFr once a week for four weeks. At 8 weeks after implantation, bone mass and its turnover were analyzed by micro CT, We use scanned the left tibia portion of the growth plate at the proximal 0.5-1.5mm, and analyzed bone mineral density, the percentage of bone tissue, trabecular number, trabecular thickness and trabecular separation. Then sacrifice to analyze their efficacy by histomorphometry. A statistically significant difference between the experimental and control groups was observed. BMSCs + PRFr transplants were shown effective in restoring bone mineral density. These findings indicated that the mixture of BMSC and PRF releasate could potentially be an effective agent in the treatment for osteoporosis