摘要:骨質疏鬆症是一種骨骼老化的現象,會造成骨質流失並容易發生骨折。本研究建立卵巢截除術(OVX)後雌激素不足的骨質疏鬆李宋迷你豬模型,進行以骨髓幹細胞(細胞數達108個)與富含血小板纖維素釋放液(20ml)對骨質疏鬆症之治療研究。幹細胞具有獨特自我更新(self-renewal)的能力,能讓其在生物體內不斷分裂,且同時保有多能(pluripotency)分化的能力,不至隨生物體老化而喪失。基於骨髓中的骨髓間葉幹細胞(bone marrow mesenchymal stem cell, BMSC)為一具有多分化能力的幹細胞,在BMSC分化成骨組織的骨化過程(osteogenesis)中,BMSC可分泌許多生長因子(growth factor)和形態原(morphogens),包括了bone morphogenetic protein (BMP)、transforming growth factor-beta (TGF-beta)、fibroblast growth factor (FGF)、platelet-derived growth factor (PDGF)及insulin-like growth (IGF)等。此外,搭配富含血小板纖維素釋放液(platelet-rich fibrin releasate, PRF releasate)含多種生長因子,包含platelet-derived growth factor (PDGF), fibroblast growth factors (FGFs), epidermal growth factor (EGF), transforming growth factor-beta (TGF-b), vascular endothelial growth factor (VEGF) and nerve growth factor (NGF)等,具有促進細胞增生與骨母細胞分化(osteoblastogenic differentiation)之能力,以建立骨質疏鬆的治療模式。在血液生化數據中分析術前及術後六、九、十二個月及治療後三個月豬血清中維生素D(Vit D),骨鹼性磷酸酶(BAP),C-端交聯端肽的I型膠原(CTx),磷及鈣的含量變化。以CTx指標為例,在生理學或病理學上骨質流失增加,代表第一型膠原蛋白的分解程度增加,血中的膠原蛋白片斷也會增加。當血清中膠原蛋白異構CTx的濃度提高,患者的骨質流失增加,而經過抑制再吸收的治療後,血清內濃度則回復到正常。目前的實驗我們發現實驗組在治療後三個月CTx有明顯的回降,而對照組在十五個月當中CTx則持續的增加。本實驗初步證實骨髓幹細胞與富含血小板纖維素釋放液有治療豬骨質疏鬆症的現象。進一步將從BMD及MRI數據評估實驗組與對照組之治療差異。
Abstract: Osteoporosis is a metabolic bone disease that causes the bone tissue to lose its normal structure and become fragile. This study established amputated ovarian surgery (OVX) estrogen deficiency after osteoporosis Lee-Sung mini pig model for the bone marrow stem cells (cells up to 108) mixed with platelet-rich fibrin releasate (20ml) on osteoporosis treatment study. It is well known that stem cells have a unique “self-renewal” ability. This enables them to continuously divide throughout the life of the organism and to maintain their pluripotency without aging. Bone marrow mesenchymal stem cells (BMSC) are multipotent stem cells capable of differentiating into various cell types which exist in bone marrow. BMSC can differentate to osteocyte when bone growth during the osteogenesis. There are a lot of growth factors and morphogens in osteogenesis, including bone morphogenetic protein (BMP)、transforming growth factor-beta (TGF-beta)、fibroblast growth factor (FGF)、platelet-derived growth factor (PDGF) and insulin-like growth (IGF). With the development of technology, we can get the concentrated platelet, platelet-rich fibrin releasate (PRF releasate), which contains a variety of growth factors, include platelet-derived growth factor (PDGF), fibroblast growth factors (FGFs), epidermal growth factor (EGF), transforming growth factor-beta (TGF-b), vascular endothelial growth factor (VEGF) and nerve growth factor (NGF), etc., that may promote osteogentic proliferation and differentiation. In this study, analysis of preoperative and postoperative six, nine, twelve months and three months after treatment pig serum vitamin D (Vit D), bone alkaline phosphatase (BAP), C-terminal cross-linked telopeptide of type I collagen (CTx), phosphorus and calcium content data changes. For CTx index in physiology or pathology increased bone loss, on behalf of the first degree of decomposition of collagen increases collagen fragments in the blood will increase. While the serum concentration of the collagen to improve heterogeneous CTx, the patient`s bone loss increases, and after re-uptake inhibiting treatment, the serum concentrations return to normal. In this experiment we found that after 3 months of treatment in experimental group had significantly CTx back down, while CTx increase continued in control group during the 15-month period. Collectively, these findings indicated that the mixture of BMSC and PRF releasate could potentially be an effective agent in the treatment of OVX. This first work thus makes it possible to plan many future applications for the PRF and bone marrow stem cells, as well in the treatment of osteoporosis, provided that its real effects are evaluated in an impartial and rigorous way.