肌肉來源細胞應用於骨組織工程之研究

Muscle-Derived Cells as the Cell Sources for Bone Tissue Engineering

吳岳修(Yueh-Hsiu Wu)

指導教授 ： 林峰輝

共同指導教授 ： 孫瑞昇(Jui-Sheng Sun)

國立臺灣大學/工學院/醫學工程學研究所/碩士(2004年)

https://doi.org/10.6342/NTU.2004.01441

摘要

目前自體移植骨來源受到限制，手術過程易造成病人傷口疼痛、化膿以及各種複雜因素，另一方面自體移植法其做法類似挖東牆補西牆，當遇到相當大的骨創傷或缺陷勢必無法治療。本研究使用之人工合成明膠/rhBMP-2複合材料，其優點為製造容易、成本低；並具有骨引導、骨誘導、以及生長因子等三大骨形成要素，足以取代自體移植骨，為相當適合之骨移植材料。

由人體取得之間質葉幹細胞 (MSCs) 或是骨母細胞，細胞量少而且生長速率相當緩慢，不容易在體外大量繁殖培養。而培養在二維平面（monolayer）的細胞容易因為非原來三維立體（3D）環境之緣故而改變其細胞型態（phenotype）。由肌肉衛星細胞（satellite cells）分離得來的肌肉來源細胞具有幹細胞自我修復新生、超強增生能力以及具備分化成不同中胚層細胞組織的條件，不論是何種年齡的個體所取出來之肌肉母細胞在體外都可以在短時間內大量培養繁殖。所需的經費以及時間都遠低於培養MSCs或是骨母細胞，充分符合組織工程及臨床手術上的需求。

本研究採用明膠和戊二醛交聯之材料作為具有骨引導作用的支架；在體外培養從大鼠（Wistar rat）舌頭取得之肌肉來源細胞取代MSCs來達到大量繁殖，將細胞以高濃度接種至明膠/rhBMP-2複合材料，誘使其分化為骨母前驅細胞，並同時具有骨誘導之能力。同時將材料培養在特定生物反應器中模擬體內的環境，並加入特定的調控因子（dexamethasone、β-glycerophosphate、L-ascorbic acid和1

關鍵字

骨組織工程 ； 明膠材料 ； 肌肉來源細胞 ； 骨形成蛋白質 ； 骨移植材料

並列摘要

Autogenous bone graft is regarded as the gold standard for bone graft implantation as it provides three necessary elements for bone regeneration and maintenance:（1）scaffolding for osteoconduction,（2）inductive agents for osteoinduction, and（3）progenitor cells for osteogenesis. However, the availability of autograft is limited and the procedure to harvest the bone tissue is associated with complications, as well as possible donor site morbidity.

Geltain/rhBMP-2 synthetic graft offers an alternative which combines three major bone formation properties in a controlled and effective way that can be used in clinical applications. On the other hand, functional osteoblasts are obtained through bone or bone marrow. These cells are limited in supply, and difficult to obtain. Recently, skeletal muscle cells are used in bone tissue engineering. The method of harvesting muscle cells is relatively easier to perform and less painful to the patient. Moreover, muscle-derived cells have been shown to differentiate into mesenchymal tissues, such as bone and muscle, as well as cartilage. In addition, bone morphogenetic protein 2 has been identified as an inducting agent that can stimulate conversion to the osteogenic pathway.

In this study, porous gelatin material are selected as a biodegradable scaffold, combining rhBMP-2 on gelatin surface as an inductive agent, and seeding with muscle-derived cells from wistar rat as progenitor cells. The resulting bone grafts were cultured in spinnar flask with

並列關鍵字

muscle-derived cells ； bone graft ； gelatin ； bone morphogenetic protein 2 ； bone tissue engineering

參考文獻

1. Alexander R. Vaccaro. The role of the osteoconductive scaffold in synthetic

2. Christopher G. Finkemeier. Bone-grafting and bone-graft substitutes. J. Bone

Joint Surg. Am 84-A:454-464 (2002)

3. Clayton R. Perry. Bone repair techniques, bone graft, and bone graft

substitutes. Clin. Orthop. 1(360):71-86 (1999)