傳統的骨骼治療技術以硬質板將受傷部位予以固定或使用骨釘來連接，並配合骨泥來填補受損的部位，這樣子的治療方式帶給病人許多的不便利。近年由於組織工程和基因治療的進步，促使著骨骼治療技術的提昇，更是病人的一大福音。在本研究中我們將結合這兩項的治療技術，發展體外非病毒式的基因治療模式，來幫助骨骼修補及再生。
　　在此利用化學載體- polyethylenimine (PEI) 將造骨蛋白質載入human neuro-teratocarcinoma (hNT) cell，改變細胞本身的功能，讓他可以分泌骨骼生長所須之造骨蛋白(Bone morphogenetic protein)，進而成為一個良好的生物幫浦(biologic pump)。但為了評估載入基因表現強度，以利用免疫細胞染色法(immunocytochemistry)配合共軛聚焦顯微鏡法(confocal microscopy) 的解析驗証BMP-2的轉殖成果。從螢光的影像來看，細胞內部確有大量造骨蛋白之存在，同時也採用西方點墨法(western blotting)來分析培養液中的蛋白質含量，結果顯示hNT cells可以表現BMP-2蛋白質而且可以分泌於培養液中。
　　本研究以體外的基因治療(ex vivo gene therapy) 方式發展骨骼修復治療模式，以非病毒式的基因修飾技術，讓hNT cells分泌造骨造骨蛋白質。未來再配合生物降解性材料，植入於骨骼的損傷的部位，以幫助組織的修補和再生。

　　Traditional bone fracture therapy requires an external fixation with bone mud at the side of wound. The disadvantages of this type of therapy is poor healing and heavy framing and assembling at wound side. With the advancement of tissue engineering and gene therapy today, bone fracture repair has become a very normative therapeutic technique, and it has ensured a rapid, effective and functional healing of complex fractures in the bone. In this study, we combined both techniques to develop an ex vivo gene therapeutic model to aid bone tissue repair.
　　A chemical vector, polyethylenimine (PEI) was used to carry bone morphogenetic protein-2 (BMP-2) gene into human neuro-teratocarcinoma (hNT) cell. BMP-2 can stimulate the regeneration of bone. In this study, immunocytochemistry and confocal microscopy were combined to examine the results after BMP-2 transfection. The fluorescent images showed that a great amount of bone morphogenetic protein existed in the cells. Western blotting was used to analyze the protein capacity in the cell culture medium. The results indicated that hNT cells can express BMP-2 protein in cytosol and generate BMP-2 in the cell culture medium.
　　In this study, a bone repair therapeutic system by ex vivo gene therapy was developed. The production of BMP-2 from hNT cells by non-viral gene-modification method was successfully demonstrated. In future, biodegradable matrix with hNT cells after transfection treatment will be implanted into bone fracture site to aid tissue regeneration and speed up the healing process.

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