摘要
本研究主要探討桿狀病毒應用在基因改質人類間葉幹細胞(mesenchymal stem cells MSC，MSC)與骨組織工程上的潛力。我們首先利用會表現骨形成蛋白(bone morphogenetic protein，BMP-2)的重組桿狀病毒(Bac-CB)轉導人類MSC，並探討其在往硬骨路徑分化及形成硬骨的潛力。在體外(in vitro)環境下，以病毒劑量MOI 40轉導過的人類MSC能夠成功地被誘導分化成為骨母細胞。在轉導過後第6天以相同的病毒劑量(MOI 40)再次轉導人類MSC，可以延長且提高BMP-2在培養基的濃度，接著根據鹼性磷酸脢(Alkaline phosphatase，ALP)活性分析，茜紅素(Alizarin red)染色分析與反轉錄聚合酶連鎖反應(reverse transcription polymerase chain reaction，RT-PCR)分析結果，證實再次轉導可以加速MSC分化進入骨母細胞。此外，再次轉導人類的MSC在植入裸鼠背部皮下2週後，H&E染色、Alizarin red染色和特殊免疫染色的分析結果顯示，MSC會分化進入骨母細胞後期，接下來細胞會逐漸礦化，並且在植入後第6週誘發異位骨生成。
在第二階段的研究，我們進一步探討桿狀病毒應用在骨缺損修復上的潛力。我們採用異種移植的方式，植入Bac-CB轉導的人類MSC實際修復免疫力健全(immunocompetent)的大鼠頭蓋骨缺損(calvarial bone defect)。組織切片染色與電腦斷層掃瞄的分析結果證實，經Bac-CB基因改質的人類MSC能夠促進骨分化與新骨生成。在植入後第1與4週，透過特殊免疫染色分析結果，仍然可以偵測到植入的人類MSC存活下來，證實了MSC本身免疫豁免(immunoprivileged)的特性，但是當植入的MSC開始聚集(aggregation)往骨母細胞分化，便會誘發免疫細胞(macrophages，CD3+和CD8+ T cells)侵入植入位置。在給予大鼠免疫抑制劑投藥，雖然能夠延長MSC在體內存活時間與促進頭蓋骨修復，但仍然無法完全抑制免疫反應與完整地修復頭蓋骨缺損。綜合上述結果，本研究證實了桿狀病毒應用在基因改質人類MSC與骨組織工程上的潛力。

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