靜磁場促進培養於聚乳酸試片似骨母細胞的生長

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題名:	靜磁場促進培養於聚乳酸試片似骨母細胞的生長 Static Magnetic Fields Promote Osteoblast-like Cell Growth on The Poly(L-lactide) Surface
作者:	馮聖偉 Sheng-Wei Feng
貢獻者:	牙醫學系碩博士班
關鍵詞:	靜磁場聚乳酸 MG63細胞 MC3T3-E1細胞生長分化 Static magnetic field Poly(lactic acid) MG63 cell MC3T3-E1 cell Growth Differentiation
日期:	2007
上傳時間:	2009-08-28 12:20:33 (UTC+8)
摘要:	口腔及顱顏手術或意外傷害若造成骨缺損時，常須以適當的骨填補物填入骨缺損，來促進骨頭癒合。近年來生物可吸收性的聚乳酸材料常被應用於顱顏骨缺損的重建，原因在於聚乳酸PLLA (Poly L-lactide)具有高度生物相容性，生物降解性，與良好的力學強度，容易加工等優點。但是聚乳酸有疏水性強、細胞的親和力差和細胞活性或增殖速度會被抑制降低等缺點，因此有許多研究利用材料表面型態的改變或利用化學方法，來改善細胞生長於聚乳酸表面的能力。本研究則利用靜磁場，以非侵入性的方法促進聚乳酸試片上的似骨母細胞生長，並且探討靜磁場可以促進聚乳酸試片上的似骨母細胞生長的假設是否正確。首先將MG63和MC3T3-E1似骨母細胞株以強度4000高斯的靜磁場連續暴露5天，靜磁場影響細胞增生與活性的作用以MTT分析，而細胞型態與細胞外基質的分泌，則利用掃描式電子顯微鏡來觀察。此外，靜磁場對培養於聚乳酸片上之似骨母細胞表現鹼性磷酸酶活性的影響亦被加以分析。在掃描式電子顯微鏡實驗中，我們發現靜磁場會促進培養於聚乳酸試片上MG63 和MC3T3-E1細胞表現更多的細胞外基質。在MTT實驗中，MG63暴露組會有較低的MTT值，但是MC3T3-E1暴露組卻有較高的MTT值。MG63細胞於靜磁場暴露1天後，暴露組的鹼性磷酸酶活性達到最高值，而且明顯比未暴露組增加1.5倍(p<0.05)。然而，MC3T3-E1細胞在靜磁場暴露第3天和第4天時，鹼性磷酸酶活性明顯比未暴露組來的低(p<0.05)。本研究結果顯示，培養於聚乳酸片上的MG63細胞，在接受靜磁場暴露之後會比控制組細胞表現出更為成熟的細胞型態與更多的分化調控因子。而靜磁場則會影響培養於聚乳酸片上MC3T3-E1細胞生長，增加整體細胞數目。 Accident-induced craniomaxillofacial injury frequently results in large bony defects which require the bone grafts to enhance the bone healing. Biodegradable polymer [Poly(L-lactide), PLLA] scaffolds are the devices which are popular used in the reconstruction of craniomaxillofacial defects because of their good biocompatibility、biodegradability and low toxicity after degradation in vivo. However, low proliferation rate and poor cell attachment were found when culturing cell on the PLLA membrane. So many geometrical and chemical strategies were undertaken to improve cells growth on the PLLA surface. In this study, static magnetic fields were non-invasively applied to our cell cultured system to improve the osteoblast-like cells growth on the PLLA membrane. Cellular models (MG63 and MC3T3-E1, osteoblast-like cells) were carried out to examine the hypothesis that static magnetic fields (SMF) affect osteoblast-like cells growth on the PLLA membrane. The cells were continuously exposed to 4000 Gauss-SMF for 5 days. The proliferation effects of SMF were tested by MTT assay. The morphologic changes and extracellular matrix release were observed by the scanning electron microscopy. In addition, the effects of SMF on the alkaline phosphatase activity levels were compared between the exposed and unexposed cells. Scanning electron microscopy studies indicated that SMF promoted MG63 cells and MC3T3-E1 cells express extracellular matrix on the PLLA membrane. The SMF exposed MG63 cells exhibited decreased MTT values at 2 and 3 days of culture. Besides, the SMF exposed MC3T3-E1 cells exhibited increased MTT values at 3 and 4 days of culture. At 1 day of observation, the alkaline phosphatase-specific activity of the SMF exposed MG63 cells demonstrated a maximum level and significantly larger than that of the unexposed cells (p<0.05) with an increasing ratio of 1.5 folds. The SMF exposed MC3T3-E1 cells express the lower alkaline phosphatase-specific activity than that of the unexposed cells at 3 and 4 days of culture (p<0.05). These results reveal that MG63 cells, seeding on the PLLA membrane, treated with SMF exhibit a more differentiated morphology and express a more differentiated regulatory marker. Furthermore, SMF can promote MC3T3-E1 cells growth on the PLLA membrane.
資料類型:	thesis
顯示於類別:	[牙醫學系] 博碩士論文

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