奈米級珍珠粉體之體外骨誘導特性

本研究使用濕式球磨機將珍珠粉研磨至奈米等級，並探討其研磨前後之差別，進一步將奈米等級的珍珠粉浸漬於模擬體液（ Simulated Body Fluid, SBF ）中，並且使用碳酸鈣及研磨前珍珠粉做對照組，結果顯示浸漬於模擬體液 4 h 後可於其表面形成氫氧基磷灰石 ( Hydroxyapatite, HA ) 鍵結，係經由溶解-沉積機制進行。經由細胞貼附試驗，成骨細胞 ( MC3T3-E1 ) 可於 1 h 內成功貼附在研磨後之珍珠粉上，經 168 h 培養後，經由定性及定量結果顯示成骨細胞依然貼附於研磨後珍珠粉。進一步使用前驅骨母細胞 ( D1 ) 細胞株進行鹼性磷酸酶 ( Alkaline Phosphatase, ALP ) 之礦化試驗，研究其體外引骨特性，定性結果顯示研磨後珍珠粉可於培養 24 h 後出現明顯的紫色沉積物，定量結果顯示經培養 24 h 後，碳酸鈣 ALP 活性含量為 0.008 ml *mM/μg，研磨前珍珠粉 ALP 活性含量為0.036 ml *mM/μg，研磨後珍珠粉 ALP 活性含量為 0.029 ml *mM/μg，研磨前珍珠粉相較於碳酸鈣增加 77%，研磨後珍珠粉較碳酸鈣增加 70%；經 168 h 培養後，研磨後珍珠粉相較於研磨前珍珠粉以及碳酸鈣增加 39% 及 33.6%，因此研磨後珍珠粉之體外骨誘導性較研磨前珍珠和碳酸鈣佳。

關鍵字

珍珠；奈米；生物活性；骨誘導

並列摘要

In this work, nano-grade pearl powder was grinded by wet ball – mill methodology. The osteo-inductive activity of ball milled pearl powder was evaluated by soaking it in simulated body fluid ( SBF ), cell attachment and mineralization assay, taking pearl powder and calcium carbonate powder as control materials at the same time. After soaking in SBF, HA particles were formed on the surface of ball milled pearl powder, and the whole process follows a dissolution and precipitation mechanism. Cell culture revealed that MC3T3-E1 cell can adhere on the surface of ball milled pearl powder after one hour incubation. After 168 hours incubation, MC3T3-E1 cell can proliferate. Qualitative of mineralization assay showed that ball milled pearl powder have the obviously purple dye after 24 hours. After 24 hours of cultivation, quantitative of mineralization assay showed that the respective alkaline phosphatase activity of calcium carbonate powder, pearl powder and ball milled pearl powder is 0.008, 0.036 and 0.029 ml *mM/μg. Pearl powder was a 77% increase compared to calcium carbonate powder. Ball milled pearl powder was a 70% increase compared to calcium carbonate powder. After 168 hours incubation, ball milled pearl powder was a 33.6% ~ 39% increase compared to pearl powder and calcium carbonate powder. It is concluded that ball milled pearl powder is a superior osteogenetic material.