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標題: Evaluation of cell seeding, differentiation and bioreactor culture for scaffolds manufactured by the fused deposition modeling
作者: 鄭家昇
Cheng, Jia sheng
關鍵字: 骨先驅細胞;osteoprogenitor cells;ALP活性;精密支架;動態培養;生物反應器;ALP activity;dynamic culture;precision scaffolds;bioreactor
出版社: 化學工程學系
In the first part of this study, the alkaline phosphatase (ALP) activity and calcification of osteoprogenitor cells after being stimulated with the differentiating reagent were measured. The ALP activity was found to increase significantly after ten days and decrease after eighteen days. The control osteoprogenitor cells without the differentiating reagent were not calcified. The cells with the differentiating reagent were calcified after ten days, and more after eighteen days, as demonstrated by the von Kossa stain.
To enhance the cell seeding efficiency in the precision scaffolds, different methods have been compared. By dropping the cell suspensions on the scaffolds, we were able to increase the seeding efficiency to ∼50%. When the precision scaffolds had a concentric circle (i.e. loose interior) structure, the seeding efficiency could reach 66%.
In the three-dimensional cell culture, the scaddolds seeded with osteoblasts were placed in a rotational bioreactor with a speed of 6 rpm. After four weeks, more cell proliferation was observed in PLLA freeze-dried scaffolds. Precision scaffolds, on the other hand, had more extracellular matrix. Based on these results, we have established a cell-precision scaffold-bioreactor model for more in-depth study in the future.

本研究首先以骨先驅細胞作為研究的主題,評估以分化劑刺激的骨先驅細胞,並鑑定分化程度。實驗結果發現在平面培養分化10天後鹼性磷酸酶ALP活性明顯上升,在10-18天ALP活性有顯著的下降;在von Kossa染色方面,骨先驅細胞未加分化劑刺激者,在培養時並沒有明顯的鈣化情形,但刺激組在10天後即有著明顯的鈣化產生,18天後更為顯著。
在三度空間細胞培養方面,以轉速6 rpm進行骨母細胞-支架複合物動態培養4週後,發現冷凍乾燥支架PLLA有較佳的細胞增生現象,而精密支架則有較多的基質分泌。依據本研究之結果可建立細胞-精密支架-生物反應器的模式。
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