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Title: | MG-63細胞在雙光子聚合粗糙表面之活體外分析 An in-vitro Analysis of MG-63 Cells on Roughened Surface Fabricated by Two Photon Polymerization |
Authors: | Subhashree Shivani 施凡妮 |
Advisor: | 王安邦(Prof.An-Bang Wang) |
Co-Advisor: | 許聿翔(Prof.Yu-Hsiang Hsu) |
Keyword: | 雙光子聚合,MG63,粗糙度梯度,無方向性之移動, Two-photon polymerization,MG63,roughness gradient,unidirectional migration, |
Publication Year : | 2018 |
Degree: | 碩士 |
Abstract: | 細胞之遷移對於傷口快速復原為一個重要之影響參數。而不同的環境也影響細胞能動性。現今研究上已有關於表面粗糙度扮演著一個重要的拓樸結構,其可以提升細胞的行為,然而對於控制沒有移動方向性之細胞仍然相當艱鉅。先前已有關於細胞遷移研究,其材料使用鈦金屬(Titanium)亦者其他金屬,其基質之硬度不同於骨頭。此外,若使用噴砂處理製造粗糙度之方式會引致尖頂形狀,將導致非均一化之粗糙度。
本研究有一系列實驗來了解關於粗糙度高以及粗糙度低之表面對於細胞在其上之移動速度、延緩、移動方向之角度與速率。我們使用快速激光雷射技術:雙光子聚合。利用生物相容性之材料(Ormocomp)使用雙光子聚合方式以製造出其模具,可得其表面之粗糙度分別為0.4 微米 與1.3 微米。奈米印製方法(Nano imprinting method)是用來去創造最終聚丙烯之基底。最終利用人類癌症骨細胞(MG63)接種於基底去研究細胞。 在表面粗糙度較高之材料顯示細胞提升其移動性並且在表面粗糙度較高之材料上,欄桿在移動方向上扮演了一個重要的角色;然而對於表面粗糙度較低之材料並沒有很有效果。根據結果,新的表面粗糙梯度會去測試細胞移動之效果,有趣的是:在沒有欄杆之結構上細胞有移動從粗糙度高至低之趨勢性。而材料結構面上之粗糙度梯度對於細胞移動扮演一個很重要之因素,在移動速度上會下降而增加其移動方向之角度。然而在放入欄杆之結構能促使細胞雙向移動。另外,當欄杆被排列成垂直表面粗糙度之梯時,其會限制細胞移動至另一處之速度,顯示了其性質對於表面粗糙度有較高之控制性。 這些設計可以進一步去探索有關於創建有表面粗糙度梯度之支架於更快速復原尤其是對於骨折之病患。 Cell migration is an important parameter for faster wound healing. Various environmental cues can affect cell motility. Research have shown that roughness is an important topological cue that can enhance cell behavior. However, controlling the unidirectional migration of the cells is still a challenge. Previous studies on cell migration used Titanium or other metals as their substrates having much different stiffness than bone. In addition, use of sandblasting method to create roughness introduced sharp peaks that could result in non-uniform roughness. In this study, a series of experiments were done on different roughness to understand the effect of higher and lower roughness on cell parameters such as migration velocity, migration delay, angle of orientation and rate of change of orientation. Uniform roughness of 0.4 µm and 1.3 µm were fabricated using rapid laser fabrication technique, two-photon polymerization (TPP) on a biocompatible material, Ormocomp to create mold. Nano imprinting method was used to create final PP substrates. Human osteosarcoma bone cells, MG63, were seeded on the substrates to analyze the cell parameters. It was demonstrated that higher roughness enhanced migration activities of the cells. Also, rails played an important role in orienting the cells in higher roughness whereas were ineffective in lower roughness. Based on these results, novel roughness gradients were designed to examine the effect on cell migration. Interestingly, cells biased their migration from higher roughness to lower roughness for no rail substrates. Area of each roughness on the gradient was also vital as larger area decreased the migration velocity of the cells and increased orientation angle. However, introduction of rails facilitated bidirectional migration. Also, when aligned perpendicularly to the gradient, rails restricted the migration of cells to one region, suggesting higher dominance on cell motility than roughness. These designs can be further explored for creating roughness gradient scaffolds for faster healing of especially fractured bones. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68034 |
DOI: | 10.6342/NTU201800024 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 應用力學研究所 |
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ntu-107-1.pdf Restricted Access | 5.54 MB | Adobe PDF |
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