模擬養分與膠原蛋白對軟骨細胞生長之影響

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姓名

林正翰(Cheng-Han Lin) 查詢紙本館藏

畢業系所

機械工程學系

論文名稱

模擬養分與膠原蛋白對軟骨細胞生長之影響
(Modeling and Simulation for the Effects of Nutrients and Collagen on Tissue Engineering Chondrocye Growth)

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摘要(中)

組織工程的目的為培養生物替代物來修復與改善人體組織與器官的缺陷。關節軟骨為結締組織的一種，由於軟骨缺乏血管與神經，因此受傷後，難以修補受傷的組織；另外細胞外基質包覆著軟骨細胞，所以健康的軟骨也不太能夠遷移至受損的區域進行修護。為了解決軟骨自我修復能力有限的缺點，因此組織工程希望能在體外培養出軟骨替代物。
本文模擬在靜態培養環境下，將軟骨細胞種植在不同厚度的支架上，培養30天，並且每3天更換一次培養液。考慮養分的擴散與消耗、細胞的隨機漫步特性、養份促進細胞的生長和膠原蛋白調控細胞的增殖，以及膠原蛋白的分泌與降解的現象，將這些現象以數學模型來表現，以探討養份對細胞生長的影響，以及膠原蛋白如何調控細胞的生長。模擬得到細胞密度與膠原蛋白含量與文獻的實驗結果一致。模擬結果發現，細胞分佈在第10天相當均勻，因為此時養份在支架的分佈也相當均勻，在第20天和第30天時，細胞在支架表面有較高的細胞密度，支架內部的細胞分佈主要受到氧氣的濃度的影響，而在支架的徑向表面，因為葡萄糖在此處的濃度較高，促使細胞在支架的徑向表面生長較佳。膠原蛋白隨著培養時間增加，其含量逐漸增加，膠原蛋白分佈與細胞分佈相似，因為膠原蛋白的分泌是受到細胞的影響。本研究也發現支架相對於培養液的厚度也影響靜態培養下細胞的生長對於養分的敏感度，種植在支架厚度較薄的細胞，細胞生長主要受到氧氣濃度的影響；而支架厚度較厚則是受到葡萄糖濃度的影響。

摘要(英)

Tissue engineering is aimed to culture biological substitutes to repair and improve the damaged human tissue and organ. Articular cartilage is a connective tissue, which is difficult to repair after injury because the tissue lacks blood vessels and nerves. Even worse is that chondrocytes are embeded within the extracellular matrix, therefore healthy chondrocytes cannot migrate to the injury site for repair. In order to solve the limited capability of cartilage repair, tissue engineering hopes to develop cartilage substitutes in vitro.
To help engineer cartilage implants, we simulate chondrocyte growth considering cells are seeded on the scaffolds of different thicknesses under static culture for 30 days, in which the medium is replaced every 3 days. We consider the phenomena of diffusion and consumption of nutrients, random walks of cells, nutrient-promoted cell growth, collagen-regulated cell proliferation, and collagen synthesis and degradation. We probe nutrient effects to cell growth, and how the collagen regulates cell growth. Results show the cell distribution is quite uniform at 10 days since the nutrient is still quite uniform in the scaffold at the moment. At 20 and 30 days, cells have higher density on the surface of the scaffold. Cell distribution within the scaffold is mainly influenced by oxygen concentration. Glucose is virtually relatively depleted but has higher concentration on the radial surface of the scaffold so that cell density is higher on the radial than on the horizontal surface. The synthesized collagen increases in amount with cultivation time. Collagen distributes similarly to cells because it is secreted by cells. Results also show that cell growth is more sensitive to oxygen variation for thinner scaffolds; on the contrary, to glucose for thicker scaffolds.

關鍵字(中)

★ 膠原蛋白
★ 葡萄糖
★ 氧氣
★ 增殖
★ 組織工程
★ 軟骨細胞
★ 擴散

關鍵字(英)

★ chondrocyte
★ diffusion
★ tissue engineering
★ proliferation
★ oxygen
★ glucose
★ collagen

論文目次

目錄
中文摘要 i
英文摘要 ii
誌謝 iii
目錄 iv
表目錄 vi
圖目錄 vii
符號說明 x
第一章緒論 ………………………………………………………… 1
1.1 前言 ……………………………………………………………… 1
1.2 文獻回顧 ………………………………………………………… 3
1.3 研究動機 ………………………………………………………… 5
第二章數學模型 …………………………………………………… 6
2.1 物理系統與基本假設 …………………………………………… 6
2.2 統御方程式 ……………………………………………………… 7
2.2.1 培養液養分濃度守恆方程式 ………………………… 7
2.2.2 支架中養分濃度守恆方程式 ………………………… 7
2.2.3 膠原蛋白守恆方程式 ………………………………… 8
2.2.4 細胞質量守恆方程式 ………………………………… 8
2.3 初始條件與邊界條件 …………………………………………… 9
2.3.1 初始條件 ……………………………………………… 9
2.3.2 邊界條件 ……………………………………………… 11
第三章數值方法 …………………………………………………… 16
3.1 COMSOL簡介 ………………………………………………… 16
3.2 網格配置 ………………………………………………………… 16
3.3 誤差與精確度 …………………………………………………… 17
3.4 最佳化參數 ……………………………………………………… 17
第四章結果與討論 ………………………………………………… 22
4.1 數學模型的驗證 ………………………………………………… 22
4.2 基本結果與討論 ………………………………………………… 23
4.2.1 氧氣濃度隨時間的變化與分佈 …………………… 23
4.2.2 葡萄糖濃度隨時間的變化與分佈 ………………… 24
4.2.3 細胞數量密度隨時間的變化與分佈 ……………… 25
4.2.4 膠原蛋白隨時間的變化與分佈 …………………… 25
4.3 參數分析 ………………………………………………………… 26
4.3.1 養分的初始濃度對細胞生長的影響 ……………… 26
4.3.2 飽和常數對細胞生長的影響 ……………………… 27
4.3.3 更換培養液的頻率對細胞生長的影響 …………… 27
4.3.4 細胞的擴散係數與養分在支架內的擴散係數對細胞生長之影響 28
4.3.5 培養液高度對細胞生長之影響 28
第五章結論與未來展望 …………………………………………… 56
參考文獻 …………………………………………………………………… 58

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指導教授

鍾志昂(Chih-Ang Chung)

審核日期

2009-7-20

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