仿肝小葉實驗室晶片檢測酒精中毒造成肝發炎 誘發嗜中性球趨化反應之研究

近年來，肝臟相關的疾病如慢性肝病、肝硬化、肝癌成為臺人死亡十大死因之一。相關治療藥物的開發、測試，一直都是醫藥科學目前發展的重點；在研發新藥過程因為臨床試驗費用昂貴且相關規定嚴格，因此開發新藥的成功率極低。隨著生物微機電技術的發展，可以達成於體外仿肝組織建構，提供有效的進行藥物測試，雖然體外測藥與實際應用於體內真實測藥情形仍存在差異性，但可提供初步且易於觀察的體外藥物測試反應的平台，此外，相關研究人員亦持續朝著更具真實性的體外肝重建模型發展。本研究設計一整合型肝藥物檢測平台。利用生醫微機電技術設計仿肝小葉電極進行體外肝臟細胞排列，藉提供不同頻率的交流電壓而產生之正負介電泳力，控制細胞被極化的程度將細胞排列成仿肝組織之形態並增加細胞和細胞之間的交互作用。此外，搭配濃度梯度流道設計與聲波震盪器產生 5 種不同的乙醛濃度影響下形成體外促使肝細胞發炎的模型，設計之檢測晶片包含上、下流道，且其中間為一層具有直徑 5μm 圓孔的多孔性 PDMS 薄膜，在上流道注入嗜中性白血球後，嗜中性球會貼附於 PDMS 薄膜上，接著因下流道體外重建肝發炎生物模型之影響而產生趨化反應。實驗結果中可以發現細胞排列成仿肝小葉後，在乙醛濃度 175μM 刺激細胞下，比起未進行細胞排列的肝小葉 IL-8 分泌高出 3 倍。嗜中性白血球也受肝發炎情形產生不同的趨化數目。此研究的目標希望以體外重建工程，建立一仿肝實驗室晶片，使其越接近於人體真實肝組織環境，可觀察酒精中毒後，肝發炎生物情形並且期望可以應用於藥物檢測的應用中。

關鍵字

肝發炎；嗜中性球；濃度梯度；聲波混合器；細胞排列

並列摘要

In recent years, liver-related diseases such as chronic liver disease, cirrhosis and liver cancer, are leading cause of disease related deaths Related drug development and testing have always been the goal of medical research. Drug development is expensive due to the several steps involved before the pre-clinical and clinical trials making it expensive and yet the success rate is very low. With the development of biological micro-electromechanical technology, we were able to construct a liver-lobule-mimetic tissue for effective drug testing. Although there are differences in the in vitro testing of drug and it’s in vivo practical application, the in vitro platform provides preliminary and easy observation which can be useful for researchers towards the development of in vitro liver reconstruction model. This research presents an integrated drug testing lab chip where in the liver-lobule-mimetic electrode pattern is used to generate in vitro liver cell pattern. Based on polarity difference of cells, positive and negative DEP force is utilized to manipulate and pattern heterogeneous cells for mimicking the inherent hepatic morphology, increasing the cell-cell interaction. The concentration gradient generator design and acoustic micromixer generates five different concentrations of acetaldehyde, which is used to induce in vitro liver inflammation and observe neutrophil migration in Alcoholic Liver Inflammation In the design of chip test platform, there are upper channel and bottom channel. Moreover, the intermediate bonding layer of a porous PDMS thin film has a hole which has a diameter of 5μm. When neutrophil injected into upper channel, it would attach to the PDMS thin film. Then bottom channel of chip established in vitro reconstruction of the biological model of liver inflammation to generate chemotactic response. The cell were treated by using acetaldehyde in the concentration of 175μM after the cell were arranged in liver lobule-mimetic. Experimental results indicate that the secretion of IL-8 is 3-fold higher compared with the liver lobule-mimetic without cell arrangement. Furthermore, neutrophil would produce different number of migration by the influence of liver inflammation. The ultimate goal of this research is to construct an in vitro liver tissue, mimicking the in vivo liver in its functioning and prove its usefulness in drug testing

並列關鍵字

Liver inflammation ； neutrophils ； concentration gradient ； acoustic mixer ； cell pattern

參考文獻

[1] K. Nishida, M. Yamato, Y. Hayashida, K. Watanabe, K. Yamamoto, E. Adachi, S. Nagai, A. Kikuchi, N. Maeda, H. Watanabe, T. Okano, and Y. Tano, "Corneal Reconstruction with Tissue-Engineered Cell Sheets Composed of Autologous Oral Mucosal Epithelium," New England Journal of Medicine, vol. 351, pp. 1187-1196, 2004.

[2] R. Taub, "Liver regeneration: from myth to mechanism," Nat Rev Mol Cell Biol, vol. 5, pp. 836-847, 2004.

[3] Y. Nahmias, R. E. Schwartz, C. M. Verfaillie, and D. J. Odde, "Laser-guided direct writing for three-dimensional tissue engineering," Biotechnology and Bioengineering, vol. 92, pp. 129-136, 2005.

[5] J. R. Fuchs, B. A. Nasseri, and J. P. Vacanti, "Tissue engineering: a 21st century solution to surgical reconstruction," Ann Thorac Surg, vol. 72, pp. 577-591, August 1, 2001 2001.

[6] K. Bhadriraju and C. S. Chen, "Engineering cellular microenvironments to improve cell-based drug testing," Drug Discovery Today, vol. 7, pp. 612-620, 2002.

國際替代計量

仿肝小葉實驗室晶片檢測酒精中毒造成肝發炎誘發嗜中性球趨化反應之研究

主題瀏覽