以片狀細胞工程重建兔角膜內皮組織

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題名:	以片狀細胞工程重建兔角膜內皮組織 Reconstruction of rabbit corneal endothelium with cell sheet engineering
作者:	徐悅琪 Hsu, Yueh-Chi
貢獻者:	醫學科學研究所曾厚
關鍵詞:	角膜內皮細胞;聚偏氟乙烯;聚麩胺酸;片狀細胞工程 corneal endothelial cell;polyvinylidene fluoride;γ-polyglutamic acid;cell sheet engineering
日期:	2016
上傳時間:	2021-11-26 19:35:05 (UTC+8)
摘要:	人類角膜內皮細胞在體內的增生能力十分有限，而角膜內皮細胞密度依年齡增加而逐漸減少，且一旦損傷則不能再生，當細胞密度下降達一定程度時，角膜內皮組織的房水屏障及排水生理功能將無法發揮，因而有可能導致角膜水腫、混濁，甚至可能視覺喪失。在目前臨床上，傳統的全層角膜移植術是以全層透明的角膜來置換混濁或病變的角膜，而大部分的角膜移植術是執行在僅角膜內皮細胞受損的病患上，若是能以體外大量培養人類演角膜內皮細胞進行受損角膜內皮組織之置換，則可以改善捐贈角膜全球捐贈角膜來源短缺之問題。本研究之目的即是以新式片狀細胞工程培養兔角膜內皮細胞層片，以開發一新穎性治療法進行眼組織重建。首先，根據電漿化學反應設計一套兩階段的材料表面改質方法，以製備聚麩胺酸(γ-polyglutamic acid)高分子接枝表面，並應用於兔角膜內皮細胞之培養。將兔角膜內皮細胞培養於聚麩胺酸/聚偏氟乙烯(polyvinylidene fluoride)薄膜上，兩周後，長滿之細胞可經由還原劑作用使片狀細胞脫著於基材。之後針對此人工片狀角膜內皮的體外性質以組織學、免疫組織化學染色以及存活率試驗進行評估是否能做為天然組織替代物，期望將來可以此一新療法未來可以應用於眼組織再生重建及臨床治療上。 Human corneal endothelial cells have no proliferative capacity in vivo, these cells decrease in number with aging, disease, or trauma. When the cell density drops below a critical level, the corneal endothelial cells can no longer pump enough water to compensate for diffusion into the cornea, resulting in stromal edema, corneal clouding and eventual vision loss. Conventional penetrating keratoplasty involves replacing the entire corneal layer to treat patients whose cornea is healthy except for damaged endothelium. However, access to donated tissue is limited worldwide resulting in critical need for new sources of corneal grafts. The number of such grafts produced by each donor eye could be increased significantly if CE cells were expanded in culture before grafting. In this study, we developed a novel therapy technique to fabricate and transplant cultured rabbit corneal endothelial cell (RCECs) sheets for corneal endothelial reconstruction. On the basis of plasma chemistry, we have designed a two-step method to prepare a γ-polyglutamic acid-grafted polyvinylidene fluoride thin film for RCEC cultivation. Confluent cell cultures were detached as a laminated sheet by reductant additive. In vitro characteristics of HCEC sheets were evaluated by histology, immunohistochemistry, and viability. We hope this work may be an optimized method to fabricate bioengineered corneal endothelium and show potential to facilitate CEC transplantation in the future.
描述:	碩士指導教授：曾厚
資料類型:	thesis
顯示於類別:	[醫學科學研究所] 博碩士論文

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