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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78157完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林?輝(Feng-Huei Lin) | |
| dc.contributor.author | Tzu-Lin Kuo | en |
| dc.contributor.author | 郭芷綝 | zh_TW |
| dc.date.accessioned | 2021-07-11T14:44:07Z | - |
| dc.date.available | 2021-10-14 | |
| dc.date.copyright | 2016-10-14 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-08-05 | |
| dc.identifier.citation | [1] Oliva MS, Schottman T, Gulati M. Turning the tide of corneal blindness. Indian J Ophthalmol 2012;60:423-7
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Transplantation of adult human or porcine corneal endothelial cells onto human recipients in vitro. Part I: cell culturing and transplantation procedure. Cornea 18, 199e206. [32]Aboalchamat, B., Engelmann, K., Bohnke, M., Eggli, P., Bednarz, J., 1999. Morphological and functional analysis of immortalized human corneal endothelial cells after transplantation. Exp. Eye Res. 69, 547e553. [33]Bohnke, M., Eggli, P., Engelmann, K., 1999. Transplantation of cultured adult human or porcine corneal endothelial cells onto human recipients in vitro. Part II: evaluation in the scanning electron microscope. Cornea 18, 207e213. [34]Chen, K.H., Azar, D., Joyce, N.C., 2001. Transplantation of adult human corneal endothelium ex vivo: a morphologic study. Cornea 20, 731e737. [35]Amano, S., 2002. Transplantation of corneal endothelial cells. Nihon Ganka Gakkai Zasshi 106, 805e835. Discussion 836. [36]Amano, S., 2003. Transplantation of cultured human corneal endothelial cells.Cornea 22, S66eS74. [37]Mimura, T., Amano, S., Usui, T., Araie, M., Ono, K., Akihiro, H., Yokoo, S., Yamagami, S., 2004a. Transplantation of corneas reconstructed with cultured adult human corneal endothelial cells in nude rats. Exp. Eye Res. 79, 231e237. [38] Mimura, T., Yamagami, S., Yokoo, S., Usui, T., Tanaka, K., Hattori, S., Irie, S., Miyata, K., Araie, M., Amano, S., 2004b. Cultured human corneal endothelial cell transplantation with a collagen sheet in a rabbit model. Invest. Ophthalmol. Vis. Sci. 45, 2992e2997. [39] Ishino, Y., Sano, Y., Nakamura, T., Connon, C.J., Rigby, H., Fullwood, N.J., Kinoshita, S., 2004. Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation. Invest. Ophthalmol. Vis. Sci. 45, 800e806. [40]Honda, N., Mimura, T., Usui, T., Amano, S., 2009. Descemet stripping automated endothelial keratoplasty using cultured corneal endothelial cells in a rabbit model. Arch. Ophthalmol. 127, 1321e1326. [41]Choi, J.S., Williams, J.K., Greven, M.,Walter, K.A., Laber, P.W., Khang, G., Soker, S., 2010. Bioengineering endothelialized neo-corneas using donor-derived corneal endothelial cells and decellularized corneal stroma. Biomaterials 31, 6738e6745. [42]Lai, J.Y., Chen, K.H., Hsiue, G.H., 2007. Tissue-engineered human corneal endothelial cell sheet transplantation in a rabbit model using functional biomaterials. Transplantation 84, 1222e1232. [43]Watanabe, R., Hayashi, R., Kimura, Y., Tanaka, Y., Kageyama, T., Hara, S., Tabata, Y., Nishida, K., 2011. A novel gelatin hydrogel carrier sheet for corneal endothelial transplantation. Tissue Eng. Part. A 17, 2213e2219. [44]Liang, Y., Liu,W., Han, B., Yang, C., Ma, Q., Zhao,W., Rong, M., Li, H., 2011. Fabrication and characters of a corneal endothelial cells scaffold based on chitosan. J. Mater. Sci. Mater. Med. 22, 175e183. [45]Watanabe R, Hayashi R, Kimura Y, Tanaka Y, Kageyama T, Hara S, et al. A novel gelatin hydrogel carrier sheet for corneal endothelial transplantation. Tissue Eng Part A. 2011;17:2213-9. [46] Yoshida J, Oshikata-Miyazaki A, Yokoo S, Yamagami S, Takezawa T, Amano S. Development and evaluation of porcine atelocollagen vitrigel membrane with a spherical curve and transplantable artificial corneal endothelial grafts. Invest Ophthalmol Vis Sci. 2014;55:4975-81. [47] Yuan F, Wang L, Lin CC, Chou CH, Li L. A cornea substitute derived from fish scale: 6-month followup on rabbit model. J Ophthalmol. 2014;2014:914542. [48] van Essen TH, Lin CC, Hussain AK, Maas S, Lai HJ, Linnartz H, et al. A fish scale-derived collagen matrix as artificial cornea in rats: properties and potential. Invest Ophthalmol Vis Sci. 2013;54:3224-33. [49] Lin CC, Ritch R, Lin SM, Ni MH, Chang YC, Lu YL, et al. A new fish scale-derived scaffold for corneal regeneration. Eur Cell Mater. 2010;19:50-7. [50] Hos D, van Essen T, Bock F, Chou C-H, Pan H-A, Lin C-C, et al. 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[55] Premix WST-1 Cell Proliferation Assay System Product Manual, Cat. #MK400 v201306, TaKaRa, Japan [56] LDH Cytotoxicity Detection Kit Product Manual, Cat. #MK401 v201404, TaKaRa, Japan [57] LIVE/DEAD® Viability/Cytotoxicity Kit for mammalian cells Product Manual, Cat. #L3224, Thermo Fisher, USA [58] Merrett K, Fagerholm P, McLaughlin CR, Dravida S, Lagali N, Shinozaki N, et al. Tissue-engineered recombinant human collagen-based corneal substitutes for implantation: performance of type I versus type III collagen. Invest Ophthalmol Vis Sci. 2008;49:3887-94. [59] Fagerholm P, Lagali NS, Ong JA, Merrett K, Jackson WB, Polarek JW, et al. Stable corneal regeneration four years after implantation of a cell-free recombinant human collagen scaffold. Biomaterials. 2014;35:2420-7. [60] N. Okumura, N. Koizumi, M. Ueno, Y. Sakamoto, H. Takahashi, H. Tsuchiya, J. Hamuro and S. Kinoshita, Am J Pathol 181 (2012) 268-277. [61] J. Yoshida, A. Oshikata-Miyazaki, S. Yokoo, S. Yamagami, T. Takezawa and S. Amano, Invest Ophthalmol Vis Sci 55 (2014) 4975-4981. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78157 | - |
| dc.description.abstract | 角膜內皮細胞因為疾病、外傷、手術操作不當等因素導致細胞數量下降，此時角膜內皮層排水功能會失償，進而導致角膜水腫混濁，甚至有失明的可能性。目前角膜內皮層受損的治療方式是以人體捐贈角膜作手術的置換，然而符合捐贈條件的角膜數量在眼庫內一直呈現短缺狀態，因此開發受損角膜內皮治療方式是相當重要且急迫的。本研究使用取自於魚鱗的膠原蛋白作為角膜內皮細胞層培養載體，並且植入前房作角膜內皮細胞層的修復，是一個極具潛力的角膜內皮細胞的治療方式。
在本研究中，經過去細胞、脫鈣、交聯、弧壓後的魚鱗利用螢光顯微鏡、電子顯微鏡、熱重分析儀來評估細胞殘留率、表面結構分析及無機質殘留率。生物相容性的評估則是使用WST-1及LDH分別對細胞活性及細胞毒性作測試。利用電子顯微鏡及ZO-1免疫螢光染色對培養在魚鱗載體上的角膜內皮細胞緊密連接進行觀察。最後將魚鱗載體連同角膜內皮細胞層植入紐西蘭大白兔前房進行體內生物相容性的評估。 結果顯示經過處理後的魚鱗膠原蛋白載體具有高透明度、高含水率及好的生物相容性，並且植入進前房後不會造成免疫反應發生。培養於魚鱗載體上的角膜內皮細胞特徵與正常角膜內皮相似。因此根據本研究的結果，魚鱗膠原蛋白符合當作角膜內皮細胞層載體的一切條件。 | zh_TW |
| dc.description.abstract | Corneal endothelial cell quantity may decline due to diseases, trauma and inappropriate surgery method and this situation will lead to corneal edema and have the possibility for patients to become blind. Nowadays, donor corneas are used to treat endothelial dysfunctions and replace damaged tissue. However, the shortage of donor corneas which can meet the conditions of the donation is always a serious problem. Therefore, developing a new corneal endothelial treatment method is important and urgent. We use collagen membrane derived from fish scale, which is a material with high potentiality, to act as a carrier for corneal endothelial cells and to implant into anterior chamber to treat the damaged endothelium.
In this study, de-cellularized, de-calcificated, cross-linking and curvature fabricated fish scale was characterized by fluorescent microscope for de-cellularized evaluation, scanning electron microscope (SEM) for microstructure examination and thermal gravitation analysis (TGA) for de-calcificated analysis. Biocompatibility was analyzed by WST-1 assay for cell viability and LDH assay for cell cytotoxicity. Besides, SEM and ZO-1 immunofluorescence were used to examine cell junction of the endothelial cells on collagen membrane. Finally, collagen membrane with endothelium layer was implanted into rabbit anterior chamber for evaluation of biocompatibility in vivo. The results show that the prepared fish scale collagen membrane has advantages of high transparency, water content and excellent biocompatibility and no immune reactions are seen in rabbit anterior chamber after implantation. In addition, the cultivated endothelial cells on the collagen membrane are similar to normal cells in vivo. According to this study, fish scale collagen membrane meets the criteria to serve as a carrier for corneal endothelial cells. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T14:44:07Z (GMT). No. of bitstreams: 1 ntu-105-R03548013-1.pdf: 2997962 bytes, checksum: 990510354f96907715013d507c61ac88 (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 口試委員會審定書 #
誌謝 i 中文摘要 ii Abstract iii 目錄 v 圖目錄 viii 表目錄 x 縮寫目錄 xi 第一章 緒論 1 1.1 前言 1 1.2 眼角膜解剖構造 1 1.3 眼角膜內皮層構造與功能 2 1.4 眼角膜內皮疾病 4 1.5 目前治療方式 5 1.6 眼角膜移植 5 第二章 理論基礎 8 2.1 組織工程 8 2.2 角膜內皮細胞層載體材料的選擇 9 2.3 魚鱗特性 9 2.4 研究目的 12 第三章 材料與方法 13 3.1 材料製備 15 3.2 魚鱗片材曲度成型 18 3.3 去細胞魚鱗片材細胞核染色 19 3.4 無機質含量分析 19 3.5 交聯劑殘存量測定 19 3.6 含水率測量 20 3.7 膠原蛋白含量測量 20 3.8 透光度測量 20 3.9 透氧率測量 21 3.10 體外生物相容性測試 21 3.10.1 細胞株培養 21 3.10.2 材料萃取液製備 21 3.10.3 WST-1 assay細胞活性測試 21 3.10.4 LDH assay細胞毒性測試 22 3.10.5 Live/Dead staining細胞存活染色 23 3.11 角膜內皮細胞培養 24 3.12 內皮細胞型態觀察 25 3.13 ZO-1免疫螢光染色 25 3.14 體內生物相容性測試 25 3.15 組織切片觀察 26 3.16 統計分析 26 第四章 實驗結果 27 4.1 魚鱗片材去細胞率 27 4.2 魚鱗片材脫鈣表面結構觀察 27 4.3 無機質熱重分析 28 4.4 交聯劑殘存量 30 4.5 膠原蛋白含量 30 4.6 含水率 31 4.7 透光度 31 4.8 透氧率 32 4.9 WST-1 assay細胞活性測試 33 4.10 LDH assay細胞毒性測試 33 4.11 Live/Dead staining細胞存活染色 34 4.12 角膜內皮細胞培養 35 4.13 內皮細胞型態觀察 36 4.14 ZO-1免疫螢光染色 37 4.15 體內生物相容性評估 38 4.16 組織切片觀察 42 第五章 討論 44 第六章 總結 46 參考文獻 47 | |
| dc.language.iso | zh-TW | |
| dc.subject | 角膜 | zh_TW |
| dc.subject | 後彈力層剝除自動角膜內皮移植術 | zh_TW |
| dc.subject | 魚鱗 | zh_TW |
| dc.subject | 細胞載體 | zh_TW |
| dc.subject | 膠原蛋白 | zh_TW |
| dc.subject | cornea | en |
| dc.subject | fish scale | en |
| dc.subject | cell carrier | en |
| dc.subject | collagen | en |
| dc.subject | DSAEK | en |
| dc.title | 以魚鱗膠原蛋白做為角膜內皮細胞層培養載體以做為後彈力層剝除自動角膜內皮移植術的手術移植體 | zh_TW |
| dc.title | Fish Scale Collagen as Feeder Layer for Corneal Endothelial Cell-sheet for DSAEK Graft | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張淑真(Shwu-Jen Chang),郭士民(Shyh-Ming Kuo),陳克紹(Ko-Shao Chen) | |
| dc.subject.keyword | 後彈力層剝除自動角膜內皮移植術,角膜,魚鱗,細胞載體,膠原蛋白, | zh_TW |
| dc.subject.keyword | DSAEK,cornea,fish scale,cell carrier,collagen, | en |
| dc.relation.page | 53 | |
| dc.identifier.doi | 10.6342/NTU201601972 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-08-05 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
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