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  1. NTU Theses and Dissertations Repository
  2. 工學院
  3. 醫學工程學研究所
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/21103
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor黃義侑(Yi-You Huang)
dc.contributor.authorJi-Feng Huangen
dc.contributor.author黃基豐zh_TW
dc.date.accessioned2021-06-08T03:27:02Z-
dc.date.copyright2020-01-16
dc.date.issued2019
dc.date.submitted2020-01-13
dc.identifier.citation1. Fu Y, Fan X, Chen P, Shao C, Lu W. 2010. Reconstruction of a Tissue-Engineered Cornea with Porcine Corneal Acellular Matrix as the Scaffold. Cells Tissues Organs 191:193-202.
2. Zhang J, Zhang C-W, Du L-Q, Wu X-Y. 2016. Acellular porcine corneal matrix as a carrier scaffold for cultivating human corneal epithelial cells and fibroblasts in vitro. International journal of ophthalmology 9:1-8.
3. INC. O. 2019. Anatomy of the Eye.
4. Waring GO, 3rd, Bourne WM, Edelhauser HF, Kenyon KR. 1982. The corneal endothelium. Normal and pathologic structure and function. Ophthalmology 89:531-590.
5. Tan DTH, Dart JKG, Holland EJ, Kinoshita S. 2012. Corneal transplantation. The Lancet 379:1749-1761.
6. Joyce NC. 2003. Proliferative capacity of the corneal endothelium. Prog Retin Eye Res 22:359-389.
7. Wilson SL, El Haj AJ, Yang Y. 2012. Control of scar tissue formation in the cornea: strategies in clinical and corneal tissue engineering. J Funct Biomater 3:642-687.
8. Kaufman HE, Clower JW. 1966. Irregularities of Bowman's Membrane ⋆. American Journal of Ophthalmology 61:227-230.
9. Eghrari AO, Riazuddin SA, Gottsch JD. 2015. Chapter Two - Overview of the Cornea: Structure, Function, and Development, p 7-23. In Hejtmancik JF, Nickerson JM (ed), Progress in Molecular Biology and Translational Science, vol 134. Academic Press.
10. Stocum DL. 2012. Chapter 3 - Regeneration of Epidermal Structures, p 43-65. In Stocum DL (ed), Regenerative Biology and Medicine (Second Edition) doi:https://doi.org/10.1016/B978-0-12-384860-4.00003-4. Academic Press, San Diego.
11. Donaghy CL VJ, Greiner MA. An May 21, 2015. Introduction to Corneal Transplantation
http://EyeRounds.org/tutorials/cornea-transplant-intro/. Accessed
12. Kinoshita S, Koizumi N, Ueno M, Okumura N, Imai K, Tanaka H, Yamamoto Y, Nakamura T, Inatomi T, Bush J, Toda M, Hagiya M, Yokota I, Teramukai S, Sotozono C, Hamuro J. 2018. Injection of Cultured Cells with a ROCK Inhibitor for Bullous Keratopathy. New England Journal of Medicine 378:995-1003.
13. Donaghy CL VJ, Greiner MA. May 21, 2015. An Introduction to Corneal Transplantation.
14. Melles GR, Ong TS, Ververs B, van der Wees J. 2006. Descemet membrane endothelial keratoplasty (DMEK). Cornea 25:987-990.
15. Williams KA, Muehlberg SM, Lewis RF, Coster DJ, on behalf of all contributors to the Australian Corneal Graft R. 1995. How successful is corneal transplantation? A report from the Australian corneal graft register. Eye 9:219-227.
16. Vladimir Mironov DNR, and Brian Derby. 2006. Tissue Engineering. Tissue Engineering doi:http://doi.org/10.1089/ten.2006.12.631:pp. 631-634.
17. Seol Y-J, Kang T-Y, Cho D-W. 2012. Solid freeform fabrication technology applied to tissue engineering with various biomaterials. Soft Matter 8:1730-1735.
18. Isaacson A, Swioklo S, Connon CJ. 2018. 3D bioprinting of a corneal stroma equivalent. Experimental eye research 173:188-193.
19. Ju C, Zhang K, Wu X. 2012. Derivation of Corneal Endothelial Cell-Like Cells from Rat Neural Crest Cells In Vitro. PLOS ONE 7:e42378.
20. Garcia CM, Kwon GP, Beebe DC. 2006. alpha-Smooth muscle actin is constitutively expressed in the lens epithelial cells of several species. Experimental eye research 83:999-1001.
21. Yamashita K, Hatou S, Inagaki E, Higa K, Tsubota K, Shimmura S. 2018. A Rabbit Corneal Endothelial Dysfunction Model Using Endothelial-Mesenchymal Transformed Cells. Scientific Reports 8:16868.
dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/21103-
dc.description.abstract眼科疾病是臨床上常見的疾病之一,常見的狀況有:「結膜炎、角膜潰傷、青光眼、飛蚊症等疾病。」內皮層細胞主要負責角膜排水的工作,能讓角膜維持透明度,使光線能順利穿過角膜進到眼球內,但最惱人的是一旦傷及角膜內皮層後,角膜無法自行再生,患者會出現水腫、角膜混濁不透光、視線模糊不清的徵兆,持續惡化之後病人逐漸喪失角膜所具有的功能,造成失明。失明後則需要接受角膜捐贈者捐贈角膜,病人進行全層角膜移植後才能重見光明,但並非每位病人都能如此幸運獲得新的角膜,現今角膜捐贈狀況可以使用供不應求來形容,因此許多研究希望能透過組織工程的方式,能在透過體外培養一片具有功能的眼角膜結構,加速病人進行角膜移植的速度。
  此研究亦希望能透過體外培養,培養出具功能性的角膜。研究內使用去細胞之豬角膜為支架,選擇豬角膜原因為結構較接近人類角膜的結構,透過培養之細胞,將細胞種於去細胞支架上進行共培養,並且觀察細胞貼附狀況、所選的支架不利於對於角膜上皮、內皮細胞貼附。
  成功將細胞種於支架後,將此支架進行全層角膜移植手術,移植至紐西蘭大白兔眼球上,透過長時間觀察,希望內皮細胞能使不透明的去細胞角膜支架轉變成透明的角膜組織,取代原先以混濁、不透光的角膜組織。
zh_TW
dc.description.abstractDisease of eyes is the most common illness clinically. There are some symptoms in common such as pink eyes, corneal ulcer, eye floaters and glaucoma. The corneal endothelium is a single layer on the inner of cornea. It has an important function which maintains the transparency of cornea. And if the corneal endothelial cells injury, it will not recover or regenerate again. Due to this factor, the cornea of patient will become edema and opacity until the patient go blind. Cornea transplant is a familiar therapy clinically, but this method is restricted to the amount of corneal donators. Therefore, there are limiting patients will complete this procedure. Nowadays, a lot of researchers want to prepare a tissue like cornea to substitute for not functioning cornea in order to boost the proportion of cornea transplant.
  The study also focuses on how to organize a functional tissue ex vivo. Acellular porcine cornea matrix is selected to be a scaffold, because the structure of acellular porcine cornea matrix is pretty approach to human cornea. It cultures with corneal endothelial cells and corneal epithelial cells. Moreover, it would be observed the circumstance which is friendly for cornea endothelial cells and epithelial cells attachment.
  And then, this scaffold was applied to carry on penetrating keratoplasty (PK) on New Zealand White; NZW. The study hopes that APCM scaffold can conduct pump function to maintain corneal transparency.
en
dc.description.provenanceMade available in DSpace on 2021-06-08T03:27:02Z (GMT). No. of bitstreams: 1
ntu-108-R06548057-1.pdf: 5280540 bytes, checksum: baaf1cc27e84f6740c0e27781c9f3692 (MD5)
Previous issue date: 2019
en
dc.description.tableofcontents圖目錄 vi
1. 眼球構造與功能(3) (圖1-1) 1
2. 眼角膜構造與功能(圖1-2) 2
3. 角膜修護手術 5
4. 合適的角膜支架 8
5. 角膜移植的風險 9
6. 角膜支架發展趨勢 10
實驗儀器 12
實驗藥品 13
實驗方法 15
1. 取豬角膜支架與去細胞方法 15
a. 配置溶液 15
b. 取豬角膜支架 15
c. 去細胞方法 15
2. 取豬角膜支架 16
3. 培養牛眼內皮細胞 16
4. 牛眼上皮細胞培養 17
5.豬角膜支架與角膜上皮、內皮細胞共培養 18
a. 去細胞後的豬角膜支架先與內皮細胞共培養 18
b. 去細胞後的豬角膜支架先與上皮細胞共培養 18
6. Live & dead cell viability assay 19
7. 組織切片( H&E染色、組織包埋 ) 19
8. PKH67、PKH26染色 19
9. DAPI染色 20
10. 角膜移植 21
11. 配置hyaluronic acid gel 21
12. PBS配置 22
13. 免疫螢光染色 22
研究動機與目的 24
實驗結果 25
1. 比較角膜去細胞前後透明度 25
2. 培養牛眼內皮細胞 26
3. 培養牛眼上皮細胞 27
4. 觀察內皮細胞於角膜支架上的貼附情形 28
5. 觀察上皮細胞於角膜支架上的貼附情形 30
6. 內皮細胞於角膜支架上進行胞毒性測試 32
7. 上皮細胞於角膜支架上進行胞毒性測試 33
8. N Cadherin免疫螢光染色 34
9. Alpha-smooth muscle actin免疫螢光染色 35
10. 將角膜支架與細胞共培養後進行全層置換手術 36
11. 將去細胞角膜支架進行全層置換手術 37
12. 比較術後加入培養液是否對於支架有影響 38
13. 角膜支架完成全層角膜置換手術後進行HE staining組織切片 40
結果討論 42
dc.language.isozh-TW
dc.subject全層角膜置換手術zh_TW
dc.subject角膜內皮細胞zh_TW
dc.subject角膜上皮細胞zh_TW
dc.subject去細胞角膜支架zh_TW
dc.subjectcorneal epithelial cellen
dc.subjectpenetrating keratoplastyen
dc.subjectcorneal endothelial cellen
dc.subjectcorneal transplanten
dc.title以兔子為模型評估去細胞豬角膜支架進行全層角膜置換手術的可行性zh_TW
dc.titleUsing decellularized porcine corneal tissue to serveas a scaffold to evaluate the feasibility of Penetrating Keratoplasty in a rabbit modelen
dc.typeThesis
dc.date.schoolyear108-1
dc.description.degree碩士
dc.contributor.oralexamcommittee黃意真(Yi-Cheng Huang),許馨云(Hsin-Yun HSU)
dc.subject.keyword角膜內皮細胞,角膜上皮細胞,去細胞角膜支架,全層角膜置換手術,zh_TW
dc.subject.keywordcorneal transplant,corneal endothelial cell,corneal epithelial cell,penetrating keratoplasty,en
dc.relation.page44
dc.identifier.doi10.6342/NTU202000069
dc.rights.note未授權
dc.date.accepted2020-01-14
dc.contributor.author-college工學院zh_TW
dc.contributor.author-dept醫學工程學研究所zh_TW
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