Development of ocular surface reconstruction using tautologies mucosal

使用粘膜同义反复进行眼表重建的发展

• 批准号: 13557145
• 负责人: SANO Yoichiro
• 金额: $ 9.02万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557145/
• 关键词: comeal transplotation; mucosa; auto graft; stem cell; amuiotic memblane; culture epithelium

PURPOSE: To determine the feasibility of using human amniotic membrane (AM) as a substrate for culturing oral epithelial cells and to investigate the possibility of using autologous cultivated oral epithelial cells in ocular surfase reconstruction. METHODS: An ocular surfase injury was created in one eye of each of eight adult albino rabbits by a lamellar keratectomy, and a conjunctival excision was performed, including and extending 5 mm outside the limbus. Oral mucosal biopsy spesimens were obtained from these eight adult albino rabbits and cultivated for 3 weeks on a denuded AM carrier. The cultivated epithelium was examined by electron microscopy (EM) and immunohistochemically labeled for several keratins. At 3 to 4 weeks after the ocular surface injury, the conjunctivalized corneal surfaces of the eight rabbits were surgically reconstructed by transplanting the autologous cultivated oral epithelial sheet had four to five layers of stratified, well-differentiated cells. EM revealed that the epithelial cells were very similar in appearance to those of normal corneal epithelium, had numbrane with hemidesmosomes. Immunohistochemistry confirmed the presence of the keratin pair 4 and 13 and keratin-3 in the cultivated oral epithelial cells. Corneas that were grafed with the cultivated oral epithelial cells on an AM carrier were clear and were all epithelialized 10 days after surgery. CONCLUSIONS: cultures of oral epithelial cells can be generated to confluence on AM expanded ex vivo from biopsy-derived oral mucosal tissue. Autologous transplantation was performed with these cuitivated oral epithelial cells onto the ocular surfaces of keratectomized rabbit eyes. Autologous transplantation of cultivated oral epithelium is a feasible method for ocular surface reconstruction

目得：目的：探讨人羊膜作为培养口腔上皮细胞的基质的可行性，以及自体培养口腔上皮细胞用于眼表面重建的可能性。方法：通过板层角膜切除术在八只成年白化病兔的每一只眼睛中产生眼表面损伤，并进行结膜切除，包括并延伸到利姆布斯外5 mm。从这8只成年白化病兔获得口腔粘膜活检标本，并在裸露的AM载体上培养3周。培养的上皮细胞用电子显微镜（EM）检查，并用化学标记法标记几种角蛋白。在眼表损伤后3 ~ 4周，通过移植具有4 ~ 5层分层的、分化良好的细胞的自体培养的口腔上皮片来手术重建8只兔的结膜角膜表面。电镜观察显示上皮细胞形态与正常角膜上皮相似，有许多细胞膜，有半桥粒。免疫组化证实了角蛋白对4和13和角蛋白-3在培养的口腔上皮细胞的存在。移植有AM载体上的培养的口腔上皮细胞的角膜是透明的，并且在手术后10天全部上皮化。结论：口腔上皮细胞的培养物可以在从活检来源的口腔粘膜组织离体扩增的AM上产生汇合。将这些培养的口腔上皮细胞自体移植到角膜切除的兔眼的眼表上。自体培养口腔上皮移植是一种可行的眼表重建方法

项目成果

Sano Y: "Non-cleavable mutant Fas ligand transfection of donor cornea abrogates ocular immune privilege"Exp. Eye. Res. 75(4). 475-483 (2002)

Sano Y：“供体角膜的不可裂解突变型 Fas 配体转染消除了眼部免疫特权”Exp。

Nakamura T, Koizumi N, Tsuzuki M, Inoki K, Sano Y, Sotozono C, Kinoshita S: "Successful regrafting of cultivated corneal epithelium using amniotic membrane as a carrier in severe ocular surface disease"Comea. 22(1). 70-71 (2003)

Nakamura T、Koizumi N、Tsuzuki M、Inoki K、Sano Y、Sotozono C、Kinoshita S：“在严重眼表疾病中使用羊膜作为载体成功移植培养角膜上皮”Comea。

Ueta M: "Immunosuppressive properties of human amniotic membrane for mixed lymphocyte reaction"Uin Exp Immunol.. 129(3). 464-470 (2002)

Ueta M：“人羊膜对混合淋巴细胞反应的免疫抑制特性”Uin Exp Nutritionol.. 129(3)。

Nakamura T: "The successful culture and autologus transplantation of rabbit oral mucosal epithelia cells on amniotic membrane"Invest Ophthalmol Vis Sci. 44(1). 106-116 (2003)

Nakamura T：“羊膜上兔口腔粘膜上皮细胞的成功培养和自体移植”Invest Ophasemol Vis Sci。

Sano Y, Yamada J, Ishino Y, Adachi W, Kawasaki S, Suzuki T, Kinoshita S, Okuyama T, Azuma N: "Non-cleavable mutant Fas ligand transfection of donor comea abrogates ocular immune privilege"Exp Eye Res. 75(4). 475-483 (2002)

Sano Y、Yamada J、Ishino Y、Adachi W、Kawasaki S、Suzuki T、Kinoshita S、Okuyama T、Azuma N：“供体角膜的不可裂解突变型 Fas 配体转染会消除眼部免疫特权”Exp Eye Res。