Ocular Surface reconstruction for cicatricial disorders using amniotic membrane.

使用羊膜进行瘢痕性疾病的眼表重建。

• 批准号: 09671819
• 负责人: SHIMAZAKI Jun
• 金额: $ 1.92万
• 依托单位: TOKYO DENTAL COLLEGE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671819/
• 关键词: fibroblasts; conjunctival epithelium; amniotic membrane; growth factors; cell proliferation; Ocular Surface; cicatricial keratoconjunctivitis; dry eye

1. Histology and preservation : Fresh human amniotic membrane (AM) had uniform thickness of collagen layer and overlying epithelial cells. The epithelial cells lost their viability up to 80% after 70 days' cryopreservation with DMSO.2. Animal modesl : Human preserved AMs were secured on bare sciera of albino rabbits and histological changes were studied. The amniotic epithelium remained for about one week postoperatively, then gradually replaced by host conjunctiva. Inflammatory cells were observed beneath the membrane in some cases.3. Clinical samples : AMs were recognized in 3 out of 14 human samples which had reoperation following AM transplantation, suggesting AMs were not degraded in some cases after transplantation.4. Growth factors : Roles of growth factors on proliferation/differentiation of ocular surface epithelia were studied using immunohitochemistry and ELISA.bFGF, HGF, and TGF-beta3 were found primarily in the cytoplasm of AM epithelium, whereas TGFbeta 1 and TGF-beta3 were distributed more diffusely. Concentrations of bFGF, HGF, TGF-betal, TGF-beta3 in AM were 5.31*3.04, 14.2*6.0, 2.45*1 .45.0.56*0.45 ng/g, respectively. The concentrations decreased to 1 7.7-23.5% after 70 days' cryopreservation.5. Effects on ocular surface cells : Addition of homogenized AMs to either cultured conjunctival epithelium or subccnjunctival fibroblasts resulted in significant increases in proliferation of these cells.6. Cultivation of conjunctival epithelium on AM : We cultivated human conjunctival epithelium on AM using SHEM and RCGM media. These cells became multilayered with intercellular complex formation.

1.组织学和保存：新鲜的人羊膜(AM)具有均匀厚度的胶原层和上皮细胞。用DMSO冷冻保存70天后，上皮细胞失去活力高达80%。2。动物模型：将人类保存的 AM 固定在白化兔子的裸鼠身上，并研究组织学变化。术后羊膜上皮保留约1周，然后逐渐被宿主结膜取代。部分病例膜下可见炎症细胞。3．临床样本：在AM移植后再次手术的14个人类样本中，有3个被识别出AMs，这表明AMs在移植后在某些情况下没有降解。4．生长因子：使用免疫组织化学和ELISA研究生长因子对眼表上皮细胞增殖/分化的作用。bFGF、HGF和TGF-β3主要存在于AM上皮细胞质中，而TGFβ1和TGF-β3分布更广泛。 AM中bFGF、HGF、TGF-β、TGF-β3的浓度分别为5.31*3.04、14.2*6.0、2.45*1.45.0.56*0.45ng/g。冻存70天后浓度降至1 7.7-23.5%。5．对眼表细胞的影响：向培养的结膜上皮或结膜下成纤维细胞中添加匀浆AM会导致这些细胞的增殖显着增加。6． AM 上结膜上皮的培养：我们使用 SHEM 和 RCGM 培养基在 AM 上培养人结膜上皮。这些细胞随着细胞间复合物的形成而变得多层。

项目成果

Ishioka M,Yagi Y,Fujita H,Shimazaki J,Tsubota K.: "Impression cytology of conjunctiva after amniotic membrane transplantation." The 22th Japan Cornea Conference.(1998)

Ishioka M，Yagi Y，Fujita H，Shimazaki J，Tsubota K.：“羊膜移植后结膜的印迹细胞学。”

Shimazaki J.: "Ocular surface reconstruction for cicatrical disorders using amniotic membrane." 264th Biannual Meeting of the Tokyo Dental College. (1998)

Shimazaki J.：“利用羊膜进行瘢痕性疾病的眼表重建。”

Shimazaki J: "Transplantation of amniotic membrane and limbal autograft for patients with tecurrent pterygium associated with symbrepharon." Br J Ophthalmol. 82. 235-240 (1998)

Shimazaki J：“为伴有 symbrepharon 的背侧翼状胬肉患者进行羊膜移植和自体角膜缘移植。”

Shimazaki J,Sato H,Shinozaki N,Tsubota K.: "Ocular surface reconstruction for cicatricial keratoconjunctivitis using amniotic membrane." 2nd Meeting of Dermotological Research. (1997)

Shimazaki J，Sato H，Shinozaki N，Tsubota K.：“使用羊膜重建瘢痕性角结膜炎的眼表。”

Shimazaki J: "Role of Basic FGF, HGF, and TGF-b in Transplantation of Human Amniotic Membrane." Br J Ophthalmol. (投稿予定).

Shimazaki J：“碱性 FGF、HGF 和 TGF-b 在人羊膜移植中的作用”。