Reconstruction of human autologous skin using three dimensional culture and its clinical application

三维培养重建人体自体皮肤及其临床应用

• 批准号: 12670807
• 负责人: ISHIKAWA Osamu
• 金额: $ 1.66万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670807/
• 关键词: three dimensional culture; fibroblast; wound model; collagen; MMP-1; systemic sclerosis; nitroglycerine; 三次元培養; 線維芽細胞; コラゲナーゼ; MMP

We have been performing basic and clinical research on the in vitro reconstruction of autologous skin using a novel three dimensional culture system in order to establish the treatment for intractable skin ulcers. In the previous study, we established the in vitro method to generate both keratinocytes and fibroblasts form the identical donor and could reconstruct the skin structure composed of those two cell types in the three dimensional culture system.In the present study, we further investigated the mRNA expressions of collagen 1A2 and 3, matrix metalloproteinase (MMP) 1 and tissue inhibitor of MMP-1 under hypoxic condition that is one of causative factors in intractable skin ulcers. We confirmed the significant decrease in mRNA expressions of collagen 1A2 and 3 as well as the significant increase in mRNA expressions of MMP-1 and those changes were restored by reoxygenation. These findings strongly suggest that hypoxic condition, usually found in a poor blood supply, may prolong or suppress the wound healing. In the clinical study using a thermography, we disclosed that a new type of nitroglycerine tape was effective to improve the peripheral circulatory disturbance in patients with systemic sclerosis, a representative disease, complicated with skin ulcers resistant to the conventional therapies.

我们一直在进行基础和临床研究，在体外重建自体皮肤使用一种新的三维培养系统，以建立治疗顽固性皮肤溃疡。在前期的研究中，我们建立了体外培养同一供体角质形成细胞和成纤维细胞的方法，并在三维培养体系中重建了由这两种细胞组成的皮肤结构，在本研究中，我们进一步研究了胶原1A 2和3的mRNA表达，基质金属蛋白酶-1（MMP-1）及其组织抑制剂在缺氧条件下的表达是难治性皮肤溃疡的致病因素之一。我们证实，胶原1A 2和3的mRNA表达的显着下降，以及MMP-1的mRNA表达的显着增加，这些变化恢复复氧。这些发现强烈地表明，通常在血液供应不良的情况下发现的缺氧条件可能会延长或抑制伤口愈合。在使用热成像法的临床研究中，我们发现一种新型的硝酸甘油胶带可有效改善系统性硬化症患者的外周循环障碍，系统性硬化症是一种代表性疾病，并发有对传统疗法有抵抗力的皮肤溃疡。

项目成果

Akimoto S, Takagi H, Abe M, Ishikawa O.: "HLA DRB1 and DQB1 genes in Japanese patients with systemic sclerosi"J Rheumtol. 27. 2940-2 (2000)

Akimoto S、Takagi H、Abe M、Ishikawa O.：“日本系统性硬化症患者的 HLA DRB1 和 DQB1 基因”J Rheumtol。

Sogabe Y, Akimoto S, Abe M, Ishikawa O, Takagi Y, Imakawa G.: "Functoins of the stratum corneum in systemic sclerosis as distinct from hypertrophic scar and keloid functions"J Dermatol Sci. (in press). (2002)

Sogabe Y、Akimoto S、Abe M、Ishikawa O、Takagi Y、Imakawa G.：“系统性硬化症中角质层的功能不同于肥厚性疤痕和疤痕疙瘩功能”J Dermatol Sci。

Akimoto S, Abe M, Ishikawa O.: "HLA DRB1 and DQB1 genes in anticentromere antibody positive with SSc and primary biliary cirrhosis"Ann Rheurn Dis. 60. 639-40 (2000)

Akimoto S、Abe M、Ishikawa O.：“SSc 和原发性胆汁性肝硬化的抗着丝粒抗体阳性的 HLA DRB1 和 DQB1 基因”Ann Rheurn Dis。

Yamanaka M, Ishikawa O.: "Hypoxic conditions decrease the mRNA expression of pro al(I) and al(III) collagens and increase matrix metalloproteinase-1 of dermal fibroblasts in three-dimensional culture"J Dermatol Sci. 24. 99-104 (2000)

Yamanaka M、Ishikawa O.：“缺氧条件会降低三维培养中真皮成纤维细胞的 pro al(I) 和 al(III) 胶原蛋白的 mRNA 表达并增加基质金属蛋白酶-1”J Dermatol Sci。

Abe M., Kan C., Zaw KK, Ishikawa O.: "Induction of matrix metalloproteinase-1 in in vitro experimaental wound model using a novel three dimensional culture system"Eur J Dermatol. 11. 112-116 (2001)

Abe M.、Kan C.、Zaw KK、Ishikawa O.：“使用新型三维培养系统在体外实验伤口模型中诱导基质金属蛋白酶-1”Eur J Dermatol。