Advanced effect of rh-FGF on properties and manufacturing conditions of cultured skin substitute

rh-FGF对培养皮肤替代品性能和生产条件的先进影响

• 批准号: 14580828
• 负责人: KUROYANAGI Yoshimitsu
• 金额: $ 1.98万
• 依托单位: KITASATO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580828/
• 关键词: fibroblasts growth factor; faibroblasts; allogeneic cultured dermal subsitute; ヒアルロン酸; コラーゲン; 血管内皮成長因子; 培養真皮

We developed the cultured dermal substitute (CDS) composed of a two-layered spongy matrix of hyaluronic acid and atelo-collagen containing fibroblasts. This type of CDS is designed to provide an effective therapy for victims with burn and ulcer wounds. The aim of this study is to evaluate the efficacy of recombinant human acidic FGF (aFGF) on the potency of fibroblasts in the two-layered spongy matrix to release vascular endothelial growth factor. At first, the effect of aFGF on the proliferation of fibroblasts in a culture flask was tested to confirm the safety and efficacy. Addition of aFGF was found to increase in cell number and decrease in cellular volume. Decrease of cellular volume does not represent an abnormal morphology but simply reflect a more rapid rate of cell division. These responses of fibroblasts with aFGF were marked but transient, because they returned the original proliferation rate and size in cellular volume after further cultivation in the medium without aFGF The CDS prepared through cultivation of fibroblasts on the two-layered spongy matrix in a medium containing aFGF. The addition of aFGF was found to increase in cell number and consequently increase the amount of VEGF released from fibroblasts.The CDS composed of the two-layered spongy matrix of hyaluronic acid and atelo-collagen containing fibroblasts was fabricated through cultivation in medium containing aFGF. This CDS is able to keep the high cell viability and potency to release VEGF that plays an important role in wound healing such as granulation tissue formation, matrix formation, and angiogeneis.

我们开发了一种由透明质酸和含纤维母细胞的间室胶原组成的双层海绵状基质真皮替代物（CDS）。这种类型的CDS旨在为烧伤和溃疡伤口的受害者提供有效的治疗。本研究的目的是评价重组人酸性FGF （aFGF）对两层海绵状基质中成纤维细胞释放血管内皮生长因子的影响。首先，在培养瓶中测试aFGF对成纤维细胞增殖的影响，以确认其安全性和有效性。aFGF的加入使细胞数量增加，细胞体积减小。细胞体积减小并不代表形态异常，而仅仅反映细胞分裂速度加快。含aFGF的成纤维细胞的这些反应是明显的，但是短暂的，因为在不含aFGF的培养基中进一步培养后，成纤维细胞的增殖速度和细胞体积大小恢复到原来的水平。发现aFGF的加入增加了细胞数量，从而增加了成纤维细胞中VEGF的释放量。通过在含aFGF的培养基中培养，制备了由透明质酸和含间系胶原的成纤维细胞组成的双层海绵状基质CDS。该CDS能够保持较高的细胞活力和释放VEGF的能力，VEGF在伤口愈合过程中发挥重要作用，如肉芽组织形成、基质形成和血管生成。

项目成果

K.Kubo, Y.Kuroyanagi: "Spongy matrix of hyaluronic acid and collagen for cultured dermal substitute"J.Artif.Organs. (in press). (2003)

K.Kubo、Y.Kuroyanagi：“用于培养真皮替代品的透明质酸和胶原蛋白的海绵状基质”J.Artif.Organs。

K.Kubo, Y.Kuroyanagi: "Development of a cultured dermal substitute composed of a spongy matrix of hyaluronic acid and atelo-collagen combined with fibroblasts : cryopreservation"Artificial Organs. in press.

K.Kubo、Y.Kuroyanagi：“开发由透明质酸和去端胶原蛋白与成纤维细胞结合的海绵状基质组成的培养真皮替代品：冷冻保存”人工器官。

黒柳能光, 久保健太郎 ほか: "同種培養真皮の製造と供給システム"熱傷(日本熱傷学会誌). 29. 28-38 (2003)

Yoshimitsu Kuroyanagi、Taro Hisa 等：“同种异体培养真皮的制造和供应系统”Burns（日本烧伤学会杂志）29. 28-38（2003）。

Kentaro Kubo, Yoshimitsu Kuroyanagi: "Characterization of a cultured dermal substitute composed of a spongy matrix of hyaluronic acid and collagen with fibroblasts."J.Artif.Orgas.. 6. 138-144 (2003)

Kentaro Kubo、Yoshimitsu Kuroyanagi：“由透明质酸和胶原蛋白与成纤维细胞组成的海绵状基质组成的培养真皮替代品的特征。”J.Artif.Orgas.. 6. 138-144 (2003)

K.Kubo, Y.Kuroyanagi: "Characterization of a cultured dermal substitute composed of a spongy matrix of hyaluronic acid and collagen with fibroblasts"J.Artif.Organs. 6. 138-144 (2003)

K.Kubo、Y.Kuroyanagi：“由透明质酸和胶原蛋白与成纤维细胞组成的海绵状基质组成的培养真皮替代品的特性”J.Artif.Organs。