Development of regenerative therapy for refractory cutaneous ulcers using multi-layered cell sheets

• Project/Area Number: 17K10759
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cardiovascular surgery
• Research Institution: Yamaguchi University
• Principal Investigator: TAKAHASHI Masaya 山口大学, 医学部, 特別医学研究員 (60634730)
• Co-Investigator(Kenkyū-buntansha): 上野 耕司 山口大学, 医学部附属病院, 助教 (30736070)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 細胞シート / 難治性皮膚潰瘍 / 再生医療

Outline of Final Research Achievements

We developed a multilayered mixed-cell sheet consisting of fibroblasts and peripheral blood mononuclear cells as a treatment for refractory cutaneous ulcers. The concentration of secreted VEGF, HGF, TGF-β1, CXCL-1, CXCL-2 in multilayered mixed-cell sheets was much higher than that in single-layered mixed-cell sheets. The supernatant in multilayered sheets enhanced angiogenic potency and fibroblast migration compared with single-layered mixed-cell sheets in an in vitro experiment. The wound healing rate in the multilayered mixed-cell sheet-treated group was higher compared with the no-treatment group (control) at the early stage of healing. Moreover, both vessel lumen area and microvessel density in tissues treated with multilayered sheets were significantly increased compared with tissues in the control group. These data suggest that multilayered sheets may be a promising therapeutic material for refractory cutaneous ulcers.

Academic Significance and Societal Importance of the Research Achievements

動物モデルにおいて単層細胞混合シート移植により一定の成果は得られたが、臨床応用へ向けては細胞シートの扱い易さの改善や治療効果の増強が必要であった。本研究で確立した積層細胞混合シートは、単層の細胞混合シートに比べ、厚みが増して安定した円形のシートとなった。さらに、積層細胞混合シートは創傷治癒に必要な成長因子やサイトカインを供給することにより血管新生と創傷治癒過程を促進し、治療効果を有することが示された。