Kato Yuka, Iwata Takanori, Washio Kaoru, Yoshida Toshiyuki, Kuroda Hozue, Morikawa Shunichi, Hamada Mariko, Ikura Kazuki, Kaibuchi Nobuyuki, Yamato Masayuki, Okano Teruo, Uchigata Yasuko
Diabetic Center, Tokyo Women's Medical University School of Medicine;
The Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University;
J Vis Exp. 2017 Aug 4(126):54539. doi: 10.3791/54539.
Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients with impeded blood flow or with large wounds might be prolonged. Cell-based therapies have appeared as a new technique for the treatment of diabetic ulcers, and cell-sheet engineering has improved the efficacy of cell transplantation. A number of reports have suggested that adipose-derived stem cells (ASCs), a type of mesenchymal stromal cell (MSC), exhibit therapeutic potential due to their relative abundance in adipose tissue and their accessibility for collection when compared to MSCs from other tissues. Therefore, ASCs appear to be a good source of stem cells for therapeutic use. In this study, ASC sheets from the epididymal adipose fat of normal Lewis rats were successfully created using temperature-responsive culture dishes and normal culture medium containing ascorbic acid. The ASC sheets were transplanted into Zucker diabetic fatty (ZDF) rats, a rat model of type 2 diabetes and obesity, that exhibit diminished wound healing. A wound was created on the posterior cranial surface, ASC sheets were transplanted into the wound, and a bilayer artificial skin was used to cover the sheets. ZDF rats that received ASC sheets had better wound healing than ZDF rats without the transplantation of ASC sheets. This approach was limited because ASC sheets are sensitive to dry conditions, requiring the maintenance of a moist wound environment. Therefore, artificial skin was used to cover the ASC sheet to prevent drying. The allogenic transplantation of ASC sheets in combination with artificial skin might also be applicable to other intractable ulcers or burns, such as those observed with peripheral arterial disease and collagen disease, and might be administered to patients who are undernourished or are using steroids. Thus, this treatment might be the first step towards improving the therapeutic options for diabetic wound healing.
人工皮肤在临床实践中已取得了显著的治疗效果。然而，对于血流受阻或伤口较大的糖尿病患者，人工皮肤治疗伤口的时间可能会延长。基于细胞的疗法已成为治疗糖尿病溃疡的一项新技术，细胞片工程提高了细胞移植的疗效。许多报告表明，脂肪来源干细胞（ASC）作为一种间充质基质细胞（MSC），由于其在脂肪组织中相对丰富，与其他组织来源的MSC相比更容易获取，因而具有治疗潜力。因此，ASC似乎是一种很好的用于治疗的干细胞来源。在本研究中，利用温度响应培养皿和含有抗坏血酸的普通培养基成功制备了正常Lewis大鼠附睾脂肪的ASC片。将ASC片移植到Zucker糖尿病脂肪大鼠（ZDF大鼠）体内，ZDF大鼠是一种2型糖尿病和肥胖的大鼠模型，其伤口愈合能力减弱。在大鼠颅后表面制造一个伤口，将ASC片移植到伤口中，并用双层人工皮肤覆盖这些片。接受ASC片移植的ZDF大鼠比未移植ASC片的ZDF大鼠伤口愈合情况更好。但这种方法存在局限性，因为ASC片对干燥条件敏感，需要维持伤口湿润的环境。因此，使用人工皮肤覆盖ASC片以防止干燥。ASC片与人工皮肤的同种异体移植也可能适用于其他难治性溃疡或烧伤，如外周动脉疾病和胶原病患者出现的溃疡或烧伤，也可能适用于营养不良或正在使用类固醇的患者。因此，这种治疗可能是改善糖尿病伤口愈合治疗选择的第一步。
