Department of Burn Surgery, The First Affiliated Hospital of Sun Yat-sen University, No.58, Zhongshan 2nd Road, Guangzhou, 510080, China.
Stem Cell Res Ther. 2019 Sep 11;10(1):154. doi: 10.1186/s13287-019-1236-7.
Autologous epidermal basal cell suspension therapy has been proven to be one of the most effective treatments for full-thickness wounds. However, we found there remain obvious defects that significantly confined the utilization and function of the epidermal basal cells (EBCs), especially the epidermal stem cells (ESCs) in it. This study investigated whether precoating fibronectin (FN) on the wound bed before spraying EBCs could overcome these defects and further explored its possible mechanisms.
In the in vitro study, EBCs were isolated from the donor skin of patients who needed skin grafting. Different concentrations of FN were used to precoat culture dishes before cell culture; the adherent efficiency, proliferation and migration ability of ESCs were analyzed and compared with traditional collagen IV precoating. In the in vivo study, Sprague-Dawley (SD) rats with full-thickness skin wounds were selected as full-thickness wounds' model. For the experiment groups, 20 μg/ml FN was precoated on the wound bed 10 min before EBC spray. The quality of wound healing was estimated by the residual wound area rate, wound healing time, and hematoxylin and eosin (H&E) staining. Expression of ESC markers, neovascular markers, inflammation markers, and collagen formation and degradation markers was elucidated by immunohistochemistry (IHC), immunofluorescence (IF), western blot (WB), and RT-qPCR analysis.
The in vitro study showed that the dishes precoated with 20 μg/ml FN had a similar adherent efficiency and colony formation rate with collagen IV, but it could improve the proliferation and migration of ESCs significantly. Similarly, in the in vivo study, precoating FN on wound bed before EBC spray also significantly promote wound healing by improving ESCs' utilization efficiency, promoting angiogenesis, decreasing inflammations, and regulating collagen formation and degradation.
FN precoating wound bed before EBC spray could significantly promote full-thickness wound healing by improving the utilization and function of the ESCs and further by promoting angiogenesis, decreasing inflammations, and regulating collagen formation and degradation.
自体表皮基底细胞悬液治疗已被证明是治疗全层创面最有效的方法之一。然而，我们发现仍存在明显缺陷，严重限制了表皮基底细胞（EBC），尤其是其中的表皮干细胞（ESC）的利用和功能。本研究旨在探讨在喷洒 EBC 前预先在创面上涂敷纤维连接蛋白（FN）是否可以克服这些缺陷，并进一步探讨其可能的机制。
在体外研究中，从需要植皮的患者供皮区分离 EBC。采用不同浓度的 FN 预先涂敷细胞培养皿，分析比较传统胶原 IV 预涂敷对 ESC 黏附效率、增殖和迁移能力的影响。在体内研究中，选择 SD 大鼠全层皮肤创面作为全层创面模型。实验组在喷洒 EBC 前 10min 将 20μg/ml FN 预先涂敷在创面上。通过残留创面面积率、创面愈合时间和苏木精-伊红（H&E）染色评估创面愈合质量。通过免疫组织化学（IHC）、免疫荧光（IF）、Western blot（WB）和 RT-qPCR 分析阐明 ESC 标志物、新生血管标志物、炎症标志物以及胶原形成和降解标志物的表达。
体外研究表明，涂敷 20μg/ml FN 的培养皿与胶原 IV 具有相似的黏附效率和集落形成率，但可显著提高 ESC 的增殖和迁移能力。同样，在体内研究中，在喷洒 EBC 前预先在创面上涂敷 FN 也可通过提高 ESCs 的利用效率、促进血管生成、减少炎症反应以及调节胶原形成和降解，显著促进创面愈合。
在喷洒 EBC 前预先在创面上涂敷 FN 可通过提高 ESC 的利用和功能，进一步通过促进血管生成、减少炎症反应以及调节胶原形成和降解，显著促进全层创面愈合。
