Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Division of Plastic Surgery, Department of Surgery, McGill University, Montreal, Quebec, Canada.
J Surg Res. 2020 Jan;245:31-44. doi: 10.1016/j.jss.2019.07.017. Epub 2019 Aug 7.
Treatment of full-thickness skin wounds with minimal scarring and complete restoration of native tissue properties still exists as a clinical challenge. A bilayer skin substitute was fabricated by coating human amniotic membrane (AM) with electrospun silk fibroin nanofibers, and its in vivo biological behavior was studied using murine full-thickness skin wound model. Donut-shaped silicon splints were utilized to prevent wound contraction in mouse skin and simulate re-epithelialization, which is the normal path of human wound healing. Skin regeneration using the bilayer scaffold was compared with AM and untreated defect after 30 d. Tissue samples were taken from healed wound areas and investigated through histopathological and immunohistochemical staining to visualize involucrin (IVL), P63, collagen I, CD31, and vascular endothelial growth factor. In addition, mRNA expression of IVL, P63, interleukin-6, and cyclooxygenase-2 was studied. The application of bilayer scaffold resulted in the best epidermal and dermal regeneration, demonstrated by histopathological examination and molecular analysis. In regenerated wounds of the bilayer scaffold group, the mRNA expression levels of inflammatory markers (interleukin-6 and cyclooxygenase-2) were downregulated, and the expression pattern of keratinocyte markers (IVL and P63) at both mRNA and protein levels was more similar to native tissue in comparison with AM and no-treatment groups. There was no significant difference in the expression level of collagen I, CD31, and vascular endothelial growth factor among different groups. Conclusively, these promising results serve as a supporting evidence for proceeding to clinical phase to examine the capacity of this bilayer scaffold for human skin regeneration.
治疗全层皮肤创面，使其瘢痕最小化，并完全恢复原有组织特性，这仍然是临床面临的挑战。本研究通过将人羊膜（AM）涂覆到静电纺丝丝素纳米纤维上来制备双层皮肤替代物，并利用鼠全层皮肤创面模型研究其体内生物学行为。使用环形硅夹板防止小鼠皮肤创面收缩，并模拟再上皮化，这是人类创面愈合的正常途径。在 30 天后，使用双层支架进行皮肤再生，并与 AM 和未处理的缺陷进行比较。从愈合的创面区域采集组织样本，并通过组织病理学和免疫组织化学染色进行研究，以可视化角蛋白 1（IVL）、P63、胶原 I、CD31 和血管内皮生长因子。此外，还研究了 IVL、P63、白细胞介素 6 和环氧化酶 2 的 mRNA 表达。通过组织病理学检查和分子分析，双层支架的应用可实现最佳的表皮和真皮再生。在双层支架组的再生创面中，炎症标志物（白细胞介素 6 和环氧化酶 2）的 mRNA 表达水平下调，角蛋白细胞标志物（IVL 和 P63）的 mRNA 和蛋白水平的表达模式与 AM 和未处理组相比更接近天然组织。不同组间胶原 I、CD31 和血管内皮生长因子的表达水平没有显著差异。综上所述，这些有前景的结果为进一步进行临床试验以检验该双层支架在人类皮肤再生中的能力提供了支持证据。
