3B's Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Zona Industrial da Gandra, S. Cláudio do Barco, 4806-909 Taipas, Guimarães, Portugal; ICVS/3B's - PT Government Associate Laboratory, Braga, Guimarães, Portugal.
3B's Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Zona Industrial da Gandra, S. Cláudio do Barco, 4806-909 Taipas, Guimarães, Portugal; ICVS/3B's - PT Government Associate Laboratory, Braga, Guimarães, Portugal.
Acta Biomater. 2014 Jul;10(7):3145-55. doi: 10.1016/j.actbio.2014.03.006. Epub 2014 Mar 18.
Skin regeneration remains a challenge, requiring a well-orchestrated interplay of cell-cell and cell-matrix signalling. Cell sheet (CS) engineering, which has the major advantage of allowing the retrieval of the intact cell layers along with their naturally organized extracellular matrix (ECM), has been poorly explored for the purpose of creating skin substitutes and skin regeneration. This work proposes the use of CS technology to engineer cellular constructs based on human keratinocytes (hKC), key players in wound re-epithelialization, dermal fibroblasts (hDFb), responsible for ECM remodelling, and dermal microvascular endothelial cells (hDMEC), part of the dermal vascular network and modulators of angiogenesis. Homotypic and heterotypic three-dimensional (3-D) CS-based constructs were developed simultaneously to target wound re-vascularization and re-epithelialization. After implantation of the constructs in murine full-thickness wounds, human cells were engrafted into the host wound bed and were present in the neotissue formed up to 14 days post-implantation. Different outcomes were obtained by varying the composition and organization of the 3-D constructs. Both hKC and hDMEC significantly contributed to re-epithelialization by promoting rapid wound closure and early epithelial coverage. Moreover, a significant increase in the density of vessels at day 7 and the incorporation of hDMEC in the neoformed vasculature confirmed its role over neotissue vacularization. As a whole, the obtained results confirmed that the proposed 3-D CS-based constructs provided the necessary cell machinery, when in a specific microenvironment, guiding both re-vascularization and re-epithelialization. Although dependent on the nature of the constructs, the results obtained sustain the hypothesis that different CS-based constructs lead to improved skin healing.
皮肤再生仍然是一个挑战，需要细胞-细胞和细胞-基质信号的良好协调。细胞片（CS）工程具有允许沿着其自然组织的细胞外基质（ECM）回收完整细胞层的主要优势，但其在用于创建皮肤替代品和皮肤再生方面的应用研究甚少。本工作提出了使用 CS 技术基于人角质形成细胞（hKC），即伤口再上皮化的关键参与者、真皮成纤维细胞（hDFb），负责 ECM 重塑，以及真皮微血管内皮细胞（hDMEC），构建基于细胞的构建体，后者是真皮血管网络的一部分，也是血管生成的调节剂。同时开发了同型和异型三维（3-D）基于 CS 的构建体，以针对伤口再血管化和再上皮化。将构建体植入小鼠全层伤口后，人细胞被植入宿主伤口床，并在植入后 14 天内存在于新形成的组织中。通过改变 3-D 构建体的组成和组织，获得了不同的结果。hKC 和 hDMEC 通过促进伤口快速闭合和早期上皮覆盖，显著促进了再上皮化。此外，第 7 天血管密度显著增加，以及 hDMEC 整合到新形成的脉管系统中，证实了其在新组织血管化中的作用。总的来说，获得的结果证实，所提出的 3-D CS 基于构建体在特定微环境中提供了必要的细胞机制，可指导再血管化和再上皮化。尽管结果取决于构建体的性质，但获得的结果支持这样的假设，即不同的基于 CS 的构建体可改善皮肤愈合。
