3B's Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine , AvePark4806-909, Taipas, Guimarães, Portugal.
Biomacromolecules. 2013 Nov 11;14(11):3997-4008. doi: 10.1021/bm4011062. Epub 2013 Oct 29.
Among the wide range of strategies to target skin repair/regeneration, tissue engineering (TE) with stem cells at the forefront, remains as the most promising route. Cell sheet (CS) engineering is herein proposed, taking advantage of particular cell-cell and cell-extracellular matrix (ECM) interactions and subsequent cellular milieu, to create 3D TE constructs to promote full-thickness skin wound regeneration. Human adipose derived stem cells (hASCs) CS were obtained within five days using both thermoresponsive and standard cell culture surfaces. hASCs-based constructs were then built by superimposing three CS and transplanted into full-thickness excisional mice skin wounds with delayed healing. Constructs obtained using thermoresponsive surfaces were more stable than the ones from standard cell culture surfaces due to the natural adhesive character of the respective CS. Both CS-generating strategies lead to prolonged hASCs engraftment, although no transdifferentiation phenomena were observed. Moreover, our findings suggest that the transplanted hASCs might be promoting neotissue vascularization and extensively influencing epidermal morphogenesis, mainly through paracrine actions with the resident cells. The thicker epidermis, with a higher degree of maturation characterized by the presence of rete ridges-like structures, as well as a significant number of hair follicles observed after transplantation of the constructs combining the CS obtained from the thermoresponsive surfaces, reinforced the assumptions of the influence of the transplanted hASCs and the importance of the higher stability of these constructs promoted by cohesive cell-cell and cell-ECM interactions. Overall, this study confirmed the potential of hASCs CS-based constructs to treat full-thickness excisional skin wounds and that their fabrication conditions impact different aspects of skin regeneration, such as neovascularisation, but mainly epidermal morphogenesis.
在针对皮肤修复/再生的众多策略中，以干细胞为前沿的组织工程（TE）仍然是最有前途的途径。本文提出了细胞片（CS）工程，利用特定的细胞-细胞和细胞-细胞外基质（ECM）相互作用以及随后的细胞环境，创建 3D TE 构建体以促进全厚皮肤伤口再生。使用热响应和标准细胞培养表面在五天内获得了人脂肪来源干细胞（hASCs）CS。然后，通过叠加三个 CS 构建 hASCs 基构建体，并将其移植到具有延迟愈合的全厚切除小鼠皮肤伤口中。由于各自 CS 的天然粘附特性，使用热响应表面获得的构建体比使用标准细胞培养表面获得的构建体更稳定。两种生成 CS 的策略都导致 hASCs 植入的延长，尽管没有观察到转分化现象。此外，我们的研究结果表明，移植的 hASCs 可能通过旁分泌作用与常驻细胞一起促进新组织血管生成，并广泛影响表皮形态发生。移植的构建体后观察到的较厚表皮，具有更高程度的成熟度，其特征为存在类似于网脊样结构的结构，以及大量的毛囊，这进一步证实了移植的 hASCs 的影响假设以及由有凝聚力的细胞-细胞和细胞-ECM 相互作用促进的这些构建体的更高稳定性的重要性。总体而言，这项研究证实了 hASCs CS 基构建体治疗全厚切除皮肤伤口的潜力，并且它们的制造条件会影响皮肤再生的不同方面，如新生血管形成，但主要是表皮形态发生。
