Sánchez-Muñoz Isabel, Granados Rosario, Holguín Holgado Purificación, García-Vela José Antonio, Casares Celia, Casares Miguel
1 Tissue Bank, Hospital Universitario de Getafe , Madrid, Spain .
Tissue Eng Part A. 2015 Jan;21(1-2):214-23. doi: 10.1089/ten.TEA.2013.0626. Epub 2014 Aug 19.
In recent years, the reconstruction of human skin by tissue engineering represents a clinical challenge and has offered a therapeutic alternative. Avascular engineered skin equivalents have been available for several years and used to treat wounds due to burns, nonhealing ulcers, and surgical excisions. They are constituted by different types of cultured cells included in a three-dimensional structure that permits cellular proliferation to create tissue substitutes. The major drawback of these artificial skin substitutes is their lack of blood supply, since the endurance and cell proliferation of the substitute depend on an adequate oxygen and nutrient supply and on toxin removal. These functions are served by the vascular system. We have produced a new model of endothelialized skin substitute that promotes the formation of capillary-like structures by seeding human umbilical vein endothelial cells (HUVECs) with dermal fibroblasts and human adipose-derived mesenchymal stem cells (hADMSCs) in a fibrin matrix. Dermal fibroblasts and hADMSCs produce extracellular matrix that stimulates cellular growth and proliferation. hADMSCs secrete significant quantities of angiogenic and antiapoptotic factors (vascular endothelial growth factor and hepatocyte growth factor), which induce in vitro differentiation of these cells into endothelial cells promoting angiogenesis and participating in tissue repair and skin regeneration processes. We obtained the artificial skin substitute with similar structure to native skin, including dermis and epidermis. We demonstrated that endothelial cells (CD31 and von Willebrand factor positive) proliferated and organized themselves into capillary-like structures within the fibrin matrix. The epidermis showed a complete epithelization by squamous cells (AE1/AE3 cytokeratin positive) with intracytoplasmic keratohyalin granules, hyperkeratosis, and parakeratosis. We have established a novel artificial skin substitute that facilitates the formation of capillary-like structures that may provide a novel therapeutic approach to different skin defects and prove to be a useful tool for regenerative medicine.
近年来，通过组织工程重建人类皮肤是一项临床挑战，同时也提供了一种治疗选择。无血管的工程皮肤替代物已经存在数年，并用于治疗烧伤、难愈合溃疡和手术切除引起的伤口。它们由包含在三维结构中的不同类型的培养细胞构成，这种结构允许细胞增殖以创建组织替代物。这些人工皮肤替代物的主要缺点是缺乏血液供应，因为替代物的耐久性和细胞增殖取决于充足的氧气和营养供应以及毒素清除。这些功能由血管系统提供。我们制备了一种新的内皮化皮肤替代物模型，通过将人脐静脉内皮细胞（HUVECs）与真皮成纤维细胞和人脂肪来源的间充质干细胞（hADMSCs）接种在纤维蛋白基质中，促进毛细血管样结构的形成。真皮成纤维细胞和hADMSCs产生刺激细胞生长和增殖的细胞外基质。hADMSCs分泌大量的血管生成和抗凋亡因子（血管内皮生长因子和肝细胞生长因子），这些因子在体外诱导这些细胞分化为内皮细胞，促进血管生成并参与组织修复和皮肤再生过程。我们获得了与天然皮肤结构相似的人工皮肤替代物，包括真皮和表皮。我们证明内皮细胞（CD31和血管性血友病因子阳性）在纤维蛋白基质内增殖并组织形成毛细血管样结构。表皮显示由鳞状细胞（AE1/AE3细胞角蛋白阳性）完全上皮化，伴有胞浆内透明角质颗粒、角化过度和不全角化。我们建立了一种新型人工皮肤替代物，它促进毛细血管样结构的形成，这可能为不同的皮肤缺陷提供一种新的治疗方法，并被证明是再生医学的一种有用工具。
