3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal; ICVS/3B's-PT Government Associate Laboratory, Braga, Guimarães, Portugal.
3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal; ICVS/3B's-PT Government Associate Laboratory, Braga, Guimarães, Portugal; The Discoveries Centre for Regenerative and Precision Medicine, Headquarters at University of Minho, Avepark, 4805-017, Barco, Guimarães, Portugal.
Mater Sci Eng C Mater Biol Appl. 2019 Oct;103:109866. doi: 10.1016/j.msec.2019.109866. Epub 2019 Jun 6.
Kidney diseases are recognized as a major health problem, which affect 10% of the population. Because currently available therapies have many limitations, some tissue engineering strategies have been emerging as promising approaches in this field. In this work, porcine kidneys were decellularized to obtain decellularized kidney extracellular matrix (dKECM). Our results demonstrate a successful protocol of decellularization characterized by the removal of nucleic acid material and preservation of collagen and glycosaminoglycans. Blends of polycaprolactone (PCL) and dKECM were prepared by electrospinning and characterized. The biological performance of the membranes was tested with a human kidney cell line (HK-2) for 7 days. It was observed that cellular metabolic activity, proliferation and protein content increased with an increase in dKECM concentrations (30, 50 and 70%). Additionally, the expression of zona occludens-1 was revealed on dKECM-containing membranes but not on pure PCL membranes. To the best of our knowledge this is the first time that natural extracellular matrix is used to mimic the kidney basement membrane as an in vitro model. This could be a valuable tool for regenerative nephrology and may have an impact on the development of kidney advanced therapies in the future.
肾脏疾病被认为是一个主要的健康问题,影响了 10%的人口。由于目前可用的治疗方法有许多局限性,一些组织工程策略已经成为该领域很有前途的方法。在这项工作中,猪肾脏被去细胞化以获得去细胞化的肾脏细胞外基质(dKECM)。我们的结果证明了一种成功的去细胞化方案,其特征是去除核酸物质并保留胶原蛋白和糖胺聚糖。通过静电纺丝制备了聚己内酯(PCL)和 dKECM 的混合物,并对其进行了表征。用人类肾细胞系(HK-2)对膜的生物学性能进行了 7 天的测试。结果表明,细胞代谢活性、增殖和蛋白质含量随着 dKECM 浓度(30、50 和 70%)的增加而增加。此外,在含有 dKECM 的膜上检测到了封闭蛋白-1 的表达,但在纯 PCL 膜上没有。据我们所知,这是第一次将天然细胞外基质用作模拟肾脏基底膜的体外模型。这可能是再生肾病学的一个有价值的工具,并可能对未来肾脏先进疗法的发展产生影响。