Ramirez J A, Estrada S, Harmsen M C, Sharma P K
University of Groningen, University Medical Center Groningen, Department of Biomaterials and Biomedical Technology, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
Tissue Engineering and Cells Therapy Group (GITTC), Cell Therapy and Biobank, Alma Mater Hospital of Antioquia, School of Medicine, University of Antioquia. Medellín, Colombia, Cra. 51a #62-42 Medellín, Colombia.
Matrix Biol Plus. 2025 Feb 22;26:100169. doi: 10.1016/j.mbplus.2025.100169. eCollection 2025 Jun.
The first organized extracellular matrix that appears during mammalian embryogenesis is a basement membrane (BM), BM is present in all adult epithelia, endothelia, muscle, nerve and fat tissues. BM is a sub-micrometer thick compact lattice of macromolecules that is maintained by the adhered cells. Systems such as collagen gels, Matrigel® or synthetic polymeric scaffolds have been proposed to mimic the BM and to study the interactions between different cell types, but all lack a structured BM. Here we aimed to obtain and characterize a natural, thin basement membrane-containing scaffold from pig urinary bladders that are subjected to blunt dissection of layers and decellularization steps, preserving the near native BM with a few layers of underlying connective tissue to maintain its structural integrity. The scanning electron microscopy, confocal multiphoton microscopy and immunohistochemistry helped confirm the presence of the BM. A veil-like network composed of thin fibers was present on top of a course network, and glycosaminoglycans, collagen and basement membrane proteins were present. The scaffold's ability to repopulation and basement membrane barrier function were further confirmed when HaCaT and MRC5 cells attached and remained respectively on the epithelial and mesenchymal side without any crossover. Cells remained viable till 2 weeks. This BM-containing scaffold allows to create models of epithelial-mesenchymal tissues through a structured basement membrane and investigate basement membrane dynamics. The basement membrane-containing scaffold was found to be isotropic under uniaxial tension with a failure strain of 0.25 allowing its use to investigate strain induced basement membrane dynamics.
哺乳动物胚胎发育过程中出现的首个有组织的细胞外基质是基底膜（BM），BM存在于所有成年上皮组织、内皮组织、肌肉组织、神经组织和脂肪组织中。BM是一种由大分子组成的亚微米厚的致密晶格结构，由黏附的细胞维持。诸如胶原凝胶、基质胶或合成聚合物支架等体系已被提出用于模拟基底膜并研究不同细胞类型之间的相互作用，但它们都缺乏结构化的基底膜。在此，我们旨在从猪膀胱中获取并表征一种天然的、含有薄基底膜的支架，该猪膀胱经过分层钝性解剖和脱细胞步骤，保留了近乎天然的基底膜以及几层下方的结缔组织以维持其结构完整性。扫描电子显微镜、共聚焦多光子显微镜和免疫组织化学有助于证实基底膜的存在。在粗糙网络之上存在由细纤维组成的面纱状网络，并且存在糖胺聚糖、胶原蛋白和基底膜蛋白。当HaCaT细胞和MRC5细胞分别附着并分别保留在上皮侧和间充质侧且没有任何交叉时，进一步证实了该支架的再增殖能力和基底膜屏障功能。细胞在2周内保持存活。这种含基底膜的支架能够通过结构化的基底膜创建上皮 - 间充质组织模型并研究基底膜动力学。发现该含基底膜的支架在单轴拉伸下呈各向同性，破坏应变为0.25，这使其可用于研究应变诱导的基底膜动力学。
