Abstract
Understanding the response of endothelial cells to aligned myotubes is important to create an appropriate environment for tissue-engineered vascularized skeletal muscle. Part of the native tissue environment is the extracellular matrix (ECM). The ECM is a supportive scaffold for cells and allows cellular processes such as proliferation, differentiation, and migration. Interstitial matrix and basal membrane both comprise proteinaceous and polysaccharide components for strength, architecture, and volume retention. Virtually all cells are anchored to their basal lamina. One of the physical factors that affects cell behavior is topography, which plays an important role on cell alignment. We tested the hypothesis that topography-driven aligned human myotubes promote and support vascular network formation as a prelude to in vitro engineered vascularized skeletal muscle. Therefore, we used a PDMS-based topography substrate to investigate the influence of pre-aligned myotubes on the network formation of microvascular endothelial cells. The aligned myotubes produced a network of collagen fibers and laminin. This network supported early stages of endothelial network formation.
Highlights
Engineered skeletal muscle substitutes are needed to treat the consequences of muscular trauma or disorders that result in loss of muscle parenchyma
Topography of aligned myotubes was measured by atomic force microscopy (AFM) to uncover the topography influence of the substrate on the structure of the myotube surface
Myotube topography resulted in nanotopography at the cell surface with aligned protruding dents of approximately 300–900 nm in width and 10–100 nm in height parallel to the length of the myotubes (Figure 2c,d)
Summary
Engineered skeletal muscle substitutes are needed to treat the consequences of muscular trauma or disorders that result in loss of muscle parenchyma. Muscle architecture is unique: it comprises of parallel-aligned myofibers held together by the structure and composition of the surrounding extracellular matrix (ECM). The extracellular matrix of muscle consists of two layers, the basal lamina, which is in close contact to the cells by binding to the integrin receptors protruding from the cellular plasma membrane, and the fibrillar reticular lamina, which surrounds the cells. The basal lamina consists of the non-fibrillar collagen type IV, laminin, and proteoglycans. This dense basal lamina, the fibrillar reticular lamina resides, which corresponds to the interstitial connective tissue comprising mainly of (fibrillar) collagens, e.g., collagen I and proteoglycans [2,3,4]. Laminin and collagen IV are the most abundant
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