Micropatterned coculture of vascular endothelial and smooth muscle cells on layered electrospun fibrous mats toward blood vessel engineering

Abstract

A major challenge in vascular engineering is the establishment of proper microenvironment to guide the spatial organization, growth, and extracellular matrix (ECM) productions of cells found in blood vessels. In the current study, micropatterned fibrous mats with distinct ridges and grooves of different width were created to load smooth muscle cells (SMCs), which were assembled by stacking on vascular endothelial cell (EC)-loaded flat fibrous mats to mimic the in vivo-like organized structure of blood vessels. SMCs were mainly distributed in the ridges, and aligned fibers in the patterned regions led to the formation of elongated cell bodies, intense actin filaments, and expressions of collagen I and α-smooth muscle actin in a parallel direction with fibers. ECs spread over the flat fibrous mats and expressed collagen IV and laminin with a cobblestone-like feature. A z-stack scanning of fluorescently stained fibrous mats indicated that SMCs effectively infiltrated into fibrous scaffolds at the depth of around 200 μm. Compared with SMCs cultured alone, the coculture with ECs enhanced the proliferation, infiltration, and cytoskeleton elongation of SMCs on patterned fibrous mats. Although the coculture of SMCs made no significant difference in the EC growth, the coculture system on patterned fibrous scaffolds promoted ECM productions of both ECs and SMCs. Thus, this patterned fibrous configuration not only offers a promising technology in the design of tissue engineering scaffolds to construct blood vessels with durable mechanical properties, but also provides a platform for patterned coculture to investigate cell-matrix and cell-cell interactions in highly organized tissues.

Chinese herb cinobufagin-reduced cancer pain is associated with increased peripheral opioids by invaded CD3/4/8 lymphocytes

“…endothelial cells and smooth muscle cells, tumor cells and endothelial cells, hepatocytes, fibroblasts and epithelial cells and corneal keratocytes and so on), to simulate the interaction between them. We use this system to study the complex interaction between cells [24–27]. …”

Multilineage Constructs for Scaffold‐Based Tissue Engineering: A Review of Tissue‐Specific Challenges

“…To imitate native interfacial areas and to create cell gradients and transitional populations, multiphasic scaffolds are investigated ( Figure A,B), with a particular interest in skin, the osteochondral and bone‐to‐ligament/tendon interfaces . Modified topographies, patterned surfaces and anisotropic scaffolds were deemed to improve the functionality of specific cell types such as smooth muscle cells or neurons . However, controlling topography and porosity of the scaffold in the context of coculture is more challenging, as cocultured cells do not necessarily present the same physiological features.…”

“…Smooth muscle cells (SMCs) are located either around the vessel wall forming a contractile vascular membrane, or in the surrounding ECM, aligned with the blood flow shear stress . Many diseases and injuries can lead to irreversibly damaged blood vessels . To achieve neovascularization in vivo, the rebuilding of smooth muscle tissues is therefore extremely important.…”

Nano- and microstructured materials for in vitro studies of the physiology of vascular cells

“…Such patterning with adhesive molecules can be realized by several techniques. Commonly used are microcontact printing [53,155] dry lift-off [58,156], dip-pen nanolithography [157–158], and block copolymer micelle nanolithography [159]. All these methods are just exemplary techniques allowing the (bio)chemical modification of structured surfaces for in vitro cell studies.…”

“…The most commonly used surface structure to study this phenomenon consists of arrays of ridges and grooves. Generally, cells orient preferably stronger along the direction of grooves and ridges, the narrower and deeper/higher these structures are [5,12,156,192,221–225]. It was observed that few tenths of nanometers in structure depth was already sufficient for some cell types to trigger contact guidance [201–202].…”