Endothelial Cell Micropatterning: Methods, Effects, and Applications

Anderson, Deirdre E.J.; Hinds, Monica T.

doi:10.1007/s10439-011-0352-z

Cited by 67 publications

(47 citation statements)

References 89 publications

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“…The lack of attachment in the non-patterned region can be attributed primarily to the BSA coating. The BSA was able to prevent most cell binding to the non-pattern region without any effect on the collagen-I substrate which is consistent with other studies of BSA coating after substrate patterning (Anderson and Hinds 2011). Despite the use of BSA coating, however, very small numbers of cells were found attached to regions without collagen; those that remained after washing showed only limited growth in all directions.…”

Section: Resultssupporting

confidence: 90%

Vacuum-assisted fluid flow in microchannels to pattern substrates and cells

et al. 2014

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Substrate and cell patterning are widely used techniques in cell biology to study cell-to-cell and cell-to-substrate interactions. Conventional patterning techniques work well only with simple shapes, small areas and selected bio-materials. This paper describes a method to distribute cell suspensions as well as substrate solutions into complex, long, closed (dead-end) polydimethylsiloxane (PDMS) microchannels using negative pressure. Our method builds upon a previous vacuum-assisted method used for micromolding (Jeon, Choi et al. 1999) and successfully patterned collagen-I, fibronectin and Sal-1 substrates on glass and polystyrene surfaces, filling microchannels with lengths up to 120 mm and covering areas up to 13 × 10 mm2. Vacuum-patterned substrates were subsequently used to culture mammalian PC12 and fibroblast cells and amphibian neurons. Cells were also patterned directly by injecting cell suspensions into microchannels using vacuum. Fibroblast and neuronal cells patterned using vacuum showed normal growth and minimal cell death indicating no adverse effects of vacuum on cells. Our method fills reversibly sealed PDMS microchannels. This enables the user to remove the PDMS microchannel cast and access the patterned biomaterial or cells for further experimental purposes. Overall, this is a straightforward technique that has broad applicability for cell biology.

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Section: Resultssupporting

confidence: 90%

Vacuum-assisted fluid flow in microchannels to pattern substrates and cells

et al. 2014

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“…EC migration, an important contributor in trans-anastomotic migration, may also be studied. Long-term culture of ECs on PVA substrates should be done to give insight into EC phenotype or function [41]. These additional assays can be used to further verify the advantage of topographical modification on EC behaviour relevant to in situ graft endothelialization.…”

Section: Discussionmentioning

confidence: 99%

In vitro and ex vivo hemocompatibility of off-the-shelf modified poly(vinyl alcohol) vascular grafts

et al. 2015

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Synthetic small diameter vascular grafts with mechanical properties of native arteries, resistance to thrombosis and capacity to stimulate in situ endothelialization are an unmet clinical need. Poly(vinyl alcohol) hydrogel (PVA) is an excellent candidate as a vascular graft due to its tunable mechanical properties. However, the hydrophilicity and bio-inertness of PVA prevents endothelialization in vivo. We hypothesize that the modification of PVA with biomolecules and topographies creates a hemocompatible environment that also enhances bioactivity. PVA modified with fibronectin, RGDS peptide, cyclicRGD (cRGD) peptide, or heparin provided cell-adhesion motifs, which were confirmed by detection of nitrogen through X-ray photoelectron spectroscopy. Protein-and peptide-modified surfaces showed a slight increase in human vascular endothelial cell proliferation over unmodified PVA. With the exception of fibronectin modification, modified surfaces showed in vitro hemocompatibility comparable with unmodified PVA. To further improve bioactivity, cRGD-PVA was combined with gratings and microlens topographies. Combined modifications of 2µm gratings or convex topography and cRGD significantly improved human vascular endothelial cell viability on PVA. In vitro hemocompatibility testing showed that topography on cRGD-PVA did not significantly trigger an increase of platelet adhesion or activation compared with unpatterned PVA. Using the more physiologically relevant ex vivo hemocompatibility testing, all PVA grafts tested showed similar platelet adhesion to ePTFE and significantly lower platelet accumulation compared to collagen-coated ePTFE grafts. The

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“…More recently, the influence of spreading area on traction forces was innovatively evaluated from the total force and the average force [66]: the total force increases and the average force decreases with larger spreading area, which attributes to changes in traction force distribution and cannot be regarded as determinate evidence. With the introduction of microcontact printing, it is possible to confine cell spreading inside the square islands on micropost arrays and thus control spreading area [3,121]. Benefiting from microcontact printing, Han et al [66] further investigated the relationship between spreading area and traction force independently and eventually determined that the total force generated by confined HPAECs (human pulmonary artery endothelial cells) is positively linear to spreading area (Fig.…”

Section: Cell Spreadingmentioning

confidence: 97%

Review of cellular mechanotransduction on micropost substrates

Geng

Wang

2015

Med Biol Eng Comput

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As physical entities, living cells can sense and respond to various stimulations within and outside the body through cellular mechanotransduction. Any deviation in cellular mechanotransduction will not only undermine the orchestrated regulation of mechanical responses, but also lead to the breakdown of their physiological function. Therefore, a quantitative study of cellular mechanotransduction needs to be conducted both in experiments and in computational simulations to investigate the underlying mechanisms of cellular mechanotransduction. In this review, we present an overview of the current knowledge and significant progress in cellular mechanotransduction via micropost substrates. In the aspect of experimental studies, we summarize significant experimental progress and place an emphasis on the coupled relationship among cellular spreading, focal adhesion and contractility as well as the influence of substrate properties on force-involved cellular behaviors. In the other aspect of computational investigations, we outline a coupled framework including the biochemically motivated stress fiber model and thermodynamically motivated adhesion model and present their predicted biomechanical responses and then compare predicted simulation results with experimental observations to further explore the mechanisms of cellular mechanotransduction. At last, we discuss the future perspectives both in experimental technologies and in computational models, as well as facing challenges in the area of cellular mechanotransduction.

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Endothelial Cell Micropatterning: Methods, Effects, and Applications

Cited by 67 publications

References 89 publications

Vacuum-assisted fluid flow in microchannels to pattern substrates and cells

Vacuum-assisted fluid flow in microchannels to pattern substrates and cells

In vitro and ex vivo hemocompatibility of off-the-shelf modified poly(vinyl alcohol) vascular grafts

Review of cellular mechanotransduction on micropost substrates

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