Peptides Bound to Silicone Membranes and 3D Microfabrication for Cardiac Cell Culture

Boateng, Samuel Y.; Lateef, Syed Salman; Crot, Carrie A.; Motlagh, Delara; Desai, Tejal A.; Samarel, Allen M.; Russell, Brenda; Hanley, Luke

doi:10.1002/1521-4095(20020318)14:6<461::aid-adma461>3.0.co;2-s

Cited by 20 publications

(9 citation statements)

References 17 publications

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“…A long-term goal of our group is to use biotechnological approaches to recapitulate both the architecture and surface chemistry (1,20), in vitro, to obtain a more biological phenotype in culture. In other studies, cells have been embedded into extracellular matrix proteins in an attempt to recapitulate some of their native environment.…”

supporting

confidence: 60%

Inhibition of fibroblast proliferation in cardiac myocyte cultures by surface microtopography

Boateng¹,

Hartman²,

Ahluwalia³

et al. 2003

American Journal of Physiology-Cell Physiology

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Cardiac myocyte cultures usually require pharmacological intervention to prevent overproliferation of contaminating nonmyocytes. Our aim is to prevent excessive fibroblast cell proliferation without the use of cytostatins. We have produced a silicone surface with 10-microm vertical projections that we term "pegs," to which over 80% of rat neonatal cardiac fibroblasts attach within 48 h after plating. There was a 50% decrease in cell proliferation by 5 days of culture compared with flat membranes (P < 0.001) and a concomitant 60% decrease (P < 0.01) in cyclin D1 protein levels, suggesting a G1/S1 cell cycle arrest due to microtopography. Inhibition of Rho kinase with 5 or 20 microM Y-27632 reduced attachment of fibroblasts to the pegs by over 50% (P < 0.001), suggesting that this signaling pathway plays an important role in the process. Using mobile and immobile 10-microm polystyrene spheres, we show that reactive forces are important for inhibiting fibroblast cell proliferation, because mobile spheres failed to reduce cell proliferation. In primary myocyte cultures, pegs also inhibit fibroblast proliferation in the absence of cytostatins. The ratio of aminopropeptide of collagen protein from fibroblasts to myosin from myocytes was significantly reduced in cultures from pegged surfaces (P < 0.01), suggesting an increase in the proportion of myocytes on the pegged surfaces. Connexin43 protein expression was also increased, suggesting improved myocyte-myocyte interaction in the presence of pegs. We conclude that this microtextured culture system is useful for preventing proliferation of fibroblasts in myocyte cultures and may ultimately be useful for tissue engineering applications in vivo.

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supporting

confidence: 60%

Inhibition of fibroblast proliferation in cardiac myocyte cultures by surface microtopography

Boateng¹,

Hartman²,

Ahluwalia³

et al. 2003

American Journal of Physiology-Cell Physiology

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“…Control over murine connective tissue cell adhesion has been achieved by selective functionalization of ridges and grooves on microfabricated polymer surfaces with fibronectin (cell adhesive) and hydrophobic organosilanes (cell repulsive) [139]. Photolithographically patterned silicone elastomers have also been used to culture cardiac myocytes in vitro [140]. Five-to 10-m-wide grooves on the surface of the silicone allow directional growth of the cells, while functionalization of the silicone surface with an RGD (Arg-Gly-Asp) peptide sequence enhanced the adhesion of the myocytes to the surface.…”

Section: Mems Technology For Tissue Engineeringmentioning

confidence: 99%

A BioMEMS Review: MEMS Technology for Physiologically Integrated Devices

et al. 2004

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“…Clark et al reported that cell-cell contact was a major determinant in the development of spontaneous activity (beating) in adult feline cardiomyocyte cultures [14]. The culture surface had a significant role, not only in the attachment [15] of the cells, but also in cell signaling. Simpson et al showed that the phenotype of cardiac myocytes was determined by the culture substrate (type I collagen) through a signaling process which involved the cardiac α-1 or β-1 integrin chain [16].…”

Section: Introductionmentioning

confidence: 99%

Growth and electrophysiological properties of rat embryonic cardiomyocytes on hydroxyl- and carboxyl-modified surfaces

Natarajan

Chun

Hickman

et al. 2008

Journal of Biomaterials Science, Polymer Edition

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Biodegradable scaffolds such as poly(lactic acid) (PLA), poly(lactic-co-glycolic acid) (PLGA) or poly(glycolic acid) (PGA) are commonly used materials in tissue engineering. The chemical composition of these scaffolds changes during degradation which provides a differential environment for the seeded cells. In this study we have developed a simple and relatively high-throughput method in order to test the physiological effects of this varying chemical environment on rat embryonic cardiac myocytes. In order to model the different degradation stages of the scaffold, glass coverslips were functionalized with 11-mercaptoundecanoic acid (MUA) and 11-mercapto-1-undecanol (MUL) as carboxyl- and hydroxyl-groups presenting surfaces, and with trimethoxysilylpropyldiethylenetriamine (DETA) and (3-aminopropyl)triethoxysilane (APTES) as controls. Embryonic cardiac myocytes formed beating islands on all tested surfaces, but the number of attached cells and beating patches was significantly lower on MUL compared to any of the other functionalized surfaces. Moreover, whole-cell patch-clamp experiments showed that the average length of action potentials generated by the beating-cardiac myocytes were significantly longer on MUL compared to the other surfaces. Our results, using our simple test system, are in basic agreement with earlier observations that utilized a complex 3D biodegradable scaffold. Thus, surface functionalization with self-assembled monolayers combined with histological/physiological testing could be a relatively high throughput method for biocompatibility studies and for the optimization of the material/tissue interface in tissue engineering.

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Peptides Bound to Silicone Membranes and 3D Microfabrication for Cardiac Cell Culture

Cited by 20 publications

References 17 publications

Inhibition of fibroblast proliferation in cardiac myocyte cultures by surface microtopography

Inhibition of fibroblast proliferation in cardiac myocyte cultures by surface microtopography

A BioMEMS Review: MEMS Technology for Physiologically Integrated Devices

Growth and electrophysiological properties of rat embryonic cardiomyocytes on hydroxyl- and carboxyl-modified surfaces

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