Photolithographic Patterning of C2C12 Myotubes using Vitronectin as Growth Substrate in Serum‐Free Medium

Molnár, Péter; Wang, Weishi; Natarajan, Anupama; Rumsey, John W.; Hickman, James J.

doi:10.1021/bp060302q

Cited by 66 publications

(55 citation statements)

References 37 publications

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“…The engagement of VN receptors favors the organization of cells into clusters at earlier time points, leading to higher levels of cell fusion and ultimately high differentiation. Additionally, VN has been previously reported to be able to sustain the adhesion, growth, and efficient myogenic differentiation of C2C12 cells 42. We postulate that the observed nonmonotonic response on the mixed coatings stems from the opposing effects on myogenic differentiation of the physical and biological changes in the biointerface properties granted by the integration of VN.…”

Section: Discussionmentioning

confidence: 76%

Vitronectin as a Micromanager of Cell Response in Material‐Driven Fibronectin Nanonetworks

et al. 2017

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Surface functionalization strategies of synthetic materials for regenerative medicine applications comprise the development of microenvironments that recapitulate the physical and biochemical cues of physiological extracellular matrices. In this context, material‐driven fibronectin (FN) nanonetworks obtained from the adsorption of the protein on poly(ethyl acrylate) provide a robust system to control cell behavior, particularly to enhance differentiation. This study aims at augmenting the complexity of these fibrillar matrices by introducing vitronectin, a lower‐molecular‐weight multifunctional glycoprotein and main adhesive component of serum. A cooperative effect during co‐adsorption of the proteins is observed, as the addition of vitronectin leads to increased fibronectin adsorption, improved fibril formation, and enhanced vitronectin exposure. The mobility of the protein at the material interface increases, and this, in turn, facilitates the reorganization of the adsorbed FN by cells. Furthermore, the interplay between interface mobility and engagement of vitronectin receptors controls the level of cell fusion and the degree of cell differentiation. Ultimately, this work reveals that substrate‐induced protein interfaces resulting from the cooperative adsorption of fibronectin and vitronectin fine‐tune cell behavior, as vitronectin micromanages the local properties of the microenvironment and consequently short‐term cell response to the protein interface and higher order cellular functions such as differentiation.

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Section: Discussionmentioning

confidence: 76%

Vitronectin as a Micromanager of Cell Response in Material‐Driven Fibronectin Nanonetworks

et al. 2017

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“…Several protein factors are used for surface functionalization and biologically mediated stimulation of cell migration, proliferation, and differentiation, as well as de novo vascularization of new tissues. They include several growth factors [47] and bone morphogenetic proteins [48] apart from cell adhesion agents such as vitronectin [49], fibronectin [50], and arginine-glycine-aspartic acid peptides [51,52]. On the other hand, engineering the roughness of surfaces where cells are to be attached and proliferate to provide appropriate mechanical stimuli is progressively being recognized as a powerful tool to modulate mammalian cell differentiation and proliferation in regenerative medicine [53,54].…”

Section: Cell Proliferation Assisted By Protein-based Nanoparticlesmentioning

confidence: 99%

Engineering Protein Based Nanoparticles for Applications in Tissue Engineering

Tatkiewicz

Seras‐Franzoso

Díez-Gil

et al. 2014

Bio‐ and Bioinspired Nanomaterials

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IntroductionBacterial inclusion bodies (IB, inclusion body) are highly pure protein deposits produced by recombinant bacteria in the size range of a few hundred nanometers [1]. The polypeptide chains that form IBs usually retain a certain degree of native-like structure, keeping their biological activity (e.g., fluorescence or enzymatic activity), and as a result are suitable for use as functional and biocompatible materials. In this chapter we characterize the relevant nanometer-scale properties of IBs and present the extent to which they can be tailored by simple approaches. As the IB size is within the range of mammalian cell micro-and nano-environment topology influencing mammalian cell proliferation, random surface decoration and patterning with IBs has a significant and positive impact on cell growth, making them promising biomaterials for tissue engineering. Despite their enormous potential as noncytotoxic nanomaterials producible by cost effective, scalable procedures, the chemical, mechanical, and nanoscale properties of IBs remain essentially unexplored.Many recombinant polypeptides produced in bacteria aggregate as IBs. These protein deposits appear as highly hydrated, chemically pure particles, as the recombinant protein itself is the main component -up to around 95% of the protein therein [2][3][4]. While in the past IBs were believed to be formed by unfolded or largely misfolded polypeptide chains and therefore biologically inert [5], more recent insights show them to be constituted by folded and biofunctional protein species [6], whose presence is allowed by a particular amyloid-like organization [7][8][9]. Therefore, IBs formed by enzymes such as β-galactosidase, D-amino acid oxidase, maltodextrin phosphorylase, sialic acid aldolase, and polyphosphate kinase [10][11][12][13] can be used as catalysts in different processes. In addition, the in vivo formation of IBs is regulated by several genes (mainly encoding proteases and chaperones), which makes the genetic manipulation of their nanometer-scale properties feasible [14,15].

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“…Our laboratory routinely uses self-assembled monolayers to modify and pattern glass surfaces for cell culture applications [8,27,[29][30][31][32]. XPS is a surface characterization technique used to assess the quality of a DETA/13F modified surface.…”

Section: Surface Characterizationmentioning

confidence: 99%

Engineered In Vitro Feed-Forward Networks

Natarajan¹

2013

J Biotechnol Biomater

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Microelectrode arrays (MEAs) are a promising new method for high throughput neuronal assays. These arrays permit non-invasive, detailed optical and multichannel electrophysiological interrogation of functional neuronal networks for drug development or neurotoxicity assessment. There has also been an effort by a number of groups to develop in vitro analogues of in vivo brain circuitry or physiological systems to serve as well defined models of in vivo tissue. However, a key hurdle in these efforts has been the ability to define and constrain the directionality of pathways within these systems. This issue is particularly relevant during the recreation of in vivo brain architectures that communicate through defined pathways, often with specific directionality. In this paper, we demonstrate a line/ gap topology that promotes the growth of axonal directionally between neurons that have been engineered into a living analogue of a feed-forward neural architecture. The effective connectivity of this architecture was estimated from neural activity measured by a multichannel microelectrode array and quantified using conditional Granger causality analysis. Plasticity was then induced to determine whether 1) LTP/LTD was supported in this novel architecture and 2) whether plasticity differed from random network controls. We show that this method promotes unidirectional feed-forward relative to opposing feedback pathways in spontaneously active networks. This study also represents the first attempt to use the Granger causality metric for the assessment of the activity of a biological neuronal network in which connectivity is highly defined.

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Photolithographic Patterning of C2C12 Myotubes using Vitronectin as Growth Substrate in Serum‐Free Medium

Cited by 66 publications

References 37 publications

Vitronectin as a Micromanager of Cell Response in Material‐Driven Fibronectin Nanonetworks

Vitronectin as a Micromanager of Cell Response in Material‐Driven Fibronectin Nanonetworks

Engineering Protein Based Nanoparticles for Applications in Tissue Engineering

Engineered In Vitro Feed-Forward Networks

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