Substratum Topography Modulates Corneal Fibroblast to Myofibroblast Transformation

Myrna, Kathern E.; Mendonsa, Rima; Russell, Paul; Pot, Simon A.; Liliensiek, Sara J.; Jester, James V.; Nealey, Paul F.; Brown, Donald J.; Murphy, Christopher J.

doi:10.1167/iovs.11-7982

Cited by 69 publications

(56 citation statements)

References 70 publications

(36 reference statements)

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“…The WGA label is clearly visible in areas of ridges and grooves, consistent with previous reports showing that the membrane spans the grooves and that the primary points of cell attachment are on the ridges (21,29,30,36,45). This further demonstrates that there is minimal fluorescent artifact from the underlying topography.…”

Section: Immunocytochemistry and Fluorescence Microscopysupporting

confidence: 90%

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Early responses of vascular endothelial cells to topographic cues

Dreier

Gasiorowski²,

Morgan

et al. 2013

American Journal of Physiology-Cell Physiology

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Vascular endothelial cells in vivo are exposed to multiple biophysical cues provided by the basement membrane, a specialized extracellular matrix through which vascular endothelial cells are attached to the underlying stroma. The importance of biophysical cues has been widely reported, but the signaling pathways that mediate cellular recognition and response to these cues remain poorly understood. Anisotropic topographically patterned substrates with nano- through microscale feature dimensions were fabricated to investigate cellular responses to topographic cues. The present study focuses on early events following exposure of human umbilical vein endothelial cells (HUVECs) to these patterned substrates. In serum-free medium and on substrates without protein coating, HUVECs oriented parallel to the long axis of underlying ridges in as little as 30 min. Immunocytochemistry showed clear differences in the localization of the focal adhesion proteins Src, p130Cas, and focal adhesion kinase (FAK) in HUVECs cultured on topographically patterned surfaces and on planar surfaces, suggesting involvement of these proteins in mediating the response to topographic features. Knockdown experiments demonstrated that FAK was not necessary for HUVEC alignment in response to topographic cues, although FAK knockdown did modulate HUVEC migration. These data identify key events early in the cellular response to biophysical stimuli.

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Section: Immunocytochemistry and Fluorescence Microscopysupporting

confidence: 90%

“…The specificity of this staining pattern on the patterned substrates was previously reported (21,29,30,36,45) and confirmed in METHODS (Fig. 1).…”

Section: Early Cell-signaling Events Triggered By Topographic Cuessupporting

confidence: 88%

Early responses of vascular endothelial cells to topographic cues

Dreier

Gasiorowski²,

Morgan

et al. 2013

American Journal of Physiology-Cell Physiology

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“…It is therefore possible that in the wound environment, native nanotopographic cues assist in stabilizing the keratocyte/fibroblast phenotype while pathologic microenvironmental alterations may be permissive for increased myofibroblast differentiation and the development of fibrosis and corneal haze. (Myrna et al, 2012) Hence, studying anti-fibrotic therapies in excimer laser ablated ex vivo corneas is a better system than in cell culture experiments as they provide pathologic microenvironmental alterations similar to in vivo conditions that may stimulate the fibroblasts to change to the myofibroblast phenotype.…”

Section: Discussionmentioning

confidence: 99%

Assessment of anti-scarring therapies in ex vivo organ cultured rabbit corneas

Sriram

Gibson

Robinson

et al. 2014

Experimental Eye Research

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The effects of a triple combination of siRNAs targeting key scarring genes was assessed using an ex vivo organ culture model of excimer ablated rabbit corneas. The central 6 mm diameter region of fresh rabbit globes was ablated to a depth of 155 microns with an excimer laser. Corneas were excised, cultured at the air-liquid interface in defined culture medium supplemented with transforming growth factor beta 1 (TGFB1), and treated with either 1% prednisolone acetate or with 22.5 μM cationic nanoparticles complexed with a triple combination of siRNAs (NP-siRNA) targeting TGFB1, TGFB Receptor (TGFBR2) and connective tissue growth factor (CTGF). Scar formation was measured using image analysis of digital images and levels of smooth muscle actin (SMA) were assessed in ablated region of corneas using qRT-PCR and immunostaining. Ex vivo cultured corneas developed intense haze-like scar in the wounded areas and levels of mRNAs for pro-fibrotic genes were significantly elevated 3 to 8 fold in wounded tissue compared to unablated corneas. Treatment with NP-siRNA or steroid significantly reduced quantitative haze levels by 55% and 68%, respectively, and reduced SMA mRNA and immunohistostaining. This ex vivo corneal culture system reproduced key molecular patterns of corneal scarring and haze formation generated in rabbits. Treatment with NP-siRNAs targeting key scarring genes or an anti-inflammatory steroid reduced corneal haze and SMA mRNA and protein.

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“…Hence, isotropic elastic substrates are not particularly suitable to investigate the effects of strain-induced local stiffening or fiber alignments that can serve as topographical cues for cell behavior (Karamichos et al, 2007;Petroll and Miron-Mendoza, 2015). Nevertheless, such cues can be introduced to isotropic substrates and were shown to influence myofibroblast activation from corneal fibroblasts and keratocytes (Myrna et al, 2012;Pot et al, 2010). Topography and elasticity of biomaterials will need to be considered when designing replacement biomaterials to support eye repair.…”

Section: Accepted Manuscriptmentioning

confidence: 99%

Myofibroblasts

Hinz¹

2016

Experimental Eye Research

330

324

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Myofibroblasts are activated in response to tissue injury with the primary task to repair lost or damaged extracellular matrix. Enhanced collagen secretion and subsequent contraction - scarring - are part of the normal wound healing response and crucial to restore tissue integrity. Due to myofibroblasts ability to repair but not regenerate, accumulation of scar tissue is always associated with reduced organ performance. This is a fair price to pay by the body for not falling apart. Whereas myofibroblasts typically vanish after successful repair, dysregulation of the normal repair process can lead to persistent myofibroblast activation, for instance by chronic inflammation or mechanical stress in the tissue. Excessive repair leads to the accumulation of stiff collagenous ECM contractures - fibrosis - with dramatic consequences for organ function. The clinical need to terminate detrimental myofibroblast activities has stimulated researchers to answer a number of essential questions: where do myofibroblasts come from, what are the factors leading to their activation, how do we discriminate myofibroblasts from other cells, what is the molecular basis for their contractile activity, and how can we stop or at least control them? This article reviews the current state of the myofibroblast literature by emphasizing their role in ocular repair and fibrosis. It appears that although the eye is quite an extraordinary organ, ocular myofibroblasts behave or misbehave just like their siblings in other organs.

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Substratum Topography Modulates Corneal Fibroblast to Myofibroblast Transformation

Cited by 69 publications

References 70 publications

Early responses of vascular endothelial cells to topographic cues

Early responses of vascular endothelial cells to topographic cues

Assessment of anti-scarring therapies in ex vivo organ cultured rabbit corneas

Myofibroblasts

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