Optimization of Corneal Epithelial Progenitor Cell Growth onBombyx moriSilk Fibroin Membranes

Scaffolds prepared from silk fibroin derived from cocoons of the domesticated silkworm mothBombyx morihave demonstrated potential to support the attachment and growth of human limbal epithelial (HLE) cellsin vitro. In this study, we attempted to further optimize protocols to promote the expansion of HLE cells onB. morisilk fibroin- (BMSF-) based scaffolds. BMSF films were initially coated with different extracellular matrix proteins and then analysed for their impact on corneal epithelial cell adhesion, cell morphology, and culture confluency. Results showed that collagen I, collagen III, and collagen IV consistently improved HCE-T cell adherence, promoted an elongated cell morphology, and increased culture confluency. By contrast, ECM coating had no significant effect on the performance of primary HLE cells cultured on BMSF films. In the second part of this study, primary HLE cells were grown on BMSF films in the presence of medium (SHEM) supplemented with keratinocyte growth factor (KGF) and the Rho kinase inhibitor, Y-27632. The results demonstrated that SHEM medium supplemented with KGF and Y-27632 dramatically increased expression of corneal differentiation markers, keratin 3 and keratin 12, whereas expression of the progenitor marker, p63, did not appear to be significantly influenced by the choice of culture medium.

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A Rho kinase inhibitor (Fasudil) suppresses TGF-β mediated autophagy in urethra fibroblasts to attenuate traumatic urethral stricture (TUS) through re-activating Akt/mTOR pathway: An in vitro study

Life Sciences ◽

10.1016/j.lfs.2020.118960 ◽

2021 ◽

Vol 267 ◽

pp. 118960

Author(s):

Huan Feng ◽

Xiaobing Huang ◽

Weihua Fu ◽

Xingyou Dong ◽

Fengxia Yang ◽

...

Keyword(s):

Urethral Stricture ◽

Kinase Inhibitor ◽

Rho Kinase ◽

In Vitro Study ◽

Mtor Pathway ◽

Vitro Study ◽

Rho Kinase Inhibitor

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Thymosin-β4 Inhibits Corneal Epithelial Cell Apoptosis after Ethanol Exposure In Vitro

Investigative Opthalmology & Visual Science ◽

10.1167/iovs.03-1002 ◽

2004 ◽

Vol 45 (4) ◽

pp. 1095 ◽

Cited By ~ 58

Author(s):

Gabriel Sosne ◽

Atif Siddiqi ◽

Michelle Kurpakus-Wheater

Keyword(s):

Epithelial Cell ◽

Cell Apoptosis ◽

Ethanol Exposure ◽

Corneal Epithelial Cell ◽

Thymosin Β4 ◽

Corneal Epithelial ◽

Epithelial Cell Apoptosis

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Immortalization and Characterizations of Rabbit Corneal Epithelial Cell Lines as Potential In Vitro Alternative Models for Evaluating the Cellular Toxicity of Ocular Drug Candidates

Advances in Ocular Toxicology ◽

10.1007/978-1-4615-5937-5_21 ◽

1997 ◽

pp. 181-191

Author(s):

C. Yao ◽

D. Wampler ◽

K. Hall ◽

D. Crouch ◽

R. Hackett ◽

...

Keyword(s):

Epithelial Cell ◽

Cell Lines ◽

Corneal Epithelial Cell ◽

Cellular Toxicity ◽

Corneal Epithelial ◽

Alternative Models ◽

Drug Candidates ◽

Epithelial Cell Lines

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Epithelial-stromal interactions: Specific stimulation of corneal epithelial cell growth in vitro by a factor(s) from cultured stromal fibroblasts

Experimental Eye Research ◽

10.1016/0014-4835(83)90008-8 ◽

1983 ◽

Vol 36 (2) ◽

pp. 231-246 ◽

Cited By ~ 36

Author(s):

Kwan Y. Chan ◽

Richard H. Haschke

Keyword(s):

Cell Growth ◽

Epithelial Cell ◽

Corneal Epithelial Cell ◽

Stromal Fibroblasts ◽

Corneal Epithelial ◽

Specific Stimulation ◽

Stimulation Of

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Hydrogel Tissue Construct-Based High-Content Compound Screening

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057110388269 ◽

2010 ◽

Vol 16 (1) ◽

pp. 120-128 ◽

Cited By ~ 8

Author(s):

Vy Lam ◽

Tetsuro Wakatsuki

Keyword(s):

Kinase Inhibitor ◽

Physiological Mechanism ◽

Rho Kinase ◽

Assay System ◽

Physiological Status ◽

Multiple Parameters ◽

Compound Screening ◽

Tissue Construct

Current pharmaceutical compound screening systems rely on cell-based assays to identify therapeutic candidates and potential toxicities. However, cells grown on 2D substrata or in suspension do not exhibit the mechanical or physiological properties of cells in vivo. To address this limitation, the authors developed an in vitro, high-throughput, 3D hydrogel tissue construct (HTC)–based assay system to quantify cell and tissue mechanical properties and multiple parameters of physiology. HTC mechanics was quantified using an automated device, and physiological status was assessed using spectroscopy-based indicators that were read on microplate readers. To demonstrate the application of this system, the authors screened 4 test compounds—rotenone (ROT), cytochalasin D (CD), 2,4-dinitrophenol (DNP), and Rho kinase inhibitor (H-1152)—for their ability to modulate HTC contractility without affecting actin integrity, mitochondrial membrane potential (MMP), or viability. All 4 compounds dose-dependently reduced HTC contractility. However, ROT was toxic, DNP dissipated MMP, and CD reduced both intracellular F-actin and viability. H-1152 was found to be the best candidate compound since it reduced HTC contractility with minimal side effects. The authors propose that their HTC-based assay system can be used to screen for compounds that modulate HTC contractility and assess the underlying physiological mechanism(s) of compound activity and toxicity.

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Animal model for angiotensin II effects in the internal anal sphincter smooth muscle: mechanism of action

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00207.2001 ◽

2002 ◽

Vol 282 (3) ◽

pp. G461-G469 ◽

Cited By ~ 13

Author(s):

Ya-Ping Fan ◽

Rajinder N. Puri ◽

Satish Rattan

Keyword(s):

Smooth Muscle ◽

Protein Kinase ◽

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Rho Kinase ◽

Ang Ii ◽

Kinase C ◽

Rho Kinase Inhibitor ◽

Isolated Smooth Muscle Cells

Effect of ANG II was investigated in in vitro smooth muscle strips and in isolated smooth muscle cells (SMC). Among different species, rat internal and sphincter (IAS) smooth muscle showed significant and reproducible contraction that remained unmodified by different neurohumoral inhibitors. The AT1antagonist losartan but not AT2 antagonist PD-123319 antagonized ANG II-induced contraction of the IAS smooth muscle and SMC. ANG II-induced contraction of rat IAS smooth muscle and SMC was attenuated by tyrosine kinase inhibitors genistein and tyrphostin, protein kinase C (PKC) inhibitor H-7, Ca2+ channel blocker nicardipine, Rho kinase inhibitor Y-27632 or p44/42mitogen-activating protein kinase (MAPK44/42) inhibitor PD-98059. Combinations of nicardipine and H-7, Y-27632, and PD-98059 caused further attenuation of the ANG II effects. Western blot analyses revealed the presence of both AT1 and AT2receptors. We conclude that ANG II causes contraction of rat IAS smooth muscle by the activation of AT1 receptors at the SMC and involves multiple intracellular pathways, influx of Ca2+, and activation of PKC, Rho kinase, and MAPK44/42.

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Long-Term Maintenance of Limbal Epithelial Progenitor Cells Using Rho Kinase Inhibitor and Keratinocyte Growth Factor

Stem Cells Translational Medicine ◽

10.5966/sctm.2012-0156 ◽

2013 ◽

Vol 2 (10) ◽

pp. 758-765 ◽

Cited By ~ 32

Author(s):

Hideyuki Miyashita ◽

Seiichi Yokoo ◽

Satoru Yoshida ◽

Tetsuya Kawakita ◽

Satoru Yamagami ◽

...

Keyword(s):

Growth Factor ◽

Progenitor Cells ◽

Kinase Inhibitor ◽

Keratinocyte Growth Factor ◽

Rho Kinase ◽

Rho Kinase Inhibitor ◽

Epithelial Progenitor Cells ◽

Term Maintenance

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Biocompatibility of Nanocellulose-Reinforced PVA Hydrogel with Human Corneal Epithelial Cells for Ophthalmic Applications

Journal of Functional Biomaterials ◽

10.3390/jfb10030035 ◽

2019 ◽

Vol 10 (3) ◽

pp. 35 ◽

Cited By ~ 2

Author(s):

Tummala ◽

Lopes ◽

Mihranyan ◽

Ferraz

Keyword(s):

Epithelial Cells ◽

Cell Viability ◽

Protein Adsorption ◽

Direct Contact ◽

Corneal Epithelial Cell ◽

Corneal Epithelial Cells ◽

Corneal Epithelial ◽

Human Corneal Epithelial Cells ◽

Hydrogel Surface

Transparent composite hydrogel in the form of a contact lens made from poly(vinyl alcohol) (PVA) and cellulose nanocrystals (CNCs) was subjected to in vitro biocompatibility evaluation with human corneal epithelial cells (HCE-2 cells). The cell response to direct contact with the hydrogels was investigated by placing the samples on top of confluent cell layers and evaluating cell viability, morphology, and cell layer integrity subsequent to 24 h culture and removal of the hydrogels. To further characterize the lens–cell interactions, HCE-2 cells were seeded on the hydrogels, with and without simulated tear fluid (STF) pre-conditioning, and cell viability and morphology were evaluated. Furthermore, protein adsorption on the hydrogel surface was investigated by incubating the materials with STF, followed by protein elution and quantification. The hydrogel material was found to have affinity towards protein adsorption, most probably due to the interactions between the positively charged lysozyme and the negatively charged CNCs embedded in the PVA matrix. The direct contact experiment demonstrated that the physical presence of the lenses did not affect corneal epithelial cell monolayers in terms of integrity nor cell metabolic activity. Moreover, it was found that viable corneal cells adhered to the hydrogel, showing the typical morphology of epithelial cells and that such response was not influenced by the STF pre-conditioning of the hydrogel surface. The results of the study confirm that PVA-CNC hydrogel is a promising ophthalmic biomaterial, motivating future in vitro and in vivo biocompatibility studies.

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