The Experts below are selected from a list of 66 Experts worldwide ranked by ideXlab platform

Christopher J Murphy - One of the best experts on this subject based on the ideXlab platform.

  • Latrunculin B and substratum stiffness regulate corneal fibroblast to myofibroblast transformation
    Experimental Eye Research, 2018
    Co-Authors: Sara M. Thomasy, Hidetaka Miyagi, Alexander T. Evashenk, Jasmyne Sermeno, Joshua T. Morgan, Geneva K Tripp, Vijay Krishna Raghunathan, Christopher J Murphy
    Abstract:

    The transformation of keratocytes and fibroblasts to myofibroblasts is important to corneal wound healing as well as formation of stromal haze. The purpose of this study was to determine the effect of latrunculin B, an actin cytoskeleton disruptor in conjunction with a fundamental Biophysical Cue, substrate stiffness, on myofibroblast transformation in vitro and in vivo. Rabbit corneal fibroblasts were cultured on substrates of differing compliance (1.5, 22, and 71 kPa) and tissue culture plastic (TCP; > 1 GPa) in media containing 0 or 10 ng/ml TGFβ1 for 72 h. Cells were treated with 0.4 μM Lat-B or DMSO for 30 min every 24 h for 72 h. RNA was collected from cells and expression of alpha-smooth muscle actin (α-SMA), keratocan, and ALDH1A1 determined using qPCR; immunocytochemistry was used to assess α-SMA protein expression. A rabbit phototherapeutic keratectomy (PTK) model was used to assess the impact of 0.1% Lat-B (n = 3) or 25% DMSO (vehicle control, n = 3) on corneal wound healing by assessment of epithelial wound size with fluorescein stain and semi-quantitative stromal haze scoring by an observer masked to treatment group as well as Fourier-domain optical coherence tomography (FD-OCT) at set time points. Statistical analysis was completed using one-way or two-way analysis of variance. Treatment with Lat-B versus DMSO resulted in significantly less αSMA mRNA (P ≤ 0.007) for RCF cells grown on 22 and 71 kPa substrates as well as TCP without or with TGFβ1, and significantly decreased α-SMA protein expression in RCFs cultured on the intermediate (22 kPa) stiffness in the absence (P = 0.028) or presence (P = 0.018) of TGFβ1. Treatment with Lat-B versus DMSO but did not significantly alter expression of keratocan or ALDH1A1 mRNA in RCFs (P > 0.05) in the absence or presence of TGFβ1, but RCFs grown on stiff hydrogels (71 kPa) had significantly more keratocan mRNA expression versus the 22 kPa hydrogel or TCP (P 

  • substratum topography modulates corneal fibroblast to myofibroblast transformation
    Investigative Ophthalmology & Visual Science, 2012
    Co-Authors: Kathern E Myrna, Rima Mendonsa, Paul Russell, Simon A Pot, Sara J Liliensiek, James V Jester, Paul F Nealey, Donald J Brown, Christopher J Murphy
    Abstract:

    Purpose. The transformation of fibroblasts to myofibroblasts is critical to corneal wound healing, stromal haze formation, and scarring. It has recently been demonstrated that the provision of biomimetic substratum topographic Cues inhibits the progression toward the myofibroblast phenotype under the influence of transforming growth factor β1 (TGF-β1). The objective of this study was to determine the effect of another fundamental Biophysical Cue, substrate compliance, on TGF-β1–induced myofibroblast transformation of primary corneal cells isolated from human and rabbit corneas.

Ajay Tijore - One of the best experts on this subject based on the ideXlab platform.

  • modulating human mesenchymal stem cell plasticity using micropatterning technique
    PLOS ONE, 2014
    Co-Authors: Ajay Tijore
    Abstract:

    In our previous work, we have reported that enforced elongation of human mesenchymal stem cells (hMSCs) through micropatterning promoted their myocardial lineage commitment. However, whether this approach is robust enough to retain the commitment when subsequently subjected to different conditions remains unsolved. This de-differentiation, if any, would have significant implication on the application of these myocardial-like hMSCs either as tissue engineered product or in stem cell therapy. Herein, we investigated the robustness of micropatterning induced differentiation by evaluating the retention of myocardial differentiation in patterned hMSCs when challenged with non-myocardial differentiation Cues. Altogether, we designed four groups of experiments; 1) Patterned hMSCs cultured in normal growth medium serving as a positive control; 2) Patterned hMSCs cultured in normal growth medium for 14 days followed by osteogenic and adipogenic media for next 7 days (to study the robustness of the effect of micropatterning); 3) Patterned hMSCs (initially grown in normal growth medium for 14 days) trypsinized and recultured in different induction media for next 7 days (to study the robustness of the effect of micropatterning without any shape constrain) and 4) Patterned hMSCs cultured in osteogenic and adipogenic media for 14 days (to study the effects of biochemical Cues versus Biophysical Cues). It was found that hMSCs that were primed to commit to myocardial lineage (Groups 2 and 3) were able to maintain myocardial lineage commitment despite subsequent culturing in osteogenic and adipogenic media. However, for hMSCs that were not primed (Group 4), the biochemical Cues seem to dominate over the Biophysical Cue in modulating hMSCs differentiation. It demonstrates that cell shape modulation is not only capable of inducing stem cell differentiation but also ensuring the permanent lineage commitment.

Guobao Chen - One of the best experts on this subject based on the ideXlab platform.

  • demineralized bone scaffolds with tunable matrix stiffness for efficient bone integration
    ACS Applied Materials & Interfaces, 2018
    Co-Authors: Mengying Liu, Guobao Chen
    Abstract:

    As a Biophysical Cue, matrix stiffness can decide the stem cell fate. However, most methods to construct three-dimensional (3D) scaffolds may change the 3D microstructure while altering their mechanical properties. In this study, demineralized bone matrix scaffolds with different compressive modulus (66.06 ± 27.83 MPa (high), 26.90 ± 13.16 MPa (medium), and 0.67 ± 0.14 MPa (low)) were constructed by controlling the decalcification duration (1 h, 12 h, and 5 days), respectively. The pore size and porosity have no significant difference between the scaffolds before and after decalcification. Cell experiments indicated that the low scaffolds could promote the osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs) in vitro. Rat subcutaneous implantation experiments further demonstrated that the low scaffolds could efficiently improve the cell infiltration, deposition of collagen fibers, and positive osteocalcin and osteopontin expression of endogenous cells as well as angiogenesis. Finally, r...

  • Demineralized Bone Scaffolds with Tunable Matrix Stiffness for Efficient Bone Integration
    2018
    Co-Authors: Mengying Liu, Guobao Chen
    Abstract:

    As a Biophysical Cue, matrix stiffness can decide the stem cell fate. However, most methods to construct three-dimensional (3D) scaffolds may change the 3D microstructure while altering their mechanical properties. In this study, demineralized bone matrix scaffolds with different compressive modulus (66.06 ± 27.83 MPa (high), 26.90 ± 13.16 MPa (medium), and 0.67 ± 0.14 MPa (low)) were constructed by controlling the decalcification duration (1 h, 12 h, and 5 days), respectively. The pore size and porosity have no significant difference between the scaffolds before and after decalcification. Cell experiments indicated that the low scaffolds could promote the osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs) in vitro. Rat subcutaneous implantation experiments further demonstrated that the low scaffolds could efficiently improve the cell infiltration, deposition of collagen fibers, and positive osteocalcin and osteopontin expression of endogenous cells as well as angiogenesis. Finally, rabbit femoral condylar defect experiments proved that the low scaffolds could significantly promote the bone repair and integration and stromal cell derived factor-1α/CXC chemokine receptor signal pathway was essential for the stiffness-mediated bone repair. These investigations provided a novel method for fabricating 3D bone grafts with different stiffness, which is also of great significance for studying the effect of stiffness on the biological behavior of MSCs in three dimensions

Guoping Chen - One of the best experts on this subject based on the ideXlab platform.

  • Influence of Cell Spreading Area on the Osteogenic Commitment and Phenotype Maintenance of Mesenchymal Stem Cells
    Scientific Reports, 2019
    Co-Authors: Yingjun Yang, Yingnan Yang, Xinlong Wang, Xiaohong Hu, Yongtao Wang, Naoki Kawazoe, Guoping Chen
    Abstract:

    Osteogenic differentiation and commitment of mesenchymal stem cells (MSCs) is a complex process that is induced and regulated by various biological factors and Biophysical Cues. Although cell spreading area, as a Biophysical Cue, has been demonstrated to play a critical role in the regulation of osteogenic differentiation of MSCs, it is unclear how it affects the maintenance of the committed phenotype after osteogenic differentiation of MSCs. In this study, poly (vinyl alcohol) was micropatterned on a tissue culture polystyrene surface, and the micropatterns were used to culture MSCs to control their cell spreading area. The influence of cell spreading area on osteogenic differentiation and maintenance of the differentiated phenotype of MSCs was investigated. MSCs with a larger spreading area showed a higher degree of osteogenic differentiation, slower loss of differentiated phenotype and slower re-expression of stem cell markers compared with MSCs with a smaller spreading area. A large cell spreading area was beneficial for osteogenic differentiation of MSCs and maintenance of their differentiated phenotype.

Amrinder S Nain - One of the best experts on this subject based on the ideXlab platform.

  • rules of contact inhibition of locomotion for cells on suspended nanofibers
    Proceedings of the National Academy of Sciences of the United States of America, 2021
    Co-Authors: Jugroop Singh, Brian A Camley, Aldwin Pagulayan, Amrinder S Nain
    Abstract:

    Contact inhibition of locomotion (CIL), in which cells repolarize and move away from contact, is now established as a fundamental driving force in development, repair, and disease biology. Much of what we know of CIL stems from studies on two-dimensional (2D) substrates that do not provide an essential Biophysical Cue-the curvature of extracellular matrix fibers. We discover rules controlling outcomes of cell-cell collisions on suspended nanofibers and show them to be profoundly different from the stereotyped CIL behavior on 2D substrates. Two approaching cells attached to a single fiber do not repolarize upon contact but rather usually migrate past one another. Fiber geometry modulates this behavior; when cells attach to two fibers, reducing their freedom to reorient, only one cell repolarizes on contact, leading to the cell pair migrating as a single unit. CIL outcomes also change when one cell has recently divided and moves with high speed-cells more frequently walk past each other. Our computational model of CIL in fiber geometries reproduces the core qualitative results of the experiments robustly to model parameters. Our model shows that the increased speed of postdivision cells may be sufficient to explain their increased walk-past rate. We also identify cell-cell adhesion as a key mediator of collision outcomes. Our results suggest that characterizing cell-cell interactions on flat substrates, channels, or micropatterns is not sufficient to predict interactions in a matrix-the geometry of the fiber can generate entirely new behaviors.

  • rules of contact inhibition of locomotion for cells on suspended nanofibers
    bioRxiv, 2020
    Co-Authors: Jugroop Singh, Brian A Camley, Amrinder S Nain
    Abstract:

    Contact inhibition of locomotion (CIL), in which cells repolarize and move away from contact, is now established as a fundamental driving force in development, repair, and disease biology. Much of what we know of CIL stems from studies on 2D substrates that fail to provide an essential Biophysical Cue - the curvature of extracellular matrix fibers. We discover rules controlling outcomes of cell-cell collisions on suspended nanofibers, and show them to be profoundly different from the stereotyped CIL behavior known on 2D substrates. Two approaching cells attached to a single fiber do not repolarize upon contact but rather usually migrate past one another. Fiber geometry modulates this behavior: when cells are attached to two fibers, reducing their freedom to reorient, only one of a pair of colliding cells repolarizes on contact, leading to the cell pair migrating as a single unit. CIL outcomes also change when one cell has recently divided and moves with high speed- cells more frequently walk past each other. In collisions with division in the two-fiber geometry, we also capture rare events where a daughter cell pushes the non-dividing cell along the fibers. Our computational model of CIL in fiber geometries reproduces the core qualitative results of the experiments robustly to model parameters. Our model shows that the increased speed of post-division cells may be sufficient to explain their increased walk-past rate. Our results suggest that characterizing cell-cell interactions on flat substrates, channels, or micropatterns is not sufficient to predict interactions in a matrix - the geometry of the fiber can generate entirely new behaviors.