Substrate Elastic Modulus Regulates the Morphology, Focal Adhesions, and α-Smooth Muscle Actin Expression of Retinal Müller Cells

Bu, Shaochong; Kuijer, Roel; Worp, Roelofje J. van der; Putten, Sander M. van; Wouters, O.; Li, Xiaorong; Hooymans, Johanna M. M.; Los, Leonoor I.

doi:10.1167/iovs.14-15969

Cited by 11 publications

(5 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The primary mechano-sensing element in the cell is the cytoskeleton, which communicates with the ECM. Specifically, cells grown on different environments have differential expression of focal adhesion proteins and actin cytoskeletal network. − In addition, perturbing the actin cytoskeleton and its associated motor, nonmuscle myosin II, hinders the differentiation process of MSCs when grown on these different substrates. ,, However, less is known about what role the microtubules play in this differentiation process in a 3D environment. In this study, we aim to understand the role of the microtubule cytoskeleton in the differentiation of MSCs to osteoblasts when grown on 3D nanofibrous scaffolds.…”

Section: Introductionmentioning

confidence: 99%

Role of Microtubules in Osteogenic Differentiation of Mesenchymal Stem Cells on 3D Nanofibrous Scaffolds

Meka

Chacko

Ravi

et al. 2017

ACS Biomater. Sci. Eng.

View full text Add to dashboard Cite

Human bone marrow mesenchymal stem cells (MSCs) cultured on three-dimensional (3D) nanofibrous scaffolds are known to undergo osteogenic differentiation even in the absence of soluble osteoinductive factors. Although this process of differentiation has been attributed to the shape that cells assume on the fibrous scaffolds, it is unclear how constriction of cell shape would contribute to the differentiation phenotype. Here, we quantitatively compared cell and nuclear morphologies of cells cultured on 3D poly(εcaprolactone) (PCL) nanofibers (NF) and two-dimensional (2D) flat films using confocal fluorescence microscopy. We discovered that while cells on the 2D films exhibited cellular and nuclear morphologies similar to those cultured on tissue culture polystyrene, cells cultured on the 3D NF showed distinct cell and nuclear morphologies, with lower areas and perimeters, but higher aspect ratios. We next tested the effect of treatment of cells with actin-depolymerizing cytochalasin D and microtubule-depolymerizing nocodazole on these morphologies. In both 2D and 3D scaffolds, actin depolymerization brought about gross changes in cell and nuclear morphologies. Remarkably, microtubule depolymerization resulted in a phenotype similar to actin depolymerization in cells cultured on 3D NF alone, indicating a significant role for the microtubule cytoskeleton in the maintenance of cell shape and structure in 3D. The morphological changes of the nucleus that were apparent upon cytoskeletal perturbation were reflected in the organization of heterochromatin in the nucleus, with MSCs on 3D alone exhibiting a differentiation phenotype. Finally, we tested the effect of cytoskeletal depolymerization on mineralization of cells. Again, we observed higher mineralization in cells cultured on 3D NF, which was lost in cells treated with either cytochalasin D or nocodazole. Taken together, our results suggest that both the actin and microtubule cytoskeletons contribute significantly toward maintenance of cell and nuclear shape in cells cultured on 3D scaffolds, and consequently to their osteogenic differentiation.

show abstract

Section: Introductionmentioning

confidence: 99%

Role of Microtubules in Osteogenic Differentiation of Mesenchymal Stem Cells on 3D Nanofibrous Scaffolds

Meka

Chacko

Ravi

et al. 2017

ACS Biomater. Sci. Eng.

View full text Add to dashboard Cite

show abstract

“…To investigate how HGF alters the phenotype of myofibroblasts, HCFs were incubated with 5 ng/mL TGF-β1 for 24 h, followed by another 24-h treatment of 20 ng/mL HGF. This resulted in an obvious drop in α-SMA mRNA level and suggested that HGF may have the potential to revert myofibroblast phenotype to fibroblast one [36]. In addition, the stiffness of TGF-β1-treated HCF was increased.…”

Section: Resultsmentioning

confidence: 99%

A Systematic Review on Cornea Epithelial-Stromal Homeostasis

Wong¹,

Poon²,

Bu³

et al. 2020

Ophthalmic Res

View full text Add to dashboard Cite

Introduction: This review aims to summarise the role of different cells, genes, proteins and lipid in regulating cornea epithelial-stromal homeostasis. Methods: We performed an Entrez PubMed literature search using keywords “human,” “cornea,” “epithelial,” “stromal,” “homeostasis,” “fibrosis response,” and “pathogenesis” on 24th of September 2019, resulting in 35 papers, of which 18 were chosen after filtering for “English language” and “published within 10 years” as well as curation for relevance by the authors. Results: The 18 selected papers showed that corneal epithelial cells, fibroblasts and telocytes, together with genes such as Klf4, Pax6 and Id found in the cells, play important roles in achieving homeostasis to maintain corneal integrity and transparency. Proteins classified as pro-fibrotic ligands and anti-fibrotic ligands are responsible for regulating cornea stromal fibrosis and extracellular matrix deposition, thus regulators of scar formation during wound healing. Anti-inflammatory ligands and wound repairing ligands are critical in eliciting protective inflammation and promoting epithelial healing, respectively. Protein receptors located on cellular membrane play a role in maintaining intercellular connections as well as corneal hydration. Discussion/Conclusion: These studies prompt development of novel therapeutic strategies such as tear drops or ointments that target certain proteins to maintain corneal homeostasis. However, more in vitro and in vivo studies are required to prove the effectiveness of exogenous administration of molecules in improving healing outcome. Hence, future investigations of the molecular pathways highlighted in this review will reveal novel therapeutic tools such as gene or cell therapy to treat corneal diseases.

show abstract

“…Müller cells, indeed, exert tractive forces on the ILM through the modulation of actin filaments, stress fibres and focal adhesion clusters. Such interactions were recently investigated in-vitro using collagen-based scaffolds of varying stiffness [39]. A stiffness increase was associated with a corresponding increase in the incorporation of actin filaments into the cytoskeleton and in the vinculin recruitment that, in its turn, stabilizes the focal adhesion promoting higher tensions.…”

Section: Discussionmentioning

confidence: 99%

Nanomechanical mapping helps explain differences in outcomes of eye microsurgery: A comparative study of macular pathologies

et al. 2019

View full text Add to dashboard Cite

Many ocular diseases are associated with an alteration of the mechanical and the material properties of the eye. These mechanically-related diseases include macular hole and pucker, two ocular conditions due to the presence of abnormal physical tractions acting on the retina. A complete relief of these tractions can be obtained through a challenging microsurgical procedure, which requires the mechanical peeling of the internal limiting membrane of the retina (ILM). In this paper, we provide the first comparative study of the nanoscale morphological and mechanical properties of the ILM in macular hole and macular pucker. Our nanoscale elastic measurements unveil a different bio-mechanical response of the ILM in the two pathologies, which correlates well to significant differences occurring during microsurgery. The results here presented pave the way to the development of novel dedicated microsurgical protocols based on the material ILM properties in macular hole or pucker. Moreover, they contribute to clarify why, despite a common aetiology, a patient might develop one disease or the other, an issue which is still debated in literature.

show abstract

Substrate Elastic Modulus Regulates the Morphology, Focal Adhesions, and α-Smooth Muscle Actin Expression of Retinal Müller Cells

Cited by 11 publications

References 52 publications

Role of Microtubules in Osteogenic Differentiation of Mesenchymal Stem Cells on 3D Nanofibrous Scaffolds

Role of Microtubules in Osteogenic Differentiation of Mesenchymal Stem Cells on 3D Nanofibrous Scaffolds

A Systematic Review on Cornea Epithelial-Stromal Homeostasis

Nanomechanical mapping helps explain differences in outcomes of eye microsurgery: A comparative study of macular pathologies

Contact Info

Product

Resources

About