Effect of Serum and Insulin Modulation on the Organization and Morphology of Matrix Synthesized by Bovine Corneal Stromal Cells

Bueno, Ericka M.; Saeidi, Nima; Melotti, Suzanna A.; Ruberti, Jeffrey W.

doi:10.1089/ten.tea.2008.0404

Cited by 10 publications

(13 citation statements)

References 84 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The effect of the combined extensional stretch and binding produces physical conditions that should promote the lateral aggregation or crystallization of polymers associated with the surface of the process. The second model is consistent with the co-alignment of corneal cell processes ("keratopodia") with collagen arrays observed by Young et al 5 and Bueno et al 76 .…”

Section: Proposed Models Of Ecm Filament Formationsupporting

confidence: 88%

Mechanical Causation of Biological Structure: Productive Pulls Produce Persistent Filaments in a Human Fibroblast Model of Matrix Development

Silverman¹,

Siadat²,

Olszewski³

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

During development, mesenchymal cells direct the elaboration of extracellular matrix that shapes the initial animal bauplan which subsequently grows to produce mechanically-competent structure. To gain insight into the processes that initiate matrix formation at the cellular level, high temporal and spatial resolution videos were obtained from a primary human corneal fibroblast (PHCF) cell culture system known to produce an organized, collagenous stroma similar to a human cornea. The images were taken over a 4-day period prior to culture confluency which permitted a clear view of the cell kinematics and any elaborated filaments. The movies reveal an active cellular system in which the PHCFs execute five types of high-velocity and high extensional strain-rate 'pulls' that produce persistent filaments. In four of the pull types, average maximum strain rates (~0.1-0.33s-1) were adequate to induce aggregation and/or crystallization in crowded biopolymer systems. The results demonstrate that PHCFs have the capacity to mechanically induce the formation of biopolymer structures intercellularly and in the path of force.

show abstract

Section: Proposed Models Of Ecm Filament Formationsupporting

confidence: 88%

Mechanical Causation of Biological Structure: Productive Pulls Produce Persistent Filaments in a Human Fibroblast Model of Matrix Development

Silverman¹,

Siadat²,

Olszewski³

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, the entire structure could not be revealed in one freeze-fracture plane, and consequently their abundance and considerable length were unappreciated. Filopodia, aligned with secreted collagen fibrils, were also a feature of bovine corneal cells cultured in vitro (40). The 3D reconstructions from our study show that in developing cornea these tubular membrane extensions travel more than 30 microns from the main body of the cell.…”

Section: Discussionmentioning

confidence: 51%

Three-dimensional aspects of matrix assembly by cells in the developing cornea

Young

Knupp

Pinali

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Cell-directed deposition of aligned collagen fibrils during corneal embryogenesis is poorly understood, despite the fact that it is the basis for the formation of a corneal stroma that must be transparent to visible light and biomechanically stable. Previous studies of the structural development of the specialized matrix in the cornea have been restricted to examinations of tissue sections by conventional light or electron microscopy. Here, we use volume scanning electron microscopy, with sequential removal of ultrathin surface tissue sections achieved either by ablation with a focused ion beam or by serial block face diamond knife microtomy, to examine the microanatomy of the cornea in three dimensions and in large tissue volumes. The results show that corneal keratocytes occupy a significantly greater tissue volume than was previously thought, and there is a clear orthogonality in cell and matrix organization, quantifiable by Fourier analysis. Three-dimensional reconstructions reveal actin-associated tubular cell protrusions, reminiscent of filopodia, but extending more than 30 μm into the extracellular space. The highly extended network of these membrane-bound structures mirrors the alignment of collagen bundles and emergent lamellae and, we propose, plays a fundamental role in dictating the orientation of collagen in the developing cornea.C onnective tissues fulfill diverse functions but conform to a general structural plan of cells surrounded by an extracellular matrix in which collagen is the main structural element (1, 2). Composition and tissue-specific organization of the component collagen fibrils are considered to be critically important for the functional properties of any particular tissue. The unique transparent quality of the cornea arises from its remarkably ordered architecture of aligned and regularly spaced fibrils with a small, consistent diameter (∼30 nm), which are arranged, not into fibers or fascicles as in most other tissues but in superimposed, flattened layers, or lamellae. Lamellae and their component collagen fibrils exhibit preferential orientations throughout the corneal thickness (3), which appear to be closely related to the biomechanical loads to which the tissue is subjected. In adult vertebrates, lamellae traverse the full diameter of the cornea for most of its thickness, and in the avian eye-the subject of most developmental studies-undergo a gradual rotation in their orientation with depth (4). Individual collagen fibrils within midstromal lamellae also appear to traverse the entire diameter of the cornea, a distance of ∼11 mm in adult human eyes. The extraordinary level of order in matrix organization within a hierarchy of fibril, lamella, and stroma overall appears to reflect a considerable level of regulatory influence presumably involving both cell activity and intermolecular interactions.Collagen fibrils exhibit a microfibrillar substructure (5-7), undergoing self-assembly spontaneously in vitro from soluble monomeric collagen in a process largely dependent upon the phy...

show abstract

“…Furthermore, this tissue was formed by hCSFs during a period of 21 days in a serum-free media formulation comparable to that previously reported as insufficient to sustain viable bovine corneal broblast for more than 1.5 weeks in culture. 52 This again indicated that the RGDS:ETTES coating functioned not only as a support for hCSF adhesion but also as an effector in tuning the cell phenotype and preventing cell death in serum-free conditions. This notion is supported by data showing that ligation of integrin aVb3, a RGD receptor expressed by hCSFs, 25 mediates cell survival aer serum withdrawal.…”

Section: Rgds:ettes 2d Coating As An Effector In the Formation Of 3d mentioning

confidence: 91%

Bioactive films produced from self-assembling peptide amphiphiles as versatile substrates for tuning cell adhesion and tissue architecture in serum-free conditions

et al. 2013

View full text Add to dashboard Cite

The development of versatile bioactive surfaces able to emulate in vivo conditions is of enormous importance to the future of cell and tissue therapy. Tuning cell behaviour on two-dimensional surfaces so that the cells perform as if they were in a natural three-dimensional tissue represents a significant challenge, but one that must be met if the early promise of cell and tissue therapy is to be fully realised. Due to the inherent complexities involved in the manufacture of biomimetic three-dimensional substrates, the scaling up of engineered tissue-based therapies may be simpler if based upon proven two-dimensional culture systems. In this work, we developed new coating materials composed of the self-assembling peptide amphiphiles (PAs) C 16 G 3 RGD (RGD) and C 16 G 3 RGDS (RGDS) shown to control cell adhesion and tissue architecture while avoiding the use of serum. When mixed with the C 16 ETTES diluent PA at 13 : 87 (mol mol À1) ratio at 1.25 Â 10 À3 M, the bioactive PAs were shown to support optimal adhesion, maximal proliferation, and prolonged viability of human corneal stromal fibroblasts (hCSFs), while improving the cell phenotype. These PAs also provided stable adhesive coatings on highly-hydrophobic surfaces composed of striated polytetrafluoroethylene (PTFE), significantly enhancing proliferation of aligned cells and increasing the complexity of the produced tissue. The thickness and structure of this highly-organised tissue were similar to those observed in vivo, comprising aligned newly-deposited extracellular matrix. As such, the developed coatings can constitute a versatile biomaterial for applications in cell biology, tissue engineering, and regenerative medicine requiring serum-free conditions.

show abstract

Effect of Serum and Insulin Modulation on the Organization and Morphology of Matrix Synthesized by Bovine Corneal Stromal Cells

Cited by 10 publications

References 84 publications

Mechanical Causation of Biological Structure: Productive Pulls Produce Persistent Filaments in a Human Fibroblast Model of Matrix Development

Mechanical Causation of Biological Structure: Productive Pulls Produce Persistent Filaments in a Human Fibroblast Model of Matrix Development

Three-dimensional aspects of matrix assembly by cells in the developing cornea

Bioactive films produced from self-assembling peptide amphiphiles as versatile substrates for tuning cell adhesion and tissue architecture in serum-free conditions

Contact Info

Product

Resources

About