scholarly journals Hierarchically Assembled Type I Collagen Fibres as Biomimetic Building Blocks of Biomedical Membranes

Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 620
Author(s):  
Jie Yin ◽  
David J. Wood ◽  
Stephen J. Russell ◽  
Giuseppe Tronci
Keyword(s):  
Type I Collagen ◽  
Network Formation ◽  
Tensile Modulus ◽  
Uv Curing ◽  
Building Blocks ◽  
Wet Spinning ◽  
Type I ◽  
Swelling Ratio ◽  
Collagen Fibres ◽  
Fibre Morphology

Wet spinning is an established fibre manufacturing route to realise collagen fibres with preserved triple helix architecture and cell acceptability for applications in biomedical membranes. However, resulting fibres still need to be chemically modified post-spinning to ensure material integrity in physiological media, with inherent risks of alteration of fibre morphology and with limited opportunities to induce fibrillogenesis following collagen fixation in the crosslinked state. To overcome this challenge, we hypothesised that a photoactive type I collagen precursor bearing either single or multiple monomers could be employed to accomplish hierarchically assembled fibres with improved processability, macroscopic properties and nanoscale organisation via sequential wet spinning and UV-curing. In-house-extracted type I rat tail collagen functionalised with both 4-vinylbenzyl chloride (4VBC) and methacrylate residues generated a full hydrogel network following solubilisation in a photoactive aqueous solution and UV exposure, whereby ~85 wt.% of material was retained following 75-day hydrolytic incubation. Wide-angle x-ray diffraction confirmed the presence of typical collagen patterns, whilst an averaged compression modulus and swelling ratio of more than 290 kPa and 1500 wt.% was recorded in the UV-cured hydrogel networks. Photoactive type I collagen precursors were subsequently wet spun into fibres, displaying the typical dichroic features of collagen and regular fibre morphology. Varying tensile modulus (E = 5 ± 1 − 11 ± 4 MPa) and swelling ratio (SR = 1880 ± 200 − 3350 ± 500 wt.%) were measured following post-spinning UV curing and equilibration with phosphate‑buffered saline (PBS). Most importantly, 72-h incubation of the wet spun fibres in PBS successfully induced renaturation of collagen-like fibrils, which were fixed following UV-induced network formation. The whole process proved to be well tolerated by cells, as indicated by a spread-like cell morphology following a 48-h culture of L929 mouse fibroblasts on the extracts of UV-cured fibres.

MAPK signaling regulates endothelial cell assembly into networks and expression of MT1-MMP and MMP-2

2005 ◽  
Vol 288 (3) ◽  
pp. C659-C668 ◽  
Author(s):  
Pamela J. Boyd ◽  
Jennifer Doyle ◽  
Eric Gee ◽  
Shelley Pallan ◽  
Tara L. Haas
Keyword(s):  
Endothelial Cells ◽  
Cellular Networks ◽  
Transcriptional Activation ◽  
Type I Collagen ◽  
Network Formation ◽  
Mapk Signaling ◽  
Mitogen Activated Protein Kinase ◽  
Type I ◽  
Mrna Levels ◽  
Collagen Lattices

Microvascular endothelial cells embedded within three-dimensional (3D) type I collagen matrixes assemble into cellular networks, a process that requires the upregulation of membrane type 1 (MT1) matrix metalloproteinase (MMP) and MMP-2. The purpose of this study was to identify the signaling pathways responsible for the transcriptional activation of MT1-MMP and MMP-2 in endothelial cells in 3D collagen lattices. We hypothesized that the 3D type I collagen induction of MT1-MMP and MMP-2 is mediated by the mitogen-activated protein kinase family of enzymes. Here, we show that 3D type I collagen elicits a persistent increase in ERK1/2 and JNK activation and a decrease in p38 activation. Inhibition of ERK1/2 or JNK disrupted endothelial network formation in 3D type I collagen lattices, whereas inhibition of p38 promoted network formation. mRNA levels of both MT1-MMP and MMP-2 were attenuated by ERK1/2 inhibition but unaffected by either JNK or p38 inhibition. By contrast, expression of constitutively active MEK was sufficient to stimulate MMP-2 production in a monolayer of endothelial cells cultured on type I collagen. These results provide evidence that signaling through both ERK1/2 and JNK regulates endothelial assembly into cellular networks but that the ERK1/2 signaling cascade specifically regulates network formation and the production of both MT1-MMP and MMP-2 genes in response to 3D type I collagen.

scholarly journals Endothelial cell crosstalk improves browning but hinders white adipocyte maturation in 3D engineered adipose tissue

2020 ◽  
Vol 12 (4) ◽  
pp. 81-89
Author(s):  
Jennifer H Hammel ◽  
Evangelia Bellas
Keyword(s):  
Endothelial Cells ◽  
Adipose Tissue ◽  
Type I Collagen ◽  
Network Formation ◽  
Uncoupling Protein ◽  
Collagen Gel ◽  
Cell Types ◽  
Vascular Network ◽  
Type I ◽  
White Adipocyte

Abstract Central to the development of adipose tissue (AT) engineered models is the supporting vasculature. It is a key part of AT function and long-term maintenance, but the crosstalk between adipocytes and endothelial cells is not well understood. Here, we directly co-culture the two cell types at varying ratios in a 3D Type I collagen gel. Constructs were evaluated for adipocyte maturation and function and vascular network organization. Further, these constructs were treated with forskolin, a beta-adrenergic agonist, to stimulate lipolysis and browning. Adipocytes in co-cultures were found to be less mature than an adipocyte-only control, shown by smaller lipid droplets and downregulation of key adipocyte-related genes. The most extensive vascular network formation was found in the 1:1 co-culture, supported by vascular endothelial growth factor (VEGF) upregulation. After forskolin treatment, the presence of endothelial cells was shown to upregulate PPAR coactivator 1 alpha (PGC-1α) and leptin, but not uncoupling protein 1 (UCP1), suggesting a specific crosstalk that enhances early stages of browning.

scholarly journals The effects of fibronectin on the adhesion and migration of chinese hamster ovary cells on collagen substrata

1981 ◽  
Vol 49 (1) ◽  
pp. 299-310 ◽  
Author(s):  
S.L. Schor ◽  
A.M. Schor ◽  
G.W. Bazill
Keyword(s):  
Chinese Hamster Ovary ◽  
Type I Collagen ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Type I ◽  
Collagen Fibres ◽  
Ovary Cells ◽  
3 Dimensional ◽  
Independent Mechanism ◽  
Dependent Mechanism

Data are presented indicating that the adhesion of Chinese hamster ovary cells (CHO) to films of denatured type I collagen occurs by a fibronectin-dependent mechanism, whereas the adhesion of these cells to 3-dimensional gels of native type I collagen fibres may occur by either a rapid, fibronectin-dependent mechanism or by a slower, fibronectin-independent mechanism, whereas the adhesion of these cells to 3-dimensional gels of native type I collagen fibres may occur by either a rapid, fibronectin-dependent mechanism or by a slower, fibronectin-independent mechanism. Data are also presented indicating that fibronectin promotes the migration of CHO cells on native type I collagen fibres.

scholarly journals Fiber Crimp Confers Matrix Mechanical Nonlinearity, Regulates Endothelial Cell Mechanosensing, and Promotes Microvascular Network Formation

2020 ◽  
Vol 142 (11) ◽  
Author(s):  
Christopher D. Davidson ◽  
Danica Kristen P. Jayco ◽  
William Y. Wang ◽  
Ariella Shikanov ◽  
Brendon M. Baker
Keyword(s):  
Type I Collagen ◽  
Network Formation ◽  
Critical Role ◽  
Type I ◽  
Matrix Remodeling ◽  
Simple Method ◽  
Nonlinear Stress ◽  
Nonlinear Mechanical ◽  
Hydrophilic Peptide ◽  
Fibrous Matrices

Abstract Mechanical interactions between cells and their surrounding extracellular matrix (ECM) guide many fundamental cell behaviors. Native connective tissue consists of highly organized, 3D networks of ECM fibers with complex, nonlinear mechanical properties. The most abundant stromal matrix component is fibrillar type I collagen, which often possesses a wavy, crimped morphology that confers strain- and load-dependent nonlinear mechanical behavior. Here, we established a new and simple method for engineering electrospun fibrous matrices composed of dextran vinyl sulfone (DexVS) with controllable crimped structure. A hydrophilic peptide was functionalized to DexVS matrices to trigger swelling of individual hydrogel fibers, resulting in crimped microstructure due to the fixed anchorage of fibers. Mechanical characterization of these matrices under tension confirmed orthogonal control over nonlinear stress–strain responses and matrix stiffness. We next examined ECM mechanosensing of individual endothelial cells (ECs) and found that fiber crimp promoted physical matrix remodeling alongside decreases in cell spreading, focal adhesion area, and nuclear localization of Yes-associated protein (YAP). These changes corresponded to an increase in migration speed, along with evidence for long-range interactions between neighboring cells in crimped matrices. Interestingly, when ECs were seeded at high density in crimped matrices, capillary-like networks rapidly assembled and contained tube-like cellular structures wrapped around bundles of synthetic matrix fibers due to increased physical reorganization of matrix fibers. Our work provides an additional level of mechanical and architectural tunability to synthetic fibrous matrices and implicates a critical role for mechanical nonlinearity in EC mechanosensing and network formation.

scholarly journals Cell migration through three-dimensional gels of native collagen fibres: collagenolytic activity is not required for the migration of two permanent cell lines

1980 ◽  
Vol 46 (1) ◽  
pp. 171-186 ◽  
Author(s):  
S.L. Schor ◽  
T.D. Allen ◽  
C.J. Harrison
Keyword(s):  
Cell Line ◽  
Type I Collagen ◽  
Chinese Hamster Ovary Cells ◽  
Three Dimensional ◽  
Chinese Hamster ◽  
Collagen Gel ◽  
Collagenolytic Activity ◽  
Type I ◽  
Collagen Fibres ◽  
Collagen Gel Matrix

Three dimensional gels of native type I collagen fibres have been used as a substratum for the growth and migration of Chinese hamster ovary cells (fibroblastoid cell line) and RPMI-3460 melanoma cells (tumorigenic cell line from Syrian hamster). Quantitative data concerning the migration of these cells from the gel surface into the 3-dimensional collagen gel matrix have been obtained. The migration of both cell types into the collagen gel matrix is not accompanied by the degradation of collagen fibres. The possible implications of these observations for tumour cell invasion in vivo are discussed.

The Interaction of von Willebrand Factor (vWf) with Collagen: Investigation of vWf-Binding Sites in the Collagen Molecule

1988 ◽  
Vol 59 (02) ◽  
pp. 186-192 ◽  
Author(s):  
C M Fitzsimmons ◽  
C G Cockburn ◽  
V Hornsey ◽  
C V Prowse ◽  
M J Barnes
Keyword(s):  
Binding Sites ◽  
Type I Collagen ◽  
Collagen Type ◽  
Wide Distribution ◽  
Helical Conformation ◽  
Carboxyl Groups ◽  
Collagen Molecule ◽  
Type I ◽  
Collagen Fibres ◽  
Arginyl Residues

SummaryFollowing fragmentation of the collagen molecule, we have examined the ability of the isolated fragments to bind vWf. In view of the importance of collagen tertiary and quaternary structure for binding, fragments were first renatured to restore triple-helical conformation and then polymerized. Results indicate the presence of specific vWf-binding sites in both the αl(I)-and α2(I)-chains of type I collagen. Cleavage of the αl(I)-chain with cyanogen bromide suggests the presence of at least four (conceivably several more) binding sites implying a wide distribution of sites along the length of the collagen type I molecule. Collagen type III appears to possess a similar wide distribution of sites. Chemical modification of specific amino acid residues indicates that interaction involves arginyl residues in collagen and carboxyl groups in vWf. Although interaction between fibronec-tin and collagen fibres also involves collagen arginyl residues and carboxyl groups in fibronectin (authors’ unpublished results), fibronectin does not compete with vWf in the binding to collagen fibres.

Effect of Liquid/Powder Ratio on the Setting, Handling and Mechanical Properties of Collagen–Apatitic Cements

2011 ◽  
Vol 493-494 ◽  
pp. 415-421 ◽  
Author(s):  
N.J. Dunne ◽  
R. O’Hara ◽  
F. Buchanan ◽  
J. Orr
Keyword(s):  
Calcium Phosphate ◽  
Type I Collagen ◽  
Mechanical Stability ◽  
Volume Fraction ◽  
Fibre Volume Fraction ◽  
Hardened Cement ◽  
Type I ◽  
Fibre Reinforcement ◽  
Compressive Properties ◽  
Collagen Fibres

Due to their insufficient mechanical stability and brittle nature, calcium phosphate cements (CPCs) have not been used for the treatment of vertebral fractures. Mechanical stability of human bone is provided by a complex interaction of type I collagen fibres and hydroxyapatite crystals. In the present study, fibre reinforcement of an apatitic calcium phosphate prepared at different liquid/powder ratio (LPR) was investigated. Different lengths of type I collagen fibres sourced from bovine Achilles tendon were used. Compressive strength and fracture behaviour were examined. Fibre addition of up to 5 wt.% had a significant influence on the compressive properties of the CPC. The mechanism of fibre reinforcement appeared to be crack bridging. Setting time and injectability of the CPC with fibre reinforcement was also investigated and decreased with fibre volume fraction. Increasing the LPR, improved the injectability and delayed the setting reaction. However, the compressive properties of the hardened cement were reduced as a consequence.

The effects of EGTA and trypsin on the serum requirements for cell attachment to collagens

1979 ◽  
Vol 40 (1) ◽  
pp. 271-279
Author(s):  
S.L. Schor
Keyword(s):  
Type I Collagen ◽  
Cell Attachment ◽  
Calf Serum ◽  
Foetal Calf Serum ◽  
Type I ◽  
Collagen Fibres ◽  
3 Dimensional ◽  
Mediate Cell ◽  
Glass Surfaces

Cells growing on plastic or glass surfaces in vitro may be brought into suspension by proteases (e.g. trypsin) or chelating agents (e.g. EGTA). Trypsin and EGTA remove different quantities and types of molecules from cell surfaces. Previous studies have revealed that when confluent cultures of either BHK or PyBHK cells are brought into suspension by exposure to trypsin, foetal calf serum (or fibronectin) is required for cell attachment to films of denature type I collagen, but not to 3-dimensional gels of native collagen fibres. In this communication the serum requirements for the attachment of BHK and PyBHK cells to collagen substrata have been examined as a function of (a) the method used to prepare the cell suspension (EGTA or trypsin), and (b) cell density. Data are presented consistent with the view that cell surface-associated fibronectin is able to mediate cell attachment directly to films of denatured collagen.

scholarly journals Wide Angle X-Ray Scattering to Study the Atomic Structure of Polymeric Fibers

Crystals ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 274 ◽  
Author(s):  
Teresa Sibillano ◽  
Alberta Terzi ◽  
Liberato De Caro ◽  
Massimo Ladisa ◽  
Davide Altamura ◽  
...  
Keyword(s):  
Textile Industry ◽  
Type I Collagen ◽  
Functional Materials ◽  
Building Blocks ◽  
Type I ◽  
Wide Angle ◽  
X Ray ◽  
Fiber Assembly ◽  
X Ray Scattering ◽  
Ray Scattering

Natural fibrillar-like macromolecules find applications in several fields, thanks to their peculiar features, and are considered perfect building blocks for natural and artificial functional materials. Indeed, fibrous proteins (such as collagen or fibroin) are commonly used in scaffold fabrication for biomedical applications, due to the high biophysical similarity with the extracellular matrix (ECM) which stimulates tissue regeneration. In the textile industry, cellulose-based fabrics are widely used in place of cotton and viscose, which both have sustainability issues related to their fabrication. With this in mind, the structural characterization of the materials at molecular scale plays a fundamental role in gaining insight into the fiber assembly process. In this work, we report on three fibers of research interest (i.e., type I collagen, silk fibroin extracted from Bombyx mori, and cellulose) to show the power of wide-angle X-ray scattering to characterize both intra- and intermolecular parameters of fibrous polymers. The latest possibilities offered in the X-ray scattering field allow one to study fibers at solid state or dispersed in solutions as well as to perform quantitative scanning X-ray microscopy of tissues entirely or partially made by fibers.

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