scholarly journals Crack Propagation Versus Fiber Alignment in Collagen Gels: Experiments and Multiscale Simulation

2015 ◽  
Vol 137 (12) ◽  
Author(s):  
Sarah M. Vanderheiden ◽  
Mohammad F. Hadi ◽  
V. H. Barocas
Keyword(s):  
Collagen Gel ◽  
Structural Complexity ◽  
Complex Loading ◽  
Multiscale Simulation ◽  
Failure Behavior ◽  
Multiscale Approach ◽  
Collagen Gels ◽  
Wide Range ◽  
Sample Width

It is well known that the organization of the fibers constituting a collagenous tissue can affect its failure behavior. Less clear is how that effect can be described computationally so as to predict the failure of a native or engineered tissue under the complex loading conditions that can occur in vivo. Toward the goal of a general predictive strategy, we applied our multiscale model of collagen gel mechanics to the failure of a double-notched gel under tension, comparing the results for aligned and isotropic samples. In both computational and laboratory experiments, we found that the aligned gels were more likely to fail by connecting the two notches than the isotropic gels. For example, when the initial notches were 30% of the sample width (normalized tip-to-edge distance = 0.7), the normalized tip-to-tip distance at which the transition occurred from between-notch failure to across-sample failure shifted from 0.6 to 1.0. When the model predictions for the type of failure event (between the two notches versus across the sample width) were compared to the experimental results, the two were found to be strongly covariant by Fisher’s exact test (p < 0.05) for both the aligned and isotropic gels with no fitting parameters. Although the double-notch system is idealized, and the collagen gel system is simpler than a true tissue, it presents a simple model system for studying failure of anisotropic tissues in a controlled setting. The success of the computational model suggests that the multiscale approach, in which the structural complexity is incorporated via changes in the model networks rather than via changes to a constitutive equation, has the potential to predict tissue failure under a wide range of conditions.

scholarly journals The selective binding and transmigration of monocytes through the junctional complexes of human endothelium.

1988 ◽  
Vol 168 (5) ◽  
pp. 1865-1882 ◽  
Author(s):  
N A Pawlowski ◽  
G Kaplan ◽  
E Abraham ◽  
Z A Cohn
Keyword(s):  
Silver Staining ◽  
Umbilical Vein ◽  
Collagen Gel ◽  
Collagen Gels ◽  
Silver Stain ◽  
Term Survival ◽  
Topographical Distribution ◽  
Junctional Complexes

Human monocytes show a high affinity for vascular endothelium both in vitro and in vivo. To explore monocyte-endothelial interaction in greater detail, we have developed a new in vitro model for growth of human endothelial cells (EC). Human umbilical vein EC (HUVEC) cultured upon collagen gels form confluent monolayers of EC that bind silver at their intercellular border similar to cells in situ. Intercellular junctional structures, both adherens and tight junctions, were identified. In contrast, HUVEC grown on plastic surfaces did not stain with silver. The silver-staining characteristic of EC-collagen monolayers was reversible and related to their in vitro maturation and senescence. Silver staining of EC borders provided a grid by which the location of monocyte binding to the luminal surface of individual EC could be assessed. Using this technique, we found that monocytes preferentially bound to the margins of EC, in approximation to the silver-staining junctions. These results suggest that EC determinants recognized by monocytes occur in a unique topographical distribution on the apical face of EC. After binding, monocytes migrated through the EC monolayers at high basal rates. The lack of penetration of collagen gels in the absence of an EC monolayer suggested the generation of EC-specific chemotactic signal(s). Monocytes were observed to pass between EC without evidence of disruption of the monolayer. Silver stain remained present during all phases of migration, and under transmission electron microscopy, junctional complexes were found proximal to monocytes that had just completed their passage through the monolayer. After orientation to the basal surface of the EC monolayer, monocytes migrated randomly into the underlying collagen gel. Monocyte adherence, penetration, migration, and long term survival can be studied under these conditions.

Prolactin effects on ion transport across cultured mouse mammary epithelium

1981 ◽  
Vol 240 (3) ◽  
pp. C110-C115 ◽  
Author(s):  
C. A. Bisbee
Keyword(s):  
Cell Cultures ◽  
Short Circuit ◽  
Collagen Gel ◽  
Mammary Epithelium ◽  
Short Circuit Current ◽  
Sodium Absorption ◽  
Collagen Gels ◽  
Mouse Mammary Epithelium

Prolactin is a known osmoregulatory hormone in lower vertebrates, and recent evidence indicates that this hormone modulates ionic concentrations in milk. In an ultrastructurally and biochemically differentiated primary cell culture system in which mouse mammary epithelium is maintained on floating collagen gels, prolactin causes an increase in short-circuit current (Isc) of monolayers of cells derived from midpregnant (24.6 to 48.0 microA . cm-2) and lactating (10.4 to 16.1 microA . cm-2) glands. Transepithelial potential differences (basal side ground) average about -12 mV and are similar to those seen in vivo. Prelactating mammary epithelial cell cultures have transepithelial resistances ranging from 374 omega . cm2 (prolactin present) to 507 omega . cm2 (prolactin absent), and lactating cell cultures have resistances averaging almost 1,000 omega . cm2. Prolactin effects require at most one day of culture maintenance in prolactin-containing medium, and the effects are not due to known contamination of prolactin preparations with arginine vasopressin or growth hormone. Medium concentrations of prolactin as low as 1 ng/ml can elicit these effects. In prelactating cell cultures not treated with prolactin, the Isc is equal to the rate of sodium absorption. Prolactin increases sodium absorption fourfold but increases Isc only twofold. Clearly, prolactin induces other active transport; neither potassium nor chloride movements can account for this additional transport. Resistance values, current-voltage plots, and permeability coefficients indicate that in vitro mammary epithelium is a moderately “tight” tissue. Comparisons with intact glands indicate that in vitro mammary epithelium closely resembles its in vivo counterpart. Floating collagen gel cultures appear suitable for elucidating transport properties in cellularly heterogeneous and structurally complex mammalian tissues.

Antibacterial Wound Healing Gels from Oligochitosan Salts

2012 ◽  
Vol 506 ◽  
pp. 31-34
Author(s):  
W. Janvikul ◽  
P. Ngamviriyavong ◽  
P. Uppanun ◽  
P. Tanjak ◽  
N. Sangjun
Keyword(s):  
Staphylococcus Epidermidis ◽  
Wound Closure ◽  
Collagen Gel ◽  
Antibacterial Activities ◽  
Dermal Fibroblasts ◽  
Collagen Gels ◽  
Collagen Gel Matrix ◽  
Growth Of Bacteria

Oligochitosan salt-based antibacterial wound gels were developed and evaluated in both in vitro and in vivo models. The antibacterial activities of the oligochitosan salts and the wound gels were investigated against Staphylococcus epidermidis RP625 and Escherichia coli ATCC 11775. The minimum inhibitory concentrations (MIC) of the oligochitosan salts were found in the range of 16-256 μg/mL. The wound gels demonstrated their in vitro activities on inhibiting the growth of bacteria. The 3-D collagen gel matrix containing human dermal fibroblasts cultured with each test gel was used as an in vitro model for the examination of cell proliferation and secretion of interleukin-8 (IL-8). The gels appeared to promote the proliferation and formation of cellular process of the fibroblasts in the 3-D collagen gels and stimulate the fibroblasts to produce more IL-8. In the in vivo model, it was noted that the gels could accelerate the wound closure process. The wounds were completely closed within 14 days.

scholarly journals Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells.

1982 ◽  
Vol 95 (1) ◽  
pp. 333-339 ◽  
Author(s):  
G Greenburg ◽  
E D Hay
Keyword(s):  
Epithelial Cells ◽  
Cell Polarity ◽  
Three Dimensional ◽  
Collagen Gel ◽  
Mesenchymal Cells ◽  
Apical Surface ◽  
Collagen Gels ◽  
Mesenchymal Transformation

This study of epithelial-mesenchymal transformation and epithelial cell polarity in vitro reveals that environmental conditions can have a profound effect on the epithelial phenotype, cell shape, and polarity as expressed by the presence of apical and basal surfaces. A number of different adult and embryonic epithelia were suspended within native collagen gels. Under these conditions, cells elongate, detach from the explants, and migrate as individual cells within the three-dimensional lattice, a previously unknown property of well-differentiated epithelia. Epithelial cells from adult and embryonic anterior lens were studied in detail. Elongated cells derived from the apical surface develop pseudopodia and filopodia characteristic of migratory cells and acquire a morphology and ultrastructure virtually indistinguishable from that of mesenchymal cells in vivo. It is concluded from these experiments that the three-dimensional collagen gel can promote dissociation, migration, and acquisition of secretory organelles by differentiated epithelial cells, and can abolish the apical-basal cell polarity characteristic of the original epithelium.

Hydration and Crosslinking Effects on the Elastic and Viscoelastic Properties of Collagen Scaffolds

2011 ◽  
Author(s):  
Bin Xu ◽  
Katherine Yanhang Zhang
Keyword(s):  
Thin Film ◽  
Solid Phase ◽  
Great Influence ◽  
Collagen Gel ◽  
Fluid Phase ◽  
The Body ◽  
Collagen Gels ◽  
Collagen Scaffolds ◽  
Fiber Network

Collagen is the most abundant protein in the body. It plays critical roles in many supporting and connecting tissues such as tendon, ligament, bone, blood vessels, skin, etc. Collagen gels prepared from commercially available collagen solutions mimic the in vivo environment and have been widely used as three-dimensional (3-D) tissue scaffolds for cell culture. Collagen thin film is the dehydrated form of collagen gel. A number of studies have examined the cell-collagen thin film interactions (1, 2). As a biphasic material, collagen scaffolds contain a solid phase which represents by collagen fiber network and an interstitial fluid phase (3). This special structure makes collagen a viscoelastic material. Viscoelasticity is related to force or energy storage, transmission and dissipation in tissues and has a great influence on the growth and development of cells (4).

scholarly journals The cxc chemokine cCAF stimulates differentiation of fibroblasts into myofibroblasts and accelerates wound closure

2002 ◽  
Vol 156 (1) ◽  
pp. 161-172 ◽  
Author(s):  
Jo Ellen Feugate ◽  
QiJing Li ◽  
Lina Wong ◽  
Manuela Martins-Green
Keyword(s):  
Granulation Tissue ◽  
Wound Closure ◽  
Smooth Muscle Actin ◽  
Collagen Gel ◽  
Angiogenic Factor ◽  
Collagen Gels ◽  
Collagen Gel Contraction ◽  
Cxc Chemokine ◽  
Pharmacological Manipulations

Chemokines are small cytokines primarily known for their roles in inflammation. More recently, however, they have been implicated in processes involved in development of the granulation tissue of wounds, but little is known about their functions during this process. Fibroblasts play key roles in this phase of healing: some fibroblasts differentiate into myofibroblasts, α-smooth muscle actin (SMA)-producing cells that are important in wound closure and contraction. Here we show that the CXC chemokine chicken chemotactic and angiogenic factor (cCAF) stimulates fibroblasts to produce high levels of α-SMA and to contract collagen gels more effectively than do normal fibroblasts, both characteristic properties of myofibroblasts. Specific inhibition of α-SMA expression resulted in abrogation of cCAF-induced contraction. Furthermore, application of cCAF to wounds in vivo increases the number of myofibroblasts present in the granulation tissue and accelerates wound closure and contraction. We also show that these effects in culture and in vivo can be achieved by a peptide containing the NH2-terminal 15 amino acids of the cCAF protein and that inhibition of α-SMA expression also results in inhibition of N-peptide–induced collagen gel contraction. We propose that chemokines are major contributors for the differentiation of fibroblasts into myofibroblasts during formation of the repair tissue. Because myofibroblasts are important in many pathological conditions, and because chemokines and their receptors are amenable to pharmacological manipulations, chemokine stimulation of myofibroblast differentiation may have implications for modulation of functions of these cells in vivo.

scholarly journals TGF-beta 1 influences the relative development of the exocrine and endocrine pancreas in vitro

Development ◽  
1994 ◽  
Vol 120 (12) ◽  
pp. 3451-3462 ◽  
Author(s):  
F. Sanvito ◽  
P.L. Herrera ◽  
J. Huarte ◽  
A. Nichols ◽  
R. Montesano ◽  
...  
Keyword(s):  
Endocrine Cells ◽  
Three Dimensional ◽  
Collagen Gel ◽  
Tgf Beta ◽  
Collagen Gels ◽  
Paracrine Factors ◽  
Fold Family ◽  
Acinar Tissue

Pancreatic rudiments from E12.5 mouse embryos undergo extensive development and differentiation when cultured in three-dimensional gels of extracellular matrix proteins for up to 12 days. Whereas collagen gels promote the formation of numerous exocrine acini and relatively small clusters of endocrine cells, in basement membrane (EHS) matrices the development of endocrine cells is dramatically favoured over that of acinar tissue. Buds embedded in a collagen gel contiguous to an EHS gel also fail to develop acini, suggesting the involvement of diffusible factor(s). Addition of cytokines to cultures of pancreatic buds in collagen gels modifies the relative proportions of the epithelial components of the gland. In the presence of EGF the proportion of the tissue occupied by ducts overrides that of acinar structures, whereas the endocrine portion of the tissue is not significantly modified. TGF-beta 1 partially mimicks the effect of EHS matrix in inhibiting the development of acinar tissue without decreasing the amount of ducts and mesenchyme; TGF-beta 1 also promotes the development of endocrine cells, in particular of insulin-containing beta cells and of cells expressing genes of the PP-fold family. These results show that cytokines can modulate the development of the pancreas and suggest a role for TGF-beta 1 in regulating the balance between the acinar and endocrine portions of the gland in vivo. More generally, they are compatible with the notion that, during organogenesis, cytokines act as paracrine factors responsible for the development and maintenance of appropriate proportions of different tissue constituents.

MMP-3 Response to Compressive Forces in vitro and in vivo

2008 ◽  
Vol 87 (7) ◽  
pp. 692-696 ◽  
Author(s):  
H.-H. Chang ◽  
C.-B. Wu ◽  
Y.-J. Chen ◽  
C.-Y. Weng ◽  
W.-P. Wong ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Tooth Movement ◽  
Interleukin 1 ◽  
Orthodontic Tooth Movement ◽  
Compressive Force ◽  
Collagen Gel ◽  
Pcr Analysis ◽  
Wide Range

During orthodontic tooth movement, bone resorption occurs at the compression site. However, the mechanism underlying resorption remains unclear. Applying compressive force to human osteoblast-like cells grown in a 3D collagen gel, we examined gene induction by using microarray and RT-PCR analysis. Among 43 genes exhibiting significant changes, cyclo-oxygenase-2, ornithine decarboxylase, and matrix metalloproteinase-3 (MMP-3) were up-regulated, whereas membrane-bound interleukin-1 receptor accessory protein was down-regulated. The MMP-3 protein increases were further confirmed by Western blot. To ascertain whether MMP-3 is up-regulated in vivo by orthodontic force, we examined human bone samples at the compressive site by realigning the angulated molars. Immunohistochemical staining revealed MMP-3 distributed along the compressive site of the bony region within 3 days of compression. Since MMP-3 participates in degradation of a wide range of extracellular matrix molecules, we propose that MMP-3 plays an important role in bone resorption during orthodontic tooth movement.

Aortic Valve Interstitial Cell Activation Does Not Occur at Low Tissue Stiffness During Embryogenesis

2012 ◽  
Author(s):  
Young Wook Chun ◽  
Joey Barnett ◽  
W. David Merryman
Keyword(s):  
Aortic Valve ◽  
Heart Valve ◽  
Cell Activation ◽  
Transforming Growth Factor ◽  
Collagen Gel ◽  
Transforming Growth Factor Β1 ◽  
Collagen Gels ◽  
Tissue Culture Polystyrene ◽  
Prosthetic Valves

An estimated 2.5 percent of the American population has heart valve (HV) disease and more than 100,000 US patients require a prosthetic valve replacement each year [1]. However, prosthetic valves can cause accelerated calcification leading to recurrence of HV disease in patients [2]. Thus, the development of a suitable tissue-engineered heart valve (TEHV) would greatly benefit patients with HV disease. Aortic valve interstitial cells (AVICs) play a crucial role in the progression of aortic valve disease as well as the maintenance of normal valve. Therefore, in order to design a suitable TEHV, these specialized cells need to be better understood. AVICs are known to synthesize ECM and express matrix degrading enzymes and their inhibitors that mediate and regulate remodeling of ECM components [3]. Interestingly, it was recently established that AVICs sense the stiffness of their surrounding ECM in vivo and are phenotypically responsive to mechanical cues with AVICs differentiating into myofibroblasts or osteoblasts, which are pathologic markers. Specifically, soft collagen gels (∼34kPa) caused less differentiation of AVICs than stiffer collagen gel (∼100kPa) [4]. However, for these experiments the AVICs were cultured on tissue culture polystyrene (TCPS) for at least one passage, and it is likely that AVICs cultured on TCPS might retain modified characteristics of AVICs in tissue prior to seed them on soft gels because of the memory to rigid substrate stiffness. Therefore, in this study, we examined the phenotype and function of AVICs on substrates that mimic ECM stiffness of adult leaflet as well as of developing embryo. In addition, we examine the effects of transforming growth factor-β1 (TGF-β1) which has been the most extensively studied cytokine initiator of fibrotic response of AVICs.

Absence of integrin alpha1beta1 in the mouse causes loss of feedback regulation of collagen synthesis in normal and wounded dermis

1999 ◽  
Vol 112 (3) ◽  
pp. 263-272 ◽  
Author(s):  
H. Gardner ◽  
A. Broberg ◽  
A. Pozzi ◽  
M. Laato ◽  
J. Heino
Keyword(s):  
Collagen Synthesis ◽  
Collagen Gel ◽  
Feedback Regulation ◽  
Skin Thickness ◽  
Collagen Gels ◽  
Cutaneous Wounds ◽  
Dermal Thickness ◽  
Feedback Regulator

Integrin alpha1beta1 is a collagen receptor predominantly found in mesenchymal tissues. Mice lacking this receptor are viable. We have previously suggested that alpha1beta1 might participate in the down-regulation of collagen gene expression observed in cells suspended inside collagen gels. The results presented here demonstrate that integrin alpha1beta1 acts as a feedback regulator of collagen synthesis both in vitro and in vivo. Firstly, alpha1 null animals show a higher rate of collagen synthesis in the dermis in vivo. Secondly, fibroblasts derived from alpha1 null cutaneous wounds show a reduced sensitivity to collagen gel induced downregulation of collagen mRNA synthesis, as compared to their wild-type counterparts. An increase in collagenase synthesis is also seen in the alpha1 null dermis and in collagen gel suspended fibroblasts. While dermal thickness is normal in the alpha1 null animals, an increase is seen in skin thickness of alpha1 null but not alpha1 heterozygote animals on a background of collagenase resistant collagen. Increased expression of both collagen and collagenase mRNA are seen in experimental granulation tissue in alpha1 null animals, but their ultimate accumulation of collagen is normal, probably due to non alpha1 dependent paracrine regulators of collagen turnover.

Sign in / Sign up

Export Citation Format

Share Document