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

Author(s):  
Bin Xu ◽  
Katherine Yanhang Zhang

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).

1988 ◽  
Vol 168 (5) ◽  
pp. 1865-1882 ◽  
Author(s):  
N A Pawlowski ◽  
G Kaplan ◽  
E Abraham ◽  
Z A Cohn

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.


1981 ◽  
Vol 240 (3) ◽  
pp. C110-C115 ◽  
Author(s):  
C. A. Bisbee

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.


1987 ◽  
Vol 58 (2) ◽  
pp. 265-276 ◽  
Author(s):  
J. D. Allen ◽  
J. M. Gawthornet

1. Merino sheep fed on a diet of chopped wheaten hay, chopped lucerne (Medicago saliva) hay and oat grain were the source of rumen contents for the study. The diet contained (mg/kg dry weight) 3.3 copper, 0.24 molybdenum and 2.8 sulphur. The effects of adding between 5 and 25 mg Mo/kg as ammonium molybdate (AM) or tetrathiomolybdate (TTM) on the distribution and forms of Cu and Mo in rumen contents were investigated in vivo and in vitro.2. Approximately 88 % of the Cu and 94% of the Mo in rumen contents were associated with the solid phase. When AM or TTM was added to rumen contents in vivo or in vitro the proportion of these elements in the solid phase was increased at the expense of the fluid phase.3. The addition of AM and TTM to rumen contents also decreased the proportion of Cu that was soluble in trichloroacetic acid (50 g/l; TCA) and increased the proportion of Cu that was not extractable by sequential treatment with TCA and neutral detergent.4. Column chromatography of neutral-detergent extracts of rumen contents revealed that TTM treatment caused Cu to be strongly bound to proteins of high molecular weight.5. Addition of sulphide to rumen contents did not result in significant changes in the distribution of Cu between the fluid and solid phases, or in the solubility of Cu in TCA.6. It is postulated that constant removal of TTM from the fluid phase via reaction with proteins and other macromolecules in the solid phase results in greater formation of TTM in vivo than would be expected from solution chemistry. The molybdo-proteins so formed are strong chelators of Cu and may be the agents responsible for the decrease in Cu absorption in animals that consume diets containing high concentrations of Mo.


2012 ◽  
Vol 506 ◽  
pp. 31-34
Author(s):  
W. Janvikul ◽  
P. Ngamviriyavong ◽  
P. Uppanun ◽  
P. Tanjak ◽  
N. Sangjun

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.


1982 ◽  
Vol 95 (1) ◽  
pp. 333-339 ◽  
Author(s):  
G Greenburg ◽  
E D Hay

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.


Author(s):  
Rita Lala ◽  
Nikita Nandvikar

Background: Acne vulgarise is an inflammatory disease involving the pathological alteration of the sebaceous glands of the body. It is not a life-threatening disease but has a great influence on lifestyle. Topical combination therapy of vitamin A and antibacterial drugs is an effective treatment for acne. Materials and Methods: The current work investigates the nanostructure lipid colloidal carrier system of Tretinoin and Clindamycin phosphate. Nanostructured lipid carriers (NLCs) were prepared by highspeed homogenization-sonication technique and characterized for physicochemical properties, permeation, in vivo anti-acne and toxicity (acute 2000 mg/Kg, repeat 1000 mg/kg) in Wistar rats.  Results: The prepared system was found to be stable, homogenous with more site retention of drugs having non-irritation and toxicity potential. The formulation showed a size of 283 nm, polydispersity index (PDI) 0.43 and Zeta potential (ZP) -37.9 mV with drug entrapment 92.0% and 66.15% for tretinoin and clindamycin respectively. Observed permeation was 18 % and 45% for Tretinoin and Clindamycin less than marketed formulation which is more focused on dermal retention of drug. No significant abnormalities and toxicological symptoms were observed for acute and repeat dose toxicity study for histopathology and haematological examinations of organs. Conclusion: Prepared NLC formulation was aimed at epidermal targeting. Based on obtained results it is concluded that developed lipid-based nanocarrier system of selected drugs showed the targeting potential for effective acne treatment. 


2002 ◽  
Vol 156 (1) ◽  
pp. 161-172 ◽  
Author(s):  
Jo Ellen Feugate ◽  
QiJing Li ◽  
Lina Wong ◽  
Manuela Martins-Green

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.


2020 ◽  
Author(s):  
Konlin Shen ◽  
Michel M. Maharbiz

AbstractThe lifetime of neural implants is strongly dependent on packaging due to the aqueous and biochemically aggressive nature of the body. Over the last decade, there has been a drive towards neuromodulatory implants which are wireless and approaching millimeter-scales with increasing electrode count. A so-far unrealized goal for these new types of devices is an in-vivo lifetime comparable to a sizable fraction of a healthy patient’s lifetime (>10-20 years). Existing, approved medical implants commonly encapsulate components in metal enclosures (e.g. titanium) with brazed ceramic inserts for electrode feedthrough. It is unclear how amenable the traditional approach is to the simultaneous goals of miniaturization, increased channel count, and wireless communication. Ceramic materials have also played a significant role in traditional medical implants due to their dielectric properties, corrosion resistance, biocompatibility, and high strength, but are not as commonly used for housing materials due to their brittleness and the difficulty they present in creating complex housing geometries. However, thin film technology has opened new opportunities for ceramics processing. Thin films derived largely from the semiconductor industry can be deposited and patterned in new ways, have conductivities which can be altered during manufacturing to provide conductors as well as insulators, and can be used to fabricate flexible substrates. In this review, we give an overview of packaging for neural implants, with an emphasis on how ceramic materials have been utilized in medical device packaging, as well as how ceramic thin film micromachining and processing may be further developed to create truly reliable, miniaturized, neural implants


Development ◽  
1994 ◽  
Vol 120 (12) ◽  
pp. 3451-3462 ◽  
Author(s):  
F. Sanvito ◽  
P.L. Herrera ◽  
J. Huarte ◽  
A. Nichols ◽  
R. Montesano ◽  
...  

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.


2015 ◽  
Vol 137 (12) ◽  
Author(s):  
Sarah M. Vanderheiden ◽  
Mohammad F. Hadi ◽  
V. H. Barocas

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.


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