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Collagen Fibrils (collagen + fibril)
Terms modified by Collagen Fibrils Selected AbstractsModeling of Bovine Type-I Collagen Fibrils: Interaction with Pickling and Retanning AgentsMACROMOLECULAR BIOSCIENCE, Issue 2 2007Rosa E. Bulo Abstract Bovine Type I collagen was investigated, building on a large scale computer model of a collagen fibril in water, and focusing on two stages of the leather manufacturing process. The effects of different salts (NaCl, CaCl2, and Na2SO4) on the swelling behavior of collagen at low pH (the pickling process) were studied. The salts suppress the swelling of the fibrils at low pH and we find specific stabilizing influences for CaCl2 and Na2SO4, due to weak Ca2+/Cl, and strong SO/lysine/arginine interactions, respectively. Using state-of-the-art sampling techniques, such as the metadynamics algorithm, to allow an efficient exploration of configuration space, we were able to investigate the effect of polyacrylate and poly(methyl acrylate) , two polymeric retanning agents , on the fibril. Both polymers interact with the ammonium groups on the surface, but polyacrylate shows significantly stronger interactions. We suggest that it is this stronger interaction that contributes to the reduced suitability of PAA as a tanning agent. [source] Rho plays a central role in regulating local cell-matrix mechanical interactions in 3D cultureCYTOSKELETON, Issue 6 2007N. Lakshman Abstract The purpose of this study was to assess quantitatively the role of the small GTPase Rho on cell morphology, f-actin organization, and cell-induced matrix remodeling in 3D culture. Human corneal fibroblasts (HTK) were infected with adenoviruses that express green fluorescent protein (GFP) or GFP-N19Rho (dominant negative Rho). One day later cells were plated inside collagen matrices and allowed to spread for 24 h. Cells were fixed and stained for f-actin. Fluorescent (for f-actin) and reflected light (for collagen fibrils) images were acquired using confocal microscopy. Fourier transform analysis was used to assess local collagen fibril alignment, and changes in cell morphology and collagen density were measured using MetaMorph. The decrease in matrix height was used as an indicator of global matrix contraction. HTK and HTK-GFP cells induced significant global matrix contraction; this was inhibited by N19Rho. HTK and HTK-GFP fibroblasts generally had a bipolar morphology and occasional intracellular stress fibers. Collagen fibrils were compacted and aligned parallel to stress fibers and pseudopodia. In contrast, HTK-GFPN19 cells were elongated, and had a more cortical f-actin distribution. Numerous small extensions were also observed along the cell body. In addition, both local collagen fibril density and alignment were significantly reduced. Rho plays a key role in regulating both the morphology and mechanical behavior of corneal fibroblasts in 3D culture. Overall, the data suggest that Rho-kinase dependent cell contractility contributes to global and local matrix remodeling, whereas Rho dependent activation of mDia and/or other downstream effectors regulates the structure and number of cell processes. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source] Tenderization of Heated Sliced Beef by Succinylated Glycerol Monostearate, a Novel Meat TenderizerJOURNAL OF FOOD SCIENCE, Issue 4 2001K. Mori ABSTRACT Succinylated glycerol monostearate (SGMS) was most effective in reducing shear-force value when used as a surfactant to tenderize meat using sheep casings as a model of intramuscular connective tissue. Roasted beef slices that had been treated with 2% granular SGMS showed a marked decrease in toughness due to the penetration of SGMS along the perimysium during heating. Collagen fibrils in the perimysium were transformed into a sheet-like structure that seemed very likely to be a complex of SGMS and thermally denatured collagen. These structural changes could account for the tenderness of the roasted beef slices. [source] Nanostructure of collagen fibrils in human nucleus pulposus and its correlation with macroscale tissue mechanicsJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 4 2010Darwesh M.K. Aladin Abstract Collagen fibrils are the main structural components of the nucleus pulposus tissue in the intervertebral discs. The structure,property relationship of the nucleus pulposus (NP) tissues is still unclear. We investigated the structure of individual collagen fibrils of the NP and evaluated its correlation with the bulk mechanical properties of the tissue. Collagen fibrils were extracted from the NP of discs retrieved from adolescents during scoliosis correction surgery, and the extracts were confirmed by SDS-PAGE. The diameters of the individual collagen fibrils were measured through atomic force microscopy, and the compressive mechanical properties of the tissues were evaluated by confined compression. The correlations between the nanoscale morphology of the collagen fibrils and the macroscale mechanical properties of the tissues were evaluated by linear regression. The SDS-PAGE results showed that the fibril extracts were largely composed of type II collagen. The mean diameter of the collagen fibrils was 92.1,±,26.54 nm; the mean swelling pressure and compressive modulus of the tissues were 6.15,±,4.3 kPa and 1.23,±,0.7 MPa, respectively. The mean fibril diameter had no linear correlation (R2,=,0.30) with the swelling pressure of the tissues. However, it had a mild linear correlation with the compressive modulus (p,=,0.023, R2,=,0.68). This is the first study, to our knowledge, to evaluate the nanostructure of the individual collagen fibrils of the nucleus pulposus and its relationship with macroscale mechanical properties of the NP tissues. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:497,502, 2010 [source] A novel flow cytometric analysis for platelet activation on immobilized von Willebrand factor or fibrillar collagenJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 2 2003S. Kao Summary., Under flow conditions, platelets adhere singly or in small aggregates on von Willebrand factor (VWF)-coated surfaces, but form large aggregates on immobilized fibrillar collagen. We developed a novel flow cytometric analysis to study the mechanisms underlying these distinct platelet deposition patterns. Flow cytometry was used to measure platelet activation after platelet adherence onto microspheres coated with either VWF or collagen fibrils. Two representative indices were calculated to quantify activated GpIIb,IIIa and P-selectin expression on adherent platelets. The signaling pathways responsible for platelet activation after interacting with fibrillar collagen were elucidated using various inhibitors. An in vitro endothelial cell wound model was also used to study the roles of VWF and fibrillar collagen in platelet deposition onto subendothelial matrixes. The adherent platelets on fibrillar collagen express more activated GpIIb,IIIa and P-selectin than those on VWF. Activation of GpIIb,IIIa and expression of P-selectin after platelet interaction with collagen occur via different intracellular signaling pathways; however, Ca2+ released from intracellular pools is common to both phenomena. Platelets were deposited singly or formed small aggregates on the endothelial cell wounded area, and this deposition pattern was dependent on VWF molecules secreted by endothelial cells and the absence of subendothelial collagen fibrils. As less activated GpIIb,IIIa and P-selectin are expressed after platelets interact with immobilized VWF alone, subsequent flowing platelet recruitment is minimal. Collagen fibrils, however, can activate adherent platelets sufficiently to promote the formation of large platelet aggregates. [source] Remodelling of collagen fibrils and proteoglycans in the zebrafish cornea during developmentACTA OPHTHALMOLOGICA, Issue 2007S AKHTAR Purpose: Collagen fibrils and proteoglycans are the main components of the corneal extracellular matrix and corneal transparency depends crucially on their proper organisation. We investigated their formation and arrangement in the developing cornea of the zebrafish, a major model of vertebrate development and genetic disease. Methods: We employed thin-section electron microscopy to investigate the ultrastructure of the zebrafish cornea at different stages of development. Results: Layering of the zebrafish cornea into an epithelium, Bowman's layer, stroma and endothelium was observed by 72 hours post-fertilization. At this stage, the stroma contained orthogonally arranged collagen fibrils and small proteoglycans. The density of proteoglycans increased gradually throughout subsequent development. In the stroma of 2 week old larvae, the collagen fibrils were organized into thin lamellae for the first time and were separated by very large, randomly distributed proteoglycans. At 4 weeks, a regular arrangement of proteoglycans around the collagen fibrils was observed for the first time and the lamellae also thickened. Conclusions: This is the first report of collagen fibril and proteoglycan development in the zebrafish cornea and it directly correlates collagen fibril and proteoglycan organisation of the zebrafish cornea with that of the human cornea. The similarities between the two species, including the possession of a Bowman layer, suggest that the zebrafish could serve as a model for the genetics of human corneal development and inherited disease. [source] Sub-micron spongiform porosity is the major ultra-structural alteration occurring in archaeological boneINTERNATIONAL JOURNAL OF OSTEOARCHAEOLOGY, Issue 6 2002G. Turner-Walker Abstract Total pore volume and pore size distribution are indicators of the degree of post-mortem modification of bone. Direct measurements of pore size distribution in archaeological bones using mercury intrusion porosimetry (HgIP) and back scattered scanning electron microscopy (BSE-SEM) reveal a common pattern in the changes seen in degraded bone as compared to modern samples. The estimates of pore size distribution from HgIP and direct measurement from the BSE-SEM images show remarkable correspondence. The coupling of these two independent approaches has allowed the diagenetic porosity changes in human archaeological bone in the >0.01 µm range to be directly imaged, and their relationship to pre-existing physiological pores to be explored. The increase in porosity in the archaeological bones is restricted to two discrete pore ranges. The smaller of these two ranges (0.007,0.1 µm) lies in the range of the collagen fibril (0.1 µm diameter) and is presumably formed by the loss of collagen, whereas the larger pore size distribution is evidence of direct microbial alteration of the bone. HgIP has great potential for the characterization of microbial and chemical alteration of bone. Copyright © 2002 John Wiley & Sons, Ltd. [source] Modeling of Bovine Type-I Collagen Fibrils: Interaction with Pickling and Retanning AgentsMACROMOLECULAR BIOSCIENCE, Issue 2 2007Rosa E. Bulo Abstract Bovine Type I collagen was investigated, building on a large scale computer model of a collagen fibril in water, and focusing on two stages of the leather manufacturing process. The effects of different salts (NaCl, CaCl2, and Na2SO4) on the swelling behavior of collagen at low pH (the pickling process) were studied. The salts suppress the swelling of the fibrils at low pH and we find specific stabilizing influences for CaCl2 and Na2SO4, due to weak Ca2+/Cl, and strong SO/lysine/arginine interactions, respectively. Using state-of-the-art sampling techniques, such as the metadynamics algorithm, to allow an efficient exploration of configuration space, we were able to investigate the effect of polyacrylate and poly(methyl acrylate) , two polymeric retanning agents , on the fibril. Both polymers interact with the ammonium groups on the surface, but polyacrylate shows significantly stronger interactions. We suggest that it is this stronger interaction that contributes to the reduced suitability of PAA as a tanning agent. [source] Combined role of type IX collagen and cartilage oligomeric matrix protein in cartilage matrix assembly: Cartilage oligomeric matrix protein counteracts type IX collagen,induced limitation of cartilage collagen fibril growth in mouse chondrocyte culturesARTHRITIS & RHEUMATISM, Issue 12 2009K. Blumbach Objective Defects in the assembly and composition of cartilage extracellular matrix are likely to result in impaired matrix integrity and increased susceptibility to cartilage degeneration. The aim of this study was to determine the functional interaction of the collagen fibril,associated proteins type IX collagen and cartilage oligomeric matrix protein (COMP) during cartilage matrix formation. Methods Primary chondrocytes from mice deficient in type IX collagen and COMP (double-deficient) were cultured in monolayer or alginate beads. Anchorage of matrix proteins, proteoglycan and collagen content, collagen crosslinks, matrix metalloproteinase activity, and mechanical properties of the matrix were measured. Electron microscopy was used to study the formation of fibrillar structures. Results In cartilage lacking both type IX collagen and COMP, matrilin 3 showed decreased matrix anchorage. Less matrilin 3 was deposited in the matrix of double-deficient chondrocytes, while larger amounts were secreted into the medium. Proteoglycans were less well retained in the matrix formed in alginate cultures, while collagen deposition was not significantly affected. Electron microscopy revealed similar cartilage collagen fibril diameters in the cultures of double-deficient and wild-type chondrocytes. In contrast, a larger fibril diameter was observed in the matrix of chondrocytes deficient in only type IX collagen. Conclusion Our results show that type IX collagen and COMP are involved in matrix assembly by mediating the anchorage and regulating the distribution of other matrix macromolecules such as proteoglycans and matrilins and have counteracting effects on collagen fibril growth. Loss of type IX collagen and COMP leads to matrix aberrations that may make cartilage more susceptible to degeneration. [source] 2142: Ulrastructural features of keratoconus cornea after cross-linking by riboflavin/UVAACTA OPHTHALMOLOGICA, Issue 2010S AKHTAR Purpose In the present studies we assess the effects of collagen cross-linking on ultrastructure organisation of the corneal stroma of keratoconus human corneas. Methods One normal, one keratoconus (KC) and three cross-linked keratoconus corneas were analysed. One was treated with standard cross linking (SXL) and two with trans-epithelial collagen cross linking (TEXL). Penetrating keratoplasty was performed three months after treatment. All samples were fixed in 2.5% glutaraldehyde containing cuprolinic blue in sodium acetate buffer and processed for electron microscopy. Results The structure of SXL corneas was very similar to normal corneas in their hemidesmosomes, basement membrane (BM), Bowman's layer (BW) and stromal lamellae that were not undulated. The architecture of TEXL corneas presented some differences. The BM was thick with degenerated hemidesmosomes. Bowman's layer was disorganised at some places and replaced by thin filaments forming pannus. There were thin undulating lamellae in anterior, middle and posterior stroma. The keratocytes were embedded between undulating lamellae. Large amounts of abnormal PGs were attached around collagen fibrils. The parallel running lamellae were very thin. In some parts of the anterior stroma collagen fibrils were oriented (running) in random directions instead of running parallel. There were some parts of the stroma which showed a normal appearance. Conclusion The present studies demonstrate that corneal cross-linking leads to modifications in keratocytes and in the organisation of collagen fibril. The morphological changes might be correlated to the process of increase in biomechanical stability although there are differences between stromal structures treated by standard and trans [source] Remodelling of collagen fibrils and proteoglycans in the zebrafish cornea during developmentACTA OPHTHALMOLOGICA, Issue 2007S AKHTAR Purpose: Collagen fibrils and proteoglycans are the main components of the corneal extracellular matrix and corneal transparency depends crucially on their proper organisation. We investigated their formation and arrangement in the developing cornea of the zebrafish, a major model of vertebrate development and genetic disease. Methods: We employed thin-section electron microscopy to investigate the ultrastructure of the zebrafish cornea at different stages of development. Results: Layering of the zebrafish cornea into an epithelium, Bowman's layer, stroma and endothelium was observed by 72 hours post-fertilization. At this stage, the stroma contained orthogonally arranged collagen fibrils and small proteoglycans. The density of proteoglycans increased gradually throughout subsequent development. In the stroma of 2 week old larvae, the collagen fibrils were organized into thin lamellae for the first time and were separated by very large, randomly distributed proteoglycans. At 4 weeks, a regular arrangement of proteoglycans around the collagen fibrils was observed for the first time and the lamellae also thickened. Conclusions: This is the first report of collagen fibril and proteoglycan development in the zebrafish cornea and it directly correlates collagen fibril and proteoglycan organisation of the zebrafish cornea with that of the human cornea. The similarities between the two species, including the possession of a Bowman layer, suggest that the zebrafish could serve as a model for the genetics of human corneal development and inherited disease. [source] Rho plays a central role in regulating local cell-matrix mechanical interactions in 3D cultureCYTOSKELETON, Issue 6 2007N. Lakshman Abstract The purpose of this study was to assess quantitatively the role of the small GTPase Rho on cell morphology, f-actin organization, and cell-induced matrix remodeling in 3D culture. Human corneal fibroblasts (HTK) were infected with adenoviruses that express green fluorescent protein (GFP) or GFP-N19Rho (dominant negative Rho). One day later cells were plated inside collagen matrices and allowed to spread for 24 h. Cells were fixed and stained for f-actin. Fluorescent (for f-actin) and reflected light (for collagen fibrils) images were acquired using confocal microscopy. Fourier transform analysis was used to assess local collagen fibril alignment, and changes in cell morphology and collagen density were measured using MetaMorph. The decrease in matrix height was used as an indicator of global matrix contraction. HTK and HTK-GFP cells induced significant global matrix contraction; this was inhibited by N19Rho. HTK and HTK-GFP fibroblasts generally had a bipolar morphology and occasional intracellular stress fibers. Collagen fibrils were compacted and aligned parallel to stress fibers and pseudopodia. In contrast, HTK-GFPN19 cells were elongated, and had a more cortical f-actin distribution. Numerous small extensions were also observed along the cell body. In addition, both local collagen fibril density and alignment were significantly reduced. Rho plays a key role in regulating both the morphology and mechanical behavior of corneal fibroblasts in 3D culture. Overall, the data suggest that Rho-kinase dependent cell contractility contributes to global and local matrix remodeling, whereas Rho dependent activation of mDia and/or other downstream effectors regulates the structure and number of cell processes. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source] Development of the corneal stroma, and the collagen,proteoglycan associations that help define its structure and functionDEVELOPMENTAL DYNAMICS, Issue 10 2008Andrew J. Quantock Abstract The cornea of the eye is a unique, transparent connective tissue. It is comprised predominantly of collagen fibrils, remarkably uniform in diameter and regularly spaced, organized into an intricate lamellar array. Its establishment involves a precisely controlled sequence of developmental events in which the embryonic cornea undergoes major structural transformations that ultimately determine tissue form and function. In this article, we will review corneal developmental dynamics from a structural perspective, consider the roles and interrelationships of collagens and proteoglycans, and comment on contemporary concepts and current challenges pertinent to developmental processes that result in an optically clear, mature cornea. Developmental Dynamics 237:2607,2621, 2008. © 2008 Wiley-Liss, Inc. [source] Biomorphic Silicon Carbide Coated with an Electrodeposition of Nanostructured Hydroxyapatite/Collagen as Biomimetic Bone Filler and Scaffold,ADVANCED ENGINEERING MATERIALS, Issue 8 2010M. Lelli Abstract The paper describes the method of preparation and chemical/physical characterization of a new biomaterial to be used as a bone substitute and bone-tissue engineering scaffold, which synergistically joins a porous bio-inspired morphology and the mechanical properties of biomorphic silicon carbide (BioSiC) with the surface bioactivity of a nanostructured hydroxyapatite/collagen biomimetic coating. FT-IR spectroscopy and XRD techniques are utilized to determine the chemical coating's composition. The morphology and size of the inorganic and protein components are investigated by TEM. The characteristic morphology of BioSiC channels and pores, which differ as a function of the transversal or longitudinal cross-section and with etching time, are investigated by SEM. Natural wood transformed into BioSiC acts as a cathode in an electrochemically assisted process that produces on its surface a biomimetic coating of hydroxyapatite nanocrystals and reconstituted type I collagen fibrils, producing an innovative apatite/collagen biomimetic porous bone filler and scaffold for tissue engineering. [source] Ultrastructural preservation of rat embryonic dental tissues after rapid fixation and dehydration under microwave irradiationEUROPEAN JOURNAL OF ORAL SCIENCES, Issue 1 2000Luciana F. Massa Adequate preservation of the cells and matrix of mineralising tissues remains difficult, as organic components and initial mineral deposits may be lost during conventional processing for electron microscopy. In this study, we have reduced significantly the processing time using microwave irradiation. Rat molar tooth germs were fixed in 4% glutaraldehyde+4% formaldehyde with 0.1 M sodium cacodylate in a laboratory microwave oven for two periods of 20 s with a maximal temperature of 37°C. After conventional washing and post-fixation, specimens were dehydrated in graded ethanols under microwave irradiation for a total of 7 min 20 s. For comparison, some specimens were processed by conventional methods. After embedding, ultrathin sections were examined by electron microscopy. In differentiating ameloblasts and odontoblasts, plasma membranes, mitochondria, rough endoplasmic reticulum, the Golgi complex, together with all other cytoplasmic organelles exhibited excellent preservation. Microtubules, microfilaments and coated vesicles were particularly evident. Crystal-like mineral deposits were conspicuously present in relation to dentine matrix vesicles and collagen fibrils as well as in enamel matrix. The matrix of forming enamel had a globular electron-lucent appearance. It is concluded that this is a rapid method which provides a preserved or even improved morphology. [source] CXC chemokine ligand 4 (Cxcl4) is a platelet-derived mediator of experimental liver fibrosis,HEPATOLOGY, Issue 4 2010Mirko Moreno Zaldivar Liver fibrosis is a major cause of morbidity and mortality worldwide. Platelets are involved in liver damage, but the underlying molecular mechanisms remain elusive. Here, we investigate the platelet-derived chemokine (C-X-C motif) ligand 4 (CXCL4) as a molecular mediator of fibrotic liver damage. Serum concentrations and intrahepatic messenger RNA of CXCL4 were measured in patients with chronic liver diseases and mice after toxic liver injury. Platelet aggregation in early fibrosis was determined by electron microscopy in patients and by immunohistochemistry in mice. Cxcl4,/, and wild-type mice were subjected to two models of chronic liver injury (CCl4 and thioacetamide). The fibrotic phenotype was analyzed by histological, biochemical, and molecular analyses. Intrahepatic infiltration of immune cells was investigated by fluorescence-activated cell sorting, and stellate cells were stimulated with recombinant Cxcl4 in vitro. The results showed that patients with advanced hepatitis C virus,induced fibrosis or nonalcoholic steatohepatitis had increased serum levels and intrahepatic CXCL4 messenger RNA concentrations. Platelets were found directly adjacent to collagen fibrils. The CCl4 and thioacetamide treatment led to an increase of hepatic Cxcl4 levels, platelet activation, and aggregation in early fibrosis in mice. Accordingly, genetic deletion of Cxcl4 in mice significantly reduced histological and biochemical liver damage in vivo, which was accompanied by changes in the expression of fibrosis-related genes (Timp-1 [tissue inhibitor of matrix metalloproteinase 1], Mmp9 [matrix metalloproteinase 9], Tgf -, [transforming growth factor beta], IL10 [interleukin 10]). Functionally, Cxcl4,/, mice showed a strongly decreased infiltration of neutrophils (Ly6G) and CD8+ T cells into the liver. In vitro, recombinant murine Cxcl4 stimulated the proliferation, chemotaxis, and chemokine expression of hepatic stellate cells. Conclusion: The results underscore an important role of platelets in chronic liver damage and imply a new target for antifibrotic therapies. (HEPATOLOGY 2010.) [source] An in vitro comparison of adhesive systems to seal pulp chamber wallsINTERNATIONAL ENDODONTIC JOURNAL, Issue 5 2004B. Ozturk Abstract Aim, To compare in vitro the sealing properties of five different dentine adhesive materials (Prime&Bond NT (PBNT); Prompt L-Pop (PLP); Clearfil SE Bond (CSEB); Scotchbond Multi Purpose Plus (SMPP); EBS-Multi (EBSM)) inside the pulp chamber. Methodology, Seventy-five recently extracted human molar teeth were used. The roof of the pulp chambers and roots were removed under water cooling. Pulp tissue was removed, and the canal orifices were sealed. The pulp chambers were then treated with 5% sodium hypochlorite (NaOCl) for 1 min. The teeth were randomly divided into five groups of 15 teeth each. Adhesive systems were applied to the pulp chamber walls according to the manufacturers' instructions. The samples were connected to Plexiglass plates, and a fluid filtration method was used for quantitative evaluation of leakage. Measurements of fluid movement were made at 2-min intervals for 8 min. The quality of seal of each specimen was measured immediately, after 24 h, 1 week and 1 month. The data were statistically analysed by repeated-measurements multivariate anova, Friedman test, Wilcoxon signed rank test, Kruskal,Wallis of one-way anova and Mann,Whitney U -tests. The pulp chamber wall with and without NaOCl and resin,dentine interfaces of specimens were observed under a scanning electron microscope (SEM). Results, The leakage values of the materials were significantly different at different measurement periods. In all groups, leakage values decreased with time (P < 0.05). PBNT and PLP had the least leakage during immediate measurements (P < 0.05). After 1 month, leakage of all adhesive systems was not significantly different (P < 0.05). SEM observation of pulp chamber walls demonstrated that the irregular dentine surface without smear layer was present in the nontreated group. However, NaOCl application removed the collagen fibrils leaving the dentine surface smooth. At resin,dentine interfaces of specimens, no hybridization zone was observed. Conclusions, None of the materials had created a perfect seal to the pulp chamber walls. PBNT and PLP had better sealing over the short term, but over the long term, there were no differences between the materials. [source] Age-related histopathological lesions in the Mongolian gerbil ventral prostate as a good model for studies of spontaneous hormone-related disordersINTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, Issue 1 2008Silvana Gisele Pegorin Campos Summary The Meriones unguiculatus (Mongolian) gerbil has demonstrated significant prostatic responses to hormonal treatments, and to drugs against human prostatic hyperplasia Spontaneous neoplasia develops in the older animals. Thirty gerbils (age 18 months) were divided into non-affected and prostatic lesion bearers and the prostate lesions were evaluated morphologically, immunohistochemically and quantitatively. The most frequent changes were in epithelial sites and, namely prostatic intraepithelial neoplasias, microinvasive carcinomas and adenocarcinomas. In the stromal compartment, cellular hyperplasia, when verified, was always associated with the sites of anomalous epithelium. Additionally, larger deposition of collagen fibrils, generating stromal fibrosis, was found in all the old gerbils analysed. The quantitative analysis showed that prostatic tissue proportions differed in altered areas, being specific for each lesion type. Isolated nuclear and nucleolar parameters were not effective in diagnosing the malign potential of lesions. However, the cellular proliferation and death indexes indicated larger cellular turnover in invasive lesions such as carcinomas. With these analyses, it could be verified that old gerbils present high propensity to develop spontaneous prostate changes and this may aid in a better understanding of the biological behaviour of human prostate cancer. [source] An interaction between opticin and heparan sulfate may provide the molecular basis for vitreoretinal adhesionINTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, Issue 4 2004V. John Hindson Introduction Opticin is a member of the extracellular matrix small leucine-rich repeat (SLRP) proteoglycan/protein family, which was originally identified in the eye associated with the collagen fibrils of the vitreous humour. A putative heparin/heparan sulfate (HS) binding motif (RKERKRR) was identified at the N-terminus of human opticin, but this is absent in the bovine form. Furthermore, the strength of attachment between the vitreous and the retina was observed to be species-dependent and related to the presence or absence of this motif. We hypothesized that opticin cross-links the collagen fibrils of the vitreous to HS proteoglycans in the inner limiting lamina (a basement membrane on the inner surface of the retina), contributing towards vitreoretinal adhesion. Materials and methods Recombinant human and bovine opticin were expressed in 293-EBNA cells and purified to apparent homogeneity. Solid phase assays and surface plasmon resonance studies were used to characterize interactions between immobilized heparin/HS and opticin. Results Solid phase and BIAcore data revealed that human opticin binds heparin/HS and binds to heparin with a dissociation constant of approximately 20 nm. By contrast bovine opticin, which lacks the basic cluster, bound severalfold less tightly. Competition studies with heparin oligosaccharides indicated that the heparin/HS binding site is greater than 6 monosaccharides in length. Heparin, HS, chondroitin sulfate A (CS-A), dermatan sulfate and hyaluronan all competed with heparin for binding to human opticin but CS-C did not. Discussion Work to date suggests that the N-terminal sequence RKERKRR contributes significantly to the binding of opticin to heparin/HS. Vitreoretinal adhesion plays a key role in a number of eye diseases and inhibitors of the opticin,HS interaction could be of therapeutic value. [source] Crimp morphology in relaxed and stretched rat Achilles tendonJOURNAL OF ANATOMY, Issue 1 2007Marco Franchi Abstract Fibrous extracellular matrix of tendon is considered to be an inextensible anatomical structure consisting of type I collagen fibrils arranged in parallel bundles. Under polarized light microscopy the collagen fibre bundles appear crimped with alternating dark and light transverse bands. This study describes the ultrastructure of the collagen fibrils in crimps of both relaxed and in vivo stretched rat Achilles tendon. Under polarized light microscopy crimps of relaxed Achilles tendons appear as isosceles or scalene triangles of different size. Tendon crimps observed via SEM and TEM show the single collagen fibrils that suddenly change their direction containing knots. The fibrils appear partially squeezed in the knots, bent on the same plane like bayonets, or twisted and bent. Moreover some of them lose their D-period, revealing their microfibrillar component. These particular aspects of collagen fibrils inside each tendon crimp have been termed ,fibrillar crimps' and may fulfil the same functional role. When tendon is physiologically stretched in vivo the tendon crimps decrease in number (46.7%) (P < 0.01) and appear more flattened with an increase in the crimp top angle (165° in stretched tendons vs. 148° in relaxed tendons, P < 0.005). Under SEM and TEM, the ,fibrillar crimps' are still present, never losing their structural identity in straightened collagen fibril bundles of stretched tendons even where tendon crimps are not detectable. These data suggest that the ,fibrillar crimp' may be the true structural component of the tendon crimp acting as a shock absorber during physiological stretching of Achilles tendon. [source] Collagen architecture and failure processes in bovine patellar cartilageJOURNAL OF ANATOMY, Issue 4 2001JACK L. LEWIS Cartilage fails by fibrillation and wearing away. This study was designed to identify the microscopic failure processes in the collagen network of bovine cartilage using scanning electron microscopy. Cartilage samples from fibrillated cartilage from the bovine patella were removed from the bone, fixed, digested to remove proteoglycans, freeze-fractured, and processed for SEM. The architecture of the collagen network in the normal cartilage was first defined, and then the failure processes were identified by examining sites of fibrillation and at crack tips. The bovine patellar cartilage was organised with a superficial layer composed of 3,5 lamina, attached to a sub-superficial tissue by angled bridging fibrils. Collagen in the sub-superficial tissue was organised in lamina oriented in the radial direction up to the transition zone. Failure of the system occurred by cracks forming in superficial layer and lamina, creating flaps of lamina that rolled up into the larger ,fronds'. Larger cracks not following the laminar planes occurred in the transition, mid, and deep zones. Failure at the crack tips in the sub-superficial tissue appeared to be by peeling of collagen fibrils, as opposed to breaking of collagen fibrils, suggesting a ,glue' bonding the collagen fibrils in a parallel fashion. Cracks propagated by breaking these bonds. This bond could be a site of disease action, since weakening of the bond would accelerate crack propagation. [source] Small-angle X-ray scattering study of intramuscular fish bone: collagen fibril superstructure determined from equidistant meridional reflectionsJOURNAL OF APPLIED CRYSTALLOGRAPHY, Issue 2 2008Christian Burger New insights into the bone collagen fibril superstructure have been obtained by novel small-angle X-ray scattering analysis. The analysis was carried out on the small-angle equidistant meridional reflections resulting from the periodic structure of collagen fibrils in their axial direction. Conventional two-dimensional analysis is difficult because of the large discrepancy of longitudinal and lateral length scales for individual fibrils, as well as their preferred orientation. The new approach represents an unapproximated analysis of the equidistant meridional reflections, which takes the exact separation of preferred orientation and fibril size effects into account. The analytical results (e.g. axial period, fibril diameter etc.) agree well with the parameters obtained from transmission electron microscopy. [source] Scanning texture analysis of lamellar bone using microbeam synchrotron X-ray radiationJOURNAL OF APPLIED CRYSTALLOGRAPHY, Issue 1 2007Wolfgang Wagermaier Texture analysis with microbeam scanning diffraction enables the local mapping of three-dimensional crystallite orientation in heterogeneous natural and synthetic materials. Cortical (compact) bone is an example of a hierarchically structured biocomposite, which is built mainly of cylindrical osteons, having a lamellar texture at the micrometre level. In this work, a combination of microbeam synchrotron X-ray texture analysis with thin sections of osteonal bone is used to measure the three-dimensional distribution of the c -axis orientation of the mineral apatite in bone with positional resolution of 1,µm. The data reduction procedure needed to go from the stereographic projection of X-ray intensity to the determination of the local orientation of mineralized collagen fibrils is described. The procedure can be applied to other mineralized tissues (such as trabecular bone and chitin) with micrometre scale and biologically controlled fibrillar texture. [source] Composite coating of bonelike apatite particles and collagen fibers on poly L-lactic acid formed through an accelerated biomimetic coprecipitation processJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2006Yun Chen Abstract Collagen and apatite were coprecipitated as a composite coating on poly L-lactic acid (PLLA) in an accelerated biomimetic process. The incubation solution contained collagen (1 g/L) and simulated body fluid with 5 times inorganic ionic concentrations as human blood plasma. The coating formed on PLLA films and scaffolds after a 24-h incubation was characterized by using energy-dispersive X-ray spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM). It was shown that the coating contained carbonated bonelike apatite and collagen, which was similar in composition to natural bone. SEM showed a complex composite coating of submicron bonelike apatite particulates combined with collagen fibrils. It is expected that such biocomposite coating may better facilitate cell interaction and osteoconductivity. This work provided an efficient process to obtain bonelike apatite/collagen composite coating, which is potentially useful in bone tissue engineering. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006 [source] Overexpression of Lysyl Hydroxylase-2b Leads to Defective Collagen Fibrillogenesis and Matrix Mineralization,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 1 2005Suchaya Pornprasertsuk Abstract Several MC3T3-E1 cell-derived clones expressing higher levels of LH2b were analyzed for their abilities to form collagen fibrils and mineralization. The clones all exhibited smaller collagen fibrils and defective matrix mineralization in vitro and in vivo, indicating a critical role of LH2b-catalyzed post-translational modifications of collagen in bone matrix formation and mineralization. Introduction: We have recently shown that lysyl hydroxylase (LH) 2b, through its action on the telopeptidyl lysine residues of collagen, regulates collagen cross-linking pathway in the osteoblastic cell line, MC3T3-E1. To further elucidate the roles of LH2b in bone physiology, the effects of overexpression of LH2b on collagen fibrillogenesis and matrix mineralization were investigated. Materials and Methods: Several MC3T3-E1-derived osteoblastic cell clones expressing higher levels of LH2b (S clones) and two controls (i.e., MC3T3-E1 cells and those transfected with an empty vector) were cultured. MALDI-TOF mass spectrometry was used to identify the LH2b. The collagen fibrillogenesis in the cultures was characterized by transmission electron microscopy, and the ability of these clones and cells to form mineralized matrix was analyzed by both in vitro and in vivo mineralization assays. Results: The diameter of collagen fibrils in the S clone cultures was markedly smaller than that of the controls. The onset of matrix mineralization in the S clones was significantly delayed, and considerably fewer mineralized nodules were formed in their cultures in comparison with the controls. When transplanted into immunodeficient mice, the S clones failed to form mineralized matrices in vivo, whereas a bone-like mineralized matrix was well formed by the controls. The diameter of the collagen fibrils and the timing/extent of matrix mineralization in vitro were inversely correlated with the level of LH2b. In vitro cell differentiation was unaffected by the LH2b overexpression. Conclusions: These results indicate a critical role of LH2b catalyzed post-translational modification of collagen (i.e., telopeptidyl lysine hydroxylation and subsequent cross-linking) in collagen matrix formation and mineralization in bone. [source] The Bone Lining Cell: Its Role in Cleaning Howship's Lacunae and Initiating Bone FormationJOURNAL OF BONE AND MINERAL RESEARCH, Issue 1 2002V. Everts Abstract In this study we investigated the role of bone lining cells in the coordination of bone resorption and formation. Ultrastructural analysis of mouse long bones and calvariae revealed that bone lining cells enwrap and subsequently digest collagen fibrils protruding from Howship's lacunae that are left by osteoclasts. By using selective proteinase inhibitors we show that this digestion depends on matrix metalloproteinases and, to some extent, on serine proteinases. Autoradiography revealed that after the bone lining cells have finished cleaning, they deposit a thin layer of a collagenous matrix along the Howship's lacuna, in close association with an osteopontin-rich cement line. Collagenous matrix deposition was detected only in completely cleaned pits. In bone from pycnodysostotic patients and cathepsin K-deficient mice, conditions in which osteoclastic bone matrix digestion is greatly inhibited, bone matrix leftovers proved to be degraded by bone lining cells, thus indicating that the bone lining cell "rescues" bone remodeling in these anomalies. We conclude that removal of bone collagen left by osteoclasts in Howship's lacunae is an obligatory step in the link between bone resorption and formation, and that bone lining cells and matrix metalloproteinases are essential in this process. [source] Study of order and dynamic processes in tendon by NMR and MRIJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 2 2007G. Navon PhD Abstract Tendons are composed of a parallel arrangement of densely packed collagen fibrils that results in unique biomechanical properties of strength and flexibility. In the present review we discuss several advanced magnetic resonance spectroscopy (MRS) and imaging (MRI) techniques that have allowed us to better understand the biophysical properties of tendons and ligaments. The methods include multiple quantum and T2 filtering combined with NMR and MRI techniques. It is shown in detail how these techniques can be used to extract a number of useful parameters: 1) the 1H- 1H and 1H- 2H dipolar interactions; 2) the proton exchange rates between water and collagen, and between water molecules; 3) the distribution of fibril orientations; and 4) the anisotropy of diffusion. It is shown that relaxation data as a function of angular dependence can be obtained in vivo using mobile NMR sensors. Finally, this article describes how double quantum filtered (DQF) MRI can be used to image and monitor the healing process in injured tendons. J. Magn. Reson. Imaging 2007. © 2007 Wiley-Liss, Inc. [source] Quantification of the graphical details of collagen fibrils in transmission electron micrographsJOURNAL OF MICROSCOPY, Issue 1 2001Y. Xia A novel 2D image analysis technique is demonstrated. Using the digitized images of articular cartilage from transmission electron microscopy (TEM), this technique performs a localized ,vector' analysis at each region that is large enough to include several or tens of collagen fibrils but small enough to provide a fine resolution for the whole tissue. For each small and localized region, the morphology of the collagen fibrils can be characterized by three quantities essential to the nature of the tissue: the concentration of the fibrils, the overall orientation of the fibrils, and the anisotropy of the fibrils. This technique is capable of providing new insight to the existing technology by assigning quantitative attributes to the qualitative graphics. The assigned quantities are sensitive to the fine structure of the collagen matrix and meaningful in the architectural nature of the collagen matrix. These quantities could provide a critical linkage between the ultrastructure of the tissue and the macroscopic behaviours of the material. In addition, coarse-graining the microscopic resolution of EM without compromising the essential features of the tissue's structure provides a direct view of the tissue's morphology and permits direct correlations and comparisons among interdisciplinary techniques. [source] Developmental changes in the ultrastructure of the lamprey lateral line nerve during metamorphosisJOURNAL OF MORPHOLOGY, Issue 7 2009S. Gelman Abstract The ultrastructure of the trunk lateral line nerve of larval and adult lampreys was studied with transmission electron microscopy. We confirmed that lampreys' lateral line nerve lacks myelin. Nevertheless, all axons were wrapped by Schwann cell processes. In the larval nerve, gaps between Schwann cells were observed, where the axolemma was covered only by a basal lamina, indicating an earlier developmental stage. In the adult nerve, glial (Schwann cell) ensheathment was mostly complete. Additionally, we observed variable ratios of axons to Schwann cells in larval and adult preparations. In the larval nerve, smaller axons were wrapped by one Schwann cell. Occasionally, a single Schwann cell surrounded two axons. Larger axons were associated with two to five Schwann cells. In the adult nerve, smaller axons were surrounded by one, but larger axons by three to eight Schwann cells. The larval epineurium contained large adipose cells, separated from each other by single fibroblast processes. This layer of adipose tissue was reduced in adult preparation. The larval perineurium was thin, and the fibroblasts, containing large amounts of glycogen granules, were arranged loosely. The adult perineurium was thicker, consisting of at least three layers of fibroblasts separated by collagen fibrils. The larval and adult endoneurium contained collagen fibrils oriented orthogonally to each other. Both larval and adult lateral line nerves possessed a number of putative fascicles weakly defined by a thin layer of perineurial fibroblasts. These results indicate that after a prolonged larval stage, the lamprey lateral line nerve is subjected to additional maturation processes during metamorphosis. J. Morphol. 2009. © 2009 Wiley-Liss, Inc. [source] Nanostructure of collagen fibrils in human nucleus pulposus and its correlation with macroscale tissue mechanicsJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 4 2010Darwesh M.K. Aladin Abstract Collagen fibrils are the main structural components of the nucleus pulposus tissue in the intervertebral discs. The structure,property relationship of the nucleus pulposus (NP) tissues is still unclear. We investigated the structure of individual collagen fibrils of the NP and evaluated its correlation with the bulk mechanical properties of the tissue. Collagen fibrils were extracted from the NP of discs retrieved from adolescents during scoliosis correction surgery, and the extracts were confirmed by SDS-PAGE. The diameters of the individual collagen fibrils were measured through atomic force microscopy, and the compressive mechanical properties of the tissues were evaluated by confined compression. The correlations between the nanoscale morphology of the collagen fibrils and the macroscale mechanical properties of the tissues were evaluated by linear regression. The SDS-PAGE results showed that the fibril extracts were largely composed of type II collagen. The mean diameter of the collagen fibrils was 92.1,±,26.54 nm; the mean swelling pressure and compressive modulus of the tissues were 6.15,±,4.3 kPa and 1.23,±,0.7 MPa, respectively. The mean fibril diameter had no linear correlation (R2,=,0.30) with the swelling pressure of the tissues. However, it had a mild linear correlation with the compressive modulus (p,=,0.023, R2,=,0.68). This is the first study, to our knowledge, to evaluate the nanostructure of the individual collagen fibrils of the nucleus pulposus and its relationship with macroscale mechanical properties of the NP tissues. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:497,502, 2010 [source] | |||||||||||||||||||