Sample records for basement membrane collagen

  1. Distribution of basement membrane type IV collagen alpha chains in ameloblastoma: an immunofluorescence study


    Nakano, K.; Siar, C. H.; Nagai, N.; Naito, I.; Sado, Y.; Nagatsuka, H; Hoh, C; Kurada, K.; Tsujigiwa, H; M. Gunduz


    Background: Type IV collagen, a heterotrimeric molecule that exists in six genetically distinct forms, alpha1(IV)-alpha6(IV) is a major structural component of basement membrane (BM) and acts as a scaffold for other BM constituents. Methods: Indirect immunofluorescence using alpha chain-specific monoclonal antibodies was employed to clarify basement membrane (BM) collagen IV distribution in two ameloblastoma, and for comparison, on oral mucosa and tooth germ. Results: Ameloblastoma BM express...

  2. Type IV Collagens and Basement Membrane Diseases: Cell Biology and Pathogenic Mechanisms. (United States)

    Mao, Mao; Alavi, Marcel V; Labelle-Dumais, Cassandre; Gould, Douglas B


    Basement membranes are highly specialized extracellular matrices. Once considered inert scaffolds, basement membranes are now viewed as dynamic and versatile environments that modulate cellular behaviors to regulate tissue development, function, and repair. Increasing evidence suggests that, in addition to providing structural support to neighboring cells, basement membranes serve as reservoirs of growth factors that direct and fine-tune cellular functions. Type IV collagens are a major component of all basement membranes. They evolved along with the earliest multicellular organisms and have been integrated into diverse fundamental biological processes as time and evolution shaped the animal kingdom. The roles of basement membranes in humans are as complex and diverse as their distributions and molecular composition. As a result, basement membrane defects result in multisystem disorders with ambiguous and overlapping boundaries that likely reflect the simultaneous interplay and integration of multiple cellular pathways and processes. Consequently, there will be no single treatment for basement membrane disorders, and therapies are likely to be as varied as the phenotypes. Understanding tissue-specific pathology and the underlying molecular mechanism is the present challenge; personalized medicine will rely upon understanding how a given mutation impacts diverse cellular functions.

  3. Attachment of cells to basement membrane collagen type IV



    Of ten different cell lines examined, three showed distinct attachment and spreading on collagen IV substrates, and neither attachment nor spreading was enhanced by adding soluble laminin or fibronectin. This reaction was not inhibited by cycloheximide or antibodies to laminin, indicating a direct attachment to collagen IV without the need of mediator proteins. Cell-binding sites were localized to the major triple-helical domain of collagen IV and required an intact triple helical conformatio...

  4. VEGF-A/Notch-Induced Podosomes Proteolyse Basement Membrane Collagen-IV during Retinal Sprouting Angiogenesis

    Directory of Open Access Journals (Sweden)

    Pirjo Spuul


    Full Text Available During angiogenic sprouting, endothelial tip cells emerge from existing vessels in a process that requires vascular basement membrane degradation. Here, we show that F-actin/cortactin/P-Src-based matrix-degrading microdomains called podosomes contribute to this step. In vitro, VEGF-A/Notch signaling regulates the formation of functional podosomes in endothelial cells. Using a retinal neovascularization model, we demonstrate that tip cells assemble podosomes during physiological angiogenesis in vivo. In the retina, podosomes are also part of an interconnected network that surrounds large microvessels and impinges on the underlying basement membrane. Consistently, collagen-IV is scarce in podosome areas. Moreover, Notch inhibition exacerbates podosome formation and collagen-IV loss. We propose that the localized proteolytic action of podosomes on basement membrane collagen-IV facilitates endothelial cell sprouting and anastomosis within the developing vasculature. The identification of podosomes as key components of the sprouting machinery provides another opportunity to target angiogenesis therapeutically.

  5. Type XV collagen in human colonic adenocarcinomas has a different distribution than other basement membrane zone proteins. (United States)

    Amenta, P S; Briggs, K; Xu, K; Gamboa, E; Jukkola, A F; Li, D; Myers, J C


    In situ carcinomas must penetrate their own basement membrane to be classified as invasive, and subsequently infiltrate surrounding connective tissue and cross vascular basement membranes to metastasize hematogenously. Accordingly, in many studies, integral basement membrane components, including type IV collagen, laminin, and heparan sulfate proteoglycan, have been localized in a spectrum of tumors to gain insight into their role in neoplasia. A number of recently identified extracellular matrix molecules and isoforms of the aforementioned proteins have been localized to the basement membrane zone, illustrating another level of biochemical heterogeneity in these structures. As the complexity of these matrices becomes more apparent, their roles in maintaining homeostasis and in tumor biology falls into question. Of the new group of collagens localized to the basement membrane zone, type XV was the first to be characterized (Cell Tissue Res, 286:493-505, 1996). This nonfibrillar collagen has a nearly ubiquitous distribution in normal human tissues via a strong association with basement membrane zones, suggesting that it functions to adhere basement membrane to the underlying stroma. To begin investigation of this protein in malignant tumors, we have localized type XV in human colonic adenocarcinomas and compared its distribution with that of type IV collagen and laminin. Collagens XV and IV and laminin were found in all normal and colonic epithelial, muscle, fat, neural, and vascular basement membrane zones, as shown previously. In moderately differentiated, invasive adenocarcinomas, laminin and type IV collagen were sometimes observed as continuous, linear deposits around some of the malignant glands, but more often they were seen in either discontinuous deposits or were completely absent. In contrast, type XV collagen was characterized as virtually absent from the basement membrane zones of malignant glandular elements in moderately differentiated tumors

  6. Basement Membrane Zone Collagens XV and XVIII/Proteoglycans Mediate Leukocyte Influx in Renal Ischemia/Reperfusion

    NARCIS (Netherlands)

    Zaferani, Azadeh; Talsma, Ditmer T.; Yazdani, Saleh; Celie, Johanna W. A. M.; Aikio, Mari; Heljasvaara, Ritva; Navis, Gerjan J.; Pihlajaniemi, Taina; van den Born, Jacob


    Collagen type XV and XVIII are proteoglycans found in the basement membrane zones of endothelial and epithelial cells, and known for their cryptic anti-angiogenic domains named restin and endostatin, respectively. Mutations or deletions of these collagens are associated with eye, muscle and microves

  7. Role of 17 beta-estradiol on type IV collagen fibers volumetric density in the basement membrane of bladder wall. (United States)

    de Fraga, Rogerio; Dambros, Miriam; Miyaoka, Ricardo; Riccetto, Cássio Luís Zanettini; Palma, Paulo César Rodrigues


    The authors quantified the type IV collagen fibers volumetric density in the basement membrane of bladder wall of ovariectomized rats with and without estradiol replacement. This study was conducted on 40 Wistar rats (3 months old) randomly divided in 4 groups: group 1, remained intact (control); group 2, submitted to bilateral oophorectomy and daily replacement 4 weeks later of 17 beta-estradiol for 12 weeks; group 3, sham operated and daily replacement 4 weeks later of sesame oil for 12 weeks; and group 4, submitted to bilateral oophorectomy and killed after 12 weeks. It was used in immunohistochemistry evaluation using type IV collagen polyclonal antibody to stain the fibers on paraffin rat bladder sections. The M-42 stereological grid system was used to analyze the fibers. Ovariectomy had an increase effect on the volumetric density of the type IV collagen fibers in the basement membrane of rat bladder wall. Estradiol replacement in castrated animals demonstrated a significative difference in the stereological parameters when compared to the castrated group without hormonal replacement. Surgical castration performed on rats induced an increasing volumetric density of type IV collagen fibers in the basement membrane of rats bladder wall and the estradiol treatment had a significant effect in keeping a low volumetric density of type IV collagen fibers in the basement membrane of rats bladder wall.

  8. Laminin and Type IV Collagen Isoform Substitutions Occur in Temporally and Spatially Distinct Patterns in Developing Kidney Glomerular Basement Membranes


    Abrahamson, Dale R.; St. John, Patricia L.; Stroganova, Larysa; Zelenchuk, Adrian; Steenhard, Brooke M.


    Kidney glomerular basement membranes (GBMs) undergo laminin and type IV collagen isoform substitutions during glomerular development, which are believed to be required for maturation of the filtration barrier. Specifically, GBMs of earliest glomeruli contain laminin α1β1γ1 and collagen α1α2α1(IV), whereas mature glomeruli contain laminin α5β2γ1 and collagen α3α4α5(IV). Here, we used confocal microscopy to simultaneously evaluate expression of different laminin and collagen IV isoforms in newb...

  9. Basement membrane proteoglycans and development

    DEFF Research Database (Denmark)

    Couchman, J R; Abrahamson, D R; McCarthy, K J


    Basement membranes contain distinct collagen, glycoprotein and proteoglycan species, and these exhibit considerable heterogeneity in isoform or type when different tissue types are compared. Additionally, many components are differentially expressed in organogenesis. We have considered the distri......Basement membranes contain distinct collagen, glycoprotein and proteoglycan species, and these exhibit considerable heterogeneity in isoform or type when different tissue types are compared. Additionally, many components are differentially expressed in organogenesis. We have considered...... the distributions in glomerulogenesis of two distinct basement membrane proteoglycans, a small heparan sulfate proteoglycan and a chondroitin sulfate proteoglycan (BM-CSPG). While the former was present in all kidney basement membranes through development, the latter was apparently regulated in distribution. BM......-CSPG was only strongly expressed in the vasculature invading late comma stage glomeruli, and later in presumptive and mature Bowman's capsule. Over the first six to eight weeks, the capillary basement membranes contained BM-CSPG, but in gradually decreasing amounts until it became completely undetectable...

  10. Fibrillar, fibril-associated and basement membrane collagens of the arterial wall: architecture, elasticity and remodeling under stress. (United States)

    Osidak, M S; Osidak, E O; Akhmanova, M A; Domogatsky, S P; Domogatskaya, A S


    The ability of a human artery to pass through 150 million liters of blood sustaining 2 billion pulsations of blood pressure with minor deterioration depends on unique construction of the arterial wall. Viscoelastic properties of this construction enable to re-seal the occuring damages apparently without direct immediate participance of the constituent cells. Collagen structures are considered to be the elements that determine the mechanoelastic properties of the wall in parallel with elastin responsible for elasticity and resilience. Collagen scaffold architecture is the function-dependent dynamic arrangement of a dozen different collagen types composing three distinct interacting forms inside the extracellular matrix of the wall. Tightly packed molecules of collagen types I, III, V provide high tensile strength along collagen fibrils but toughness of the collagen scaffold as a whole depends on molecular bonds between distinct fibrils. Apart of other macromolecules in the extracellular matrix (ECM), collagen-specific interlinks involve microfilaments of collagen type VI, meshwork-organized collagen type VIII, and FACIT collagen type XIV. Basement membrane collagen types IV, XV, XVIII and cell-associated collagen XIII enable transmission of mechanical signals between cells and whole artery matrix. Collagen scaffold undergoes continuous remodeling by decomposition promoted with MMPs and reconstitution from newly produced collagen molecules. Pulsatile stress-strain load modulates both collagen synthesis and MMP-dependent collagen degradation. In this way the ECM structure becomes adoptive to mechanical challenges. The mechanoelastic properties of the arterial wall are changed in atherosclerosis concomitantly with collagen turnover both type-specific and dependent on the structure. Improving the feedback could be another approach to restore sufficient blood circulation.

  11. Laminin and type IV collagen isoform substitutions occur in temporally and spatially distinct patterns in developing kidney glomerular basement membranes. (United States)

    Abrahamson, Dale R; St John, Patricia L; Stroganova, Larysa; Zelenchuk, Adrian; Steenhard, Brooke M


    Kidney glomerular basement membranes (GBMs) undergo laminin and type IV collagen isoform substitutions during glomerular development, which are believed to be required for maturation of the filtration barrier. Specifically, GBMs of earliest glomeruli contain laminin α1β1γ1 and collagen α1α2α1(IV), whereas mature glomeruli contain laminin α5β2γ1 and collagen α3α4α5(IV). Here, we used confocal microscopy to simultaneously evaluate expression of different laminin and collagen IV isoforms in newborn mouse GBMs. Our results show loss of laminin α1 from GBMs in early capillary loop stages and continuous linear deposition of laminin bearing the α5 chain thereafter. In contrast, collagen α1α2α1(IV) persisted in linear patterns into late capillary loop stages, when collagen α3α4α5(IV) first appeared in discontinuous, non-linear patterns. This patchy pattern for collagen α3α4α5(IV) continued into maturing glomeruli where there were lengths of linear, laminin α5-positive GBM entirely lacking either isoform of collagen IV. Relative abundance of laminin and collagen IV mRNAs in newborn and 5-week-old mouse kidneys also differed, with those encoding laminin α1, α5, β1, β2, and γ1, and collagen α1(IV) and α2(IV) chains all significantly declining at 5 weeks, but α3(IV) and α4(IV) were significantly upregulated. We conclude that different biosynthetic mechanisms control laminin and type IV collagen expression in developing glomeruli.

  12. Comprehensive Characterization of Glycosylation and Hydroxylation of Basement Membrane Collagen IV by High-Resolution Mass Spectrometry. (United States)

    Basak, Trayambak; Vega-Montoto, Lorenzo; Zimmerman, Lisa J; Tabb, David L; Hudson, Billy G; Vanacore, Roberto M


    Collagen IV is the main structural protein that provides a scaffold for assembly of basement membrane proteins. Posttranslational modifications such as hydroxylation of proline and lysine and glycosylation of lysine are essential for the functioning of collagen IV triple-helical molecules. These modifications are highly abundant posing a difficult challenge for in-depth characterization of collagen IV using conventional proteomics approaches. Herein, we implemented an integrated pipeline combining high-resolution mass spectrometry with different fragmentation techniques and an optimized bioinformatics workflow to study posttranslational modifications in mouse collagen IV. We achieved 82% sequence coverage for the α1 chain, mapping 39 glycosylated hydroxylysine, 148 4-hydroxyproline, and seven 3-hydroxyproline residues. Further, we employed our pipeline to map the modifications on human collagen IV and achieved 85% sequence coverage for the α1 chain, mapping 35 glycosylated hydroxylysine, 163 4-hydroxyproline, and 14 3-hydroxyproline residues. Although lysine glycosylation heterogeneity was observed in both mouse and human, 21 conserved sites were identified. Likewise, five 3-hydroxyproline residues were conserved between mouse and human, suggesting that these modification sites are important for collagen IV function. Collectively, these are the first comprehensive maps of hydroxylation and glycosylation sites in collagen IV, which lay the foundation for dissecting the key role of these modifications in health and disease.

  13. Fibrosis is not just fibrosis - basement membrane modelling and collagen metabolism differs between hepatitis B- and C-induced injury

    DEFF Research Database (Denmark)

    Nielsen, M J; Karsdal, Morten A; Kazankov, K


    . AIM: To investigate whether differences in extracellular matrix (ECM) composition of the liver during fibrogenesis in two seemingly similar types of viral hepatitis could be reflected by differences in ECM turnover. METHODS: Utilising a cross-sectional design, we measured specific ECM protein...... fragments in plasma from 197 chronic hepatitis B (CHB) patients and 403 chronic hepatitis C (CHC) patients matched for inflammation grade and fibrosis stage. Markers of matrix metalloprotease degraded type I, III, IV and VI collagen (C1M, C3M, C4M, C6M) and type III and IV collagen formation (Pro-C3, P4NP7S...... and fibrosis only in CHC. Basement membrane collagen fragments P4NP7S and C4M were significantly higher in matched activity and fibrosis cohorts within CHB vs CHC. CONCLUSION: The main parameters to determine extracellular matrix biomarker levels are inflammation, fibrosis, and type of viral insult. Compared...

  14. Comparative analysis of fibrillar and basement membrane collagen expression in embryos of the sea urchin, Strongylocentrotus purpuratus. (United States)

    Suzuki, H R; Reiter, R S; D'Alessio, M; Di Liberto, M; Ramirez, F; Exposito, J Y; Gambino, R; Solursh, M


    The time of appearance and location of three distinct collagen gene transcripts termed 1 alpha, 2 alpha, and 3 alpha, were monitored in the developing S. purpuratus embryo by in situ hybridization. The 1 alpha and 2 alpha transcripts of fibrillar collagens were detected simultaneously in the primary (PMC) and secondary (SMC) mesenchyme cells of the late gastrula stage and subsequently expressed in the spicules and gut associated cells of the pluteus stage. The 3 alpha transcripts of the basement membrane collagen appeared earlier than 1 alpha and 2 alpha, and were first detected in the presumptive PMC at the vegetal plate of the late blastula stage. The PMC exhibited high expression of 3 alpha at the mesenchyme blastula stage, but during gastrulation the level of expression was reduced differentially among the PMC. In the late gastrula and pluteus stages, both PMC and SMC expressed 3 alpha mRNA, and thus at these stages all three collagen genes displayed an identical expression pattern by coincidence. This study thus provides the first survey of onset and localization of multiple collagen transcripts in a single sea urchin species.

  15. Assessment of proteolytic degradation of the basement membrane: a fragment of type IV collagen as a biochemical marker for liver fibrosis

    DEFF Research Database (Denmark)

    Veidal, Sanne S.; Karsdal, Morten A.; Nawrocki, Arkadiusz


    by proteases produces small fragments, so-called neoepitopes, which are released systemically. Technologies investigating MMP-generated fragments of collagens may provide more useful information than traditional serological assays that crudely measure total protein. In the present study, we developed an ELISA......Collagen deposition and an altered matrix metalloproteinase (MMP) expression profile are hallmarks of fibrosis. Type IV collagen is the most abundant structural basement membrane component of tissue, which increases 14-fold during fibrogenesis in the liver. Proteolytic degradation of collagens...... for the quantification of a neoepitope generated by MMP degradation of type IV collagen and evaluated the association of this neoepitope with liver fibrosis in two animal models....

  16. Type IV Collagen Controls the Axogenesis of Cerebellar Granule Cells by Regulating Basement Membrane Integrity in Zebrafish. (United States)

    Takeuchi, Miki; Yamaguchi, Shingo; Yonemura, Shigenobu; Kakiguchi, Kisa; Sato, Yoshikatsu; Higashiyama, Tetsuya; Shimizu, Takashi; Hibi, Masahiko


    Granule cells (GCs) are the major glutamatergic neurons in the cerebellum, and GC axon formation is an initial step in establishing functional cerebellar circuits. In the zebrafish cerebellum, GCs can be classified into rostromedial and caudolateral groups, according to the locations of their somata in the corresponding cerebellar lobes. The axons of the GCs in the caudolateral lobes terminate on crest cells in the dorsal hindbrain, as well as forming en passant synapses with Purkinje cells in the cerebellum. In the zebrafish mutant shiomaneki, the caudolateral GCs extend aberrant axons. Positional cloning revealed that the shiomaneki (sio) gene locus encodes Col4a6, a subunit of type IV collagen, which, in a complex with Col4a5, is a basement membrane (BM) component. Both col4a5 and col4a6 mutants displayed similar abnormalities in the axogenesis of GCs and retinal ganglion cells (RGCs). Although type IV collagen is reported to control axon targeting by regulating the concentration gradient of an axonal guidance molecule Slit, Slit overexpression did not affect the GC axons. The structure of the BM surrounding the tectum and dorsal hindbrain was disorganized in the col4a5 and col4a6 mutants. Moreover, the abnormal axogenesis of the caudolateral GCs and the RGCs was coupled with aberrant BM structures in the type IV collagen mutants. The regrowth of GC axons after experimental ablation revealed that the original and newly formed axons displayed similar branching and extension abnormalities in the col4a6 mutants. These results collectively suggest that type IV collagen controls GC axon formation by regulating the integrity of the BM, which provides axons with the correct path to their targets.

  17. Type IV Collagen Controls the Axogenesis of Cerebellar Granule Cells by Regulating Basement Membrane Integrity in Zebrafish.

    Directory of Open Access Journals (Sweden)

    Miki Takeuchi


    Full Text Available Granule cells (GCs are the major glutamatergic neurons in the cerebellum, and GC axon formation is an initial step in establishing functional cerebellar circuits. In the zebrafish cerebellum, GCs can be classified into rostromedial and caudolateral groups, according to the locations of their somata in the corresponding cerebellar lobes. The axons of the GCs in the caudolateral lobes terminate on crest cells in the dorsal hindbrain, as well as forming en passant synapses with Purkinje cells in the cerebellum. In the zebrafish mutant shiomaneki, the caudolateral GCs extend aberrant axons. Positional cloning revealed that the shiomaneki (sio gene locus encodes Col4a6, a subunit of type IV collagen, which, in a complex with Col4a5, is a basement membrane (BM component. Both col4a5 and col4a6 mutants displayed similar abnormalities in the axogenesis of GCs and retinal ganglion cells (RGCs. Although type IV collagen is reported to control axon targeting by regulating the concentration gradient of an axonal guidance molecule Slit, Slit overexpression did not affect the GC axons. The structure of the BM surrounding the tectum and dorsal hindbrain was disorganized in the col4a5 and col4a6 mutants. Moreover, the abnormal axogenesis of the caudolateral GCs and the RGCs was coupled with aberrant BM structures in the type IV collagen mutants. The regrowth of GC axons after experimental ablation revealed that the original and newly formed axons displayed similar branching and extension abnormalities in the col4a6 mutants. These results collectively suggest that type IV collagen controls GC axon formation by regulating the integrity of the BM, which provides axons with the correct path to their targets.

  18. Laminins in basement membrane assembly. (United States)

    Hohenester, Erhard; Yurchenco, Peter D


    The heterotrimeric laminins are a defining component of all basement membranes and self-assemble into a cell-associated network. The three short arms of the cross-shaped laminin molecule form the network nodes, with a strict requirement for one α, one β and one γ arm. The globular domain at the end of the long arm binds to cellular receptors, including integrins, α-dystroglycan, heparan sulfates and sulfated glycolipids. Collateral anchorage of the laminin network is provided by the proteoglycans perlecan and agrin. A second network is then formed by type IV collagen, which interacts with the laminin network through the heparan sulfate chains of perlecan and agrin and additional linkage by nidogen. This maturation of basement membranes becomes essential at later stages of embryo development.

  19. MMP Mediated Degradation of Type IV Collagen Alpha 1 and Alpha 3 Chains Reflects Basement Membrane Remodeling in Experimental and Clinical Fibrosis - Validation of Two Novel Biomarker Assays

    DEFF Research Database (Denmark)

    Sand, Jannie Marie; Larsen, Lise Skakkebæk; Hogaboam, Cory;


    Fibrosis is characterized by excessive tissue remodeling resulting from altered expression of various growth factors, cytokines and proteases. We hypothesized that matrix metalloproteinase (MMP) mediated degradation of type IV collagen, a main component of the basement membrane, will release...... peptide fragments (neo-epitopes) into the circulation. Here we present the development of two competitive enzyme-linked immunosorbent assays (ELISAs) for assessing the levels of specific fragments of type IV collagen α1 (C4M12a1) and α3 (C4M12a3) chains in serum as indicators of fibrosis....

  20. Basement membrane proteoglycans are of epithelial origin in rodent skin

    DEFF Research Database (Denmark)

    Yamane, Y; Yaoita, H; Couchman, J R


    proteoglycan and rat and mouse perlecan. While the isolated rat epidermis was shown to completely lack rat basement membrane chondroitin sulfate proteoglycan and rat basement membrane heparan sulfate proteoglycans, including perlecan, immunofluorescence staining of tissue sections from the grafted sites......-epidermal junction and hair follicle epithelium are of epidermal (epithelial) origin in vivo. Stratified rat keratinocytes cultured on a collagen matrix at the air-liquid interface showed the synthesis of perlecan, laminin 1, and type IV collagen in basement membranes, but not clearly detectable basement membrane...

  1. Contribution of alpha3(IV)alpha4(IV)alpha5(IV) Collagen IV to the Mechanical Properties of the Glomerular Basement Membrane (United States)

    Gyoneva, Lazarina

    The glomerular basement membrane (GBM) is a vital part of the blood-urine filtration barrier in the kidneys. In healthy GBMs, the main tension-resisting component is alpha3(IV)alpha4(IV)alpha5(IV) type IV collagen, but in some diseases it is replaced by other collagen IV isoforms. As a result, the GBM becomes leaky and disorganized, ultimately resulting in kidney failure. Our goal is to understanding the biomechanical aspects of the alpha3(IV)alpha4(IV)alpha5(IV) chains and how their absence could be responsible for (1) the initial injury to the GBM and (2) progression to kidney failure. A combination of experiments and computational models were designed for that purpose. A model basement membrane was used to compare experimentally the distensibility of tissues with the alpha3(IV)alpha4(IV)alpha5(IV) chains present and missing. The experiments showed basement membranes containing alpha3(IV)alpha4(IV)alpha5(IV) chains were less distensible. It has been postulated that the higher level of lateral cross-linking (supercoiling) in the alpha3(IV)alpha4(IV)alpha5(IV) networks contributes additional strength/stability to basement membranes. In a computational model of supercoiled networks, we found that supercoiling greatly increased the stiffness of collagen IV networks but only minimally decreased the permeability, which is well suited for the needs of the GBM. It is also known that the alpha3(IV)alpha4(IV)alpha5(IV) networks are more protected from enzymatic degradation, and we explored their significance in GBM remodeling. Our simulations showed that the more protected network was needed to prevent the system from entering a dangerous feedback cycle due to autoregulation mechanisms in the kidneys. Overall, the work adds to the evidence of biomechanical differences between the alpha3(IV)alpha4(IV)alpha5(IV) networks and other collagen IV networks, points to supercoiling as the main source of biomechanical differences, discusses the suitability of alpha3(IV)alpha4(IV

  2. Basement membrane zone remodeling during appendageal development in human fetal skin. The absence of type VII collagen is associated with gelatinase-A (MMP2) activity. (United States)

    Karelina, T V; Bannikov, G A; Eisen, A Z


    Epithelial cell adhesion, migration, and differentiation are controlled by interactions at the basement membrane zone (BMZ). Type VII collagen is the major collagenous component of anchoring fibrils that are essential for the attachment of the epidermis to the dermis. Gelatinase A (MMP-2) is believed to be necessary for the degradation of type VII collagen. In this study we have examined the in vivo distribution of type VII collagen and gelatinase A (Gel A) in the developing human epidermis and its appendages. At 13-15 wk of gestation a marked decrease in type VII collagen immunoreactivity was seen in the BMZ surrounding invading appendageal buds; however, type VII collagen mRNA was strongly expressed in the budding epidermal keratinocytes adjacent to the BMZ. At these stages, Gel A-positive mesenchymal-like cells were found scattered throughout the stroma with numerous Gel A-containing cells in direct contact with the developing appendageal buds. In situ zymography was used to show Gel A-activity in vivo. Gel A-mediated lysis was present at the interface between the appendageal buds and the underlying BMZ. By 20-25 wk of gestational age, immunostaining for type VII collagen protein was absent from the BMZ surrounding the distal portion of invading appendageal epithelial cords of both hair follicles and sweat glands. In contrast, type VII collagen mRNA was present in the basal keratinocytes adjacent to the BMZ surrounding the distal portion of these invading appendageal epithelial cords. At these stages Gel A-positive cells were present in the stroma directly adjacent to the distal portion of developing appendageal cords that lacked type VII collagen. In situ zymography showed zones of Gel A-mediated stromal lysis at the distal portion of developing appendageal cords. Interestingly, no differences were seen in the distribution of type IV collagen in the BMZ of both budding and resting fetal epidermis. These observations suggest that the absence of type VII collagen

  3. The major basement membrane components localize to the chondrocyte pericellular matrix--a cartilage basement membrane equivalent?

    DEFF Research Database (Denmark)

    Kvist, Alexander J.; Nyström, Alexander; Hultenby, Kjell;


    In this study, we demonstrate that articular cartilage chondrocytes are surrounded by the defining basement membrane proteins laminin, collagen type IV, nidogen and perlecan, and suggest that these form the functional equivalent of a basement membrane. We found by real-time PCR that mouse chondro...... to the progression of degenerative joint disorders....

  4. Cross-reactivity of cell-mediated immunity between interstitial (type I) and basement membrane (type IV) collagens



    In the present study, we demonstrate delayed-type hypersensitivity (DTH) to homologous type I collagen that cross-reacts with type IV collagen. Mice immunized with native or denatured type I collagens and challenged with these same antigens or native type IV collagen develop a peak DTH response on day 7. Challenge with denatured type IV collagen or collagenase-treated type IV collagen failed to elicit DTH in type I collagen-sensitized mice. Type I collagen-sensitized spleen cells adoptively t...

  5. Superficial Dermal Fibroblasts Enhance Basement Membrane and Epidermal Barrier Formation in Tissue-Engineered Skin: Implications for Treatment of Skin Basement Membrane Disorders



    Basement membrane is a highly specialized structure that binds the dermis and the epidermis of the skin, and is mainly composed of laminins, nidogen, collagen types IV and VII, and the proteoglycans, collagen type XVIII and perlecan, all of which play critical roles in the function and resilience of skin. Both dermal fibroblasts and epidermal keratinocytes contribute to the development of the basement membrane, and in turn the basement membrane and underlying dermis influence the development ...

  6. Molecular cloning of a cDNA encoding the porcine type XVII collagen noncollagenous 16 A domain and localization of the domain to the upper part of porcine skin basement membrane zone. (United States)

    Xu, Luting; Olivry, Thierry; Chan, Lawrence S


    Bullous pemphigoid is an autoimmune blistering human skin disease mediated by immunoglobulin (Ig)G autoantibodies targeting skin basement membrane component type XVII collagen, a transmembrane protein. Also designated BP180 and BPAG2, type XVII collagen is an extracellular matrix element essential for the connection between the epidermis and the underlying dermis. In addition to being a target antigen in the human disease bullous pemphigoid, type XVII collagen is also targeted by autoantibodies of canine, feline, equine and porcine patients suffering from a similar blistering skin disease. Previously, enzyme-linked immunosorbent assay and Western blot analyses have shown that autoantibodies from pigs affected with bullous pemphigoid recognize the human NC16A domain of type XVII collagen. To facilitate the development of porcine model of bullous pemphigoid, we isolated cDNA encoding the porcine type XVII collagen NC16A domain using a reverse transcription-polymerase chain reaction technique. The amino acids deduced from the NC16A cDNA showed 61% identity with the sequence of human NC16A. An antibody generated against a 20-amino acid peptide within the porcine NC16A localized the NC16A epitope to the upper part of porcine skin basement membrane zone. Our data provide further information of the porcine bullous pemphigoid target antigen and may help investigators for their further studies of this disease.

  7. Expression of basement membrane antigens in spindle cell melanoma. (United States)

    Prieto, V G; Woodruff, J M


    Spindle cell melanoma (SCM) is an uncommon form of melanoma that may be confused histologically with other tumors, including malignant peripheral nerve sheath tumors (MPNST). Tumors with neural differentiation and melanocytic nevi may both show basement membrane immunohistochemically and at the ultrastructural level. However, most ultrastructural studies of melanoma have failed to demonstrate well formed basement membrane around tumor cells. The presence of basement membrane has been used by some authors as evidence favoring MPNST, as opposed to SCM. To evaluate this distinction immunohistochemically, 22 primary and metastatic cutaneous melanomas having a spindle cell component (SCM) were studied using monoclonal antibodies against laminin and Type IV collagen. S100 protein and HMB45 antigen expression were also studied. All but one of the SCM were reactive for S100 protein in at least 25% of the cells. Thirteen of 20 tumors (65%) were focally reactive with HMB45. Laminin was expressed in 42% of the tumors (only membranous pattern in 3; cytoplasmic and membranous in 5). Seventeen tumors (77%) expressed type IV collagen (only membranous pattern in 7; cytoplasmic and membranous pattern in 10). Laminin and type IV collagen, known components of basement membrane, are often found in SCM. Therefore, their detection cannot be used to distinguish SCM from MPNST.

  8. The nature and biology of basement membranes. (United States)

    Pozzi, Ambra; Yurchenco, Peter D; Iozzo, Renato V


    Basement membranes are delicate, nanoscale and pliable sheets of extracellular matrices that often act as linings or partitions in organisms. Previously considered as passive scaffolds segregating polarized cells, such as epithelial or endothelial cells, from the underlying mesenchyme, basement membranes have now reached the center stage of biology. They play a multitude of roles from blood filtration to muscle homeostasis, from storing growth factors and cytokines to controlling angiogenesis and tumor growth, from maintaining skin integrity and neuromuscular structure to affecting adipogenesis and fibrosis. Here, we will address developmental, structural and biochemical aspects of basement membranes and discuss some of the pathogenetic mechanisms causing diseases linked to abnormal basement membranes.

  9. MMP mediated degradation of type IV collagen alpha 1 and alpha 3 chains reflects basement membrane remodeling in experimental and clinical fibrosis--validation of two novel biomarker assays.

    Directory of Open Access Journals (Sweden)

    Jannie Marie Sand

    Full Text Available OBJECTIVES: Fibrosis is characterized by excessive tissue remodeling resulting from altered expression of various growth factors, cytokines and proteases. We hypothesized that matrix metalloproteinase (MMP mediated degradation of type IV collagen, a main component of the basement membrane, will release peptide fragments (neo-epitopes into the circulation. Here we present the development of two competitive enzyme-linked immunosorbent assays (ELISAs for assessing the levels of specific fragments of type IV collagen α1 (C4M12a1 and α3 (C4M12a3 chains in serum as indicators of fibrosis. METHODS: Fragments of type IV collagen cleaved in vitro by MMP-12 were identified by mass spectrometry, and two were chosen for ELISA development due to their unique sequences. The assays were evaluated using samples from a carbon tetrachloride (CCl₄ rat model of liver fibrosis and from patients with idiopathic pulmonary fibrosis (IPF or chronic obstructive pulmonary disease (COPD. RESULTS: Two technically robust ELISAs were produced using neo-epitope specific monoclonal antibodies. Mean serum C4M12a1 levels were significantly elevated in CCl₄-treated rats compared with controls in weeks 12, 16, and 20, with a maximum increase of 102% at week 16 (p < 0.0001. Further, C4M12a1 levels correlated with the total collagen content of the liver in CCl₄-treated rats (r = 0.43, p = 0.003. Mean serum C4M12a3 levels were significantly elevated in patients with mild, moderate, and severe IPF, and COPD relative to healthy controls, with a maximum increase of 321% in COPD (p < 0.0001. CONCLUSIONS: Two assays measuring C4M12a1 and C4M12a3 enabled quantification of MMP mediated degradation of type IV collagen in serum. C4M12a1 was elevated in a pre-clinical model of liver fibrosis, and C4M12a3 was elevated in IPF and COPD patients. This suggests the use of these assays to investigate pathological remodeling of the basement membrane in different organs. However, validations in

  10. Immunohistochemical localization of basement membrane components during hair follicle morphogenesis

    DEFF Research Database (Denmark)

    Westgate, G E; Shaw, D A; Harrap, G J


    Specific antisera were used to investigate the distributions of several basement membrane zone (BMZ) components, namely, bullous pemphigoid antigen (BPA), heparan sulfate proteoglycan (HSPG), laminin, and type IV collagen, during the development of hair follicles in late embryo rats. BPA was not ......Specific antisera were used to investigate the distributions of several basement membrane zone (BMZ) components, namely, bullous pemphigoid antigen (BPA), heparan sulfate proteoglycan (HSPG), laminin, and type IV collagen, during the development of hair follicles in late embryo rats. BPA...... of the elongating follicle. HSPG was associated with the basal cell layer prior to the appearance of hair follicle primordia and became BMZ-associated before birth but after follicle buds were first observed. HSPG was also found to be associated with the basal cell surfaces in the epidermis, but not in the hair...... follicle. Laminin and type IV collagen were continually present in epidermal and follicular BMZ both before and during development of hair follicles and were later present in the dermal papilla matrix. From these observations we conclude that (1) laminin and type IV collagen are functionally important...


    Directory of Open Access Journals (Sweden)

    Ana Maria Abreu Velez


    Full Text Available Autoimmune mucocutaneous blistering diseases (ABDs represent a group of conditions that manifest with blisters on the skin and/or mucous membranes. Bullous pemphigoid (BP is the most common autoimmune mucocutaneous blistering disease. In BP, the location of the blisters is subepidermal and the oral involvement is rare. Variants of BP have been described, including pemphigoid vegetans; however, this disease is not completely characterized. The majority of ABDs have blisters and/or vesicles, that are often pruritic, and manifest autoantibodies to diverse proteins. These proteins include 1 hemidesmosomal plaque proteins(ie, BP230, plectins, 2 transmembrane proteins such as BP180 and α6β4-integrin, which are connected via laminin 332 to type VII collagen and 3 currently uncharacterized 105 kDa and 200 kDa molecules. Other ABDs include drug-induced linear IgA disease, bullous systemic lupus erythematosus (BSLE, dermatitis herpetiformis (DH, cicatricial pemphigoid (CP; also termed mucous membrane pemphigoid, lichen planus pemphigoides (LPP, pemphigoid gestationis (PG, herpes gestationis(HG, chronic bullous dermatosis of childhood (CBDC and the localized forms of CP, such as Brunsting-Perry pemphigoid. The diagnosis of ABDs requires clinical data; skin biopsies (in 10% buffered formalin for hematoxylin and eosin (H&E examination and skin biopsies(in Michel’s transport medium for direct immunofluorescence (DIF. In many ABDs, the histopathologic findings demonstrate a subepidermal vesicle or bulla with a luminal inflammatory infiltrate of neutrophils, eosinophils and/or lymphocytes. In many ABDs, an extensive perivascular and interstitial inflammatory infiltrate is also noted subjacent to the blister in the upper dermis. Normal skin adjacent to an ABD plaque is often excellent for DIF results. Many ABD biopsies reveal autoantibody deposition at the lesional basement membrane zone (BMZ; IgG, IgM, IgA, other immunoglobulins, complement components and

  12. ROCK1-directed basement membrane positioning coordinates epithelial tissue polarity. (United States)

    Daley, William P; Gervais, Elise M; Centanni, Samuel W; Gulfo, Kathryn M; Nelson, Deirdre A; Larsen, Melinda


    The basement membrane is crucial for epithelial tissue organization and function. However, the mechanisms by which basement membrane is restricted to the basal periphery of epithelial tissues and the basement membrane-mediated signals that regulate coordinated tissue organization are not well defined. Here, we report that Rho kinase (ROCK) controls coordinated tissue organization by restricting basement membrane to the epithelial basal periphery in developing mouse submandibular salivary glands, and that ROCK inhibition results in accumulation of ectopic basement membrane throughout the epithelial compartment. ROCK-regulated restriction of PAR-1b (MARK2) localization in the outer basal epithelial cell layer is required for basement membrane positioning at the tissue periphery. PAR-1b is specifically required for basement membrane deposition, as inhibition of PAR-1b kinase activity prevents basement membrane deposition and disrupts overall tissue organization, and suppression of PAR-1b together with ROCK inhibition prevents interior accumulations of basement membrane. Conversely, ectopic overexpression of wild-type PAR-1b results in ectopic interior basement membrane deposition. Significantly, culture of salivary epithelial cells on exogenous basement membrane rescues epithelial organization in the presence of ROCK1 or PAR-1b inhibition, and this basement membrane-mediated rescue requires functional integrin β1 to maintain epithelial cell-cell adhesions. Taken together, these studies indicate that ROCK1/PAR-1b-dependent regulation of basement membrane placement is required for the coordination of tissue polarity and the elaboration of tissue structure in the developing submandibular salivary gland.

  13. Molecular sieve of the rat glomerular basement membrane: a transmission electron microscopic study of enzyme-treated specimens.

    Directory of Open Access Journals (Sweden)



    Full Text Available Isolated rat glomerular basement membrane was treated with elastase and observed by transmission electron microscopy. The treatment with elastase revealed the fundamental structure of the glomerular basement membrane quite clearly, and enabled the observation of a sieve structure within the glomerular basement membrane. This sieve structure may play a major role in the filtration of blood as well as in the production of urine. Treatment with antibody showed that the sieve was mainly constituted of type IV collagen.

  14. The Acinar Cage: Basement Membranes Determine Molecule Exchange and Mechanical Stability of Human Breast Cell Acini.

    Directory of Open Access Journals (Sweden)

    Aljona Gaiko-Shcherbak

    Full Text Available The biophysical properties of the basement membrane that surrounds human breast glands are poorly understood, but are thought to be decisive for normal organ function and malignancy. Here, we characterize the breast gland basement membrane with a focus on molecule permeation and mechanical stability, both crucial for organ function. We used well-established and nature-mimicking MCF10A acini as 3D cell model for human breast glands, with ether low- or highly-developed basement membrane scaffolds. Semi-quantitative dextran tracer (3 to 40 kDa experiments allowed us to investigate the basement membrane scaffold as a molecule diffusion barrier in human breast acini in vitro. We demonstrated that molecule permeation correlated positively with macromolecule size and intriguingly also with basement membrane development state, revealing a pore size of at least 9 nm. Notably, an intact collagen IV mesh proved to be essential for this permeation function. Furthermore, we performed ultra-sensitive atomic force microscopy to quantify the response of native breast acini and of decellularized basement membrane shells against mechanical indentation. We found a clear correlation between increasing acinar force resistance and basement membrane formation stage. Most important native acini with highly-developed basement membranes as well as cell-free basement membrane shells could both withstand physiologically relevant loads (≤ 20 nN without loss of structural integrity. In contrast, low-developed basement membranes were significantly softer and more fragile. In conclusion, our study emphasizes the key role of the basement membrane as conductor of acinar molecule influx and mechanical stability of human breast glands, which are fundamental for normal organ function.

  15. Rat hair follicle dermal papillae have an extracellular matrix containing basement membrane components

    DEFF Research Database (Denmark)

    Couchman, J R


    Dermal papillae are small mesenchymally derived zones at the bases of hair follicles which have an important role in hair morphogenesis in the embryo and control of the hair growth cycle in postnatal mammals. The cells of the papilla are enmeshed in a dense extracellular matrix which undergoes...... extensive changes in concert with the hair cycle. Here it is shown that this matrix in anagen pelage follicles of postnatal rats contains an abundance of basement membrane components rather than dermal components such as interstitial collagens. In particular, type IV collagen, laminin, and basement membrane...

  16. Superficial dermal fibroblasts enhance basement membrane and epidermal barrier formation in tissue-engineered skin: implications for treatment of skin basement membrane disorders. (United States)

    Varkey, Mathew; Ding, Jie; Tredget, Edward E


    Basement membrane is a highly specialized structure that binds the dermis and the epidermis of the skin, and is mainly composed of laminins, nidogen, collagen types IV and VII, and the proteoglycans, collagen type XVIII and perlecan, all of which play critical roles in the function and resilience of skin. Both dermal fibroblasts and epidermal keratinocytes contribute to the development of the basement membrane, and in turn the basement membrane and underlying dermis influence the development and function of the epidermal barrier. Disruption of the basement membrane results in skin fragility, extensive painful blistering, and severe recurring wounds as seen in skin basement membrane disorders such as epidermolysis bullosa, a family of life-threatening congenital skin disorders. Currently, there are no successful strategies for treatment of these disorders; we propose the use of tissue-engineered skin as a promising approach for effective wound coverage and to enhance healing. Fibroblasts and keratinocytes isolated from superficial and deep dermis and epidermis, respectively, of tissue from abdominoplasty patients were independently cocultured on collagen-glycosaminoglycan matrices, and the resulting tissue-engineered skin was assessed for functional differences based on the underlying specific dermal fibroblast subpopulation. Tissue-engineered skin with superficial fibroblasts and keratinocytes formed a continuous epidermis with increased epidermal barrier function and expressed higher levels of epidermal proteins, keratin-5, and E-cadherin, compared to that with deep fibroblasts and keratinocytes, which had an intermittent epidermis. Further, tissue-engineered skin with superficial fibroblasts and keratinocytes formed better basement membrane, and produced more laminin-5, nidogen, collagen type VII, compared to that with deep fibroblasts and keratinocytes. Overall, our results demonstrate that tissue-engineered skin with superficial fibroblasts and keratinocytes

  17. Distribution of individual components of basement membrane in human colon polyps and adenocarcinomas as revealed by monoclonal antibodies

    DEFF Research Database (Denmark)

    Ljubimov, A V; Bartek, J; Couchman, J R


    -membrane components (laminin, entactin/nidogen, collagen type IV and large heparan sulfate proteoglycan), as well as to keratin 8. In all adenocarcinomas, including mucinous, basement membranes were altered more at the invasive front than in the parenchyma. The degree of this alteration was inversely correlated...... by the presence of fibrillar deposits of basement-membrane components, mainly of collagen type IV and/or heparan sulfate proteoglycan. This reaction was never observed in polyps and may be derived from myofibroblasts reported to accumulate in colon cancer stroma. The combined use of antibodies to basement...

  18. The bi-functional organization of human basement membranes. (United States)

    Halfter, Willi; Monnier, Christophe; Müller, David; Oertle, Philipp; Uechi, Guy; Balasubramani, Manimalha; Safi, Farhad; Lim, Roderick; Loparic, Marko; Henrich, Paul Bernhard


    The current basement membrane (BM) model proposes a single-layered extracellular matrix (ECM) sheet that is predominantly composed of laminins, collagen IVs and proteoglycans. The present data show that BM proteins and their domains are asymmetrically organized providing human BMs with side-specific properties: A) isolated human BMs roll up in a side-specific pattern, with the epithelial side facing outward and the stromal side inward. The rolling is independent of the curvature of the tissue from which the BMs were isolated. B) The epithelial side of BMs is twice as stiff as the stromal side, and C) epithelial cells adhere to the epithelial side of BMs only. Side-selective cell adhesion was also confirmed for BMs from mice and from chick embryos. We propose that the bi-functional organization of BMs is an inherent property of BMs and helps build the basic tissue architecture of metazoans with alternating epithelial and connective tissue layers.

  19. The bi-functional organization of human basement membranes.

    Directory of Open Access Journals (Sweden)

    Willi Halfter

    Full Text Available The current basement membrane (BM model proposes a single-layered extracellular matrix (ECM sheet that is predominantly composed of laminins, collagen IVs and proteoglycans. The present data show that BM proteins and their domains are asymmetrically organized providing human BMs with side-specific properties: A isolated human BMs roll up in a side-specific pattern, with the epithelial side facing outward and the stromal side inward. The rolling is independent of the curvature of the tissue from which the BMs were isolated. B The epithelial side of BMs is twice as stiff as the stromal side, and C epithelial cells adhere to the epithelial side of BMs only. Side-selective cell adhesion was also confirmed for BMs from mice and from chick embryos. We propose that the bi-functional organization of BMs is an inherent property of BMs and helps build the basic tissue architecture of metazoans with alternating epithelial and connective tissue layers.

  20. The spatial organization of Descemet's membrane-associated type IV collagen in the avian cornea



    The organization of type IV collagen in the unconventional basement membrane of the corneal endothelium (Descemet's membrane) was investigated in developing chicken embryos using anti-collagen mAbs. Both immunofluorescence histochemistry and immunoelectron microscopy were performed. In mature embryos (greater than 15 d of development), the type IV collagen of Descemet's membrane was present as an array of discrete aggregates of amorphous material at the interface between Descemet's membrane a...

  1. Basement membrane chondroitin sulfate proteoglycan alterations in a rat model of polycystic kidney disease

    DEFF Research Database (Denmark)

    Ehara, T; Carone, F A; McCarthy, K J;


    Alterations in basement membrane components, notably proteoglycans, in a rat model of polycystic kidney disease have been investigated. Rats were fed phenol II (2-amino-4-hydroxyphenyl-5-phenyl thiazole) for 4 days and then changed to normal diet for a 7-day recovery period. Marked dilation...... of distal tubules and collecting ducts was observed by 4 days with phenol II treatment, but the morphology returned to normal after 7 days of subsequent normal diet. Staining of tissue sections with two mouse monoclonal antibodies to a recently described basement membrane chondroitin sulfate proteoglycan...... membrane heparan sulfate proteoglycan core protein related to perlecan did not diminish but rather stained affected tubules intensely, whereas laminin, on the other hand, was apparently diminished in the basement membranes of the cystic tubules. Type IV collagen staining did not change through disease...

  2. Basement membrane-specific chondroitin sulfate proteoglycan is abnormally associated with the glomerular capillary basement membrane of diabetic rats

    DEFF Research Database (Denmark)

    McCarthy, K J; Abrahamson, D R; Bynum, K R;


    We have previously reported the production of monoclonal antibodies (MAb) recognizing the core protein of a basement membrane-specific chondroitin sulfate proteoglycan (BM-CSPG). Using immunohistochemical techniques, we have shown that BM-CSPG is present in almost every basement membrane, one...

  3. Accelerating repaired basement membrane after bevacizumab treatment on alkali-burned mouse cornea

    Directory of Open Access Journals (Sweden)

    Koon-Ja Lee


    Full Text Available To understand the corneal regeneration induced by bevacizumab,we investigated the structure changes of stroma andbasement membrane regeneration. A Stick soaked in 0.5 NNaOH onto the mouse cornea and 2.5 mg/ml of bevacizumabwas delivered into an alkali-burned cornea (2 μl by subconjunctivalinjections at 1 hour and 4 days after injury. At 7 daysafter injury, basement membrane regeneration was observedby transmission electron microscope. Uneven and thin epithelialbasement membrane, light density of hemidesmosomes,and edematous collagen fibril bundles are shown in thealkali-burned cornea. Injured epithelial basement membraneand hemidesmosomes and edematous collagen fibril bundlesresulting from alkali-burned mouse cornea was repaired bybevacizumab treatment. This study demonstrates that bevacizumabcan play an important role in wound healing in thecornea by accelerating the reestablishment of basementmembrane integrity that leads to barriers for scar formation.[BMB Reports 2013; 46(4: 195-200

  4. Coarctation induces alterations in basement membranes in the cardiovascular system

    DEFF Research Database (Denmark)

    Lipke, D W; McCarthy, K J; Elton, T S;


    A coarctation hypertensive rat model was used to examine the effects of elevated blood pressure on basement membrane component synthesis by cardiac myocytes and aorta using immunohistochemistry and Northern blot analysis. Carotid arterial pressure increased immediately on coarctation, and left...

  5. Expression of VLA-integrins and their related basement membrane ligands in gingiva from patients of various periodontitis categories

    DEFF Research Database (Denmark)

    Gürses, N.; Thorup, Alis Karabulut; Reibel, J.;


    integrins, basement membrane, gingiva, periodontitis, periodontal disease activity immunofluorescence......integrins, basement membrane, gingiva, periodontitis, periodontal disease activity immunofluorescence...

  6. Still more complexity in mammalian basement membranes

    DEFF Research Database (Denmark)

    Erickson, A C; Couchman, J R


    laminins, entactin-1/nidogen-1, Type IV collagen, and proteoglycans. However, within the past few years this complexity has increased as new components are described. The entactin/nidogen (E/N) family has expanded with the recent description of a new isoform, E/N-2/osteonidogen. Agrin and Type XVIII...... collagen have been reclassified as heparan sulfate proteoglycans (HSPGs), expanding the repertoire of HSPGs in the BM. The laminin family has become more diverse as new alpha-chains have been characterized, increasing the number of laminin isoforms. Interactions between BM components are now appreciated...

  7. Drosophila laminins act as key regulators of basement membrane assembly and morphogenesis. (United States)

    Urbano, Jose M; Torgler, Catherine N; Molnar, Cristina; Tepass, Ulrich; López-Varea, Ana; Brown, Nicholas H; de Celis, Jose F; Martín-Bermudo, Maria D


    Laminins are heterotrimeric molecules found in all basement membranes. In mammals, they have been involved in diverse developmental processes, from gastrulation to tissue maintenance. The Drosophila genome encodes two laminin alpha chains, one beta and one Gamma, which form two distinct laminin trimers. So far, only mutations affecting one or other trimer have been analysed. In order to study embryonic development in the complete absence of laminins, we mutated the gene encoding the sole laminin beta chain in Drosophila, LanB1, so that no trimers can be made. We show that LanB1 mutant embryos develop until the end of embryogenesis. Electron microscopy analysis of mutant embryos reveals that the basement membranes are absent and the remaining extracellular material appears disorganised and diffuse. Accordingly, abnormal accumulation of major basement membrane components, such as Collagen IV and Perlecan, is observed in mutant tissues. In addition, we show that elimination of LanB1 prevents the normal morphogenesis of most organs and tissues, including the gut, trachea, muscles and nervous system. In spite of the above structural roles for laminins, our results unravel novel functions in cell adhesion, migration and rearrangement. We propose that while an early function of laminins in gastrulation is not conserved in Drosophila and mammals, their function in basement membrane assembly and organogenesis seems to be maintained throughout evolution.

  8. Basement membrane reconstruction in human skin equivalents is regulated by fibroblasts and/or exogenously activated keratinocytes

    NARCIS (Netherlands)

    El Ghalbzouri, A; Jonkman, MF; Dijkman, R; Ponec, M


    This study was undertaken to examine the role fibroblasts play in the formation of the basement membrane (BM) in human skin equivalents. For this purpose, keratinocytes were seeded on top of fibroblast-free or fibroblast-populated collagen matrix or de-epidermized dermis and cultured in the absence

  9. Hypoplastic basement membrane of the lens anlage in the inheritable lens aplastic mouse (lap mouse). (United States)

    Aso, S; Baba, R; Noda, S; Ikuno, S; Fujita, M


    Adult homozygous lap mice show various eye abnormalities such as aphakia, retinal disorganization, and dysplasia of the cornea and anterior chamber. In the fetal eye of a homozygous lap mouse, the lens placode appears to develop normally. However, the lens vesicle develops abnormally to form a mass of cells without a cavity, and the mass vanishes soon afterward. Apoptotic cell death is associated with the disappearance of the lens anlage. We examined the basement membranes of the lens anlage of this mutant by immunohistochemical methods under light microscopy using antibodies against basement membrane components of the lens anlage, type IV collagen, fibronectin, laminin, heparan sulfate proteoglycan, and entactin and by transmission electron microscopy. Immunohistochemistry showed the distribution and intensity of antibody binding to the lens anlage to be almost the same for each these antibodies regardless of the stage of gestation or whether the anlagen were from normal BALB/c or lap mice. Thus, positive continuous reactions were observed around the exterior region of the lens anlage from day 10 of gestation for type IV collagen, fibronectin, laminin, heparan sulfate proteoglycan antibodies, and at least from day 11of gestation for entactin antibody. The basement membrane lamina densa of both normal and lap mice was shown by electron microscopy to be discontinuous at days 10 and 10.5 of gestation. However, by day 11 the lamina densa was continuous in the lens anlagen of normal mice but still discontinuous in the lap mice. By day 12 of gestation, the lamina densa had thickened markedly in normal mice, whereas in lap mice it remained discontinuous and its thinness indicated hypoplasia. These results indicate that, while all basement components examined are produced and deposited in the normal region of the lens anlage in the lap mouse, the basement membrane is, for some reason, imperfectly formed. The time at which hypoplasia of the basement membrane was observed

  10. Lysyl Hydroxylase 3 Localizes to Epidermal Basement Membrane and Is Reduced in Patients with Recessive Dystrophic Epidermolysis Bullosa.

    Directory of Open Access Journals (Sweden)

    Stephen A Watt

    Full Text Available Recessive dystrophic epidermolysis bullosa (RDEB is caused by mutations in COL7A1 resulting in reduced or absent type VII collagen, aberrant anchoring fibril formation and subsequent dermal-epidermal fragility. Here, we identify a significant decrease in PLOD3 expression and its encoded protein, the collagen modifying enzyme lysyl hydroxylase 3 (LH3, in RDEB. We show abundant LH3 localising to the basement membrane in normal skin which is severely depleted in RDEB patient skin. We demonstrate expression is in-part regulated by endogenous type VII collagen and that, in agreement with previous studies, even small reductions in LH3 expression lead to significantly less secreted LH3 protein. Exogenous type VII collagen did not alter LH3 expression in cultured RDEB keratinocytes and we show that RDEB patients receiving bone marrow transplantation who demonstrate significant increase in type VII collagen do not show increased levels of LH3 at the basement membrane. Our data report a direct link between LH3 and endogenous type VII collagen expression concluding that reduction of LH3 at the basement membrane in patients with RDEB will likely have significant implications for disease progression and therapeutic intervention.

  11. Pericapillary basement membrane thickening in human skeletal muscles. (United States)

    Baum, Oliver; Bigler, Marius


    The basement membrane (BM) surrounding capillaries in skeletal muscles varies physiologically in thickness according to age, physical fitness, and anatomical site in humans. Furthermore, the pericapillary BM thickness (CBMT) increases pathophysiologically during several common disease states, including peripheral arterial disease and diabetes mellitus. This review on CBM thickening in human skeletal muscles is two pronged. First, it addresses the advantages/disadvantages of grid- and tablet-based measuring and morphometric techniques that are implemented to assess the CBMT on transmission electron micrographs. Second, it deals with the biology of CBM thickening in skeletal muscles, particularly its possible causes, molecular mechanisms, and functional impact. CBM thickening is triggered by several physical factors, including diabetes-associated glycation, hydrostatic pressure, and inflammation. Increased biosynthesis of type IV collagen expression or repetitive cycles in pericyte or endothelial cell degeneration/proliferation appear to be most critical for CBM accumulation. A thickened CBM obviously poses a greater barrier for diffusion, lowers the microvascular elasticity, and impedes transcytosis of inflammatory cells. Our own morphometric data reveal the CBM enlargement to be not accompanied by the pericyte coverage. Owing to an overlap or redundancy in the capillary supply, CBM thickening in skeletal muscles might not be such a devastating occurrence as in organs with endarterial circulation (e.g., kidney and retina). CBM growth in skeletal muscles can be reversed by training or administration of antidiabetic drugs. In conclusion, CBM thickening in skeletal muscles is a microvascular remodeling process by which metabolic, hemodynamic, and inflammatory forces are integrated together and which could play a hitherto underestimated role in etiology/progression of human diseases.

  12. Abnormal glomerular basement membrane in idiopathic multicentric osteolysis

    NARCIS (Netherlands)

    Bakker, SJL; Vos, GD; Verschure, PDMM; Mulder, AH; Tiebosch, TMG


    The primary cause of nephropathy in idiopathic multicentric osteolysis is as yet unknown. We report a young girl with idiopathic multicentric osteolysis and nephropathy. An abnormal glomerular basement membrane was the only abnormality found in a renal biopsy taken 2 years before the development of

  13. Ultrastructure of basement membranes in developing shark tooth. (United States)

    Sawada, T; Inoue, S


    Based on studies of the tooth of largely mammalian species, the dental basement membranes are shown to be specialized for various roles significant in the development and maintenance of the tooth. Comparative studies with the nonmammalian tooth will facilitate further clarification of the mechanisms of mammalian tooth formation. In this study, basement membranes of the shark tooth in successive developmental stages was ultrastructurally examined for elucidation of their roles in odontogenesis. Teeth of a shark, Cephaloscyllium umbratile, were processed for thin section electron microscopy. Throughout the developmental stages the lamina densa of the basement membrane was made up of a fine network of "cords," irregular anastomosing strands known to be the major component of mammalian basement membranes. In the presecretory stage of the shark tooth, dental papilla cells were immobilized for their differentiation into odontoblasts by means of the binding of their processes to numerous narrow extensions of the lamina densa of the inner dental epithelium. In the secretory stage, a number of cords of the widened lamina densa were extended towards and bound to tubular vesicles of the forming enameloid. During the mineralization stage, fragments of the degrading enameloid matrix appeared to be moving through the lamina densa to the epithelial cells for processing. In the maturation stage, half of the lamina densa facing the enameloid was mineralized forming an advancing edge of mineralization of the enameloid. It provided strong binding and smooth transition of organic to mineral phase which may allow transportation of substances across the phases for enameloid maturation in a way similar to that reported in the mammalian tooth. These observations indicate that basement membranes of the developing shark tooth, as those in the mammalian tooth, play various roles, including anchoring, firm binding, and possible mediation of the transport of substances that are known to be

  14. Ultrastructural localization of the core protein of a basement membrane-specific chondroitin sulfate proteoglycan in adult rat skin

    DEFF Research Database (Denmark)

    McCarthy, K J; Horiguchi, Y; Couchman, J R


    Basement membranes are complex extracellular matrices present at epithelial/mesenchymal interfaces of tissues. The dermal-epidermal junction has been shown to contain numerous components, some of the most well known being laminin, types IV and VII collagens, heparan sulfate proteoglycan, fibronec......Basement membranes are complex extracellular matrices present at epithelial/mesenchymal interfaces of tissues. The dermal-epidermal junction has been shown to contain numerous components, some of the most well known being laminin, types IV and VII collagens, heparan sulfate proteoglycan...... primarily within the basal lamina, apparently concentrated in the lamina densa. In addition, some of the proteoglycan was also present beneath the lamina densa, associated with the reticular lamina collagen fibrils....

  15. Basement membrane abnormalities in human eyes with diabetic retinopathy

    DEFF Research Database (Denmark)

    Ljubimov, A V; Burgeson, R E; Butkowski, R J


    discontinuously for laminin-1, entactin/nidogen, and alpha3-alpha4 Type IV collagen, in contrast to non-DR corneas. Major BM alterations were found in DR retinas compared to normals and non-DR diabetics. The inner limiting membrane (retinal BM) of DR eyes had accumulations of fibronectin (including cellular......) and Types I, III, IV (alpha1-alpha2), and V collagen. The BM zone of new retinal blood vessels in neovascularized areas accumulated tenascin and Type XII collagen, whereas normal, diabetic, and adjacent DR retinas showed only weak and irregular staining. In preretinal membranes, perlecan, bamacan, and Types...... VI, VIII, XII, and XIV collagen were newly identified. Diabetic BM thickening appears to involve qualitative alterations of specific BM markers at an advanced disease stage, with the appearance of DR....

  16. Rat mesangial cells in vitro synthesize a spectrum of proteoglycan species including those of the basement membrane and interstitium

    DEFF Research Database (Denmark)

    Thomas, G J; Shewring, L; McCarthy, K J;


    Accumulation of extracellular matrix within the mesangium is an important event in the development of glomerular disease. In this report we have used indirect immunofluorescence to positively identify a number of constituents of the mesangial matrix synthesized by rat mesangial cells (RMC) in vitro...... including laminin, fibronectin, type IV collagen and the basement membrane heparan sulphate proteoglycan (BM-HSPG) known as perlecan. In addition, using Mab 2B5 we demonstrate that RMC synthesize a specific basement membrane chondroitin sulfate (BM-CSPG), a matrix component that in normal animals...... is localized in the mesangium but is not found in the pericapillary glomerular basement membrane (GBM). Further characterization of the proteoglycans synthesized by RMC in vitro revealed: (i) a second large CSPG, identified as versican; (ii) two small dermatan sulphate proteoglycans identified as biglycan...

  17. Regulation of the basement membrane by epithelia generated forces (United States)

    Tanner, Kandice


    Tumor metastasis involves a progressive loss of tissue architecture and dissolution of structural boundaries between the epithelium and connective tissue. The basement membrane (BM), a specialized network of extracellular matrix proteins forms a barrier that physically restricts pre-invasive lesions such that they remain as local insults. The BM is not a static structure, but one that is constantly regenerated and remodeled in the adult organism. Matrix organization also regulates cell function. Thus alterations in the balance of synthesis, remodeling and proteolytic degradation of the extracellular matrix proteins may contribute to a loss of structural integrity. However, the de novo assembly and maintenance of the complex structural properties of in vivo basement membranes remain elusive. Here, this paper highlights the current understanding on the structural properties and the establishment of the BM, and discusses the potential role of self-generated forces in adult tissue remodeling and the maintenance of the BM as a malignancy suppressor.

  18. Antiglomerular basement membrane antibody-crescentic glomerulonephritis complicating chronic bronchiectasis. (United States)

    Enríquez, R; Cabezuelo, J B; Sirvent, A E; Andrada, E; Amorós, F; Orti, C


    A 68-year-old woman with chronic bronchiectasis presented with haematuria and severe oligoanuric renal failure with no other serious systemic manifestation. Antiglomerular basement membrane (anti-GBM) antibodies and anti-myeloperoxidase antibodies were positive. Renal biopsy revealed anti-GBM crescentic glomerulonephritis. A conservative approach was followed and the patient is stable on chronic haemodialysis 6 months later. To the authors' knowledge, there has only been one previous report of anti-GBM disease complicating bronchiectasis.

  19. Basement membrane chondroitin sulfate proteoglycans: localization in adult rat tissues

    DEFF Research Database (Denmark)

    McCarthy, K J; Couchman, J R


    Heparan sulfate proteoglycans have been described as the major proteoglycan component of basement membranes. However, previous investigators have also provided evidence for the presence of chondroitin sulfate glycosaminoglycan in these structures. Recently we described the production...... and characterization of core protein-specific monoclonal antibodies (MAb) against a chondroitin sulfate proteoglycan (CSPG) present in Reichert's membrane, a transient extra-embryonic structure of rodents. This CSPG was also demonstrated to be present in adult rat kidney. We report here the tissue distribution...... sulfate proteoglycans previously described....

  20. Scaffold-forming and Adhesive Contributions of Synthetic Laminin-binding Proteins to Basement Membrane Assembly. (United States)

    McKee, Karen K; Capizzi, Stephanie; Yurchenco, Peter D


    Laminins that possess three short arms contribute to basement membrane assembly by anchoring to cell surfaces, polymerizing, and binding to nidogen and collagen IV. Although laminins containing the alpha4 and alpha5 subunits are expressed in alpha2-deficient congenital muscular dystrophy, they may be ineffective substitutes because they bind weakly to cell surfaces and/or because they lack the third arm needed for polymerization. We asked whether linker proteins engineered to bind to deficient laminins that provide such missing activities would promote basement membrane assembly in a Schwann cell model. A chimeric fusion protein (alphaLNNd) that adds a short arm terminus to laminin through the nidogen binding locus was generated and compared with the dystrophy-ameliorating protein miniagrin (mAgrin) that binds to the laminin coiled-coil dystroglycan and sulfatides. alphaLNNd was found to mediate laminin binding to collagen IV, to bind to galactosyl sulfatide, and to selectively convert alpha-short arm deletion-mutant laminins LmDeltaalphaLN and LmDeltaalphaLN-L4b into polymerizing laminins. This protein enabled polymerization-deficient laminin but not an adhesion-deficient laminin lacking LG domains (LmDeltaLG) to assemble an extracellular matrix on Schwann cell surfaces. mAgrin, on the other hand, enabled LmDeltaLG to form an extracellular matrix on cell surfaces without increasing accumulation of non-polymerizing laminins. These gain-of-function studies reveal distinct polymerization and anchorage contributions to basement membrane assembly in which the three different LN domains mediate the former, and the LG domains provide primary anchorage with secondary contributions from the alphaLN domain. These findings may be relevant for an understanding of the pathogenesis and treatment of laminin deficiency states.

  1. Synthesis and deposition of basement membrane proteins by primary brain capillary endothelial cells in a murine model of the blood-brain barrier

    DEFF Research Database (Denmark)

    Thomsen, Maj Schneider; Birkelund, Svend; Burkhart, Annette;


    basement membrane proteins such as laminin-411, laminin-511, collagen IV [α1(IV)2 α2(IV)], agrin, perlecan, and nidogen 1 and 2 in vitro. Increased expression of the laminin α5 subunit correlated to the addition of BBB inducing factors (hydrocortisone, Ro 20-1724, and pCPT-cAMP), whereas increased...... expression of collagen IV α1 primarily correlated to increased levels of cAMP. In conclusion, BCECs cultured in vitro coherently form a BBB and express basement membrane proteins as a feature of maturation. This article is protected by copyright. All rights reserved....

  2. Scaffold-forming and Adhesive Contributions of Synthetic Laminin-binding Proteins to Basement Membrane Assembly*S⃞


    McKee, Karen K.; Capizzi, Stephanie; Yurchenco, Peter D.


    Laminins that possess three short arms contribute to basement membrane assembly by anchoring to cell surfaces, polymerizing, and binding to nidogen and collagen IV. Although laminins containing the α4 and α5 subunits are expressed in α2-deficient congenital muscular dystrophy, they may be ineffective substitutes because they bind weakly to cell surfaces and/or because they lack the third arm needed for polymerization. We asked whether linker proteins engineered to bind...

  3. Effects of radiation on the permeability of human basement membranes (United States)

    Fan, B.-T.; Achour, S.; Simmonet, F.; Guerin, D.


    The influence of radiation on the permeability properties of human basement membrane was investigated by measuring the diffusion rate of several organic compounds (glycine, proline, glucose, urea and insulin) through human anterior lens capsules. The basement membranes borne an γ-irradiation treatment change significantly their permeability vis-a-vis studied organic substances. This modification in physico-chemical properties is probably due to the radiation, which alters or degrades the complex structure (or architecture) of basement membranes. Moreover the change in permeability is dependent upon the diffusing compounds. An increase in diffusion has been observed for glucose, glycine and urea. However for insulin and proline, a decrease in diffusion rate was observed. L'influence de radiation sur la perméabilité de la membrane basale a été étudiée par la mesure de la vitesse de diffusion de plusieurs composés organiques d'intérêt biologique (glycine, proline, glucose, urée et insuline) à travers la lame basale antérieure du cristallin de l'oil humain. Les membranes basales qui sont traitées avec l'irradiation γ changent significativement leur perméabilité vis-à-vis des substances organiques. Ce changement de propriétés physico-chimiques est probablement dû à l'altération ou la dégradation de la structure (ou de l'architecture) de la membrane basale entraînée par l'irradiation. De plus, la modification de la perméabilité de la membrane basale est dépendante des composés diffusants. Une augmentation de la vitesse de diffusion a été observée pour le glucose, le glycine et l'urée. Par contre, dans les cas de l'insuline et de la proline, on a observé une diminution de la vitesse de diffusion.

  4. Mechanical Stretch on Human Skin Equivalents Increases the Epidermal Thickness and Develops the Basement Membrane.

    Directory of Open Access Journals (Sweden)

    Eijiro Tokuyama

    Full Text Available All previous reports concerning the effect of stretch on cultured skin cells dealt with experiments on epidermal keratinocytes or dermal fibroblasts alone. The aim of the present study was to develop a system that allows application of stretch stimuli to human skin equivalents (HSEs, prepared by coculturing of these two types of cells. In addition, this study aimed to analyze the effect of a stretch on keratinization of the epidermis and on the basement membrane. HSEs were prepared in a gutter-like structure created with a porous silicone sheet in a silicone chamber. After 5-day stimulation with stretching, HSEs were analyzed histologically and immunohistologically. Stretch-stimulated HSEs had a thicker epidermal layer and expressed significantly greater levels of laminin 5 and collagen IV/VII in the basal layer compared with HSEs not subjected to stretch stimulation. Transmission electron microscopy revealed that the structure of the basement membrane was more developed in HSEs subjected to stretching. Our model may be relevant for extrapolating the effect of a stretch on the skin in a state similar to an in vivo system. This experimental system may be useful for analysis of the effects of stretch stimuli on skin properties and wound healing and is also expected to be applicable to an in vitro model of a hypertrophic scar in the future.

  5. Immunohistochemical expression of basement membrane proteins of verrucous carcinoma of the oral mucosa. (United States)

    Arduino, Paolo G; Carrozzo, Marco; Pagano, Marco; Broccoletti, Roberto; Scully, Crispian; Gandolfo, Sergio


    Squamous cell carcinoma (SCC) of the oral cavity is an extremely invasive tumour of stratified squamous epithelium that spreads throughout degradation of the basement membrane (BM) and extra-cellular matrix. Oral verrucous carcinoma (VC) is a rare low-grade variant of oral SCC that penetrates into the subepithelial connective tissue. It also has a different clinical behaviour from classical oral SCC. We investigated the immunohistochemical expression of laminin, laminin-5, collagen IV and fibronectin in VC, severe epithelial dysplasia (SED) and SCC in order to analyse if the pattern of these molecules expression contributes to the differences in the biological behaviour of these diseases. The staining pattern of laminin was less intensive in SCC compared with SED and VC, and collagen IV expression was increased in VC compared with SED. Discontinuities of laminin, collagen IV and fibronectin were more evident in SED than in VC. This study indicates that VC has a biological behaviour different from SED or SCC, observable by immunohistochemistry in the BM zone.

  6. Laminin isoforms in endothelial and perivascular basement membranes. (United States)

    Yousif, Lema F; Di Russo, Jacopo; Sorokin, Lydia


    Laminins, one of the major functional components of basement membranes, are found underlying endothelium, and encasing pericytes and smooth muscle cells in the vessel wall. Depending on the type of blood vessel (capillary, venule, postcapillary venule, vein or artery) and their maturation state, both the endothelial and mural cell phenotype vary, with associated changes in laminin isoform expression. Laminins containing the α4 and α5 chains are the major isoforms found in the vessel wall, with the added contribution of laminin α2 in larger vessels. We here summarize current data on the precise localization of these laminin isoforms and their receptors in the different layers of the vessel wall, and their potential contribution to vascular homeostasis.

  7. Production of monoclonal antibodies to human glomerular basement membrane.

    Directory of Open Access Journals (Sweden)



    Full Text Available Using the technique of somatic cell fusion, we produced monoclonal antibodies to collagenase-digested human glomerular basement membrane (GBM. Fourteen monoclonal antibodies which reacted with normal human kidney in indirect immunofluorescence (IIF studies were produced. An analysis of the binding patterns indicated that the antigens recognized could be divided into six broad groups. Monoclonal antibody B3-H10 (Group 1 reacted with only GBM in a fine granular pattern. A5-B12 and B5-C2 (Group 2 reacted with GBM and peritubular capillary in a linear pattern. B2-A12 (Group 3 reacted with only epithelial cells. Al-C9 and A4-E2 (Group 4 showed a mesangial pattern in glomerulus and a lineal pattern in tubular basement membrane (TBM, Bowman's capsule and peritubular capillary. A1-E1, A1-E11, A2-E6, A3-B6, A4-F8 and B5-H2 (Group 5 recognized determinants common to GBM, TBM, Bowman's capsule and/or peritubular capillary. A3-F1 and B5-E10 (Group 6 reacted with TBM and Bowman's capsule. The staining pattern of B3-H10 (Group 1 was characteristic because it was not linear, but finely granular along the GBM. The staining pattern of B2-A12 (Group 3 was also characteristic because only epithelial cells were stained, and processes of epithelial cells were observed as fine fibrils. To the best of our knowledge, these two types of monoclonal antibodies have not been reported previously.

  8. MT1-MMP-mediated basement membrane remodeling modulates renal development

    Energy Technology Data Exchange (ETDEWEB)

    Riggins, Karen S.; Mernaugh, Glenda [Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Su, Yan; Quaranta, Vito [Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Koshikawa, Naohiko; Seiki, Motoharu [Division of Cancer Cell Research, Institute of Medical Science, The University of Tokyo, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Pozzi, Ambra [Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Research Medicine, Veterans Affairs Hospital, Nashville, TN 37232 (United States); Zent, Roy, E-mail: [Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Research Medicine, Veterans Affairs Hospital, Nashville, TN 37232 (United States)


    Extracellular matrix (ECM) remodeling regulates multiple cellular functions required for normal development and tissue repair. Matrix metalloproteinases (MMPs) are key mediators of this process and membrane targeted MMPs (MT-MMPs) in particular have been shown to be important in normal development of specific organs. In this study we investigated the role of MT1-MMP in kidney development. We demonstrate that loss of MT1-MMP leads to a renal phenotype characterized by a moderate decrease in ureteric bud branching morphogenesis and a severe proliferation defect. The kidneys of MT1-MMP-null mice have increased deposition of collagen IV, laminins, perlecan, and nidogen and the phenotype is independent of the MT-1MMP target, MMP-2. Utilizing in vitro systems we demonstrated that MTI-MMP proteolytic activity is required for renal tubule cells to proliferate in three dimensional matrices and to migrate on collagen IV and laminins. Together these data suggest an important role for MT1-MMP in kidney development, which is mediated by its ability to regulate cell proliferation and migration by proteolytically cleaving kidney basement membrane components.

  9. The borderline: Basement membranes and the transition from premalignant to malignant neoplasia

    NARCIS (Netherlands)

    F.T.B. Bosman (Fré)


    textabstractIn this paper, the use of immunohistochemistry for the analysis of basement membrane components and related extracellular matrix proteins in human cancer is reviewed. Basement membranes in cancer are dynamic structures that are constantly degraded but also deposited, in close collaborati

  10. Enhanced assembly of basement membrane matrix by endodermal cells in response to fibronectin substrata

    DEFF Research Database (Denmark)

    Austria, M R; Couchman, J R


    Basement membranes are complex extracellular matrices contributing to the regulation of growth, migration and differentiation of many cell types. However, little is known about the mechanisms regulating the deposition and assembly of basement membrane from its constituents. We have investigated t...

  11. Role of the basement membrane in regulation of cardiac electrical properties. (United States)

    Yang, Huaxiao; Borg, Thomas K; Wang, Zhonghai; Ma, Zhen; Gao, Bruce Z


    In the heart muscle, each adult cardiomyocyte is enclosed by a basement membrane (BM). This innermost extracellular matrix is a layered assembly of laminin, collagen IV, glycoproteins, and proteoglycans. In this study, the role of the BM network in regulation of the electrical properties of neonatal cardiomyocytes (NCMs) cultured on an aligned collagen I gel was investigated using a multielectrode array (MEA). A laminin antibody was added to the culture medium for 48-120 h to conjugate newly secreted laminin. Then, morphology of the NCMs on an MEA was monitored using a phase contrast microscope, and the BM network that was immunocytostained for laminin was imaged using a fluorescence microscope. When the BM laminin was absent in this culture model, dramatic changes in NCM morphology were observed. Simultaneously, the MEA-recorded cardiac field potential showed changes compared to that from the control groups: The period of contraction shortened to 1/2 of that from the control groups, and the waveform of the calcium influx shifted from a flat plateau to a peak-like waveform, indicating that the electrical properties of the NCMs were closely related to the components and distribution of the BM network.

  12. Chitosan facilitates structure formation of the salivary gland by regulating the basement membrane components. (United States)

    Yang, Tsung-Lin; Hsiao, Ya-Chuan


    Tissue structure is important for inherent physiological function and should be recapitulated during tissue engineering for regenerative purposes. The salivary gland is a branched organ that is responsible for saliva secretion and regulation. The salivary glands develop from epithelial-mesenchymal interactions, and depend on the support of the basement membrane (BM). Chitosan-based biomaterials have been demonstrated to be competent in facilitating the formation of salivary gland tissue structure. However, the underlying mechanisms have remained elusive. In the developing submandibular gland (SMG), the chitosan effect was found to diminish when collagen and laminin were removed from cultured SMG explants. Chitosan increased the expression of BM components including collagen, laminin, and heparan sulfate proteoglycan, and also facilitated BM components and the corresponding receptors to be expressed in tissue-specific patterns beneficial for SMG branching. The chitosan effect decreased when either laminin components or receptors were inhibited, as well when the downstream signaling was blocked. Our results revealed that chitosan promotes salivary glands branching through the BM. By regulating BM components and receptors, chitosan efficiently stimulated downstream signaling to facilitate salivary gland branching. The present study revealed the underlying mechanism of the chitosan effect in engineering SMG structure formation.

  13. Basement membrane and interstitial proteoglycans produced by MDCK cells correspond to those expressed in the kidney cortex

    DEFF Research Database (Denmark)

    Erickson, A C; Couchman, J R


    ) cells has been utilized. Proteoglycans were prepared from conditioned medium by DEAE anion exchange chromatography. The eluted PGs were treated with heparitinase or chondroitinase ABC (cABC), separately or combined, followed by SDS-PAGE. Western blot analysis, using antibodies specific for various PG...... core proteins or CS stubs generated by cABC treatment, revealed that both basement membrane and interstitial PGs are secreted by MDCK cells. HSPGs expressed by MDCK cells are perlecan, agrin, and collagen XVIII. Various CSPG core proteins are made by MDCK cells and have been identified as biglycan...

  14. Clinical and genetic features of X-linked Alport syndrome in men positive for the collagen Ⅳ a5 chain in epidermal basement membrane%皮肤基底膜Ⅳ型胶原α5链染色正常的男性X连锁Alport综合征基因型和表型分析

    Institute of Scientific and Technical Information of China (English)

    张琰琴; 丁洁; 王芳; 张宏文; 肖慧捷; 姚勇; 钟旭辉; 管娜; 刘晓宇


    目的 分析皮肤基底膜Ⅳ型胶原α5[α5(Ⅳ)]链染色正常的男性x连锁Alport综合征(XLAS)的基因突变和临床特点.方法 回顾性分析1998年1月至2014年12月北京大学第一医院儿科诊断为Alport综合征的725个家系,其中男性XLAS患者450例,通过入选标准:(1)有皮肤基底膜Ⅳ型胶原α5链染色检查,(2)存在COI4A5基因突变,筛选出α5(Ⅳ)染色正常及α5(Ⅳ)染色阴性的男性XLAS患者,比较、分析两组基因突变和临床特点的差异.组间比较采用Mann-Whitney检验或x2检验.结果 共140例男性XLAS患者既进行了皮肤基底膜Ⅳ型胶原α5链染色检查又检测到了COI4A5基因突变.其中α5(Ⅳ)染色正常组18例,α5(Ⅳ)染色阴性组122例.两组患者就诊中位数年龄分别为11.0、7.2岁(Z=-1.839,P=0.066),就诊时24h尿蛋白定量中位数分别是1.50、0.57 g/d(Z=-1.212,P=0.226),合并听力减退28%、53%(x2=3.619,P=0.067),进展至终末期肾脏病(ESRD)的分别为4、12例(x2=2.377,P=0.128),ESRD发生的中位数年龄分别是31.0、16.6岁(Z=-2.554,P=0.011),COL4A5基因错义突变分别为12例(67%)、63例(52%)(x2=1.424,P=0.313).结论 皮肤基底膜α5(Ⅳ)染色正常的男性XLAS患者与α5(Ⅳ)染色阴性的男性XLAS患者在尿蛋白定量、听力减退发生率、COL4A5基因型差别不明显,但α5(Ⅳ)染色正常组患者发生ESRD的年龄明显较晚.%Objective To analyze the clinical and genetic features of X-linked Alport syndrome (XLAS) in men positive for the collagen α5 (Ⅳ) chain in epidermal basement membrane.Method This was a retrospective study.Totally 725 families were diagnosed as Alport syndrome in Department of Pediatrics of Peking University First Hospital during January 1998 to December 2014, among them 450 patients were males with XLAS.Patients who met both of the following two criteria were included in this study.(1) Patients underwent α5 (Ⅳ) chain staining in the epidermal basement membrane.(2

  15. Basement membrane proteoglycans in glomerular morphogenesis: chondroitin sulfate proteoglycan is temporally and spatially restricted during development

    DEFF Research Database (Denmark)

    McCarthy, K J; Bynum, K; St John, P L;


    We previously reported the presence of a basement membrane-specific chondroitin sulfate proteoglycan (BM-CSPG) in basement membranes of almost all adult tissues. However, an exception to this ubiquitous distribution was found in the kidney, where BM-CSPG was absent from the glomerular capillary......, the present study used light and electron microscopic immunohistochemistry to examine the distribution of BM-CSPG and basement membrane heparan sulfate proteoglycan (BM-HSPG) during prenatal and postnatal renal development in the rat. Our results show that the temporal and spatial pattern of expression of BM...

  16. FOS-1 promotes basement-membrane removal during anchor-cell invasion in C. elegans. (United States)

    Sherwood, David R; Butler, James A; Kramer, James M; Sternberg, Paul W


    Cell invasion through basement membranes is crucial during morphogenesis and cancer metastasis. Here, we genetically dissect this process during anchor-cell invasion into the vulval epithelium in C. elegans. We have identified the fos transcription factor ortholog fos-1 as a critical regulator of basement-membrane removal. In fos-1 mutants, the gonadal anchor cell extends cellular processes normally toward vulval cells, but these processes fail to remove the basement membranes separating the gonad from the vulval epithelium. fos-1 is expressed in the anchor cell and controls invasion cell autonomously. We have identified ZMP-1, a membrane-type matrix metalloproteinase, CDH-3, a Fat-like protocadherin, and hemicentin, a fibulin family extracellular matrix protein, as transcriptional targets of FOS-1 that promote invasion. These results reveal a key genetic network that controls basement-membrane removal during cell invasion.

  17. Perlecan and basement membrane-chondroitin sulfate proteoglycan (bamacan) are two basement membrane chondroitin/dermatan sulfate proteoglycans in the Engelbreth-Holm-Swarm tumor matrix

    DEFF Research Database (Denmark)

    Couchman, J R; Kapoor, R; Sthanam, M;


    The presence of proteoglycans bearing galactosaminoglycan chains has been reported, but none has been identified previously in the matrix of the Engelbreth-Holm-Swarm tumor, which is a source of several basement membrane components. This tumor matrix contains perlecan, a large, low buoyant density...... heparan sulfate proteoglycan, widespread in many basement membranes and connective tissues. We now identify two distinct proteoglycan species from this tumor source, which are substituted with galactosaminoglycans and which show basement membrane localization by immunohistochemistry. One species...... is perlecan but, in addition to being present as a heparan sulfate proteoglycan, it is also present as a hybrid molecule, with dermatan sulfate chains. A minor population of perlecan apparently lacks heparan sulfate chains totally, and some of this is substituted with chondroitin sulfate. The second species...

  18. Anti-glomerular basement membrane disease superimposed on membranous nephropathy: a case report and review of the literature

    Directory of Open Access Journals (Sweden)

    Nivera Noel


    Full Text Available Abstract Introduction Anti-glomerular basement membrane disease is a rare autoimmune disorder characterized by pulmonary hemorrhage, crescentic glomerulonephritis and the presence of circulating anti-glomerular basement membrane antibodies. The simultaneous occurrence of both anti-glomerular basement membrane disease and membranous nephropathy is rare. Case presentation A 59-year-old Hispanic man presented with acute onset of nausea and vomiting and was found to have renal insufficiency. Work-up included a kidney biopsy, which revealed anti-glomerular basement membrane disease with underlying membranous nephropathy. He was treated with emergent hemodialysis, intravenous corticosteroids, plasmapheresis, and cyclophosphamide without improvement in his renal function. Conclusion Simultaneous anti-glomerular basement membrane disease and membranous nephropathy is very rare. There have been 16 previous case reports in the English language literature that have been associated with a high mortality and morbidity, and a very high rate of renal failure resulting in hemodialysis. Co-existence of membranous nephropathy and anti-glomerular basement membrane disease may be immune-mediated, although the exact mechanism is not clear.

  19. Active Peptide-Conjugated Chitosan Matrices as an Artificial Basement Membrane

    Directory of Open Access Journals (Sweden)

    Kentaro Hozumi


    Full Text Available The basement membrane, a thin extracellular matrix, plays a critical role in tissue development and repair. Laminins are the major component of basement membrane and have diverse biological activities. We have identified various cell-adhesive peptides from laminins and their specific cell surface receptors. Polysaccharides, including chitosan, have been used as scaffolds, which regulate cellular functions for tissue engineering. We have developed laminin-derived active peptide-chitosan matrices as functional scaffolds. The biological activity of the peptides was enhanced when the peptides were conjugated to a chitosan matrix, suggesting that the peptide-chitosan matrix approach has an advantage for an active biomaterial. Further, the laminin peptide-chitosan matrices have the potential to mimic the basement membrane and are useful for tissue engineering as an artificial basement membrane.

  20. Anti-glomerular basement membrane glomerulonephritis and thrombotic microangiopathy in first degree relatives; a case report


    Idorn Thomas; Schejbel Lone; Rydahl Casper; Heaf James; Jølvig Karen; Bergstrøm Marie; Garred Peter; Kamper Anne-Lise


    Abstract Background Anti-glomerular basement membrane glomerulonephritis and thrombotic microangiopathy are rare diseases with no known coherence. Case Presentation A daughter and her biological mother were diagnosed with pregnancy-induced thrombotic microangiopathy and anti-glomerular basement membrane glomerulonephritis, respectively. Both developed end-stage renal disease. Exploration of a common aetiology included analyses of HLA genotypes, functional and genetic aspects of the complement...

  1. Expression of basement membrane components through morphological changes in the hair growth cycle

    DEFF Research Database (Denmark)

    Couchman, J R; Gibson, W T


    The amount and distribution of fibronectin associated with hair follicles was found to vary during the hair growth cycle in the rat. Immunocytochemical staining of follicles in mid-late anagen (the growth stage) revealed the presence of fibronectin in the dermal papilla matrix, in the basement...... membrane separating this from the epithelial cells of the hair bulb, and in the basement membrane and connective tissue sheath which underly the cells of the outer root sheath. Early in catagen, the transitional stage, staining of the dermal papilla matrix disappeared. Fibronectin persisted in the basement...

  2. A Review on the Potential Role of Basement Membrane Laminin in the Pathogenesis of Psoriasis. (United States)

    McFadden, J P; Kimber, I


    We have previously reviewed alterations to basement membrane laminin in psoriasis and how disruption of this layer could lead to at least some of the pathological changes observed. We here postulate that basement membrane laminin is the key antigen in driving psoriasis, inducing a T cell-mediated autoimmune response. For laminin to be considered as the key autoantigen in psoriasis, it would be reasonable to expect the following to be demonstrable: (1) that autoantigens are present in psoriatic inflammation; (2) that basement membrane laminin is perturbed in involved and uninvolved skin, and that some of the pathological changes associated with psoriasis could be predicted as a sequel to this; (3) that disruption of the basement membrane is among the earliest events in the evolution of psoriatic lesions; (4) that as streptococcal pharyngitis is the most clearly defined event to trigger or exacerbate psoriasis, then a T cell-mediated autoimmune response to laminin should be anticipated as a potential sequelae to streptococcal pharyngitis; (5) that T cells in psoriasis can be shown to react to peptides with homology to laminin; (6) that HLACw6, as the most closely related gene associated with psoriasis and which is involved in antigen expression, should be preferentially expressed within lesional psoriasis towards the basement membrane, together with other proximal associated immune activity; and (7) that there is some association between antilaminin pemphigoid, a humorally mediated autoimmune disease to skin basement membrane laminin, and psoriasis. We here review the data relevant to each of these requirements.

  3. Influence of collagen source on fibrillar architecture and properties of vitrified collagen membranes. (United States)

    Majumdar, Shoumyo; Guo, Qiongyu; Garza-Madrid, Marcos; Calderon-Colon, Xiomara; Duan, Derek; Carbajal, Priscilla; Schein, Oliver; Trexler, Morgana; Elisseeff, Jennifer


    Collagen vitrigel membranes are transparent biomaterials characterized by a densely organized, fibrillar nanostructure that show promise in the treatment of corneal injury and disease. In this study, the influence of different type I collagen sources and processing techniques, including acid-solubilized collagen from bovine dermis (Bov), pepsin-solubilized collagen from human fibroblast cell culture (HuCC), and ficin-solubilized collagen from recombinant human collagen expressed in tobacco leaves (rH), on the properties of the vitrigel membranes was evaluated. Postvitrification carbodiimide crosslinking (CX) was also carried out on the vitrigels from each collagen source, forming crosslinked counterparts BovXL, HuCCXL, and rHXL, respectively. Collagen membrane ultrastructure and biomaterial properties were found to rely heavily on both collagen source and crosslinking. Bov and HuCC samples showed a random fibrillar organization of collagen, whereas rH vitrigels showed remarkable regional fibril alignment. After CX, light transmission was enhanced in all groups. Denaturation temperatures after CX increased in all membranes, of which the highest increase was seen in rH (14.71°C), suggesting improved thermal stability of the collagen fibrils in the membranes. Noncrosslinked rH vitrigels may be reinforced through CX to reach levels of mechanical strength and thermal stability comparable to Bov.

  4. Does Tensile Rupture of Tumor Basement Membrane Mark the Onset of Cancer Metastasis? (United States)

    Prakash, Sai


    Recognizing a conceptual analogy from polymer physics and reasoning via induction, we infer the plausibility that a malignant tumor (carcinoma) grows in size until a threshold determined by its mechanochemical state in relation to its microenvironment whence, peripheral cells undergo epithelial-to-mesenchymal transitions (EMT) facilitating metastasis. This state is equated to the tensile yielding/rupture of the proteolytically-weakened basement membrane (BM) that encapsulates the growing neoplasm. BMs are typically constituted of tri-continuous hydrogel networks of collagen-IV, laminin, and interstitial fluid, with connector proteins such as nidogens, and perlecans. We test this postulate by formulating a theoretical model based on continuum fluid-solid mechanics, diffusion, and biochemical kinetics of energy metabolism. Herein, a prototypical, viscous tumor spheroid grows radially, consuming metabolic nutrients while being constrained by an elastic BM ca. 0.5-2 microns-thick, and cell adhesion molecules (CAMs), chiefly cadherins and integrins. The model is computationally analyzed via Comsol®. Results validate the a priori conjecture, and predict subsequent crack-tip stresses shifting strains on the CAMs from compressive to tensile, that might also indicate mechanotransduced switches in their conformations, such as from non-invasive, adhesive E-cadherins to invasive, non-adhesive N-cadherin phenotypes. Grant from Brady Urological Institute, JHMI.

  5. Integrating Activities of Laminins that Drive Basement Membrane Assembly and Function. (United States)

    Yurchenco, Peter D


    Studies on extracellular matrix proteins, cells, and genetically modified animals have converged to reveal mechanisms of basement membrane self-assembly as mediated by γ1 subunit-containing laminins, the focus of this chapter. The basic model is as follows: A member of the laminin family adheres to a competent cell surface and typically polymerizes followed by laminin binding to the extracellular adaptor proteins nidogen, perlecan, and agrin. Assembly is completed by the linking of nidogen and heparan sulfates to type IV collagen, allowing it to form a second stabilizing network polymer. The assembled matrix provides structural support, anchoring the extracellular matrix to the cytoskeleton, and acts as a signaling platform. Heterogeneity of function is created in part by the isoforms of laminin that vary in their ability to polymerize and to interact with integrins, dystroglycan, and other receptors. Mutations in laminin subunits, affecting expression or LN domain-specific functions, are a cause of human diseases that include those of muscle, nerve, brain, and kidney.

  6. Vascular basement membranes as pathways for the passage of fluid into and out of the brain. (United States)

    Morris, Alan W J; Sharp, Matthew MacGregor; Albargothy, Nazira J; Fernandes, Rute; Hawkes, Cheryl A; Verma, Ajay; Weller, Roy O; Carare, Roxana O


    In the absence of conventional lymphatics, drainage of interstitial fluid and solutes from the brain parenchyma to cervical lymph nodes is along basement membranes in the walls of cerebral capillaries and tunica media of arteries. Perivascular pathways are also involved in the entry of CSF into the brain by the convective influx/glymphatic system. The objective of this study is to differentiate the cerebral vascular basement membrane pathways by which fluid passes out of the brain from the pathway by which CSF enters the brain. Experiment 1: 0.5 µl of soluble biotinylated or fluorescent Aβ, or 1 µl 15 nm gold nanoparticles was injected into the mouse hippocampus and their distributions determined at 5 min by transmission electron microscopy. Aβ was distributed within the extracellular spaces of the hippocampus and within basement membranes of capillaries and tunica media of arteries. Nanoparticles did not enter capillary basement membranes from the extracellular spaces. Experiment 2: 2 µl of 15 nm nanoparticles were injected into mouse CSF. Within 5 min, groups of nanoparticles were present in the pial-glial basement membrane on the outer aspect of cortical arteries between the investing layer of pia mater and the glia limitans. The results of this study and previous research suggest that cerebral vascular basement membranes form the pathways by which fluid passes into and out of the brain but that different basement membrane layers are involved. The significance of these findings for neuroimmunology, Alzheimer's disease, drug delivery to the brain and the concept of the Virchow-Robin space are discussed.

  7. Entactin: ultrastructural localization of an ubiquitous basement membrane glycoprotein in mouse skin

    DEFF Research Database (Denmark)

    Horiguchi, Y; Fine, J D; Ljubimov, A V;


    Entactin is a recently described sulfated glycoprotein component of mouse endodermal cell-derived extracellular matrix and is present in a number of basement membranes. It has been ultrastructurally localized to both lamina densa and adjacent epithelial cell membranes in rodent kidney. In the pre...

  8. The deformation matrix theory of basement membrane: a study of water flow through elastic and rigid filaments in the rat. (United States)

    Fisher, R F


    1. When the volume of water per unit time which flows through natural elastic basement membrane is divided by the applied pressure, the value-the hydraulic conductivity-is not constant but decreases as pressure increases. In contrast when the same membrane is tanned with glutaraldehyde and rendered inelastic, the hydraulic conductivity is constant at all pressures. 2. Over a pressure range of 0-6.7 kPa equivalent to a membrane stress of 0-195 kPa in natural elastic membrane the hydraulic conductivity (Lp) can be related by the linear equation Lp = Lp.0 + apP where P is the hydraulic pressure, Lp.0 is the initial hydraulic conductivity and ap is a constant which is the decreased hydraulic conductivity per unit pressure (correlation coefficient 0.764. P less than 0.001). 3. The initial conductivity of the basement membrane of the crystalline lens of the adult rat (lens capsule) was 47.6 +/- 7.3 x 10(-12) m s-1 Pa-1 while the decrease in hydraulic conductivity per unit increase in pressure was -3.38 x 10(-15) m s-1 Pa-2. 4. Following tanning with glutaraldehyde the hydraulic conductivity was constant at 27.4 +/- 4.0 x 10(-12) m s-1 Pa-1. 5. A change in the configuration of the superhelices of the filaments of type IV collagen which form the framework of basement membrane is termed. 'The deformation matrix theory' and can satisfactorily account for the changes in hydraulic conductivity of both natural and tanned membrane. 6. In natural membrane the filaments deform easily and the pitch of the filament superhelices is increased by axial stress induced by pressure. The filaments straighten and become compacted together and the hydraulic permeability is thereby decreased. 7. In tanned membrane the filaments become more rigid and axial stress barely deforms them: moreover the pitch of the filament superhelices is decreased so that the filaments become more closely coiled and compacted together. Because of these changes the hydraulic conductivity is reduced as compared with

  9. Intercellular deposits of basement membrane material in active human pituitary adenomas detected by immunostaining for laminin and electron microscopy

    DEFF Research Database (Denmark)

    Holck, S; Wewer, U M; Albrechtsen, R


    Thirty-eight human pituitary adenomas (24 endocrine active and 14 endocrine inactive tumors) were studied immunohistochemically for the presence of the basement membrane component, laminin, and ultrastructurally for the presence of basement membrane. Immunoreactivity of laminin delineated staining...... of epithelial and endothelial basement membranes, the reaction product being confined mostly to the perivascular zones. Moreover, a hitherto undescribed presence of intercellular laminin-positive droplets was observed in ten of the active adenomas (nine patients with hyperprolactinemia and/or acromegalia...... matrix, indicating a mutual dependence between excessive hormone extrusion and an increase of "misplaced" deposits of basement membrane components, e.g., laminin....

  10. Basement membrane changes in breast cancer detected by immunohistochemical staining for laminin

    DEFF Research Database (Denmark)

    Albrechtsen, R; Nielsen, M; Wewer, U


    The distribution of the basement membrane glycoprotein laminin was studied by the immunoperoxidase technique in benign and malignant human breast tissue and in axillary lymph nodes from patients with breast cancer. An antiserum prepared against rat laminin was used. The specificity of this antise......The distribution of the basement membrane glycoprotein laminin was studied by the immunoperoxidase technique in benign and malignant human breast tissue and in axillary lymph nodes from patients with breast cancer. An antiserum prepared against rat laminin was used. The specificity...... by laminin staining, but they were thinner and discontinuous. The poorly differentiated carcinomas lacked organized basement membranes detectable by laminin staining. Our studies suggest that staining for laminin may be a useful adjunct test for detection of micrometatases in lymph nodes. The correlation...

  11. Co-deposition of basement membrane components during the induction of murine splenic AA amyloid

    DEFF Research Database (Denmark)

    Lyon, A W; Narindrasorasak, S; Young, I D


    Past studies have demonstrated that during murine AA amyloid induction there is co-deposition of the AA amyloid peptide and the basement membrane form of heparan sulfate proteoglycan. The synthesis and accumulation of heparan sulfate proteoglycan does not usually occur in the absence of other bas...... enhancing factor induction of amyloid, the period when amyloid is first detected. These observations raise the possibility that an abnormality in basement membrane metabolism is a very early event, and potentially plays an integral part in the process of AA amyloidogenesis....

  12. Heterogeneous distribution of a basement membrane heparan sulfate proteoglycan in rat tissues

    DEFF Research Database (Denmark)

    Couchman, J R


    in immunohistochemical studies on frozen tissue sections from many rat organs. However, there was no reactivity with some basement membranes, notably those of several smooth muscle types and cardiac muscle. In addition, it was found that pancreatic acinar basement membranes also lacked the HSPG type recognized...... HSPG from the murine Engelbreth-Holm swarm tumor. It was, however, confirmed that only a single population of antibodies was present in the serum. Despite the presence of similar epitopes on these two proteoglycans of different hydrodynamic properties, it was apparent that the PYS-2 HSPG represents...

  13. A model of strain-dependent glomerular basement membrane maintenance and its potential ramifications in health and disease. (United States)

    Barocas, Victor H; Dorfman, Kevin D; Segal, Yoav


    A model is developed and analyzed for type IV collagen turnover in the kidney glomerular basement membrane (GBM), which is the primary structural element in the glomerular capillary wall. The model incorporates strain dependence in both deposition and removal of the GBM, leading to an equilibrium tissue strain at which deposition and removal are balanced. The GBM thickening decreases tissue strain per unit of transcapillary pressure drop according to the law of Laplace, but increases the transcapillary pressure drop required to maintain glomerular filtration. The model results are in agreement with the observed GBM alterations in Alport syndrome and thin basement membrane disease, and the model-predicted linear relation between the inverse capillary radius and inverse capillary thickness at equilibrium is consistent with published data on different mammals. In addition, the model predicts a minimum achievable strain in the GBM based on the geometry, properties, and mechanical environment; that is, an infinitely thick GBM would still experience a finite strain. Although the model assumptions would be invalid for an extremely thick GBM, the minimum achievable strain could be significant in diseases, such as Alport syndrome, characterized by focal GBM thickening. Finally, an examination of reasonable values for the model parameters suggests that the oncotic pressure drop-the osmotic pressure difference between the plasma and the filtrate due to large molecules-plays an important role in setting the GBM strain and, thus, leakage of protein into the urine may be protective against some GBM damage.

  14. Effects of radiation on the permeability of human basement membranes; Effets des radiations sur la permeabilite de membranes basales humaines

    Energy Technology Data Exchange (ETDEWEB)

    Fan, B.T. [Paris-7 Univ., ITODYS, UPRES-A 7086 CNRS, 75 (France); Achour, S. [Paris-7 Univ., 75 (France). Unite de Recheche Chimie et Pharmacologie; Simmonet, F. [CEA Saclay, 91 - Gif-sur-Yvette (France). INSTN, Institut National des Sciences et Techniques Nucleaires; Guerin, D. [Clinique d`Aulnay, 93 - Aulnay-sous-Bois (France)


    The influence of radiation on the permeability properties of human basement membrane was investigated by measuring the diffusion rate of several organic compounds (glycine, proline, glucose, urea and insulin) through human anterior lens capsules. The basement membranes borne an {gamma}-irradiation treatment change significantly their permeability vis-a-vis studied organic substances. This modification in physico-chemical properties is probably due to the radiation, which alters or degrades the complex structure (or architecture) of basement membranes. Moreover the change in permeability is dependent upon the diffusing compounds. An increase in diffusion has been observed for glucose, glycine and urea. However for insulin and proline, a decrease in diffusion rate was observed. (authors) 21 refs.

  15. Permeation of macromolecules into the renal glomerular basement membrane and capture by the tubules (United States)

    Lawrence, Marlon G.; Altenburg, Michael K.; Sanford, Ryan; Willett, Julian D.; Bleasdale, Benjamin; Ballou, Byron; Wilder, Jennifer; Li, Feng; Miner, Jeffrey H.; Berg, Ulla B.; Smithies, Oliver


    How the kidney prevents urinary excretion of plasma proteins continues to be debated. Here, using unfixed whole-mount mouse kidneys, we show that fluorescent-tagged proteins and neutral dextrans permeate into the glomerular basement membrane (GBM), in general agreement with Ogston's 1958 equation describing how permeation into gels is related to molecular size. Electron-microscopic analyses of kidneys fixed seconds to hours after injecting gold-tagged albumin, negatively charged gold nanoparticles, and stable oligoclusters of gold nanoparticles show that permeation into the lamina densa of the GBM is size-sensitive. Nanoparticles comparable in size with IgG dimers do not permeate into it. IgG monomer-sized particles permeate to some extent. Albumin-sized particles permeate extensively into the lamina densa. Particles traversing the lamina densa tend to accumulate upstream of the podocyte glycocalyx that spans the slit, but none are observed upstream of the slit diaphragm. At low concentrations, ovalbumin-sized nanoparticles reach the primary filtrate, are captured by proximal tubule cells, and are endocytosed. At higher concentrations, tubular capture is saturated, and they reach the urine. In mouse models of Pierson’s or Alport’s proteinuric syndromes resulting from defects in GBM structural proteins (laminin β2 or collagen α3 IV), the GBM is irregularly swollen, the lamina densa is absent, and permeation is increased. Our observations indicate that size-dependent permeation into the lamina densa of the GBM and the podocyte glycocalyx, together with saturable tubular capture, determines which macromolecules reach the urine without the need to invoke direct size selection by the slit diaphragm. PMID:28246329

  16. Anti-DNA autoantibodies initiate experimental lupus nephritis by binding directly to the glomerular basement membrane in mice. (United States)

    Krishnan, Meera R; Wang, Congmiao; Marion, Tony N


    The strongest serological correlate for lupus nephritis is antibody to double-stranded DNA, although the mechanism by which anti-DNA antibodies initiate lupus nephritis is unresolved. Most recent reports indicate that anti-DNA must bind chromatin in the glomerular basement membrane or mesangial matrix to form glomerular deposits. Here we determined whether direct binding of anti-DNA antibody to glomerular basement membrane is critical to initiate glomerular binding of anti-DNA in experimental lupus nephritis. Mice were co-injected with IgG monoclonal antibodies or hybridomas with similar specificity for DNA and chromatin but different IgG subclass and different relative affinity for basement membrane. Only anti-DNA antibodies that bound basement membrane bound to glomeruli, activated complement, and induced proteinuria whether injected alone or co-injected with a non-basement-membrane-binding anti-DNA antibody. Basement membrane-binding anti-DNA antibodies co-localized with heparan sulfate proteoglycan in glomerular basement membrane and mesangial matrix but not with chromatin. Thus, direct binding of anti-DNA antibody to antigens in the glomerular basement membrane or mesangial matrix may be critical to initiate glomerular inflammation. This may accelerate and exacerbate glomerular immune complex formation in human and murine lupus nephritis.

  17. Macrophage Chemotaxis in Anti-tubular Basement Membrane-Induced Interstitial Nephritis in Guinea Pigs

    NARCIS (Netherlands)

    Kennedy, Thomas L.; Merrow, Martha; Phillips, S. Michael; Norman, Michael; Neilson, Eric G.


    Interstitial renal lesions containing T cells and macrophages develop after 14 days in guinea pigs immunized to produce anti-tubular basement membrane-induced interstitial nephritis. We serially examined the renal venous and systemic arterial sera from such animals to determine if chemotactic factor

  18. The clinical utility of reticular basement membrane thickness measurements in asthmatic children

    NARCIS (Netherlands)

    van Mastrigt, Esther; Vanlaeken, Leonie; Heida, Fardou; Caudri, Daan; de Jongste, Johan C.; Timens, Wim; Rottier, Bart L.; de Krijger, Ronald R.; Pijnenburg, Marielle W.


    Objective: Reticular basement membrane (RBM) thickness is one of the pathological features of asthma and can be measured in endobronchial biopsies. We assessed the feasibility of endobronchial biopsies in a routine clinical setting and investigated the clinical value of RBM thickness measurements fo

  19. Peroxynitrous acid induces structural and functional modifications to basement membranes and its key component, laminin

    DEFF Research Database (Denmark)

    Degendorfer, Georg; Chuang, Christine Y.; Hammer, Astrid;


    Basement membranes (BM) are specialized extracellular matrices underlying endothelial cells in the artery wall. Laminin, the most abundant BM glycoprotein, is a structural and biologically active component. Peroxynitrous acid (ONOOH), a potent oxidizing and nitrating agent, is formed in vivo at s...

  20. Cdc42 expression in keratinocytes is required for the maintenance of the basement membrane in skin

    DEFF Research Database (Denmark)

    Wu, Xunwei; Quondamatteo, Fabio; Brakebusch, Cord


    , structure and number of hemidesomosomes were not significantly changed in the Cdc42 mutant skin compared with the control mice and no blister formation was observed in mutant skin. These data indicate that Cdc42 in keratinocytes is important for maintenance of the basement membrane of skin....

  1. Ultrastructure of basement membranes in monkey and shark teeth at an early stage of development. (United States)

    Sawada, Takashi


    The basement membrane, which separates the inner enamel epithelium from the dental papilla in the early stages of tooth development, is known to play a significant role in odontogenesis. In this review article, this basement membrane was described in detail based on our recent findings with the use of high-resolution electron microscopy. Tooth germs of a monkey (Macaca fuscata) and a shark (Cephaloscyllium umbratile) were processed for thin-section observations. During the early stage of development, the basement membrane of the inner enamel (dental) epithelium was composed of a lamina lucida, lamina densa, and much wider lamina fibroreticularis. At higher magnification, the lamina densa in both species was made up of a fine network of cords, which are generally the main constituents of the basement membranes. In the monkey tooth, the lamina fibroreticularis was rich in fibrils, which were now characterized as basotubules, 10-nm-wide microfibril-like structures. The space between the basotubules was filled with a cord network that extended from the lamina densa. Dental papilla cell processes were inserted into the lamina fibroreticularis, and their surface was closely associated with numerous parallel basotubules via 1.5- to 3-nm-wide filaments. In the shark tooth during its early stage of development, the basotubules were absent in the lamina fibroreticularis and only narrow extensions, 60-90 nm wide and 1-2 microm long, of the cord network of the lamina densa were present. The dental papilla cells were immobilized by means of the binding of their processes to the extensions. These results indicate that basement membranes in both monkey and shark teeth at early stage of development are specialized for functions as anchoring and firm binding, which are essential for the successful differentiation of the odontoblasts.

  2. Release kinetics of prolyl hydroxylase inhibitors from collagen barrier membranes. (United States)

    Hamid, Omar; Pensch, Manuela; Agis, Hermann


    Collagen barrier membranes are used in guided tissue regeneration to support healing. This strategy, however, relies on the healing capacity of the tissue. Pharmacological inhibitors of prolyl hydroxylases can support regeneration by enhancing angiogenesis and are therefore a promising tool for periodontology. Here we evaluate the release kinetics of the prolyl hydroxylase inhibitors dimethyloxalylglycine and L-mimosine from collagen barrier membranes. Dimethyloxalylglycine and L-mimosine were lyophilized onto the collagen barrier membranes. The morphology of the collagen barrier membranes was analysed using scanning electron microscopy. The release of prolyl hydroxylase inhibitors was assessed by colorimetric and spectroscopic methods. Their ability to induce a cellular response was assessed in bioassays with gingival and periodontal ligament fibroblasts based on vascular endothelial growth factor production, proliferation, and metabolic activity of the cells. We found that loading of collagen barrier membranes with prolyl hydroxylase inhibitors did not change the overall membrane morphology. Assessment of the release kinetics by direct measurements and based on vascular endothelial growth factor production showed that supernatants obtained from the collagen barrier membranes in the first 6 hours had a sufficient level of prolyl hydroxylase inhibitors to induce vascular endothelial growth factor production. A similar kinetic was found when cell proliferation was assessed. Changes in metabolic activity did not reach the level of significance in the MTT assay. In conclusion, collagen barrier membranes can release prolyl hydroxylase inhibitors thereby increasing the pro-angiogenic capacity of periodontal cells in vitro. These findings provide the basis for preclinical studies to evaluate the regenerative capacity of prolyl hydroxylase inhibitors in periodontology and oral surgery.

  3. Mechanical qualification of collagen membranes used in dentistry

    Directory of Open Access Journals (Sweden)

    Emanuela Ortolani


    Full Text Available AIM. The aim of this work is the qualification of commercially available collagen membranes in a comparative manner. The natural origin of collagen makes standardization difficult. Nevertheless, through dimensional and mechanical measures it is possible to mechanically qualify collagen membranes, and compare them. METHODS. Three commercially available collagen membranes used in Guided Bone Regeneration (GBR and in Guided Tissue Regeneration (GTR techniques, namely Bio-Gide, Collprotect and Jason, were chosen for the comparison. Quasi-static (tensile tests and time-dependent (stress relaxation test mechanical tests together with a functional test (tear test were done to determine the responses of collagen membranes under different loading conditions. RESULTS. The tested membranes exhibited different behaviours, different deformability values and thickness, Jason being the thinnest and Bio-Gide the thickest. Similar differences were also observed in terms of surface density. DISCUSSION. Even though clinical observations were not within the aim of this study, our findings indicate that a better understanding of the correlation between mechanical properties and thickness could lead to a more rational design and use of these membranes in the face of specific clinical cases.

  4. Impact of ischemia-reperfusion on extracellular matrix processing and structure of the basement membrane of the heart.

    Directory of Open Access Journals (Sweden)

    Alexander Lauten

    Full Text Available PURPOSE: Acute ischemic injury is a strong inductor of cardiac remodelling, resulting in structural changes of the extracellular matrix (ECM and basement membrane (BM. In a large animal model of ischemia-reperfusion (I/R we investigated the post-ischemic liberation of the collagen-IV-fragments Tumstatin (TUM; 28 kDa-fragment of collagen-IV-alpha-3, Arresten (ARR; 26 kDa-fragment of collagen-IV-alpha-1 and Endorepellin (LG3, 85 kDa-fragment of perlecan which are biologically active in angiogenesis and vascularization in the post-ischemic myocardium. METHODS AND RESULTS: In this blinded study, 30 pigs were randomized to 60 min of global I/R at either 4°C or 32°C or served as control. Three transmyocardial tissue samples were collected prior to ischemia and within 30 min and 150 min of reperfusion. Tissue content of TUM, ARR and LG3 was analyzed by western blotting and immunostaining. Within 150 min of mild hypothermic I/R a significantly increased tissue content of ARR (0.17±0.14 vs. 0.56±0.56; p = 0.001 and LG3 (1.13±0.34 vs. 2.51±1.71, p11fold elevation of creatine kinase (2075±2595 U/l vs. 23248±6551 U/l; p<0.001 in the coronary sinus plasma samples. Immunostaining demonstrated no changes for ARR and LG3 presentation irrespective of temperature. In contrast, TUM significantly decreased in the BM surrounding cardiomyocytes and capillaries after mild and deep hypothermic I/R, thus representing structural alterations of the BM in these groups. CONCLUSION: The study demonstrates an early temperature-dependent processing of Col-IV as major component of the BM of cardiomyocytes and vascular endothelium. These observations support the protective effects of deep hypothermia during I/R. Furthermore, the results suggest an increased structural remodelling of the myocardial basement membrane with potential functional impairment during mild hypothermic I/R which may contribute to the progression to post-ischemic heart failure.

  5. Quantitative Proteome Analysis Reveals Increased Content of Basement Membrane Proteins in Arteries from Patients with Type 2 Diabetes and Lower Levels among Metformin Users

    DEFF Research Database (Denmark)

    Rørdam Preil, Simone; Kristensen, Lars P; Beck, Hans C;


    BACKGROUND: -The increased risk of cardiovascular diseases (CVD) in type 2 diabetes has been extensively documented, but the origins of the association remain largely unknown. We sought to determine changes in protein expressions in arterial tissue from patients with type 2 diabetes and moreover...... hypothesized that metformin intake influences the protein composition. METHODS AND RESULTS: -We analyzed non-atherosclerotic repair arteries gathered at coronary by-pass operations from 30 patients with type 2 diabetes, as well as from 30 age- and gender-matched non-diabetic individuals. Quantitative proteome...... analysis was done by iTRAQ-labelling and LC-MS/MS analysis on individual arterial samples. The amounts of the basement membrane (BM) components, alpha-1- and alpha-2- type IV collagen, gamma-1- and beta-2-laminin were significantly increased in patients with diabetes. Moreover, the expressions of basement...

  6. Heparan sulfate proteoglycans made by different basement-membrane-producing tumors have immunological and structural similarities

    DEFF Research Database (Denmark)

    Wewer, U M; Albrechtsen, R; Hassell, J R


    Using immunological assays, we determined the relationship between the heparan sulfate proteoglycans produced by two different murine basement-membrane-producing tumors, i.e., the mouse Engelbreth-Holm-Swarm (EHS) tumor and the L2 rat yolk-sac tumor. Antibodies prepared against the heparan sulfate...... mainly heparan sulfate (75%) along with smaller amounts of chondroitin sulfate (19%), whereas the L2 rat yolk-sac tumor produced mainly chondroitin sulfate (76%) with smaller amounts of heparan sulfate (21%). We conclude that these two murine basement-membrane-producing tumors elaborate...... proteoglycans obtained from these two sources immunoprecipitated the same precursor protein with a molecular mass of 400,000 daltons from 35S-methionine pulse-labeled cells of both tumors. Immunohistochemistry showed the heparan sulfate proteoglycan to be distributed in the extracellular matrix and also...

  7. Collagen-apatite nanocomposite membranes for guided bone regeneration. (United States)

    Song, Ju-Ha; Kim, Hyoun-Ee; Kim, Hae-Won


    Collagen-apatite nanocomposite is regarded as a potential biomaterial because of its composition and structure, which are similar to those of human hard tissues. However, there have been few investigations of its mechanical and biological benefits in direct comparison with a collagen equivalent. Herein, we successfully produced a biomedical membrane made of a nanocomposite, and systemically evaluated the mechanical, chemical, and biological properties of the nanocomposite in comparison with those of pure collagen. The results showed that significant improvements were achieved by the nanocomposite approach, particularly in terms of the mechanical strength and chemical stability. The present findings point to the potential usefulness of the collagen-apatite nanocomposite membrane in the field of guided bone regeneration (GBR).

  8. Tissue fibrocytes in patients with mild asthma: A possible link to thickness of reticular basement membrane?

    Directory of Open Access Journals (Sweden)

    Bjermer Leif


    Full Text Available Abstract Background Myofibroblasts, proposed as being derived from circulating fibrocytes, are considered to be important cells in thickening of the basement membrane in patients with asthma. We have studied the correlation of tissue fibrocyte levels to basement membrane thickness and the presence of fibrocytes in bronchoalveolar lavage fluid (BALF in steroid-naive patients with mild asthma and controls. Methods Patients with mild asthma (n = 9 were recruited and divided into two categories based on whether or not fibroblast-like cells could be established from BALF. Non-asthmatic healthy subjects (n = 5 were used as controls. Colocalization of the fibrocyte markers CD34, CD45RO, procollagen I, and α-smooth muscle actin (α-SMA were identified in bronchial biopsies from patients and controls by confocal microscopy. Kruskall-Wallis method was used to calculate statistical significance and Spearman coefficient of rank correlation was used to assess the degree of association. Results In patients with BALF fibroblasts, a 14-fold increase of tissue cells expressing CD34/CD45RO/α-SMA and a 16-fold increase of tissue cells expressing CD34/procollagen I was observed when compared to controls (p Conclusion These findings indicate a correlation between recruited fibrocytes in tissue and thickness of basement membrane. Fibroblast progenitor cells may therefore be important in airway remodeling in steroid-naive patients with mild asthma.

  9. Subepithelial basement membrane thickness in patients with normal colonic mucosal appearance in colonoscopy:Results from southern Turkey

    Institute of Scientific and Technical Information of China (English)

    Fazilet Kayaselcuk; Ender Serin; Yǖksel Gumurdulu; Birol Ozer; Ilhan Tuncer; Sedat Boyacioglu


    AIM: Our aims were to determine the normal limits of subepithelial basement membrane (SEBM) thickness in order to more accurately diagnose collagenous colitis in the population from southern Turkey and to investigate into links between SEBM thickness and age, and sex.METHODS: The study included 100 patients (mean age 50.0±13.3 years; male, 34; female, 66) with miscellaneous gastrointestinal symptoms, and normal colonic mucosal appearance in colonoscopic evaluation. Biopsies were taken from five different regions of the colon. SEBM was measured with a calibrated eyepiece on specimens prepared with specific stains for collagen. Intensity of inflammatory cells was graded semiquantitatively. Differences in SEBM thickness among the different colon regions, and relationships between SEBM thickness and age, sex, and density of inflammatory cells were statistically evaluated.RESULTS: The cecum and rectum showed the largest amounts of infiltrate. None of the specimens showed histologic findings of collagenous colitis. The SEBM thicknesses measured for each case ranged from 3-20 μm. The biggest thickness was observed in rectal mucosa (median value: 10 μm).Cecum and ascending colon showed similar SEBM thickness (median value: 5 μm). SEBM thickness was not correlated with patient age or sex, but was positively correlated with the intensity of inflammatory cells in each colon segment.CONCLUSION: In this patient group from southern Turkey,SEBM was thickest in the rectum. Our results indicate that,in this population, SEBM thickness is not correlated with age or sex, but is positively correlated with severity of inflammation. The findings also support the concept that measuring SEBM thickness at one segment in the colon is inadequate and may be misleading.

  10. Cytotoxicity of bovine and porcine collagen membranes in mononuclear cells. (United States)

    Moura, Camilla Christian Gomes; Soares, Priscilla Barbosa Ferreira; Carneiro, Karine Fernandes; Souza, Maria Aparecida de; Magalhães, Denildo


    This study compared the cytotoxicity and the release of nitric oxide induced by collagen membranes in human mononuclear cells. Peripheral blood was collected from each patient and the separation of mononuclear cells was performed by Ficoll. Then, 2x10(5) cells were plated in 48-well culture plates under the membranes in triplicate. The polystyrene surface was used as negative control. Cell viability was assessed by measuring mitochondrial activity (MTT) at 4, 12 and 24 h, with dosage levels of nitrite by the Griess method for the same periods. Data had non-normal distribution and were analyzed by the Kruskal-Wallis test (pporcine membrane induced a higher release of nitrite compared with the control and bovine membrane, respectively (pporcine collagen membrane induces an increased production of proinflammatory mediators by mononuclear cells in the first hours of contact, decreasing with time.

  11. Regeneration of the epidermis and basement membrane of the planarian Dugesia japonica after total-body x irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Hori, I.


    Fresh-water planarians were studied to examine effects of x rays on regeneration of the epidermis and basement membrane. During early stages of regeneration, free rhabdite-forming cells were associated with the wound epidermis and recruited it. In later stages, however, a gradual degeneration occurred in the epidermis and cells undergoing epithelization decreased in number. Eventually epidermal cells on the wound surface appeared necrotic as evidenced by pyknotic nuclei and vacuolized dense cytoplasm. The entire basement membrane could not be reconstituted in any stage after wounding though its precursor-like material was secreted in the interspace between epidermis and parenchyma. Morphological changes in extracellular products and in the cells surrounding the products suggest that epidermal cells which have covered the wound surface synthesize precursors of the basement membrane. Possible factors of a characteristic perturbation in epithelization and basement membrane formation after total-body irradiation are discussed.

  12. Human skin basement membrane-associated heparan sulphate proteoglycan: distinctive differences in ultrastructural localization as a function of developmental age

    DEFF Research Database (Denmark)

    Horiguchi, Y; Fine, J D; Couchman, J R


    Recent studies have demonstrated that skin basement membrane components are expressed within the dermo-epidermal junction in an orderly sequence during human foetal development. We have investigated the ultrastructural localization of basement membrane-related antigens in human foetal skin...... was identical to that observed in neonatal and adult human skin. These findings demonstrate that active remodelling of the dermo-epidermal junction occurs during at least the first two trimesters, and affects not only basement membrane-associated structures but also specific antigens....... at different developmental ages using two monoclonal antibodies to a well-characterized basement membrane-associated heparan sulphate proteoglycan. A series of foetal skin specimens (range, 54-142 gestational days) were examined using an immunoperoxidase immunoelectron microscopic technique. In specimens...

  13. Collagen based barrier membranes for periodontal guided bone regeneration applications. (United States)

    Sheikh, Zeeshan; Qureshi, Javairia; Alshahrani, Abdullah M; Nassar, Heba; Ikeda, Yuichi; Glogauer, Michael; Ganss, Bernhard


    Certain cell populations within periodontal tissues possess the ability to induce regeneration, provided they have the opportunity to populate the wound or defect. Guided regeneration techniques have been investigated for regenerating periodontal tissues and such therapies usually utilize barrier membranes. Various natural and synthetic barrier membranes have been fabricated and tested to prevent epithelial and connective tissue cells from invading while allowing periodontal cells to selectively migrate into the defect. This paper focuses on the literature relevant to the use and potential of resorbable collagen membranes in GBR procedures, sites of periodontal and intrabony defects, in cases of socket and alveolar ridge preservation and at implant sites. The results of their use in GBR procedures has shown them to be effective and comparable with non-resorbable membranes with regards to clinical attachment gain, probing depth reduction and defect bone filling. They have also shown to prevent epithelial ingrowth into the defect space during the initial wound healing phase postsurgically. Collagen membranes have also been used for root coverage and GBR procedures and have shown good success rates comparable to subepithelial connective tissue grafts and expanded-polytetrafluoroethylene (e-PTFE) membranes. The future for periodontal tissue engineering is very exciting with the use of barrier membranes expected to continue playing a critical role. However, long-term clinical trials are required to further evaluate and confirm the efficacy of the available collagen barrier membranes for periodontal and bone regeneration use.

  14. Furcation therapy with bioabsorbable collagen membrane: a clinical trial. (United States)

    Pruthi, Vijay K; Gelskey, Shirley C; Mirbod, Sayed M


    This study compared the effectiveness of 2 barrier membranes, expanded polytetrafluoroethylene (e-PTFE) and collagen, in treating Class II furcation defects of mandibular molars in humans. Seventeen nonsmoking subjects with no history of systemic disease each presenting with Class II furcation defects in 2 mandibular molars were selected and underwent initial therapy. At the time of the surgery and at 8-month follow-up, soft-tissue measurements consisting of the gingival index, vertical and horizontal probing depth, recession and clinical attachment level were obtained at the midfurcation level. At the time of membrane placement and at 12-month re-entry, horizontal midfurcation probing depth and hard-tissue measurement of vertical fill (from the crown to the depth of the pocket) were also obtained. According to the surgical protocol, both membranes were completely covered with a coronally positioned flap, and in all cases healing was uneventful. Data were analyzed first by comparing baseline measurements (at surgery) with measurements at 8-month follow-up and 12-month re-entry for both e-PTFE and collagen membranes according to repeated-measures analysis of variance. The changes from surgery to follow-up and re-entry were then compared between the 2 treatment modalities with paired Wilcoxon rank-sum tests. No statistically significant differences were found between e-PTFE and collagen membranes with respect to gingival index, reduction in probing depth, gain in clinical attachment or filling of the horizontal defect. However, the improvement in vertical fill at 12-month re-entry was more substantial for the teeth treated with collagen membrane than those treated with e-PTFE (p collagen is a beneficial material for regenerative therapy of Class II furcation defects in humans, yielding results that are similar to or better than (vertical fill) those for e-PTFE membrane.

  15. Mass transfer of large molecules through collagen and collagen-silica hybrid membranes (United States)

    Jofre-Lora, Pedro

    Diabetes is a growing concern in the United States and around the world that must be addressed through new treatment options. Current standard treatment options of diabetes are limiting and have tremendous impacts on patient's lives. Emerging therapies, such as the implantation of encapsulated islets, are promising treatment options, but have not yet materialized due to unsolved problems with material properties. Hybrid silica-collagen membranes address some of these unsolved problems and are a promising material for cell encapsulation. However, the mass transfer properties of large molecules, such as insulin, TNF-alpha, IL1beta, and other important proteins in the etiology of diabetes, through these hybrid membranes are poorly characterized. In order to begin characterizing these properties, a device was constructed to accurately and efficiently measure the mass transfer of other similar large molecules, fluorescein isothiocyanate dextrans (FITC-dextran), through collagen-silica hybrid membranes. The device was used to measure diffusion coefficients of 4, 20, 40, and 150 kDa FITC-dextrans through non-silicified and silicified samples of 200 and 1000 Pa porcine skin collagen. Diffusion coefficients were found to be in the 10-7-10-6 cm2s -1 range, which is in agreement with previously published data for similar molecules through similar hydrogels. The effects of collagen stiffness, FITC-dextran molecular weight, and silicification treatment on diffusion were investigated. It was found that collagen stiffness and FITC-dextran molecular weight had a negative correlation with diffusion, whereas silicification treatment had no global impact on diffusion. The device created, and the results of this preliminary investigation, can be used to develop collagen-silica hybrid membranes as an alternative material for cell encapsulation in a forward-design manner.

  16. Basement membrane heparan sulfate proteoglycan from the L2 rat yolk sac carcinoma

    DEFF Research Database (Denmark)

    Fenger, M; Wewer, U; Albrechtsen, R


    Heparan sulfate proteoglycan from the L2 rat yolk sac carcinoma has been purified and partially characterized. The proteoglycan has an apparent Mr of 750 000, 35% of which represents the core protein. The core protein seems to be homogeneous, whereas the heparan sulfate chains are heterogeneous w...... with an Mr of about 50 000-70 000, with 30% of the glucosamine being N-sulfated. Antibodies raised against the core protein of the heparan sulfate proteoglycan reacted with basement membranes of various rat and human tissue....

  17. An unusual case of anti-glomerular basement membrane disease presenting with nephrotic syndrome. (United States)

    Okafor, Chidi C; Balogun, Rasheed A; Bourne, David T; Alhussain, Turki O; Abdel-Rahman, E M


    Anti-glomerular basement membrane (anti-GBM) disease is a vasculitic disease characterized by acute kidney injury, oliguria, hematuria and proteinuria. Proteinuria is rarely in the nephrotic range. A case of anti-GBM disease with proteinuria of 22.5 g/day is discussed. Immunofluorescence showed strong linear IgG deposits while electron microscopy showed widespread visceral epithelial cell foot cell process effacement. No electron dense immune complex-type deposits were identified. Pathology findings were not suggestive of simultaneous presentation of anti-GBM disease and other diseases associated with nephrotic range proteinuria. Anti-GBM disease should be considered in a comprehensive differential diagnosis of severe proteinuria.

  18. Basement membrane heparan sulfate proteoglycan from the L2 rat yolk sac carcinoma

    DEFF Research Database (Denmark)

    Fenger, M; Wewer, U; Albrechtsen, R


    Heparan sulfate proteoglycan from the L2 rat yolk sac carcinoma has been purified and partially characterized. The proteoglycan has an apparent Mr of 750 000, 35% of which represents the core protein. The core protein seems to be homogeneous, whereas the heparan sulfate chains are heterogeneous...... with an Mr of about 50 000-70 000, with 30% of the glucosamine being N-sulfated. Antibodies raised against the core protein of the heparan sulfate proteoglycan reacted with basement membranes of various rat and human tissue....

  19. The Alteration of the Epidermal Basement Membrane Complex of Human Nevus Tissue and Keratinocyte Attachment after High Hydrostatic Pressurization

    Directory of Open Access Journals (Sweden)

    Naoki Morimoto


    Full Text Available We previously reported that human nevus tissue was inactivated after high hydrostatic pressure (HHP higher than 200 MPa and that human cultured epidermis (hCE engrafted on the pressurized nevus at 200 MPa but not at 1000 MPa. In this study, we explore the changes to the epidermal basement membrane in detail and elucidate the cause of the difference in hCE engraftment. Nevus specimens of 8 mm in diameter were divided into five groups (control and 100, 200, 500, and 1000 MPa. Immediately after HHP, immunohistochemical staining was performed to detect the presence of laminin-332 and type VII collagen, and the specimens were observed by transmission electron microscopy (TEM. hCE was placed on the pressurized nevus specimens in the 200, 500, and 1000 MPa groups and implanted into the subcutis of nude mice; the specimens were harvested at 14 days after implantation. Then, human keratinocytes were seeded on the pressurized nevus and the attachment was evaluated. The immunohistochemical staining results revealed that the control and 100 MPa, 200 MPa, and 500 MPa groups were positive for type VII collagen and laminin-332 immediately after HHP. TEM showed that, in all of the groups, the lamina densa existed; however, anchoring fibrils were not clearly observed in the 500 or 1000 MPa groups. Although the hCE took in the 200 and 500 MPa groups, keratinocyte attachment was only confirmed in the 200 MPa group. This result indicates that HHP at 200 MPa is preferable for inactivating nevus tissue to allow its reuse for skin reconstruction in the clinical setting.

  20. Agar/collagen membrane as skin dressing for wounds

    Energy Technology Data Exchange (ETDEWEB)

    Bao Lei; Yang Wei; Mao Xuan; Mou Shansong; Tang Shunqing [Biomedical Engineering Institute, Jinan University, Guangzhou (China)], E-mail:, E-mail:


    Agar, a highly hydrophilic polymer, has a special gel property and favorable biocompatibility, but moderate intension strength in an aqueous condition and a low degradation rate. In order to tailor both properties of mechanical intension and degradation, type I collagen was composited with agar in a certain ratio by drying at 50 {sup 0}C or by a freeze-dry process. Glutaraldehyde was chosen as a crosslinking agent, and the most favorable condition for crosslinking was that the weight ratio of agar to glutaraldehyde was 66.7 and the pH value about 5. Dynamic mechanical analysis results showed that the single agar membrane had a modulus value between 640 MPa and 1064 MPa, but it was between 340 MPa and 819 MPa after being composited with type I collagen. It was discovered under an optical microscope that the pores were interconnected in the composite scaffolds instead of the honeycomb-like pores in a single type I collagen scaffold or the laminated gaps in a single agar scaffold. The results of an acute toxicity test disclosed that the composites were not toxic to mice although the composites were crosslinked with a certain concentration of glutaraldehyde. The results of gross examinations showed that when the composite membranes or scaffolds were applied to a repair rabbit skin lesion, the composites had a good repair effect without infection, liquid exudation or visible scar in the lesion covered with them. But in the control group, the autologous skin showed necrosis and there were a lot of scar tissues in the lesion site. H and E staining results showed that the repair tissue was similar to the normal one and very few scaffolds or membranes were left without degradation after 2 or 3 weeks. In conclusion, it is proved that type I collagen increases the toughness of the agar membrane, and the agar/type I collagen composites are promising biomaterials as wound dressings for healing burns or ulcers.

  1. Anti-glomerular basement membrane glomerulonephritis and thrombotic microangiopathy in first degree relatives; a case report

    Directory of Open Access Journals (Sweden)

    Idorn Thomas


    Full Text Available Abstract Background Anti-glomerular basement membrane glomerulonephritis and thrombotic microangiopathy are rare diseases with no known coherence. Case Presentation A daughter and her biological mother were diagnosed with pregnancy-induced thrombotic microangiopathy and anti-glomerular basement membrane glomerulonephritis, respectively. Both developed end-stage renal disease. Exploration of a common aetiology included analyses of HLA genotypes, functional and genetic aspects of the complement system, ADAMTS13 activity and screening for autoantibodies. The daughter was heterozygous carrier of the complement factor I G261D mutation, previously described in patients with membranoproliferative glomerulonephritis and atypical haemolytic uremic syndrome. The mother was non-carrier of this mutation. They shared the disease associated complement factor H silent polymorphism Q672Q (79602A>G. Conclusion An unequivocal functional or molecular association between these two family cases was not found suggesting that the patients probably share another, so far undiagnosed and unknown, predisposing factor. It seems highly unlikely that two infrequent immunologic diseases would occur by unrelated pathophysiological mechanisms within first degree relatives.

  2. Validation of glomerular basement membrane thickness changes with aging in minimal change disease. (United States)

    Sato, Shigeru; Sasaki, Yoshihiro; Adachi, Akiko; Ghazizadeh, Mohammad


    Measurement of the normal range of glomerular basement membrane (GBM) thickness by electron microscopy is required for the diagnosis of thin basement membrane disease or diabetic nephropathy; however, this measurement is influenced by aging. The aim of this study was to introduce a simple histogram plotting method for the validation of the results of the GBM thickness measurements by the accepted arithmetic mean ± SD method. We examined renal biopsy specimens obtained from 19 patients (10 males and 9 females) with minimal change disease, ranging in age from 3 to 70 years. Renal tissue samples obtained at autopsy from a male baby (3 months old) with no renal disease were also examined. For each case, GBM thicknesses at 10-15 evenly distributed points per glomerular loop were directly measured and the arithmetic mean ± SD was calculated. Subsequently, the arithmetic mean ± SD for each group of cases classified by age into 4 groups, i.e. babyhood (3 months old), childhood (3-11 years old), adulthood (12-57 years old), and old age (60-70 years old), was determined. On the other hand, a histogram of the frequency of GBM points measured against thickness was plotted to determine the distribution pattern and the range of measurements in each age group. The histogram plot showed 4 clearly divided modes for GBM thickness. Comparison of the results obtained by the 2 methods revealed a significant correlation indicating the feasibility of the histogram plotting method as a useful adjunct to validate GBM thickness measurements.

  3. Nephritogenic lupus antibodies recognize glomerular basement membrane-associated chromatin fragments released from apoptotic intraglomerular cells. (United States)

    Kalaaji, Manar; Mortensen, Elin; Jørgensen, Leif; Olsen, Randi; Rekvig, Ole Petter


    Antibodies to dsDNA represent a classification criterion for systemic lupus erythematosus. Subpopulations of these antibodies are involved in lupus nephritis. No known marker separates nephritogenic from non-nephritogenic anti-dsDNA antibodies. It is not clear whether specificity for glomerular target antigens or intrinsic antibody-affinity for dsDNA or nucleosomes is a critical parameter. Furthermore, it is still controversial whether glomerular target antigen(s) is constituted by nucleosomes or by non-nucleosomal glomerular structures. Previously, we have demonstrated that antibodies eluted from murine nephritic kidneys recognize nucleosomes, but not other glomerular antigens. In this study, we determined the structures that bind nephritogenic autoantibodies in vivo by transmission electron microscopy, immune electron microscopy, and colocalization immune electron microscopy using experimental antibodies to dsDNA, to histones and transcription factors, or to laminin. The data obtained are consistent and point at glomerular basement membrane-associated nucleosomes as target structures for the nephritogenic autoantibodies. Terminal deoxynucleotidyl-transferase-mediated dUTP nick end-labeling or caspase-3 assays demonstrate that lupus nephritis is linked to intraglomerular cell apoptosis. The data suggest that nucleosomes are released by apoptosis and associate with glomerulus basement membranes, which may then be targeted by pathogenic anti-nucleosome antibodies. Thus, apoptotic nucleosomes may represent both inducer and target structures for nephritogenic autoantibodies in systemic lupus erythematosus.

  4. Association of anti-glomerular basement membrane antibody disease with dermatomyositis and psoriasis: case report

    Directory of Open Access Journals (Sweden)

    Natália Pereira Machado

    Full Text Available CONTEXT: Anti-glomerular basement membrane (anti-GBM antibody syndrome is characterized by deposition of anti-GBM antibodies on affected tissues, associated with glomerulonephritis and/or pulmonary involvement. This syndrome has been described in association with other autoimmune disorders, but as far as we know, it has not been described in association with dermatomyositis and psoriasis. CASE REPORT: A 51-year-old man with a history of dermatomyositis and vulgar psoriasis presented with a condition of sensitive-motor polyneuropathy of the hands and feet, weight loss of 4 kg, malaise and fever. On admission, he had been making chronic use of cyclosporin and antihypertensive drugs for three months because of mild arterial hypertension. Laboratory tests showed anemia and leukocytosis, elevated serum urea and creatinine and urine presenting proteinuria, hematuria, leukocyturia and granular casts. The 24-hour proteinuria was 2.3 g. Renal biopsy showed crescentic necrotizing glomerulonephritis with linear immunoglobulin G (IgG deposits on the glomerular basement membrane by means of direct immunofluorescence, which were suggestive of anti-GBM antibodies. The patient was then treated initially with methylprednisolone and with monthly cyclophosphamide in the form of pulse therapy.

  5. Using absorbable collagen membranes for guided tissue regeneration, guided bone regeneration, and to treat gingival recession. (United States)

    Wang, H L; Carroll, W J


    This article reviews the role of barrier membranes in guided tissue regeneration (GTR) and guided bone regeneration (GBR), including the advantages of using absorbable barrier membranes in GTR and GBR and the unique properties of collagen membranes. The indications and contraindications for using collagen membranes for these procedures are examined, and successful cases are presented. Finally, the role of collagen membranes in the future of regenerative therapy is considered.

  6. Isolation and partial characterization of antigens from basement membranes and streptococcal cell membrane (SCM) employing anti-SCM monoclonal antibody. (United States)

    Zelman, M E; Lange, C F


    Monoclonal antibodies (mAb) against streptococcal cell membrane (SCM) antigen were used to identify specific cross-reactive peptides prepared by trypsin digestion of purified glomerular basement membrane (GBM) and lung basement membrane (LBM). Anti-SCM mAb-coupled HPLC columns were used to affinity isolate soluble LBM, GBM, and SCM antigens which then were sized by HPLC. Alternatively, SCM, GBM, and LBM digests were subjected to an initial separation by HPLC into component polypeptides, followed by affinity purification and ELISA of these fractions using anti-SCM mAb. Comparison of the antigenic reactivities by ELISA of the sized polypeptides on a nanomolar basis permitted the estimation of their individual relative epitope densities. The results for SCM antigens showed increasing epitope density with increasing molecular size, which suggests that intact SCM consists of repeating epitopes. Low mol. wt GBM polypeptides in nanogram amounts inhibited mAb binding to SCM, indicating that these small GBM polypeptides may similarly contain more than a single cross-reactive epitope. The identification of these cross-reactive epitopes in LBM and GBM has important implications for the etiology of post-streptococcal sequelae.

  7. Laminin, a noncollagenous component of epithelial basement membranes synthesized by a rat yolk sac tumor

    DEFF Research Database (Denmark)

    Wewer, U; Albrechtsen, R; Ruoslahti, E


    Laminin, a glycoprotein antigenically similar or identical to a component of epithelial basement membranes, was identified as a major component of the abundant extracellular matrix synthesized by an experimentally induced rat yolk sac tumor. Immunocytochemical staining revealed laminin in cultured...... polypeptides with molecular weights of approximately 200,000 and 400,000. These comigrated with the polypeptides of mouse laminin isolated previously. The yolk sac tumor tissue grown in vivo contained laminin in the tumor cells and in the extracellular material as evidenced by immunofluorescence...... membranes in rat tissues in a manner indistinguishable from antilaminin. The presence of laminin in rat yolk sac cells, the presumed origin of our yolk sac tumor, was studied in some detail. Laminin was found to be present in normal cells of the visceral as well as the parietal yolk sac layer...

  8. Breaches of the pial basement membrane are associated with defective dentate gyrus development in mouse models of congenital muscular dystrophies. (United States)

    Li, Jing; Yu, Miao; Feng, Gang; Hu, Huaiyu; Li, Xiaofeng


    A subset of congenital muscular dystrophies (CMDs) has central nervous system manifestations. There are good mouse models for these CMDs that include POMGnT1 knockout, POMT2 knockout and Large(myd) mice with all exhibiting defects in dentate gyrus. It is not known how the abnormal dentate gyrus is formed during the development. In this study, we conducted a detailed morphological examination of the dentate gyrus in adult and newborn POMGnT1 knockout, POMT2 knockout, and Large(myd) mice by immunofluorescence staining and electron microscopic analyses. We observed that the pial basement membrane overlying the dentate gyrus was disrupted and there was ectopia of granule cell precursors through the breached pial basement membrane. Besides these, the knockout dentate gyrus exhibited reactive gliosis in these mouse models. Thus, breaches in the pial basement membrane are associated with defective dentate gyrus development in mouse models of congenital muscular dystrophies.

  9. Immunomodulatory effects of amniotic membrane matrix incorporated into collagen scaffolds. (United States)

    Hortensius, Rebecca A; Ebens, Jill H; Harley, Brendan A C


    Adult tendon wound repair is characterized by the formation of disorganized collagen matrix which leads to decreases in mechanical properties and scar formation. Studies have linked this scar formation to the inflammatory phase of wound healing. Instructive biomaterials designed for tendon regeneration are often designed to provide both structural and cellular support. In order to facilitate regeneration, success may be found by tempering the body's inflammatory response. This work combines collagen-glycosaminoglycan scaffolds, previously developed for tissue regeneration, with matrix materials (hyaluronic acid and amniotic membrane) that have been shown to promote healing and decreased scar formation in skin studies. The results presented show that scaffolds containing amniotic membrane matrix have significantly increased mechanical properties and that tendon cells within these scaffolds have increased metabolic activity even when the media is supplemented with the pro-inflammatory cytokine interleukin-1 beta. Collagen scaffolds containing hyaluronic acid or amniotic membrane also temper the expression of genes associated with the inflammatory response in normal tendon healing (TNF-α, COLI, MMP-3). These results suggest that alterations to scaffold composition, to include matrix known to decrease scar formation in vivo, can modify the inflammatory response in tenocytes. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1332-1342, 2016.

  10. Three-dimensional architecture of rat glomerular basement membrane by ultra-high resolution scanning electron microscopy.

    Directory of Open Access Journals (Sweden)



    Full Text Available We demonstrated the ultrastructure of rat glomerular basement membrane (GBM by ultra-high resolution scanning electron microscopy. GBM prepared by sonication methods and conductive-staining could be observed without metal coating at magnifications as high as 400,000 times. The GBM showed an irregular meshwork structure composed of various strands and pores. The width of the strands ranged from 6 to 15 nm, and the diameter of pores ranged from 6 to 50 nm. The present study confirmed our molecular sieve theory of the basement membrane.

  11. The central role of vascular extracellular matrix and basement membrane remodeling in metabolic syndrome and type 2 diabetes: the matrix preloaded

    Directory of Open Access Journals (Sweden)

    Tyagi Suresh C


    Full Text Available Abstract The vascular endothelial basement membrane and extra cellular matrix is a compilation of different macromolecules organized by physical entanglements, opposing ionic charges, chemical covalent bonding, and cross-linking into a biomechanically active polymer. These matrices provide a gel-like form and scaffolding structure with regional tensile strength provided by collagens, elasticity by elastins, adhesiveness by structural glycoproteins, compressibility by proteoglycans – hyaluronans, and communicability by a family of integrins, which exchanges information between cells and between cells and the extracellular matrix of vascular tissues. Each component of the extracellular matrix and specifically the capillary basement membrane possesses unique structural properties and interactions with one another, which determine the separate and combined roles in the multiple diabetic complications or diabetic opathies. Metabolic syndrome, prediabetes, type 2 diabetes mellitus, and their parallel companion (atheroscleropathy are associated with multiple metabolic toxicities and chronic injurious stimuli. The adaptable quality of a matrix or form genetically preloaded with the necessary information to communicate and respond to an ever-changing environment, which supports the interstitium, capillary and arterial vessel wall is individually examined.

  12. Manufacture and scanning electron microscopic observation of human dermis collagen membrane

    Institute of Scientific and Technical Information of China (English)


    @@ Introduction Collagen is a kind of biomacromolecule and can be used as cover material for burn wounds. In this article,we report the scanning electron microscopic observation of human dermis collagen membrane prepared by three methods.

  13. Antiglomerular basement membrane antibody-mediated glomerulonephritis after intranasal cocaine use. (United States)

    Peces, R; Navascués, R A; Baltar, J; Seco, M; Alvarez, J


    We report a case of rapidly progressive glomerulonephritis due to antiglomerular basement membrane (anti-GBM) antibodies that progressed to end-stage renal disease in a 35-year-old man who used intranasal cocaine on an occasional basis. In contrast to many prior reports of acute renal failure occurring with cocaine-associated rhabdomyolysis, this patient did not have any evidence of acute muscle damage and myoglobin release. Circulating anti-GBM antibodies and renal biopsy with linear IgG and C3 deposits confirmed the diagnosis of anti-GBM disease. The possibility of anti-GBM must be considered in the differential diagnosis of acute renal failure in cocaine addicts. This unusual combination raises complex questions regarding the pathogenesis of this type of renal injury.

  14. Cell invasion through basement membrane: the anchor cell breaches the barrier. (United States)

    Hagedorn, Elliott J; Sherwood, David R


    Cell invasion through basement membrane (BM) is a specialized cellular behavior critical to many normal developmental events, immune surveillance, and cancer metastasis. A highly dynamic process, cell invasion involves a complex interplay between cell-intrinsic elements that promote the invasive phenotype, and cell-cell and cell-BM interactions that regulate the timing and targeting of BM transmigration. The intricate nature of these interactions has made it challenging to study cell invasion in vivo and model in vitro. Anchor cell invasion in Caenorhabditis elegans is emerging as an important experimental paradigm for comprehensive analysis of BM invasion, revealing the gene networks that specify invasive behavior and the interactions that occur at the cell-BM interface.

  15. Transplacental transmission of antibodies to tubular basement membrane in guinea-pigs with autoimmune tubulointerstitial nephritis. (United States)

    Albini, B; Milgrom, M; Noble, B; Albini, C; Ossi, E; Andres, G A


    The offspring of female guinea-pigs with tubulo-interstitial nephritis were studied for possible passive transfer of disease. Whereas no immune deposits were seen on or before day 30 of gestation, IgG was detected in the tubular basement membrane (TBM) of fetuses at and after day 44. Serum of offspring contained antibodies to TBM, albeit in much lower titres than found in circulation of the mother guinea-pigs. No histopathological changes were seen in fetal kidneys. Thus, autoantibodies induced by heteroimmunization of pregnant guinea-pigs may be transmitted to offspring in the last third of the gestation period and can bind to fetal TBM. However, this transfer of antibodies does not cause disease.

  16. In vivo turnover of the basement membrane and other heparan sulfate proteoglycans of rat glomerulus

    Energy Technology Data Exchange (ETDEWEB)

    Beavan, L.A.; Davies, M.; Couchman, J.R.; Williams, M.A.; Mason, R.M.


    The metabolic turnover of rat glomerular proteoglycans in vivo was investigated. Newly synthesized proteoglycans were labeled during a 7-h period after injecting sodium (35S)sulfate intraperitoneally. At the end of the labeling period a chase dose of sodium sulfate was given. Subsequently at defined times (0-163 h) the kidneys were perfused in situ with 0.01% cetylpyridinium chloride in phosphate-buffered saline to maximize the recovery of 35S-proteoglycans. Glomeruli were isolated from the renal cortex and analyzed for 35S-proteoglycans by autoradiographic, biochemical, and immunochemical methods. Grain counting of autoradiographs revealed a complex turnover pattern of 35S-labeled macromolecules, commencing with a rapid phase followed by a slower phase. Biochemical analysis confirmed the biphasic pattern and showed that the total population of (35S)heparan sulfate proteoglycans had a metabolic half-life (t1/2) of 20 and 60 h in the early and late phases, respectively. Heparan sulfate proteoglycans accounted for 80% of total 35S-proteoglycans, the remainder being chondroitin/dermatan sulfate proteoglycans. Whole glomeruli were extracted with 4% 3-((cholamidopropyl)dimethy-lammonio)-1-propanesulfonate-4 M guanidine hydrochloride, a procedure which solubilized greater than 95% of the 35S-labeled macromolecules. Of these 11-13% was immunoprecipitated by an antiserum against heparan sulfate proteoglycan which, in immunolocalization experiments, showed specificity for staining the basement membrane of rat glomeruli. Autoradiographic analysis showed that 18% of total radioactivity present at the end of the labeling period was associated with the glomerular basement membrane.

  17. Collagen and chitosan membranes from alternative sources: evaluation of their potential for Tissue Engineering applications



    Natural polymers such as collagen and chitosan possess physical, chemical and biological characteristics that make them good candidates as extracellular matrix scaffolds with potential applications in Tissue Engineering. In the present work, collagen and chitosan biopolymer membranes made from waste material, were evaluated for dermal fibroblasts cell culture. Several membrane compositions were analyzed, including 100% collagen, 100% chitosan, 8:2, 2:8, 6:4, 4:6 collagen-chitosan, obtained fr...

  18. Normal and tumor-derived myoepithelial cells differ in their ability to interact with luminal breast epithelial cells for polarity and basement membrane deposition

    Energy Technology Data Exchange (ETDEWEB)

    Gudjonsson, Thorarinn; Ronnov-Jessen, Lone; Villadsen, Rene; Rank, Fritz; Bissell, Mina J.; Petersen, Ole William


    The signals that determine the correct polarity of breast epithelial structures in vivo are not understood. We have shown previously that luminal epithelial cells can be polarized when cultured within a reconstituted basement membrane gel. We reasoned that such cues in vivo may be given by myoepithelial cells. Accordingly, we used an assay where luminal epithelial cells are incorrectly polarized to test this hypothesis. We show that culturing human primary luminal epithelial cells within collagen-I gels leads to formation of structures with no lumina and with reverse polarity as judged by dual stainings for sialomucin, epithelial specific antigen or occludin. No basement membrane is deposited, and {beta}4-integrin staining is negative. Addition of purified human myoepithelial cells isolated from normal glands corrects the inverse polarity, and leads to formation of double-layered acini with central lumina. Among the laminins present in the human breast basement membrane (laminin-1, -5 and -10/11), laminin-1 was unique in its ability to substitute for myoepithelial cells in polarity reversal. Myoepithelial cells were purified also from four different breast cancer sources including a biphasic cell line. Three out of four samples either totally lacked the ability to interact with luminal epithelial cells, or conveyed only correction of polarity in a fraction of acini. This behavior was directly related to the ability of the tumor myoepithelial cells to produce {alpha}-1 chain of laminin. In vivo, breast carcinomas were either negative for laminin-1 (7/12 biopsies) or showed a focal, fragmented deposition of a less intensely stained basement membrane (5/12 biopsies). Dual staining with myoepithelial markers revealed that tumorassociated myoepithelial cells were either negative or weakly positive for expression of laminin-1, establishing a strong correlation between loss of laminin-1 and breast cancer. We conclude that the double-layered breast acinus may be

  19. Insecticidal Activity of a Basement Membrane-Degrading Protease against Heliothis virescens (Fabricius) and Acyrthosiphon pisum (Harris) (United States)

    ScathL is a cathepsin L-like cysteine protease derived from the flesh fly Sarcophaga peregrina that functions in basement membrane (BM) remodeling during insect development. A recombinant baculovirus expressing ScathL (AcMLF9.ScathL) kills larvae of the tobacco budworm, Heliothis virescens, signific...

  20. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane

    Energy Technology Data Exchange (ETDEWEB)



    An essential feature of mammary gland differentiation during pregnancy is the formation of alveoli composed of polarized epithelial cells, which, under the influence of lactogenic hormones, secrete vectorially and sequester milk proteins. Previous culture studies have described either organization of cells polarized towards lumina containing little or no demonstrable tissue-specific protein, or establishment of functional secretory cells exhibiting little or no glandular architecture. In this paper, we report that tissue-specific vectorial secretion coincides with the formation of functional alveoli-like structures by primary mammary epithelial cells cultured on a reconstituted basement membrane matrix (derived from Engelbreth-Holm-Swarm murine tumour). Morphogenesis of these unique three-dimensional structures was initiated by cell-directed remodelling of the exogenous matrix leading to reorganization of cells into matrixensheathed aggregates by 24 h after plating. The aggregates subsequently cavitated, so that by day 6 the cells were organized into hollow spheres in which apical cell surfaces faced lumina sealed by tight junctions and basal surfaces were surrounded by a distinct basal lamina. The profiles of proteins secreted into the apical (luminal) and basal (medium) compartments indicated that these alveoli-like structures were capable of an appreciable amount of vectorial secretion. Immunoprecipitation with a broad spectrum milk antiserum showed that more than 80% of caseins were secreted into the lumina, whereas iron-binding proteins (both lactoferrin and transferrin) were present in comparable amounts in each compartment. Thus, these mammary cells established protein targeting pathways directing milk-specific proteins to the luminal compartment. A time course monitoring secretory activity demonstrated that establishment of tissue-specific vectorial secretion and increased total and milk protein secretion coincided with functional alveolar

  1. Deletion of PPAR-γ in immune cells enhances susceptibility to antiglomerular basement membrane disease

    Directory of Open Access Journals (Sweden)

    Cristen Chafin


    Full Text Available Cristen Chafin2, Sarah Muse2, Raquel Hontecillas5, Josep Bassaganya-Riera5, David L Caudell2, Samuel K Shimp III4, M Nichole Rylander4, John Zhang6, Liwu Li3, Christopher M Reilly1,21Virginia College of Osteopathic Medicine, 2Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA; 3Department of Biological Sciences, 4Department of Mechanical Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA; 5Nutritional Immunology and Molecular Medicine Laboratory, Virginia Bioinformatics Institute, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA; 6Medical University of SC, Charleston, SC, USAAbstract: Activation of the nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPAR-γ has been shown to be immunoregulatory in autoimmune diseases by inhibiting production of a number of inflammatory mediators. We investigated whether PPAR-γ gene deletion in hematopoietic cells would alter disease pathogenesis in the antiglomerular basement membrane (anti-GBM mouse model. PPAR-γ+/+ and PPAR-γ-/- mice were immunized with rabbit antimouse GBM antibodies and lipopolysaccharide and evaluated for two weeks. Although both the PPAR-γ+/+ and PPAR-γ-/- mice had IgG deposition in the glomerulus and showed proteinuria two weeks after injection, glomerular and tubulointerstitial disease in PPAR-γ-/- mice were significantly more severe compared with the PPAR-γ+/+ animals. We observed that the PPAR-γ-/- mice had decreased CD4+CD25+ regulatory T cells and an increased CD8+:CD4+ ratio as compared with the PPAR-γ+/+ mice, suggesting that PPAR-γ has a role in the regulation of T cells. Furthermore, plasma interleukin-6 levels were significantly increased in the PPAR-γ-/- mice at two weeks as compared with the PPAR-γ+/+ animals. Taken together, these studies show that

  2. Numerical analysis of viscous flow through fibrous media: a model for glomerular basement membrane permeability. (United States)

    Palassini, M; Remuzzi, A


    Viscous flow through fibrous media is characterized macroscopically by the Darcy permeability (KD). The relationship between KD and the microscopic structure of the medium has been the subject of experimental and theoretical investigations. Calculations of KD based on the solution of the hydrodynamic flow at fiber scale exist in literature only for two-dimensional arrays of parallel fibers. We considered a fiber matrix consisting of a three-dimensional periodic array of cylindrical fibers with uniform radius (r) and length connected in a tetrahedral structure. According to recent ultrastructural studies, this array of fibers can represent a model for the glomerular basement membrane (GBM). The Stokes flow through the periodic array was simulated using a Galerkin finite element method. The dimensionless ratio K* = KD/r2 was determined for values of the fractional solid volume (phi) in the range 0.005 equation only for phi > 0.4. Among the other theoretical analysis considered, only that of Spielman and Goren (Environ. Sci. Technol. 2: 279-287, 1968) gives satisfactory agreement in the whole range of phi considered. These results can be useful to model combined transport of water and macromolecules through the GBM for the estimation of the radius and length of extracellular protein fibrils.

  3. Vascular Basement Membrane-derived Multifunctional Peptide, a Novel Inhibitor of Angiogenesis and Tumor Growth

    Institute of Scientific and Technical Information of China (English)

    Jian-Guo CAO; Shu-Ping PENG; Li SUN; Hui LI; Li WANG; Han-Wu DENG


    Vascular basement membrane-derived multifunctional peptide (VBMDMP) gene (fusion gene of the human immunoglobulin G3 upper hinge region and two tumstatin-derived fragments) obtained by chemical synthesis was cloned into vector pUC 19, and introduced into the expression vector pGEX-4T-1 to construct a prokaryotic expression vector pGEX-4T-1-VBMDMP. Recombinant VBMDMP produced in Escherichia coli has been shown to have significant activity of antitumor growth and antimetastasis in Lewis lung carcinoma transplanted into mouse C57B1/6. In the present study, we have studied the ability of rVBMDMP to inhibit endothelial cell tube formation and proliferation, to induce apoptosis in vitro, and to suppress tumor growth in vivo. The experimental results showed that rVBMDMP potently inhibited proliferation of human endothelial (HUVEC-12) cells and human colon cancer (SW480) cells in vitro, with no inhibition of proliferation in Chinese hamster ovary (CHO-K1) cells. rVBMDMP also significantly inhibited human endothelial cell tube formation and suppressed tumor growth of SW480 cells in a mouse xenograft model. These results suggest that rVBMDMP is a powerful therapeutic agent for suppressing angiogenesis and tumor growth.

  4. Cadherin 11 Involved in Basement Membrane Damage and Dermal Changes in Melasma. (United States)

    Kim, Nan-Hyung; Choi, Soo-Hyun; Lee, Tae Ryong; Lee, Chang-Hoon; Lee, Ai-Young


    Basement membrane (BM) disruption and dermal changes (elastosis, collagenolysis, vascular ectasia) have been reported in melasma. Although ultraviolet (UV) irradiation can induce these changes, UV is not always necessary for melasma development. Cadherin 11 (CDH11), which is upregulated in some melasma patients, has previously been shown to stimulate melanogenesis. Because CDH11 action requires cell-cell adhesion between fibroblasts and melanocytes, BM disruption in vivo should facilitate this. The aim of this study was to examine whether CDH11 overexpression leads to BM disruption and dermal changes, independent of UV irradiation. Immunohistochemistry/immunofluorescence, real-time PCR, Western blotting, and zymography suggested that BM disruption/dermal changes and related factors were present in the hyperpigmented skin of CDH11-upregulated melasma patients and in CDH11-overexpressing fibroblasts/keratinocytes. The opposite was seen in CDH11-knockdown cells. UV irradiation of the cultured cells did not increase CDH11 expression. Collectively, these data demonstrate that CDH11 overexpression could induce BM disruption and dermal changes in melasma, regardless of UV exposure.

  5. Anti-glomerular basement membrane glomerulonephritis in an HIV positive patient: case report

    Directory of Open Access Journals (Sweden)

    Eduardo José Bellotto Monteiro


    Full Text Available We report on a case of a patient with HIV infection, diagnosed 18 months prior to the development of an anti-glomerular basement membrane (anti-GBM rapidly progressive glomerulonephritis; this is probably the first report of such an association. A 30-year-old white man presented with elevation of serum creatinine (1.3 - 13.5 mg/dL within one month. At admission, the urinalysis showed proteinuria of 7.2 g/L and 8,000,000 erythrocytes/mL. Renal biopsy corresponded to a crescentic diffuse proliferative glomerulonephritis mediated by anti-GBM, and serum testing for anti-GBM antibodies was positive; antinuclear antibodies (ANA and anti-neutrophilic cytoplasmic antibodies (ANCA were also positive. The patient underwent hemodyalisis and was treated with plasmapheresis, cyclophosphamide and prednisone. The association described here is not casual, as crescentic glomerulonephritis is not common in HIV-positive patients, anti-GBM glomerulonephritis is rare and anti-GBM antibodies are frequently observed in HIV-positive subjects when compared to the overall population. Based on the current case and on the elevated frequency of the positivity for such antibodies in this group of patients, it is advisable to be aware of the eventual association between these two conditions and to promote an active search for anti-GBM antibodies and early diagnosis of eventual urinary abnormalities in HIV-positive subjects, considering the severity of anti-GBM glomerulonephritis.

  6. Biodegradation of differently cross-linked collagen membranes: an experimental study in the rat.

    NARCIS (Netherlands)

    Rothamel, D.; Schwarz, F.; Sager, M.; Herten, M. van; Sculean, A.; Becker, J.M.


    The aim of the present study was to compare the biodegradation of differently cross-linked collagen membranes in rats. Five commercially available and three experimental membranes (VN) were included: (1) BioGide (BG) (non-cross-linked porcine type I and III collagens), (2) BioMend (BM), (3) BioMendE

  7. De novo deposition of laminin-positive basement membrane in vitro by normal hepatocytes and during hepatocarcinogenesis

    DEFF Research Database (Denmark)

    Albrechtsen, R; Wewer, U M; Thorgeirsson, S S


    De novo formation of laminin-positive basement membranes was found to be a distinct morphologic feature of diethylnitrosamine/phenobarbital-induced hepatocellular carcinomas of the rat. The first appearance of extracellularly located laminin occurred in the preneoplastic liver lesions...... (corresponding to neoplastic nodules), and this feature became successively more prominent during the course of hepatocellular carcinoma development. Most groups of tumor cells were surrounded by laminin-positive basement membrane material. The laminin-positive material was also deposited along the sinusoids......, a location where no laminin was seen in normal rat liver. The amount of extractable laminin from hepatocellular carcinomas was significantly higher (approximately 100 ng per mg tissue) than that of normal liver tissue (less than 20 ng per mg). In vitro experiments demonstrated that normal and preneoplastic...

  8. Distribution of two basement membrane proteoglycans through hair follicle development and the hair growth cycle in the rat

    DEFF Research Database (Denmark)

    Couchman, J R; King, J L; McCarthy, K J


    The distribution of two distinct populations of basement membrane proteoglycans has been monitored through hair growth development in the rat embryo and subsequent hair growth cycle. An antiserum against a small heparan sulfate proteoglycan uniformly stained the dermal-epidermal junction...... of embryonic rats throughout the period of hair follicle formation. On the other hand, monoclonal antibodies recognizing a basement membrane-specific chondroitin sulfate proteoglycan only weakly stained 16-d embryo dermal-epidermal junction, but strong staining was associated with hair follicle buds...... as they developed. Through the hair growth cycle, it was found that the heparan sulfate proteoglycan persisted around the follicles, while the chondroitin sulfate proteoglycan decreased in amount through catagen until it was undetectable at the base and dermal papilla of the telogen follicle. As anagen commenced...

  9. Defective muscle basement membrane and lack of M-laminin in the dystrophic dy/dy mouse

    DEFF Research Database (Denmark)

    Xu, H; Christmas, P; Wu, X R;


    M-laminin is a major member of the laminin family of basement membrane proteins. It is prominently expressed in striated muscle and peripheral nerve. M-laminin is deficient in patients with the autosomal recessive Fukuyama congenital muscular dystrophy but is normal in patients with the sex......-linked Duchenne and Becker muscular dystrophies. We have examined M-laminin expression in mice with autosomal recessive muscular dystrophy caused by the mutation dy. The heavy chain of M-laminin was undetectable in skeletal muscle, heart muscle, and peripheral nerve by immunofluorescence and immunoblotting...... in homozygous dystrophic dy/dy mice but was normal in heterozygous and wild-type nondystrophic mice. Immunofluorescence confirmed the presence of other major basement membrane proteins in the dystrophic mice. Very low levels of M-laminin heavy chain mRNA were detected by Northern blotting of muscle and heart...

  10. Evaluation of epigallocatechin-3-gallate (EGCG) cross-linked collagen membranes and concerns on osteoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Chu, Chenyu; Deng, Jia [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Xiang, Lin; Wu, Yingying [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Oral Implantology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Wei, Xiawei [State Key Laboratory of Biotherapy and Laboratory for Aging Research, West China Hospital, Sichuan University, and Collaborative Innovation Center for Biotherapy, Chengdu, Sichuan 610041 (China); Qu, Yili [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Oral Implantology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Man, Yi, E-mail: [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Department of Oral Implantology, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China)


    Collagen membranes have ideal biological and mechanical properties for supporting infiltration and proliferation of osteoblasts and play a vital role in guided bone regeneration (GBR). However, pure collagen can lead to inflammation, resulting in progressive bone resorption. Therefore, a method for regulating the level of inflammatory cytokines at surgical sites is paramount for the healing process. Epigallocatechin-3-gallate (EGCG) is a component extracted from green tea with numerous biological activities including an anti-inflammatory effect. Herein, we present a novel cross-linked collagen membrane containing different concentrations of EGCG (0.0064%, 0.064%, and 0.64%) to regulate the level of inflammatory factors secreted by pre-osteoblast cells; improve cell proliferation; and increase the tensile strength, wettability, and thermal stability of collagen membranes. Scanning electron microscope images show that the surfaces of collagen membranes became smoother and the collagen fiber diameters became larger with EGCG treatment. Measurement of the water contact angle demonstrated that introducing EGCG improved membrane wettability. Fourier transform infrared spectroscopy analyses indicated that the backbone of collagen was intact, and the thermal stability was significant improved in differential scanning calorimetry. The mechanical properties of 0.064% and 0.64% EGCG-treated collagen membranes were 1.5-fold greater than those of the control. The extent of cross-linking was significantly increased, as determined by a 2,4,6-trinitrobenzenesulfonic acid solution assay. The Cell Counting Kit-8 (CCK-8) and live/dead assays revealed that collagen membrane cross-linked by 0.0064% EGCG induced greater cell proliferation than pure collagen membranes. Additionally, real-time polymerase chain reaction and enzyme-linked immunosorbent assay results showed that EGCG significantly affected the production of inflammatory factors secreted by MC3T3-E1 cells. Taken together, our

  11. IgE basement membrane zone antibodies induce eosinophil infiltration and histological blisters in engrafted human skin on SCID mice. (United States)

    Zone, John J; Taylor, Ted; Hull, Christopher; Schmidt, Linda; Meyer, Laurence


    Bullous pemphigoid (BP) is characterized by the deposition of IgG in the basement membrane zone, infiltration of eosinophils, and blister formation. The purpose of this study was to evaluate a potential role of IgE basement membrane antibodies in the histological findings of BP. LABD97 is a component of the shed ectodomain of bullous pemphigoid antigen 2. We have developed an IgE hybridoma to LABD97 antigen. This hybridoma was injected subcutaneously in SCID mice with engrafted human skin. A subcutaneous hybridoma secreting IgE antibodies developed. An IgE mouse hybridoma to trinitrophenyl was used as a control. Human grafts and mouse skin were examined grossly over 21 days, histologically, and immunopathologically at day 21 after injection of the hybridoma. A visible subcutaneous tumor developed in 10-14 days. Erythema and intense scratching developed 2-3 days before the tumor in test mice, but not in controls. At day 21, 16/16 test mice developed intense eosinophil infiltration and degranulation of the human mast cells within the grafts and 13/16 developed histological, but not clinically visible, basement membrane blisters. Human skin grafts of control mice and normal mouse skin on the test mice and control mice did not develop any histological abnormalities. IgE antibodies to LABD97 recapitulate the histological inflammatory process seen in BP.

  12. A Case of Fibrillary Glomerulonephritis Associated with Thrombotic Microangiopathy and Anti-Glomerular Basement Membrane Antibody

    Directory of Open Access Journals (Sweden)

    Akishi Momose


    Full Text Available We present the first report of a case of fibrillary glomerulonephritis (FGN associated with thrombotic microangiopathy (TMA and anti-glomerular basement membrane antibody (anti-GBM antibody. A 54-year-old man was admitted to our hospital for high fever and anuria. On the first hospital day, we initiated hemodialysis for renal dysfunction. Laboratory data revealed normocytic-normochromic anemia with schistocytes in the peripheral smear, thrombocytopenia, increased serum lactate dehydrogenase, decreased serum haptoglobin, and negative results for both direct and indirect Coombs tests. Based on these results, we diagnosed TMA. Assays conducted several days later indicated a disintegrin-like and metalloprotease with a thrombospondin motif 13 (ADAMTS13 activity of 31.6%, and ADAMTS13 inhibitors were negative. We started plasma exchange using fresh frozen plasma and steroid pulse therapy. Anti-GBM antibody was found to be positive. Renal biopsy showed FGN. Blood pressure rose on the 46th hospital day, and mild convulsions developed. Based on magnetic resonance imaging of the head, the patient was diagnosed with reversible posterior leukoencephalopathy syndrome. Hypertension persisted despite administration of multiple antihypertensive agents, and the patient experienced a sudden generalized seizure. Computed tomography of the head showed multiple cerebral hemorrhages. However, his blood pressure subsequently decreased and the platelet count increased. TMA remitted following 36 plasma exchange sessions, but renal function was not restored, and maintenance hemodialysis was continued. The patient was discharged on the 119th day of hospitalization. In conclusion, it was shown that TMA, FGN and anti-GBM antibody were closely related.

  13. Putative role of basement membrane for dentinogenesis in the mesenchyme of murine dental papillae in vitro. (United States)

    Kikuchi, H; Amano, H; Yamada, S


    In a new culture-conditioning system of agar-coated mesenchyme of isolated incisor dental papillae, dentinogenesis has been induced adjacent to an agar substratum that functions as a foothold for cell immobilisation. To elucidate the role of the basement membrane (BM) in dentinogenesis, we have examined the way in which dentinogenesis depends upon BM components or transforming growth factor (TGF)-beta1 in this system. At the mesenchymal-epithelial junction of odontogenic organs (cut incisor tooth germs), TGF-beta1 visibly increased in the BM during incubation. In isolated dental papillae, BM components were synthesised and deposited at aligned peripheral cells of the explants, together with an increasing amount of TGF-beta1. These components were not assembled into extracellular matrix (ECM)-absorbed agar adjacent to explants, although dentinogenesis proceeded in the presence of pericellular BM components associated with TGF-beta1. When signalling via TGF-beta type II receptors was blocked, neither ECM production nor dentinogenesis was observed but explants partially detached from the agar surface, presumably as a result of the suppressed production of ECM, since attachment was retained by pre-coating explants with artificial matrices. Rescue experiments showed that TGF-beta1 regulated dentinogenesis through ECM production. With regard to BM components, inducible dentinogenesis was Arg-Gly-Asp (RGD)-dependent. Thus, pericellular BM components associated with TGF-beta1 and an ECM-absorbed agar substratum, which affects dentinogenesis, synergistically play a role similar to that of BM components in vivo. The BM therefore serves as a structural meshwork that acts as a foothold for cell immobilisation; its components act as ligands for RGD-dependent cell adhesion and it stores TGF-beta1, which regulates ECM production.

  14. Long-term outcome of anti-glomerular basement membrane antibody disease treated with immunoadsorption.

    Directory of Open Access Journals (Sweden)

    Peter Biesenbach

    Full Text Available Anti-glomerular basement membrane (GBM antibody disease may lead to acute crescentic glomerulonephritis with poor renal prognosis. Current therapy favours plasma exchange (PE for removal of pathogenic antibodies. Immunoadsorption (IAS is superior to PE regarding efficiency of antibody-removal and safety. Apart from anecdotal data, there is no systemic analysis of the long-term effects of IAS on anti-GBM-disease and antibody kinetics.To examine the long-term effect of high-frequency IAS combined with standard immunosuppression on patient and renal survival in patients with anti-GBM-disease and to quantify antibody removal and kinetics through IAS.Retrospective review of patients treated with IAS for anti-GBM-antibody disease confirmed by biopsy and/or anti-GBM-antibodies.University Hospital of Vienna, Austria.10 patients with anti-GBM-disease treated with IAS.Patient and renal survival, renal histology, anti-GBM-antibodies.Anti-GBM-antibodies were reduced by the first 9 IAS treatments (mean number of 23 to negative levels in all patients. Renal survival was 40% at diagnosis, 70% after the end of IAS, 63% after one year and 50% at the end of observation (mean 84 months, range 9 to 186. Dialysis dependency was successfully reversed in three of six patients. Patient survival was 90% at the end of observation.IAS efficiently eliminates anti-GBM-antibodies suggesting non-inferiority to PE with regard to renal and patient survival. Hence IAS should be considered as a valuable treatment option for anti-GBM-disease, especially in patients presenting with a high percentage of crescents and dialysis dependency due to an unusual high proportion of responders.

  15. The effect of asthma on the perimeter of the airway basement membrane. (United States)

    Elliot, John G; Budgeon, Charley A; Harji, Salima; Jones, Robyn L; James, Alan L; Green, Francis H


    When comparing the pathology of airways in individuals with and without asthma, the perimeter of the basement membrane (Pbm) is used as a marker of airway size, as it is independent of airway smooth muscle shortening or airway collapse. The extent to which the Pbm is itself altered in asthma has not been quantified. The aim of this study was to compare the Pbm from the same anatomical sites in postmortem lungs from subjects with (n = 55) and without (n = 30) asthma (nonfatal or fatal). Large and small airways were systematically sampled at equidistant "levels" from the apical segment of the left upper lobes and anterior and basal segments of the left lower lobes of lungs fixed in inflation. The length of the Pbm was estimated from cross sections of airway at each relative level. Linear mixed models were used to investigate the relationships between Pbm and sex, age, height, smoking status, airway level, and asthma group. The final model showed significant interactions between Pbm and airway level in small (<3 mm) airways, in subjects having asthma (P < 0.0001), and by sex (P < 0.0001). No significant interactions for Pbm between asthma groups were observed for larger airways (equivalent to a diameter of ∼3 mm and greater) or smoking status. Asthma is not associated with remodeling of the Pbm in large airways. In medium and small airways, the decrease in Pbm in asthma (≤20%) would not account for the published differences in wall area or area of smooth muscle observed in cases of severe asthma.

  16. Enzymatic degradation of collagen-guided tissue regeneration membranes by periodontal bacteria. (United States)

    Sela, Michael N; Kohavi, David; Krausz, Emanuela; Steinberg, Doron; Rosen, Graciela


    Bacterial infection in the vicinity of guided tissue regeneration barrier membranes was shown to have a negative effect on the clinical outcomes of this increasingly used technique. Several oral and specifically periodontal bacteria were shown to adhere to such membranes in vivo and in vitro with a higher affinity to membranes constructed from collagen. The present study examined the role of periodontal bacteria and their enzymes in the degradation of commercially used collagen membranes. Degradation of two collagen membranes [Biomend (Calcitek, Colla-Tec Inc., Plainsboro, NJ) and Bio-Gide (Geistlich Biomaterials, Wolhousen, Switzerland)] labeled by fluorescein isothiocyanate was examined by measuring soluble fluorescence. Porphyromonas gingivalis, Treponema denticola and Actinobacillus actinomycetemcomitans and their enzymes were evaluated. Collagenase from Clostridium hystolyticum was used as a positive control. While whole cells of P. gingivalis were able to degrade both types of membranes, T. denticola could degrade Bio-Gide membranes only and A. actinomycetemcomitans whole cells could degrade none of the membranes. Fractionation of P. gingivalis cells revealed that cell membrane associated proteases were responsible for the degradation of the two collagen membranes. In T. denticola, the purified major phenylalanine protease was found to be responsible for the degradation of Bio-Gide membranes. These results suggest that proteolytic bacterial enzymes may take part in the degradation of collagen barrier membranes used for guided tissue regeneration.

  17. Chitosan-Coated Collagen Membranes Promote Chondrocyte Adhesion, Growth, and Interleukin-6 Secretion

    Directory of Open Access Journals (Sweden)

    Nabila Mighri


    Full Text Available Designing scaffolds made from natural polymers may be highly attractive for tissue engineering strategies. We sought to produce and characterize chitosan-coated collagen membranes and to assess their efficacy in promoting chondrocyte adhesion, growth, and cytokine secretion. Porous collagen membranes were placed in chitosan solutions then crosslinked with glutaraldehyde vapor. Fourier transform infrared (FTIR analyses showed elevated absorption at 1655 cm-1 of the carbon–nitrogen (N=C bonds formed by the reaction between the (NH2 of the chitosan and the (C=O of the glutaraldehyde. A significant peak in the amide II region revealed a significant deacetylation of the chitosan. Scanning electron microscopy (SEM images of the chitosan-coated membranes exhibited surface variations, with pore size ranging from 20 to 50 µm. X-ray photoelectron spectroscopy (XPS revealed a decreased C–C groups and an increased C–N/C–O groups due to the reaction between the carbon from the collagen and the NH2 from the chitosan. Increased rigidity of these membranes was also observed when comparing the chitosan-coated and uncoated membranes at dried conditions. However, under wet conditions, the chitosan coated collagen membranes showed lower rigidity as compared to dried conditions. Of great interest, the glutaraldehyde-crosslinked chitosan-coated collagen membranes promoted chondrocyte adhesion, growth, and interleukin (IL-6 secretion. Overall results confirm the feasibility of using designed chitosan-coated collagen membranes in future applications, such as cartilage repair.

  18. In vivo laser confocal microscopy findings in patients with map-dot-fingerprint (epithelial basement membrane dystrophy

    Directory of Open Access Journals (Sweden)

    Kobayashi A


    Full Text Available Akira Kobayashi, Hideaki Yokogawa, Kazuhisa SugiyamaDepartment of Ophthalmology, Kanazawa University Graduate School of Medical Science, Kanazawa, JapanBackground: The purpose of this study was to investigate pathological changes of the corneal cell layer in patients with map-dot-fingerprint (epithelial basement membrane dystrophy by in vivo laser corneal confocal microscopy.Methods: Two patients were evaluated using a cornea-specific in vivo laser scanning confocal microscope (Heidelberg Retina Tomograph 2 Rostock Cornea Module, HRT 2-RCM. The affected corneal areas of both patients were examined. Image analysis was performed to identify corneal epithelial and stromal deposits correlated with this dystrophy.Results: Variously shaped (linear, multilaminar, curvilinear, ring-shape, geographic highly reflective materials were observed in the “map” area, mainly in the basal epithelial cell layer. In “fingerprint” lesions, multiple linear and curvilinear hyporeflective lines were observed. Additionally, in the affected corneas, infiltration of possible Langerhans cells and other inflammatory cells was observed as highly reflective Langerhans cell-like or dot images. Finally, needle-shaped materials were observed in one patient.Conclusion: HRT 2-RCM laser confocal microscopy is capable of identifying corneal microstructural changes related to map-dot-fingerprint corneal dystrophy in vivo. The technique may be useful in elucidating the pathogenesis and natural course of map-dot-fingerprint corneal dystrophy and other similar basement membrane abnormalities.Keywords: cornea, confocal microscopy, map-dot-fingerprint dystrophy, epithelial basement membrane dystrophy, Heidelberg Retina Tomograph 2 Rostock Cornea Module (HRT 2-RCM

  19. Comparative analysis of collagen membranes for the treatment of implant dehiscence defects. (United States)

    Oh, Tae-Ju; Meraw, Stephen J; Lee, Eun-Ju; Giannobile, William V; Wang, Hom-Lay


    Guided bone regeneration (GBR) evolved from the concept of guided tissue regeneration (GTR) and has been used for reconstructing sites with bone deficiencies associated with dental implants. For GBR, the use of absorbable collagen membranes has been increasing, but, at present, scientific information on the use of collagen membranes for GBR is limited. This study was aimed to clinically and histomorphometrically compare two collagen membranes, Bio-Gide(R) and BioMend ExtendTM, for the treatment of implant dehiscence defects in eight mongrel dogs. Implant dehiscence defects were surgically created in edentulous ridges, followed by the placement of three endosseous implants bilaterally in the mandible. Each implant dehiscence defect was randomly assigned to one of three treatment groups: (1) control (no membrane), (2) porcine dermis collagen barrier (Bio-Gide) or (3) bovine tendon collagen barrier (BioMend Extend). Dogs were sacrificed at 4 and 16 weeks (four dogs each) after treatment. Histomorphometric analysis included percentage linear bone fill (LF), new bone-to-implant contact (BIC) and area of new bone fill (BF). The results of the study revealed no significant differences among groups for any parameter at 4 weeks. However, at 16 weeks, more LF, BIC, and BF were noted in the membrane-treated groups than controls. BioMend Extend-treated defects demonstrated significantly greater BIC than control (P Membrane exposure occurred in 9 out of 15 sites examined, resulting in significantly less LF and BIC than the sites without membrane exposure (P collagen membranes may significantly enhance bone regeneration, manifested at late stage (16 weeks) of healing; and (2) space maintenance and membrane coverage were the two most important factors affecting GBR using bioabsorbable collagen membranes.

  20. Anti-Glomerular Basement Membrane Disease Combined with IgA Nephropathy Complicated with Reversible Posterior Leukoencephalopathy Syndrome: An Unusual Case (United States)

    Ge, Ya-ting; Liao, Jin-lan; Liang, Wei; Xiong, Zu-ying


    Patient: Male, 24 Final Diagnosis: Crescentic glomerulonephritis (type I) with IgA nephropathy Symptoms: Headache • gross hematuria • nocturia • seizures Medication: Cyclophosphamide Clinical Procedure: Dignosis to treatment Specialty: Nephrology Objective: Rare co-existance of disease or pathology Background: Anti-glomerular basement membrane disease (anti-GBM disease) is an autoimmune glomerulonephritis disease that is characterized by IgG linear deposition along the non-collagen domain of α3 chains of type IV collagen on the GBM. Although anti-GBM disease accompanied with IgA linear deposition along GBMs was discussed previously in some papers, anti-GBM disease combined with IgA granular deposition in the mesangial area, especially complicated with reversible posterior leukoencephalopathy syndrome (RPLS), was rarely reported. RPLS is usually caused by hypertensive encephalopathy, renal decompensation, fluid retention, and adverse effects of immunosuppressive drugs. Case Report: A male patient with the chief complaints of headache, gross hematuria, and nocturia was referred to our hospital. Based on renal biopsy, the diagnosis was finally confirmed as anti-GBM disease combined with IgA nephropathy and, the patient received comprehensive treatment, including cyclophosphamide (CTX), which led to symptom improvement. Two days after the third impulse CTX was given, he suddenly experienced headache and dizziness, which eventually developed into a tonic-clonic seizure. RPLS was identified by cranial magnetic resonance imaging (MRI) with reversible neuroimaging. After diazepam and antihypertension management, seizures were controlled. RPLS, a neurological complication, was found in anti-GBM disease with IgA nephropathy during our immunosuppressants therapy for the first time. Conclusions: It is worth paying more attention to patients with rapidly progressive glomerulonephritis (RPGN), as they might be complicated with RPLS during intravenous administration of CTX

  1. Collagen mRNA levels changes during colorectal cancer carcinogenesis

    DEFF Research Database (Denmark)

    Skovbjerg, Hanne; Anthonsen, Dorit; Lothe, Inger M B;


    BACKGROUND: Invasive growth of epithelial cancers is a complex multi-step process which involves dissolution of the basement membrane. Type IV collagen is a major component in most basement membranes. Type VII collagen is related to anchoring fibrils and is found primarily in the basement membrane....... In addition, corresponding tissue was examined from healthy volunteers (n = 20). mRNA levels were normalized to beta-actin. Immunohistochemical analysis of the distributions of type IV and type VII collagens were performed on normal and affected tissues from colorectal cancer patients. RESULTS: The alpha1(IV......). The level of alpha 6(IV) was 5-fold lower in colorectal cancer tissue as compared to healthy individuals (p collagen was visualized by immunohistochemical staining. CONCLUSION: Our results suggest that the down-regulation of alpha 6(IV) mRNA coincides...

  2. In Vitro and In Vivo Study of a Novel Porcine Collagen Membrane for Guided Bone Regeneration

    Directory of Open Access Journals (Sweden)

    Eisner Salamanca


    Full Text Available For years, in order to improve bone regeneration and prevent the need of a second stage surgery to remove non-resorbable membranes, biological absorbable membranes have gradually been developed and applied in guided tissue regeneration (GTR. The present study’s main objective was to achieve space maintenance and bone regeneration using a new freeze-dried developed porcine collagen membrane, and compare it with an already commercial collagen membrane, when both were used with a bovine xenograft in prepared alveolar ridge bone defects. Prior to surgery, the membrane’s vitality analysis showed statistically significant higher cell proliferation in the test membrane over the commercial one. In six beagle dogs, commercial bone xenograft was packed in lateral ridge bone defects prepared in the left and right side and then covered with test porcine collagen membrane or commercial collagen membrane. Alveolar height changes were measured. Histomorphometric results, in vitro and in vivo properties indicated that the new porcine collagen membrane is biocompatible, enhances bone xenograft osteoconduction, and reduces the alveolar ridge height reabsorption rate.

  3. Detection of basement membrane components on the surface of acellular porcine cornea%脱细胞猪角膜表面基底膜成分的检测

    Institute of Scientific and Technical Information of China (English)

    林旭初; 金岩; 惠延年


    BACKGROUND: The previous experiments have suggested that the acellular porcine cornea stroma (APCS) has a good compatibility and can support the growth of keratocytes and skin epithelial cellsOBJECTIVE:To detect whether the important structure for the grwth of corneal cells basement membrane canbe preserved on the surface of the APCSMETHODS: Fluorescent antibody was used to detect the basement membrane component (laminin and collagen IV) by immunohistochemistry Fluorescence microscopy was observed whether the basement membrane could be preserved on the surface oftheAPCS RESULTS AND CONCLUSION:Immunofluorescence staining showed that larnimn and collagen IV positively expressedthe APCS can preserve the natural basement membrane component of the cornea which is beneficial to the growth of comeal epithelial cells%背景:前期实验显示脱细胞猪角膜具有良好的组织相容性,可以支持角膜细胞和皮肤上皮细胞的生长.目的:检测脱细胞猪角膜是否保存了利于角膜上皮细胞生长的重要组织结构-基底膜.方法:利用荧光抗体对脱细胞猪角膜表面的基底膜成分(层粘蛋白和Ⅳ型胶原)进行免疫组织化学检测,荧光显微镜下观察脱细胞猪角膜表面是否保存了基底膜成分.结果与结论:免疫荧光染色显示脱细胞猪角膜前基质表面层粘蛋白和Ⅳ型胶原呈阳性表达,与新鲜猪角膜表面基底膜的荧光表达相同,表明脱细胞猪角膜保存了利于角膜上皮细胞生长的基底膜.

  4. Targeted Expression of Stromelysin-1 in Mammary Gland Provides Evidence for a Role of Proteinases in Branching Morphogenesis and the Requirement for an Intact Basement Membrane for Tissue-specific Gene Expression

    Energy Technology Data Exchange (ETDEWEB)

    Sympson, Carolyn J; Talhouk, Rabih S; Alexander, Caroline M; Chin, Jennie R; Cliff, Shirley M; Bissell, Mina J; Werb, Zena


    The extracellular matrix (ECM) is an important regulator of the differentiated phenotype of mammary epithelial cells in culture. Despite the fact that ECM-degrading enzymes have been implicated in morphogenesis and tissue remodeling, there is little evidence for a direct role for such regulation in vivo. We generated transgenic mice that express autoactivated isoforms of the matrix metalloproteinase stromelysin-1, under the control of the whey acidic protein gene promoter, to examine the effect of inappropriate expression of this enzyme. Stromelysin-1 is implicated as the primary player in the loss of basement membrane and loss of function in the mammary gland during involution. The transgene was expressed at low levels in mammary glands of virgin female mice, leading to an unexpected phenotype: The primary ducts had supernumerary branches and showed precocious development of alveoli that expressed beta-casein at levels similar to that of an early- to mid-pregnant gland. Lactating glands showed high levels of transgene expression, with accumulation at the basement membrane, and a decrease in laminin and collagen IV, resulting in a loss of basement membrane integrity; this was accompanied by a dramatic alteration of alveolar morphology, with decreased size and shrunken lumina containing little beta-casein. During pregnancy, expression of endogenous whey acidic protein and beta-casein was reduced in transgenic glands, confirming the observed dependence of milk protein transcription of ECM in mammary epithelial cells in culture. These data provide direct evidence that stromelysin-1 activity can be morphogenic for mammary epithelial cells, inducing hyperproliferation and differentiation in virgin animals, and that its lytic activity can, indeed, disrupt membrane integrity and reduce mammary-specific function. We conclude that the balance of ECM-degrading enzymes with their inhibitors, and the associated regulation of ECM structure, is crucial for tissue-specific gene

  5. [Ultrastructure of glomerular podocyts in the incipient phase of minimal change nephrotic syndrome with thin basement membrane disease]. (United States)

    Ogawa, Ryo; Miyoshi, Ken-ichi; Nagao, Tomoaki; Jotoku, Masanori; Irita, Jun; Okura, Takafumi; Higaki, Jitsuo


    An 80-year-old woman was referred to the Division of Nephrology at Ehime University Hospital because of leg edema in December 2010. She had been treated with 300 mg of tocopherol for scleroderma since 2007 and treated with 9 mg of prednisolone (PSL) for autoimmune hearing loss since 2010. Due to the occurrence of mild hematuria (5-9/HPF), proteinuria (0.9 g/day) and an increased serum creatinine level (1.31 mg/dL), a renal biopsy was performed. Light microscopy (LM) showed minor abnormality in the glomeruli, and immunohistology showed the absence of deposits of immunoglobulins and complements. Electron microscopy (EM) showed a thin glomerular basement membrane with a limited level of podocyte abnormalities. Due to the findings of intimal thickening of interlobular arteries and subcapsular accumulation of global sclerosis on LM, she was diagnosed with nephrosclerosis and thin basement membrane disease. Four weeks later, her leg edema had increased considerably and urinary protein had increased to 12.4 g/day. The second biopsy showed similar findings in LM and IF as the first biopsy, but EM revealed diffuse foot process effacement. She was diagnosed with minimal change nephrotic syndrome (MCNS) and treated with methylprednisolone pulse therapy followed by 40 mg of oral PSL. Her urinary protein had completely disappeared 6 weeks later. Complete remission with PSL treatment indicates that urinary protein at first renal biopsy was due to MCNS. Our case exhibited podocyte features in the incipient phase of human MCNS.

  6. Basement Membrane Mimics of Biofunctionalized Nanofibers for a Bipolar-Cultured Human Primary Alveolar-Capillary Barrier Model. (United States)

    Nishiguchi, Akihiro; Singh, Smriti; Wessling, Matthias; Kirkpatrick, Charles J; Möller, Martin


    In vitro reconstruction of an alveolar barrier for modeling normal lung functions and pathological events serve as reproducible, high-throughput pharmaceutical platforms for drug discovery, diagnosis, and regenerative medicine. Despite much effort, the reconstruction of organ-level alveolar barrier functions has failed due to the lack of structural similarity to the natural basement membrane, functionalization with specific ligands for alveolar cell function, the use of primary cells and biodegradability. Here we report a bipolar cultured alveolar-capillary barrier model of human primary cells supported by a basement membrane mimics of fully synthetic bifunctional nanofibers. One-step electrospinning process using a bioresorbable polyester and multifunctional star-shaped polyethylene glycols (sPEG) enables the fabrication of an ultrathin nanofiber mesh with interconnected pores. The nanofiber mesh possessed mechanical stability against cyclic expansion as seen in the lung in vivo. The sPEGs as an additive provide biofunctionality to fibers through the conjugation of peptide to the nanofibers and hydrophilization to prevent unspecific protein adsorption. Biofunctionalized nanofiber meshes facilitated bipolar cultivation of endothelial and epithelial cells with fundamental alveolar functionality and showed higher permeability for molecules compared to microporous films. This nanofiber mesh for a bipolar cultured barrier have the potential to promote growth of an organ-level barrier model for modeling pathological conditions and evaluating drug efficacy, environmental pollutants, and nanotoxicology.

  7. [Use of native and cross-linked collagen membranes for guided tissue and bone regeneration]. (United States)

    Schwarz, Frank; Sager, Martin; Rothamel, Daniel; Herten, Monika; Sculean, Anton; Becker, Jürgen


    A material which is used as a barrier for GBR/GTR procedures has to satisfy several physicochemical characteristics such as biocompatibility, tissue integration, barrier function, and dimensional stability. Recently, many investigations reported on the use of products derived from type I and type III porcine or bovine collagen. Collagen membranes are predominantly resorbed by enzymatic activity (protease and collagenase). To decrease resorption, various physical and chemical cross-linking techniques have been used. Although nowadays cross-linking of collagen seems to be a commonly used procedure, its impact on physicochemical properties of the membrane is still unknown. The aim of the present literature review is to evaluate the potential use of different collagen membranes for GBR/GTR procedures.

  8. Ultrastructural immunocytochemical localization of chondroitin sulfate proteoglycan in Bruch's membrane of the rat

    DEFF Research Database (Denmark)

    Lin, W L; Essner, E; McCarthy, K J


    Two monoclonal antibodies (Mab 4D5 and 2D6) raised against the core protein of a basement membrane chondroitin sulfate proteoglycan from Reichert's membrane of the rat, were used for ultrastructural immunoperoxidase localization of this protein in Bruch's membrane of the rat. Immunoreactivity for...... for both antibodies was found in the basal lamina (basement membrane) of the choriocapillary endothelium and retinal pigment epithelium, in collagen fibers in the collagenous zones, and surrounding the elastic layer....

  9. Collagen membrane alleviates peritendinous adhesion in the rat Achilles tendon injury model

    Institute of Scientific and Technical Information of China (English)

    ZHAO Huan; GUAN Hong-geng; GU Jun; LUO Zong-ping; ZHANG Wen; CHEN Bing; GU Qiao-li


    Background Tendon adhesion is one of the most common causes of disability following tendon surgery.Therefore,prevention of peritendinous adhesion after surgical repair of tendon is a major challenge.The aim of this study was to explore the possible application of a collagen membrane for the prevention or attenuation of peritendinous adhesions.Methods Sprague-Dawley (SD) rat Achilles tendon was cut and sutured by a modified Kessler's technique with or without the collagen membrane wrapped.Macroscopic,morphological and biomechanical evaluations were applied to examine the recovery of the injured tendon at 4 and 8 weeks after surgery.Results The surgery group wrapped by collagen membranes had a better outcome than the group with surgery repair only.In the collagen membrane-treated group,less adhesion appeared,stronger tensile strength was detected,and more tendon fibers and collagen I expression were observed morphologically.Conclusion Wrapping the tendon with a collagen membrane may be an efficient approach for tendon repair and preventing tendon adhesion after its ruptures.

  10. Improving the mechanical properties of collagen-based membranes using silk fibroin for corneal tissue engineering. (United States)

    Long, Kai; Liu, Yang; Li, Weichang; Wang, Lin; Liu, Sa; Wang, Yingjun; Wang, Zhichong; Ren, Li


    Although collagen with outstanding biocompatibility has promising application in corneal tissue engineering, the mechanical properties of collagen-based scaffolds, especially suture retention strength, must be further improved to satisfy the requirements of clinical applications. This article describes a toughness reinforced collagen-based membrane using silk fibroin. The collagen-silk fibroin membranes based on collagen [silk fibroin (w/w) ratios of 100:5, 100:10, and 100:20] were prepared by using silk fibroin and cross-linking by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. These membranes were analyzed by scanning electron microscopy and their optical property, and NaCl and tryptophan diffusivity had been tested. The water content was found to be dependent on the content of silk fibroin, and CS10 membrane (loading 10 wt % of silk fibroin) performed the optimal mechanical properties. Also the suture experiments have proved CS10 has high suture retention strength, which can be sutured in rabbit eyes integrally. Moreover, the composite membrane proved good biocompatibility for the proliferation of human corneal epithelial cells in vitro. Lamellar keratoplasty shows that CS10 membrane promoted complete epithelialization in 35 ± 5 days, and their transparency is restored quickly in the first month. Corneal rejection reaction, neovascularization, and keratoconus are not observed. The composite films show potential for use in the field of corneal tissue engineering.

  11. Effect of a collagen membrane enriched with fibronectin on guided tissue regeneration in dogs. (United States)

    Kurtis, Bülent; Balos, Köksal; Oygür, Tülin


    The present study was planned to assess the capacity of a resorbable collagen membrane enriched with fibronectin to prevent the apical migration of epithelium and to facilitate new attachment and new bone. Experimental osseous dehiscence defects were produced on the bilateral labial aspect of mandibular 2nd, 3rd and 4th premolar teeth in six mongrel dogs. Guided tissue regeneration therapy using collagen membranes, which were rehydrated with fibronectin solution, was performed on one quadrant (group A). In the contralateral quadrant, the same collagen membranes, but rehydrated only with saline (group B), were placed over the bony defects. The third premolar teeth, which were treated by open-flap debridement, served as control (group C). Flaps were positioned slightly coronally and sutured; sutures were removed 10 days later. The dogs were killed 30 days after reconstructive therapy. Tissue blocks containing the experimental and control teeth were excised, demineralized in EDTA, and embedded in paraffin. Histological and histometric evaluation revealed that all groups demonstrated similar effects on preventing the down-growth of epithelium and formation of new cementum and new bone. Collagen membranes were tolerated well within the tissues, and membrane remnants were identified at 30 days. In summary, this study indicated that in this dog model similar healing results could be achieved with a bovine type I collagen membrane with or without fibronectin solution and open-flap debridement.

  12. cDNA cloning of the basement membrane chondroitin sulfate proteoglycan core protein, bamacan: a five domain structure including coiled-coil motifs

    DEFF Research Database (Denmark)

    Wu, R R; Couchman, J R


    Basement membranes contain several proteoglycans, and those bearing heparan sulfate glycosaminoglycans such as perlecan and agrin usually predominate. Most mammalian basement membranes also contain chondroitin sulfate, and a core protein, bamacan, has been partially characterized. We have now...... obtained cDNA clones encoding the entire bamacan core protein of Mr = 138 kD, which reveal a five domain, head-rod-tail configuration. The head and tail are potentially globular, while the central large rod probably forms coiled-coil structures, with one large central and several very short interruptions....... This molecular architecture is novel for an extracellular matrix molecule, but it resembles that of a group of intracellular proteins, including some proposed to stabilize the mitotic chromosome scaffold. We have previously proposed a similar stabilizing role for bamacan in the basement membrane matrix...

  13. Characterization and in vitro release studies of tetracycline and rolitetracycline imobilized on anionic collagen membranes

    Directory of Open Access Journals (Sweden)

    Gilberto Goissis


    Full Text Available This work reports the covalent immobilization of tetracycline and rolitetracycline over anionic collagen membranes and the drug release studies as an effort to develop a two stage drug release based on diffusion (fast release and on the rate of membrane biodegradation (slow release. Independent from casting conditions antibiotics incorporated by dispersion were released in the range from 80 to 100% within 7 hours in concentrations significantly higher than those described for the prevention of bacterial growth. Antibiotic release within this period was predominantly diffusion controlled. Covalent immobilization by a modified azide procedure occurred with preservation of collagen structure independently from pH of casting and reaction conditions. Its expected that anionic collagen membranes with dispersed and covalently bound rolitetracycline or tetracycline, in association with conventional therapy, may significantly reduce membrane induced infections observed post-implantation, one of the major problem associated with periodontal ligaments reconstruction by the Guided Tissue Regeneration procedure.

  14. Comparative study of two collagen membranes for guided tissue regeneration therapy in periodontal intrabony defects: a randomized clinical trial


    Chung, Young-Mi; Lee, Jue-Yeon; Jeong, Seong-Nyum


    Purpose The purpose of this study was to assess and compare the clinical and radiographic outcomes of guided tissue regeneration therapy for human periodontal intrabony defects using two different collagen membranes: a porous nonchemical cross-linking collagen membrane (NC) and a bilayer collagen membrane (BC). Methods Thirty subjects were randomly assigned and divided into the following 3 groups: a test group (NC+BM), in which a NC was used with xenograft bone mineral (BM), a positive contro...

  15. Sequential development of pulmonary hemorrhage with MPO-ANCA complicating anti-glomerular basement membrane antibody-mediated glomerulonephritis. (United States)

    Peces, R; Rodríguez, M; Pobes, A; Seco, M


    We report a case of rapidly progressive glomerulonephritis caused by anti-glomerular basement membrane (anti-GBM) antibodies that progressed to end-stage renal disease in a 67-year-old woman with diabetes. Intensive combined immunosuppressive therapy with methylprednisolone bolus, oral prednisone, and cyclophosphamide led to negativity of anti-GBM antibodies but was not able to restore renal function. After 28 months of hemodialysis, the patient suddenly presented with pulmonary hemorrhage. In this setting, high levels of myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody (ANCA) and negative anti-GBM antibodies were found. Therapy with oral prednisone and cyclophosphamide led to resolution of pulmonary hemorrhage and negativity of MPO-ANCA.

  16. Creatinine clearance, urinary excretion of glomerular basement membrane antigens and renal histology in congenital nephrotic syndrome of Finnish type. (United States)

    Huttunen, N P


    The endogenous creatinine clearance and urinary excretion rate of glomerular basement membrane (GBM) antigens were followed from 2 to 19 months in fifteen patients with congenital nephrotic syndrome (CNF). The quantitative examination of renal morphology was made on fourteen of these patients. Creatinine clearance increased during the first few months of life and thereafter gradually decreased. The urinary excretion rate of GBM antigens rose during the course of the disease. The creatinine clearance did not correlate significantly with glomerular fibrosis but it did correlate with tubular atrophy and interstitial fibrosis. The urinary excretion of GBM antigens correlated significantly with glomerular and interstitial fibrosis and with tubular atrophy. It is concluded that there is a clear progress in the disease and the renal histological changes probably are caused by accumulation of GBM material in glomeruli.

  17. Guided tissue regeneration-based root coverage utilizing collagen membranes: technique and case reports. (United States)

    Wang, Hom-Lay; Al-Shammari, Khalaf F


    Gingival recession defects have traditionally been treated with various grafting procedures. Recently, guided tissue regeneration with collagen membranes has shown promising results. This article reviews the rationale, indications, contraindications, and clinical methods for the use of bioabsorbable collagen membrane barriers. Several properties make collagen membranes attractive candidates for use as barriers in guided tissue regeneration-based root coverage procedures. These include the inhibition of epithelial migration and promotion of new connective tissue attachment; the ability to aggregate platelets, thereby facilitating wound stabilization and maturation; the promotion of cellular migration and wound closure; the elimination of the need for reentry surgery; and the ability to augment tissue thickness. Cases are presented to illustrate the surgical principles and techniques.

  18. Symposium: Role of the extracellular matrix in mammary development. Regulation of milk protein and basement membrane gene expression: The influence of the extracellular matrix

    Energy Technology Data Exchange (ETDEWEB)

    Aggeler, J.; Park, C.S.; Bissell, M.J.


    Synthesis and secretion of milk proteins ({alpha}-casein, {beta}-casein, {gamma}-casein, and transferrin) by cultured primary mouse mammary epithelial cells is modulated by the extracellular matrix. In cells grown on released or floating type I collagen gels, mRNA for {beta}-casein and transferrin is increased as much as 30-fold over cells grown on plastic. Induction of {beta}-casein expression depends strongly on the presence of lactogenic hormones, especially prolactin, in the culture. When cells are plated onto partially purified reconstituted basement membrane, dramatic changes in morphology and milk protein gene expression are observed. Cells cultured on the matrix for 6 to 8 d in the presence of prolactin, insulin, and hydrocortisone form hollow spheres and duct-like structures that are completely surrounded by matrix. The cells lining these spheres appear actively secretory and are oriented with their apices facing the lumen. Hybridization experiments indicate that mRNA for {beta}-casein can be increased as much as 70-fold in these cultures. Because > 90% of the cultured cells synthesize immunoreactive {beta}-casein, as compared with only 40% of cells in the late pregnant gland, the matrix appears to be able to induce protein expression in previously silent cells. Synthesis of laminin and assembly of a mammary-specific basal lamina by cells cultured on different extracellular matrices also appears to depend on the presence of lactogenic hormones. These studies provide support for the concept of dynamic reciprocity in which complex interactions between extracellular matrix and the cellular cytoskeleton contribute to the induction and maintenance of tissue-specific gene expression in the mammary gland.

  19. Elastase, but not proteinase 3 (PR3), induces proteinuria associated with loss of glomerular basement membrane heparan sulphate after in vivo renal perfusion in rats

    NARCIS (Netherlands)

    Heeringa, P; VanDenBorn, J; Brouwer, E; Dolman, KM; Klok, PA; Huitema, MG; Limburg, PC; Bakker, MAH; Berden, JHM; Daha, MR; Kallenberg, CGM


    Elastase, but not PR3, induces proteinuria associated with loss of glomerular basement membrane (GEM) heparan sulphate after in vivo renal perfusion in rats. PR3 and elastase are cationic neutral serine proteinases present in the azurophilic granules of polymorphonuclear leucocytes. Release of these


    NARCIS (Netherlands)



    Histones can mediate the binding of DNA and anti-DNA to the glomerular basement membrane (GBM). Zn ELISA histone/DNA/anti-DNA complexes are able to bind to heparan sulfate (HS), an intrinsic constituent of the GBM. We questioned whether histone containing immune complexes are able to bind to the GBM

  1. 19-DEJ-1, a hemidesmosome-anchoring filament complex-associated monoclonal antibody. Definition of a new skin basement membrane antigenic defect in junctional and dystrophic epidermolysis bullosa

    DEFF Research Database (Denmark)

    Fine, J D; Horiguchi, Y; Couchman, J R


    A murine monoclonal antibody (19-DEJ-1) was recently produced that recognizes a unique antigenic epitope of human skin basement membrane localized to the midlamina lucida exclusively in those areas bordered by overlying hemidesmosomes. To determine whether the antigen defined by 19-DEJ-1 is norma...

  2. Distribution, ultrastructural localization, and ontogeny of the core protein of a heparan sulfate proteoglycan in human skin and other basement membranes

    DEFF Research Database (Denmark)

    Horiguchi, Y; Couchman, J R; Ljubimov, A V;


    A variety of heparan sulfate proteoglycans (HSPG) have been identified on cell surfaces and in basement membrane (BM). To more fully characterize HSPG in human skin BM, we used two monoclonal antibodies (MAb) directed against epitopes of the core protein of a high molecular weight HSPG isolated...

  3. Plasma membrane overgrowth causes fibrotic collagen accumulation and immune activation in Drosophila adipocytes (United States)

    Zang, Yiran; Wan, Ming; Liu, Min; Ke, Hongmei; Ma, Shuangchun; Liu, Lu-Ping; Ni, Jian-Quan; Carlos Pastor-Pareja, José


    Many chronic diseases are associated with fibrotic deposition of Collagen and other matrix proteins. Little is known about the factors that determine preferential onset of fibrosis in particular tissues. Here we show that plasma membrane (PM) overgrowth causes pericellular Collagen accumulation in Drosophila adipocytes. We found that loss of Dynamin and other endocytic components causes pericellular trapping of outgoing Collagen IV due to dramatic cortex expansion when endocytic removal of PM is prevented. Deposits also form in the absence of negative Toll immune regulator Cactus, excess PM being caused in this case by increased secretion. Finally, we show that trimeric Collagen accumulation, downstream of Toll or endocytic defects, activates a tissue damage response. Our work indicates that traffic imbalances and PM topology may contribute to fibrosis. It also places fibrotic deposits both downstream and upstream of immune signaling, consistent with the chronic character of fibrotic diseases. DOI: PMID:26090908

  4. Proteomic analysis of urinary exosomes from patients of early IgA nephropathy and thin basement membrane nephropathy. (United States)

    Moon, Pyong-Gon; Lee, Jeong-Eun; You, Sungyong; Kim, Taek-Kyun; Cho, Ji-Hoon; Kim, In-San; Kwon, Tae-Hwan; Kim, Chan-Duck; Park, Sun-Hee; Hwang, Daehee; Kim, Yong-Lim; Baek, Moon-Chang


    To identify biomarker candidates associated with early IgA nephropathy (IgAN) and thin basement membrane nephropathy (TBMN), the most common causes presenting isolated hematuria in childhood, a proteomic approach of urinary exosomes from early IgAN and TBMN patients was introduced. The proteomic results from the patients were compared with a normal group to understand the pathophysiological processes associated with these diseases at the protein level. The urinary exosomes, which reflect pathophysiological processes, collected from three groups of young adults (early IgAN, TBMN, and normal) were trypsin-digested using a gel-assisted protocol, and quantified by label-free LC-MS/MS, using an MS(E) mode. A total of 1877 urinary exosome proteins, including cytoplasmic, membrane, and vesicle trafficking proteins, were identified. Among the differentially expressed proteins, four proteins (aminopeptidase N, vasorin precursor, α-1-antitrypsin, and ceruloplasmin) were selected as biomarker candidates to differentiate early IgAN from TBMN. We confirmed the protein levels of the four biomarker candidates by semi-quantitative immunoblot analysis in urinary exosomes independently prepared from other patients, including older adult groups. Further clinical studies are needed to investigate the diagnostic and prognostic value of these urinary markers for early IgAN and TBMN. Taken together, this study showed the possibility of identifying biomarker candidates for human urinary diseases using urinary exosomes and might help to understand the pathophysiology of early IgAN and TBMN at the protein level.

  5. A direct contact between astrocyte and vitreous body is possible in the rabbit eye due to discontinuities in the basement membrane of the retinal inner limiting membrane

    Directory of Open Access Journals (Sweden)

    A. Haddad


    Full Text Available Different from most mammalian species, the optic nerve of the rabbit eye is initially formed inside the retina where myelination of the axons of the ganglion cells starts and vascularization occurs. Astrocytes are confined to these regions. The aforementioned nerve fibers known as medullated nerve fibers form two bundles that may be identified with the naked eye. The blood vessels run on the inner surface of these nerve fiber bundles (epivascularization and, accordingly, the accompanying astrocytes lie mostly facing the vitreous body from which they are separated only by the inner limiting membrane of the retina. The arrangement of the astrocytes around blood vessels leads to the formation of structures known as glial tufts. Fragments (N = 3 or whole pieces (N = 3 of the medullated nerve fiber region of three-month-old male rabbits (Orictolagus cuniculus were fixed in glutaraldehyde followed by osmium tetroxide, and their thin sections were examined with a transmission electron microscope. Randomly located discontinuities (up to a few micrometers long of the basement membrane of the inner limiting membrane of the retina were observed in the glial tufts. As a consequence, a direct contact between the astrocyte plasma membrane and vitreous elements was demonstrated, making possible functional interactions such as macromolecular exchanges between this glial cell type and the components of the vitreous body.

  6. Role of LARP6 and nonmuscle myosin in partitioning of collagen mRNAs to the ER membrane.

    Directory of Open Access Journals (Sweden)

    Hao Wang

    Full Text Available Type I collagen is extracellular matrix protein composed of two α1(I and one α2(I polypeptides that fold into triple helix. Collagen polypeptides are translated in coordination to synchronize the rate of triple helix folding to the rate of posttranslational modifications of individual polypeptides. This is especially important in conditions of high collagen production, like fibrosis. It has been assumed that collagen mRNAs are targeted to the membrane of the endoplasmic reticulum (ER after translation of the signal peptide and by signal peptide recognition particle (SRP. Here we show that collagen mRNAs associate with the ER membrane even when translation is inhibited. Knock down of LARP6, an RNA binding protein which binds 5' stem-loop of collagen mRNAs, releases a small amount of collagen mRNAs from the membrane. Depolimerization of nonmuscle myosin filaments has a similar, but stronger effect. In the absence of LARP6 or nonmuscle myosin filaments collagen polypeptides become hypermodified, are poorly secreted and accumulate in the cytosol. This indicates lack of coordination of their synthesis and retro-translocation due to hypermodifications and misfolding. Depolimerization of nonmuscle myosin does not alter the secretory pathway through ER and Golgi, suggesting that the role of nonmuscle myosin is primarily to partition collagen mRNAs to the ER membrane. We postulate that collagen mRNAs directly partition to the ER membrane prior to synthesis of the signal peptide and that LARP6 and nonmuscle myosin filaments mediate this process. This allows coordinated initiation of translation on the membrane bound collagen α1(I and α2(I mRNAs, a necessary step for proper synthesis of type I collagen.

  7. The collagen receptor uPARAP/Endo180 in tissue degradation and cancer (Review)

    DEFF Research Database (Denmark)

    Carlsen Melander, Eva Maria; Jürgensen, Henrik J; Madsen, Daniel H


    as well as interstitial collagen to lysosomal degradation. This capacity, shared only with the mannose receptor from the same protein family, endows uPARAP/Endo180 with a critical role in development and homeostasis, as well as in pathological disruptions of the extracellular matrix structure. Important......The collagen receptor uPARAP/Endo180, the product of the MRC2 gene, is a central component in the collagen turnover process governed by various mesenchymal cells. Through the endocytosis of collagen or large collagen fragments, this recycling receptor serves to direct basement membrane collagen...

  8. Biodegradation of differently cross-linked collagen membranes: an experimental study in the rat. (United States)

    Rothamel, Daniel; Schwarz, Frank; Sager, Martin; Herten, Monika; Sculean, Anton; Becker, Jürgen


    The aim of the present study was to compare the biodegradation of differently cross-linked collagen membranes in rats. Five commercially available and three experimental membranes (VN) were included: (1) BioGide (BG) (non-cross-linked porcine type I and III collagens), (2) BioMend (BM), (3) BioMendExtend (BME) (glutaraldehyde cross-linked bovine type I collagen), (4) Ossix (OS) (enzymatic-cross-linked bovine type I collagen), (5) TutoDent (TD) (non-cross-linked bovine type I collagen, and (6-8) VN(1-3) (chemical cross-linked porcine type I and III collagens). Specimens were randomly allocated in unconnected subcutaneous pouches separated surgically on the back of 40 wistar rats, which were divided into five groups (2, 4, 8, 16, and 24 weeks), including eight animals each. After 2, 4, 8, 16, and 24 weeks of healing, the rats were sacrificed and explanted specimens were prepared for histologic and histometric analysis. The following parameters were evaluated: biodegradation over time, vascularization, tissue integration, and foreign body reaction. Highest vascularization and tissue integration was noted for BG followed by BM, BME, and VN(1); TD, VN(2), and VN(3) showed prolongated, while OS exhibited no vascularization. Subsequently, biodegradation of BG, BM, BME and VN(1) was faster than TD, VN(2), and VN(3). OS showed only a minute amount of superficial biodegradation 24 weeks following implantation. Biodegradation of TD, BM, BME, VN(2), and VN(3) was associated with the presence of inflammatory cells. Within the limits of the present study, it was concluded that cross-linking of bovine and porcine-derived collagen types I and III was associated with (i) prolonged biodegradation, (ii) decreased tissue integration and vascularization, and (iii) in case of TD, BM, BME, VN(2), and VN(3) foreign body reactions.

  9. Bone neoformation in defects treated with platelet-rich fibrin membrane versus collagen membrane: a histomorphometric study in rabbit femurs.

    Directory of Open Access Journals (Sweden)

    Edwin Meza


    Full Text Available The aim of the present research was to compare bone neoformation in bone defects treated with platelet-rich fibrin (PRF and collagen membrane (CM at 3 and 5 weeks. For this purpose, two bone defects with a width of 4 mm and depth of 6 mm were created in the left distal femur diaphysis of New Zealand rabbits (n=12. The subjects were randomly allocated into two groups. One of the defects was covered with a platelet-rich fibrin membrane (Centrifuged resorbable autologous blood biopolymer without biochemical modification or a collagen membrane (gold standard - Neo Mem. The second defect was left uncovered (NC. The rabbits were sacrificed after 3 and 5 weeks (3 rabbits per period. The femur was completely removed and processed histomophometrically. The bone neformation analysis was performed using a differential point-counting method. Data was statistically analyzed (ANOVA, Tukey. The histomorphometric results showed that bone neformation in the defects treated with PRF at 3 weeks was equivalent to the CM (p<0.05. After 5 weeks, bone neformation obtained with PRF was higher than the control group and lower compared with the CM (p<0.05. The conclusion of the present study is that bone neformation in defects treated with PRF showed lower histomorphometric results compared with the one obtained with the collagen membrane and higher when compared with the control defects.

  10. Synthesis and localization of two sulphated glycoproteins associated with basement membranes and the extracellular matrix

    DEFF Research Database (Denmark)

    Hogan, B L; Taylor, A; Kurkinen, M;


    Two sulphated glycoproteins (sgps) of apparent molecular weight (Mr) 180,000 and 150,000, are synthesized by murine PYS and PF HR9 parietal endoderm and Swiss 3T3 cells. The Mr 150,000 sgp has a similar chemical structure to the sulphated glycoprotein, C, synthesized and laid down in Reichert......'s membrane by mouse embryo parietal endoderm cells (Hogan, B. L.M., A. Taylor, and A.R. Cooper, 1982, Dev. Biol., 90:210-214). Both the Mr 180,000 and 150,000 sgps are deposited in the detergent-insoluble matrix of cultured cells, but they do not apparently undergo any disulphide-dependent intermolecular...

  11. 皮肤中基底膜的结构与功能%The structure and functions of skin basement membrane

    Institute of Scientific and Technical Information of China (English)

    张丽丽; 李贤玉; 穆荣; 范春晖(综述); 北垣雅人; 高须; 惠美子(审校)


    Basement membranes (BE) are a kind of extracellular matrices in every tissue of the whole human body and play the most important role as keeping cellular functions and structures. It is the most important that BE in skin functionally connects and separates with epidermal and dermis containing of various proteins,such as type Ⅳ and Ⅶ collagen,laminins,nidogen and perlecan etc.Those interacted with themselves to form networks to support diverse bio functions in BE. And BE connects between epidermis and dermis signaling etc.It will lead the circulation smoothly among epidermis-basement membrane-dermis that healthy BE contributes diverse bio-functions,so that BE could maintain integrity and healthy of skin.Sunlight exposure may predominantly changes BE,such as sunburn, photoaging and photo-related skin cancers. On the other hand, some studies showed that various herbal extracts could protect BE well.It will be a new approach to develop functional cosmetics that how to utilize those active herbal ingredients.%基底膜是一类细胞外基质,存在于人体中所有的器官组织中,承担着重要的功能。皮肤的基底膜是表皮与真皮间重要的承接结构,主要成分有Ⅳ型和Ⅶ型胶原蛋白、层粘连蛋白、巢蛋白、串珠素等,通过它们交互形成的网状结构,构成了基底膜丰富多样的生物学功能,主要为表皮-真皮的连接功能、信号传导功能及渗透屏障功能等。健康的基底膜发挥多样生物学功能,使表皮-基底膜-真皮三者之间能够顺畅循环,保持皮肤的健康完整性。暴露于阳光下可引起基底膜的改变,可以引发多种肌肤烦恼,例如晒伤、光老化、皮肤癌等。研究发现一些植物提取物可以保护基底膜,如何运用这些活性成分对研究开发功能性化妆品可以提供更多的发展途径。

  12. Acute podocyte injury is not a stimulus for podocytes to migrate along the glomerular basement membrane in zebrafish larvae (United States)

    Siegerist, Florian; Blumenthal, Antje; Zhou, Weibin; Endlich, Karlhans; Endlich, Nicole


    Podocytes have a unique 3D structure of major and interdigitating foot processes which is the prerequisite for renal blood filtration. Loss of podocytes leads to chronic kidney disease ending in end stage renal disease. Until now, the question if podocytes can be replaced by immigration of cells along the glomerular basement membrane (GBM) is under debate. We recently showed that in contrast to former theories, podocytes are stationary in the zebrafish pronephros and neither migrate nor change their branching pattern of major processes over 23 hours. However, it was still unclear whether podocytes are able to migrate during acute injury. To investigate this, we applied the nitroreductase/metronidazole zebrafish model of podocyte injury to in vivo two-photon microscopy. The application of metronidazole led to retractions of major processes associated with a reduced expression of podocyte-specific proteins and a formation of subpodocyte pseudocyst. Electron microscopy showed that broad areas of the capillaries became denuded. By 4D in vivo observation of single podocytes, we could show that the remaining podocytes did not walk along GBM during 24 h. This in vivo study reveals that podocytes are very stationary cells making regenerative processes by podocyte walking along the GBM very unlikely. PMID:28252672

  13. Complement and Humoral Adaptive Immunity in the Human Choroid Plexus: Roles for Stromal Concretions, Basement Membranes, and Epithelium. (United States)

    Moore, G R Wayne; Laule, Cornelia; Leung, Esther; Pavlova, Vladimira; Morgan, B Paul; Esiri, Margaret M


    The choroid plexus (CP) provides a barrier to entry of toxic molecules from the blood into the brain and transports vital molecules into the cerebrospinal fluid. While a great deal is known about CP physiology, relatively little is known about its immunology. Here, we show immunohistochemical data that help define the role of the CP in innate and adaptive humoral immunity. The results show that complement, in the form of C1q, C3d, C9, or C9neo, is preferentially deposited in stromal concretions. In contrast, immunoglobulin (Ig) G (IgG) and IgA are more often found in CP epithelial cells, and IgM is found in either locale. C4d, IgD, and IgE are rarely, if ever, seen in the CP. In multiple sclerosis CP, basement membrane C9 or stromal IgA patterns were common but were not specific for the disease. These findings indicate that the CP may orchestrate the clearance of complement, particularly by deposition in its concretions, IgA and IgG preferentially via its epithelium, and IgM by either mechanism.

  14. Basement membrane and vascular remodelling in smokers and chronic obstructive pulmonary disease: a cross-sectional study

    Directory of Open Access Journals (Sweden)

    Muller H Konrad


    Full Text Available Abstract Background Little is known about airway remodelling in bronchial biopsies (BB in smokers and chronic obstructive pulmonary disease (COPD. We conducted an initial pilot study comparing BB from COPD patients with nonsmoking controls. This pilot study suggested the presence of reticular basement membrane (Rbm fragmentation and altered vessel distribution in COPD. Methods To determine whether Rbm fragmentation and altered vessel distribution in BB were specific for COPD we designed a cross-sectional study and stained BB from 19 current smokers and 14 ex-smokers with mild to moderate COPD and compared these to 15 current smokers with normal lung function and 17 healthy and nonsmoking subjects. Results Thickness of the Rbm was not significantly different between groups; although in COPD this parameter was quite variable. The Rbm showed fragmentation and splitting in both current smoking groups and ex-smoker COPD compared with healthy nonsmokers (p Conclusions Airway remodelling in smokers and mild to moderate COPD is associated with fragmentation of the Rbm and altered distribution of vessels in the airway wall. Rbm fragmentation was also present to as great an extent in ex-smokers with COPD. These characteristics may have potential physiological consequences.

  15. Skin Basement Membrane: The Foundation of Epidermal Integrity—BM Functions and Diverse Roles of Bridging Molecules Nidogen and Perlecan

    Directory of Open Access Journals (Sweden)

    Dirk Breitkreutz


    Full Text Available The epidermis functions in skin as first defense line or barrier against environmental impacts, resting on extracellular matrix (ECM of the dermis underneath. Both compartments are connected by the basement membrane (BM, composed of a set of distinct glycoproteins and proteoglycans. Herein we are reviewing molecular aspects of BM structure, composition, and function regarding not only (i the dermoepidermal interface but also (ii the resident microvasculature, primarily focusing on the per se nonscaffold forming components perlecan and nidogen-1 and nidogen-2. Depletion or functional deficiencies of any BM component are lethal at some stage of development or around birth, though BM defects vary between organs and tissues. Lethality problems were overcome by developmental stage- and skin-specific gene targeting or by cell grafting and organotypic (3D cocultures of normal or defective cells, which allows recapitulating BM formation de novo. Thus, evidence is accumulating that BM assembly and turnover rely on mechanical properties and composition of the adjacent ECM and the dynamics of molecular assembly, including further “minor” local components, nidogens largely functioning as catalysts or molecular adaptors and perlecan as bridging stabilizer. Collectively, orchestration of BM assembly, remodeling, and the role of individual players herein are determined by the developmental, tissue-specific, or functional context.

  16. Efficient differentiation of embryonic stem cells into hepatic cells in vitro using a feeder-free basement membrane substratum.

    Directory of Open Access Journals (Sweden)

    Nobuaki Shiraki

    Full Text Available The endoderm-inducing effect of the mesoderm-derived supportive cell line M15 on embryonic stem (ES cells is partly mediated through the extracellular matrix, of which laminin α5 is a crucial component. Mouse ES or induced pluripotent stem cells cultured on a synthesized basement membrane (sBM substratum, using an HEK293 cell line (rLN10-293 cell stably expressing laminin-511, could differentiate into definitive endoderm and subsequently into pancreatic lineages. In this study, we investigated the differentiation on sBM of mouse and human ES cells into hepatic lineages. The results indicated that the BM components played an important role in supporting the regional-specific differentiation of ES cells into hepatic endoderm. We show here that knockdown of integrin β1 (Itgb1 in ES cells reduced their differentiation into hepatic lineages and that this is mediated through Akt signaling activation. Moreover, under optimal conditions, human ES cells differentiated to express mature hepatocyte markers and secreted high levels of albumin. This novel procedure for inducing hepatic differentiation will be useful for elucidating the molecular mechanisms controlling lineage-specific fates during gut regionalization. It could also represent an attractive approach to providing a surrogate cell source, not only for regenerative medicine, but also for pharmaceutical and toxicologic studies.

  17. Reorganization of endothelial cord-like structures on basement membrane complex (Matrigel): involvement of transforming growth factor beta 1. (United States)

    Kuzuya, M; Kinsella, J L


    The formation of capillary-like network structures by cultured vascular endothelial cells on reconstituted basement membrane matrix, Matrigel, models endothelial cell differentiation, the final step of angiogenesis (Kubota et al., 1988; Grant et al., 1989). When endothelial cells derived from bovine aorta and brain capillaries were plated on Matrigel, DNA synthesis was suppressed and a network of capillary-like structures rapidly formed in 8-12 h. With time, the network broke down, resulting in dense cellular cords radiating from multiple cellular clusters in 16-24 h. Finally, multicellular aggregates of cells were formed as the network underwent further retraction. Network regression was prevented when either dithiothreitol (DTT) or anti-TGF-beta 1 antibodies were added during the assay. The addition of exogenous TGF-beta 1 promoted the regression of endothelial cells into the clusters. This response to TGF-beta 1 was blocked by potent serine threonine protein kinase inhibitors, H-7 and HA100. TGF-beta 1 was released from polymerized Matrigel by incubation with Dulbecco's modified eagle's medium (DMEM) in the absence of cells. The Matrigel-conditioned DMEM inhibited endothelial DNA synthesis even in the presence of anti-TGF-beta 1 antibodies. These results suggest that TGF-beta 1 and possibly other soluble factors from Matrigel may be important for differentiation and remodeling of endothelial cells in a capillary network with possible implications for wound healing and development.

  18. Different collagen types define two types of idiopathic epiretinal membranes


    Kritzenberger, Michaela; Junglas, Benjamin; Framme, Carsten; Helbig, Horst; Gabel, Veit-Peter; Fuchshofer, Rudolf; Ernst R. Tamm; Hillenkamp, Jost


    Abstract Aims: To identify differences in extracellular matrix contents between idiopathic epiretinal membranes (IEM) of cellophane macular reflex (CMRM) or preretinal macular fibrosis (PMFM) type. Methods and results: IEM were analyzed by light and quantitative transmission electron microscopy, immunohistochemistry, and Western blotting. Substantial differences between CMRM and PMFM were observed regarding the nature of extracellular fibrils. In CMRM, the fibrils were thin with...

  19. Increased initiation and growth of tumor cell lines, cancer stem cells and biopsy material in mice using basement membrane matrix protein (Cultrex or Matrigel) co-injection. (United States)

    Fridman, Rafael; Benton, Gabriel; Aranoutova, Irina; Kleinman, Hynda K; Bonfil, R Daniel


    This protocol requires 2-4 h and presents a method for injecting tumor cells, cancer stem cells or dispersed biopsy material into subcutaneous or orthotopic locations within recipient mice. The tumor cells or biopsy are mixed with basement membrane matrix proteins (CultrexBME or Matrigel) at 4 °C and then injected into recipient animals at preferred anatomical sites. Tumor cells can also be co-injected with additional cell types, such as fibroblasts, stromal cells, endothelial cells and so on. Details are given on appropriate cell numbers, handling and concentration of the basement membrane proteins, recipient animals, injection location and techniques. This procedure enables the growth of tumors from cells or biopsy material (tumor graft) with greater efficiency of take and growth, and with retention of the primary tumor phenotype based on histology. Co-injection with additional cell types provides more physiological models of human cancers for use in drug screening and studying cancer biology.

  20. [Healing of osseous defects by guided bone regeneration using ribose cross linked collagen membranes]. (United States)

    Tal, H


    The ultimate goal of periodontal therapy has long been the complete regeneration of the periodontal attachment apparatus. Guided Tissue Regeneration (GTR) and Guided Bone Regeneration (GBR) are two regenerative procedures which converted this goal from a dream to reality. In search of a biocompatible resorbable tissue barrier, collagen, being a natural protein and a weak antigen, has attracted much interest and became the focus of much intention during the 80's and the 90's. The understanding that cross linking of collagen with aldehyde sugars, especially ribose, produces collagen which is highly resistant to resorption in vivo led to the development of a "natural" Crossed-Linked Collagen Barrier (CB-SX). Animal and Human studies have shown that the newly developed membrane is biocompatible, remains intact in the tissues 6 months and more, and results in impressive guided tissue/bone regeneration. Spontaneous early exposure of the membrane is common but the healing potential of the resulted tissue dehiscence is favorable with no tendency for bacterial infection. The commercial version of the CB-SX is especially suitable for GBR procedures; it is highly recommended that the gingival flaps involved will properly be released, will lack tension, and be thoroughly sutured.

  1. Treatment of gingival recession with collagen membrane and DFDBA: a histometric study in dogs

    Directory of Open Access Journals (Sweden)

    Elizabeth Pimentel Rosetti


    Full Text Available In a previous study, we evaluated the findings related to the use of resorbable collagen membranes in humans along with DFDBA (demineralized freeze-dried bone allograft. The aim of this subsequent study was to histometrically evaluate in dogs, the healing response of gingival recessions treated with collagen membrane + DFDBA (Guided Tissue Regeneration, GTR compared to a coronally positioned flap (CPF. Two types of treatment were randomly carried out in a split-mouth study. Group 1 was considered as test (GTR: collagen membrane + DFDBA, whereas Group 2 stood for the control (only CPF. The dogs were given chemical bacterial plaque control with 0.2% chlorhexidine digluconate during a 90-day repair period. Afterwards, the animals were killed to obtain biopsies and histometric evaluation of the process of cementum and bone formation, epithelial migration and gingival level. A statistically significant difference was found between groups with a larger extension of neoformed cementum (GTR = 32.72%; CPF = 18.82%; p = 0.0004, new bone (GTR = 23.20%; CPF = 09.90%; p = 0.0401 and with a smaller area of residual gingival recession in the test group (GTR = 50.69%; CPF = 59.73%; p = 0.0055 compared to the control group. The only item assessed that showed no statistical difference was epithelial proliferation on the root surface, with means of 15.14% for the GTR group and 20.34% for the CPF group (p = 0.0890. Within the limits of this study we concluded that the treatment of gingival recession defects with GTR, associating collagen membrane with DFDBA, showed better outcomes in terms of a larger extension of neoformed cementum and bone, as well as in terms of a smaller proportion of residual recessions.

  2. Downregulation of a newly identified laminin, laminin-3B11, in vascular basement membranes of invasive human breast cancers. (United States)

    Mori, Taizo; Kariya, Yoshinobu; Komiya, Eriko; Higashi, Shouichi; Miyagi, Yohei; Sekiguchi, Kiyotoshi; Miyazaki, Kaoru


    Laminins present in the basement membranes (BM) of blood vessels are involved in angiogenesis and other vascular functions that are critical for tumor growth and metastasis. Two major vascular laminins, the α4 (laminin-411/421) and α5 (laminin-511/521) types, have been well characterized. We recently found a third type of vascular laminin, laminin-3B11, consisting of the α3B, β1 and γ1 chains, and revealed its biological activity. Laminin-3B11 potently stimulates vascular endothelial cells to extend lamellipodial protrusions. To understand the roles of laminin-3B11 in blood vessel functions and tumor growth, we examined localization of the laminin α3B chain in normal mammary glands and breast cancers, in comparison with the α4 and α5 laminins. In the immunohistochemical analysis, the α3B laminin was co-localized with the α4 and α5 laminins in the BM of venules and capillaries of normal breast tissues, but α3B was scarcely detected in vessels near invasive breast carcinoma cells. In contrast, the α4 laminin was overexpressed in capillaries of invasive carcinomas, where a large number of macrophages were found. The α5 laminin appeared to be weakly downregulated in cancer tissues, especially in capillary vessels. Furthermore, our in vitro analysis indicated that TNF-α significantly suppressed the laminin α3B expression in vascular endothelial cells, while it, as well as IL-1β and TGF-α, upregulated the α4 expression. These results suggest that Lm3B11/3B21 may be required for normal mature vessels and interfere with tumor angiogenesis.

  3. Linear IgA bullous disease with possible immunoreactivity to the basement membrane zone and dermal blood vessels

    Directory of Open Access Journals (Sweden)

    Ana Maria Abreu Velez


    Full Text Available Introduction: Linear IgA bullous dermatosis (LAD is an immunobullous disorder, in which IgA antibodies are deposited along the basement membrane zone (BMZ of the skin in a linear pattern. The cause of this disease is unknown, but the eruption may occur more commonly in association with certain medications. Case report: A 61 year old woman presented with blisters in the axillae and legs, with pain, itching and swelling. She was taking many medications for other conditions such diabetes and obesity. Tense blisters were seen, primarily on the legs and accompanied by some ankle swelling. Methods: Skin biopsies for hematoxylin and eosin (H&E examination, as well as for direct immunofluorescence (DIF, and immunohistochemistry (IHC studies were performed. Results: The H&E examination revealed a subepidermal blister, with small numbers of lymphocytes, neutrophils and eosinophils noted within the blister lumen. The dermis also displayed a mild, superficial, perivascular infiltrate of lymphocytes and histiocytes; eosinophils and neutrophils were also noted. DIF and IHC studies confirmed the diagnosis of linear IgA (LAD at the BMZ. However, in addition to immunoglobulin A, we also observed deposits of IgA, IgM, IgG, IgD, Kappa, Lambda, Complement/C3c, C1q, fibrinogen and albumin around upper dermal blood vessels. Conclusions: LAD has been most commonly associated with medication intake; the most common DIF immune response is the presence of linear IgA at the BMZ. However, here we found additional reactivity to against dermal blood vessels. Because the patient is affected by diabetes mellitus, it is difficult to know if the observed vascular reactivity was associated with the diabetes or solely an immune reaction to the vessels. Based on our findings, we encourage searching for vascular reactivity in cases of LAD.

  4. First Identification of a Triple Corneal Dystrophy Association: Keratoconus, Epithelial Basement Membrane Corneal Dystrophy and Fuchs' Endothelial Corneal Dystrophy

    Directory of Open Access Journals (Sweden)

    Cosimo Mazzotta


    Full Text Available Purpose: To report the observation of a triple corneal dystrophy association consisting of keratoconus (KC, epithelial basement membrane corneal dystrophy (EBMCD and Fuchs' endothelial corneal dystrophy (FECD. Methods: A 55-year-old male patient was referred to our cornea service for blurred vision and recurrent foreign body sensation. He reported bilateral recurrent corneal erosions with diurnal visual fluctuations. He underwent corneal biomicroscopy, Scheimpflug tomography, in vivo HRT confocal laser scanning microscopy and genetic testing for TGFBI and ZEB1 mutations using direct DNA sequencing. Results: Biomicroscopic examination revealed the presence of subepithelial central and paracentral corneal opacities. The endothelium showed a bilateral flecked appearance, and the posterior corneal curvature suggested a possible concomitant ectatic disorder. Corneal tomography confirmed the presence of a stage II KC in both eyes. In vivo confocal laser scanning microscopy revealed a concomitant bilateral EBMCD with hyperreflective deposits in basal epithelial cells, subbasal Bowman's layer microfolds and ridges with truncated subbasal nerves as pseudodendritic elements. Stromal analysis revealed honeycomb edematous areas, and the endothelium showed a strawberry surface configuration typical of FECD. The genetic analysis resulted negative for TGFBI mutations and positive for a heterozygous mutation in exon 7 of the gene ZEB1. Conclusion: This is the first case reported in the literature in which KC, EBMCD and FECD are present in the same patient and associated with ZEB1 gene mutation. The triple association was previously established by means of morphological analysis of the cornea using corneal Scheimpflug tomography and in vivo HRT II confocal laser scanning microscopy.

  5. Utilizing collagen membranes for guided tissue regeneration-based root coverage. (United States)

    Wang, Hom-Lay; Modarressi, Marmar; Fu, Jia-Hui


    Gingival recession is a common clinical problem that can result in hypersensitivity, pain, root caries and esthetic concerns. Conventional soft tissue procedures for root coverage require an additional surgical site, thereby causing additional trauma and donor site morbidity. In addition, the grafted tissues heal by repair, with formation of long junctional epithelium with some connective tissue attachment. Guided tissue regeneration-based root coverage was thus developed in an attempt to overcome these limitations while providing comparable clinical results. This paper addresses the biologic foundation of guided tissue regeneration-based root coverage, and describes the indications and contraindications for this technique, as well as the factors that influence outcomes. The step-by-step clinical techniques utilizing collagen membranes are also described. In comparison with conventional soft tissue procedures, the benefits of guided tissue regeneration-based root coverage procedures include new attachment formation, elimination of donor site morbidity, less chair-time, and unlimited availability and uniform thickness of the product. Collagen membranes, in particular, benefit from product biocompatibility with the host, while promoting chemotaxis, hemostasis, and exchange of gas and nutrients. Such characteristics lead to better wound healing by promoting primary wound coverage, angiogenesis, space creation and maintenance, and clot stability. In conclusion, collagen membranes are a reliable alternative for use in root coverage procedures.

  6. Clinical and biometric evaluation of collagen membrane application for covering exposed root surfaces

    Directory of Open Access Journals (Sweden)

    A. Mieremadi


    Full Text Available This study is aimed to evaluate the effect of covering exposed root surface by collagen membrane and to compare this method with coronoally advanced flap covering. 26 teeth in 20 patients (12m/8f with Miller1 gingival recession were selected. 14 teeth were allocated into test group and 12 teeth in control group. Following first phase of treatment, clinical parameters including the height and width of gingival recession, attachment height, gingival crevice depth and keratinized tissue width were measured. In test group, coronally advanced flap was prepared and gingistat (as space maintainer and collagen membrane was applied. For control group, coronally advanced flap method was applied without any further action. The measurements were done in 1,2 and 3 months post surgery. In test group, the reported root coverage was 71% while in control group was 57% which represents statistically significant difference. From first to third month post surgically, the average gingival recession in test group was 0.21 mm which was considered as creeping attachment phenomenon while in control group gingival height was increased 0.14 mm. the average width of recession and crevice depth decrease was not significantly different between the groups. On the other hand, The average attachment gain and keratinized tissue increase was significantly varied among the groups. In general, it can be concluded that application of collagen membrane as well as gingastat space maintainer can bring about satisfactory results in covering exposed root surfaces.

  7. Dynamic interplay between the collagen scaffold and tumor evolution

    DEFF Research Database (Denmark)

    Egeblad, Mikala; Rasch, Morten G; Weaver, Valerie M


    and remodeling of the ECM network regulate tissue tension, generate pathways for migration, and release ECM protein fragments to direct normal developmental processes such as branching morphogenesis. Collagens are major components of the ECM of which basement membrane type IV and interstitial matrix type I...... are the most prevalent. Here we discuss how abnormal expression, proteolysis and structure of these collagens influence cellular functions to elicit multiple effects on tumors, including proliferation, initiation, invasion, metastasis, and therapy response....

  8. Collagen IV in normal skin and in pathological processes

    Directory of Open Access Journals (Sweden)

    Ana Maria Abreu-Velez


    Full Text Available Context: Type IV collagen is a type of collagen found primarily in the skin within the basement membrane zone. The type IV collagen C4 domain at the C-terminus is not removed in post-translational processing, and the fibers are thus link head-to-head, rather than in a parallel fashion. Also, type IV collagen lacks a glycine in every third amino-acid residue necessary for the tight collagen helix. Thus, the overall collagen-IV conformation is structurally more pliable and kinked, relative to other collagen subtypes. These structural features allow collagen IV to form sheets, which is the primary structural form found in the cutaneous basal lamina. There are six human genes associated with collagen IV, specifically COL4A1, COL4A2, COL4A3, COL4A4, COL4A5 and COL4A6. The aim of this review is to highlight the significance of this protein in normal skin, and in selected diseases. Results: The alpha 3 protein constituent of type IV collagen is thought to be the antigen implicated in Goodpasture′s syndrome, wherein the immune system attacks the basement membranes of the renal glomeruli and pulmonary alveoli. In addition, mutations to the genes coding for type IV collagen lead to the Alport syndrome. Furthermore, autoantibodies directed against denatured human type IV collagen have been described in rheumatoid arthritis, scleroderma, and SLE. Structural studies of collagen IV have been utilized to differentiate between subepidermal blistering diseases, including bullous pemphigoid, acquired epidermolysis bullosa, anti-epiligrin cicatricial pemphigoid, and bullous lupus erythematosus. Collagen IV is also of importance in wound healing and in embryogenesis. Conclusions: Pathological studies have demonstrated that minor structural differences in collagen IV can lead to distinct, clinically different diseases.

  9. Comparable efficacy of silk fibroin with the collagen membranes for guided bone regeneration in rat calvarial defects


    Kim, Jwa-Young; Yang, Byoung-Eun; Ahn, Jin-Hee; Park, Sang O; Shim, Hye-Won


    PURPOSE Silk fibroin (SF) is a new degradable barrier membrane for guided bone regeneration (GBR) that can reduce the risk of pathogen transmission and the high costs associated with the use of collagen membranes. This study compared the efficacy of SF membranes on GBR with collagen membranes (Bio-Gide®) using a rat calvarial defect model. MATERIALS AND METHODS Thirty-six male Sprague Dawley rats with two 5 mm-sized circular defects in the calvarial bone were prepared (n=72). The study groups...

  10. Evaluation of biodegradation and biocompatibility of collagen/chitosan/alkaline phosphatase biopolymeric membranes

    Indian Academy of Sciences (India)



    The aim of this study was to develop a new variant of membranes based on collagen (COL), chitosan (CHI) and alkaline phosphatase (ALP) immobilized and cross-linking with glutaraldehyde (GA) at different concentrations. The biodegradation in the presence of collagenase was investigated. Biocompatibility was evaluated by MTT assay using a mouse fibroblast cell culture type NCTC (clone 929). Non-cross-linked samples were biocompatible and membranes cross-linked with low concentrations of GA (0.04, 0.08%) were also iocompatible. However, high concentrations of GA lead to a decreased biocompatibility. The adsorption behaviour of Ca$^{2+}$ ions to all membraneswere evaluated using the Freundlich isotherms. Haemolytic studies were performed in order to consider their applications in biomineralization process. By the addition of collagen and ALP to chitosan, the haemolytic indexdecreases, the COL–CHI–ALP membrane being in the non-haemolytic domain, while the COL–CHI–ALP–GA membrane has a haemolytic index greater than 2, and is slightly haemolytic.

  11. A case of subepidermal blistering disease with autoantibodies to multiple laminin subunits who developed later autoantibodies to alpha-5 chain of type IV collagen associated with membranous glomerulonephropathy. (United States)

    Sueki, Hirohiko; Sato, Yoshinori; Ohtoshi, Shinpei; Nakada, Tokio; Yoshimura, Ashio; Tateishi, Chiharu; Borza, Dorin-Bogdan; Fader, William; Ghohestani, Reza F; Hirako, Yoshiaki; Koga, Hiroshi; Ishii, Norito; Tsuchisaka, Atsunari; Qian, Hua; Li, Xiaoguang; Hashimoto, Takashi


    We report a 68-year-old Japanese female patient with subepidermal blistering disease with autoantibodies to multiple laminins, who subsequently developed membranous glomerulonephropathy. At skin disease stage, immunofluorescence demonstrated IgG anti-basement membrane zone antibodies reactive with dermal side of NaCl-split skin. Immunoblotting of human dermal extract, purified laminin-332, hemidesmosome-rich fraction and laminin-521 trimer recombinant protein (RP) detected laminin γ-1 and α-3 and γ-2 subunits of laminin-332. Three years after skin lesions disappeared, nephrotic symptoms developed. Antibodies to α-3 chain of type IV collagen (COL4A3) were negative, thus excluding the diagnosis of Goodpasture syndrome. All anti-laminin antibodies disappeared. Additional IB and ELISA studies of RPs of various COL4 chains revealed reactivity with COL4A5, but not with COL4A6 or COL4A3. Although diagnosis of anti-laminin γ-1 (p200) pemphigoid or anti-laminin-332-type mucous membrane pemphigoid could not be made, this case was similar to previous cases with autoantibodies to COL4A5 and/or COL4A6.

  12. Fabrication and Properties of Poly(vinylalcohol)-glycosaminoglycantype I Collagen Composite Membrane as Tissue Regeneration Scaffolds

    Institute of Scientific and Technical Information of China (English)

    LI Qin-hua; LIN Dong-qing


    The objective of this paper is to design a porous polyvinyl alcohol (PVA) based on composite membrane with certain mechanical strength and biocompatibilities serving as tissue regenerative scaffolds. PVA-glycosaminoglycan (GAG)-type I collagen (COL) composite membrane was fabricated by PVA with different molecular weight (Mw) and alcoholysis degree (AD) being blended with certain amounts of GAG and COL and dried at 38℃for 24 h. The water content of the composite membranes were from 61.9%to 95.1%and swelling ratio ranged from 123.6%to 621.7%. Scanning electron micro-scope (SEM) analysis proved that PVA-GAG-COL composite membrane has porous and homogenous structure. Biocompatibility test results showed that the composite membrane was nontoxic, which could promote adhesion and proliferation of fibroblasts on the com-posite membrane. In conclusion, PVA-GAG-COL composite membrane with high water content and swelling ratio, suitable mechanical strength and good biocompatibility, has potential in tissue engineering and regenerative medicine.

  13. An Immunohistochemical Study of Retinal Collagen IV Expression during Pre- and Postnatal Postnatal Periods in Balb/c Mice


    Mehdi Jalali; Mohammad Reza Nikravesh; Abbas Ali Moeen; Mohammad Hassan Karimfar; Shahin Saidi Nejat; Shabnam Mohammadi; Houshang Rafighdoost


    Objective: Basement membranes are specialized extracellular matrices which playimportant roles such as cell regulation, proliferation and migration. Collagen fibers,especially type IV, are the most important basement membrane constituents. As retinais one of the target organs in diabetes mellitus, and nephropathy is a major cause ofend stage renal and retinal diseases resulting in increased morbidity and mortality,early diagnosis leads to better treatment. Hence, in this investigation, the ap...

  14. Discoidin Domain Receptor 2 Mediates Collagen-Induced Activation of Membrane-Type 1 Matrix Metalloproteinase in Human Fibroblasts. (United States)

    Majkowska, Iwona; Shitomi, Yasuyuki; Ito, Noriko; Gray, Nathanael S; Itoh, Yoshifumi


    Membrane-Type 1 Matrix Metalloproteinase (MT1-MMP) is a membrane-bound MMP that is highly expressed in cells with invading capacity including fibroblasts and invasive cancer cell. A potential physiological stimulus for MT1-MMP expression is fibrillar collagen, and it has been shown that it upregulates both MT1-MMP gene and functions in various cell types. However, the mechanisms of collagen-mediated MT1-MMP activation is not clearly understood. In this study we identified discoidin domain receptor 2 (DDR2) as a crucial receptor that mediates this process in human fibroblasts. Knocking down DDR2, but not β1 integrin subunit, a common subunit for all collagen-binding integrins, inhibited collagen-induced activation of proMMP-2 and upregulation of MT1-MMP at the gene and protein level. Interestingly DDR2 knockdown or pharmacological inhibition of DDR2 also inhibited MT1-MMP-dependent cellular degradation of collagen film, suggesting that cell surface collagen degradation by MT1-MMP involves DDR2-mediated collagen signalling. This DDR2-mediated mechanism is only present in non-transformed mesenchymal cells, as collagen-induced MT1-MMP activation in HT1080 fibrosarcoma cells and MT1-MMP function in MDA-MB231 breast cancer cells were not affected by DDR kinase inhibition. DDR2 activation was found to be noticeably more effective when cells were stimulated by collagen without non-helical telopeptides region compared to intact collagen fibrils. Those data suggest that DDR2 is a microenvironmental sensor that regulates fibroblasts migration in collagen-rich environment.

  15. Fras1, a basement membrane-associated protein mutated in Fraser syndrome, mediates both the initiation of the mammalian kidney and the integrity of renal glomeruli


    Pitera JE, Scambler PJ, Woolf AS.


    FRAS1 is mutated in some individuals with Fraser syndrome (FS) and the encoded protein is expressed in embryonic epidermal cells, localizing in their basement membrane (BM). Syndactyly and cryptophthalmos in FS are sequelae of skin fragility but the bases for associated kidney malformations are unclear. We demonstrate that Fras1 is expressed in the branching ureteric bud (UB), and that renal agenesis occurs in homozygous Fras1 null mutant blebbed (bl) mice on a C57BL6J background. In vivo, th...

  16. In vitro aging of mineralized collagen-based composite as guided tissue regeneration membrane

    Energy Technology Data Exchange (ETDEWEB)

    Pan, S.X. [Department of Prothodontics, School of Stomatology, Peking University, Beijing 100875 (China)]. E-mail:; Li, Y. [Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China); Feng, H.L. [Department of Prothodontics, School of Stomatology, Peking University, Beijing 100875 (China); Bai, W. [Department of Prothodontics, School of Stomatology, Peking University, Beijing 100875 (China); Gu, Y.Y. [Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China)


    The technique of guided tissue regeneration (GTR) has been developed for the regeneration of periodontal tissues, bone around natural teeth and dental implants. The aim of this study is to investigate the biodegradability and mechanic behavior of a novel mineralized nano-hydroxyapatite/collagen/poly (lactic acid) (nHAC/PLA) composite as GTR membrane in vitro. The elastic modulus and maximum tensile strength of GTR film samples with different nHAC/PLA ratio were measured to get an optimal nHAC/PLA ratio. Thermogravimetric analysis was conducted to evaluate the change of the inorganic component in the samples during the process of in vitro aging. Morphology of samples was checked by using scanning electron microscopy. On the basis of the above results, it can be concluded that the GTR membranes maintained integrity and the original appearance throughout the 1-month in vitro aging. There is an active dissolution and deposition process of crystals which is propitious to the bone formation on the surface of the composite membrane. The optimal nHAC/PLA ratio of the novel membrane is 0.4:1. For a longer period of bone repair, PLA with higher molecular weight should be chosen as the scaffold for the GTR membrane.

  17. β2 and γ3 laminins are critical cortical basement membrane components: ablation of Lamb2 and Lamc3 genes disrupts cortical lamination and produces dysplasia. (United States)

    Radner, Stephanie; Banos, Charles; Bachay, Galina; Li, Yong N; Hunter, Dale D; Brunken, William J; Yee, Kathleen T


    Cortical development is dependent on the timely production and migration of neurons from neurogenic sites to their mature positions. Mutations in several receptors for extracellular matrix (ECM) molecules and their downstream signaling cascades produce dysplasia in brain. Although mutation of a critical binding site in the gene that encodes the ECM molecule laminin γ1 (Lamc1) disrupts cortical lamination, the ECM ligand(s) for many ECM receptors have not been demonstrated directly in the cortex. Several isoforms of the heterotrimeric laminins, all containing the β2 and γ3 chain, have been isolated from the brain, suggesting they are important for CNS function. Here, we report that mice homozygous null for the laminin β2 and γ3 chains exhibit cortical laminar disorganization. Mice lacking both of these laminin chains exhibit hallmarks of human cobblestone lissencephaly (type II, nonclassical): they demonstrate severe laminar disruption; midline fusion; perturbation of Cajal-Retzius cell distribution; altered radial glial cell morphology; and ectopic germinal zones. Surprisingly, heterozygous mice also exhibit laminar disruption of cortical neurons, albeit with lesser severity. In compound null mice, the pial basement membrane is fractured, and the distribution of a key laminin receptor, dystroglycan, is altered. These data suggest that β2 and γ3-containing laminins play an important dose-dependent role in development of the cortical pial basement membrane, which serves as an attachment site for Cajal-Retzius and radial glial cells, thereby guiding neural development.

  18. Study on Effect Difference between Guizhi Decoction and Huanglianjiedu Decoction on Immuno - inflammatory Factors and Myocardial Basement Membrane in Spontaneous Diabetic Rats%桂枝汤和黄连解毒汤对 GK 大鼠心肌炎症因子及基膜影响的差异研究

    Institute of Scientific and Technical Information of China (English)

    姜萍; 戴玲玲; 王雪; 李晓


    目的:观察桂枝汤及黄连解毒汤对自发性糖尿病大鼠(GK 大鼠)心肌基膜厚度、心肌核因子-κB(NF -κB)、Ⅳ型胶原表达的作用,探讨有关炎症损伤的机制。方法以 Wistar 大鼠10只作为正常对照组,40只 GK 大鼠随机分为 GK 对照组、二甲双胍组、黄连解毒汤组、桂枝汤组,分别以相应药物灌胃给药12周,检测各组血糖、NF -κB、Ⅳ型胶原阳性表达程度及基膜厚度。结果 GK大鼠血糖升高,NF -κB 阳性表达增强,Ⅳ型胶原蛋白表达增强,基底膜厚度增加。二甲双胍、黄连解毒汤均可降低 GK 大鼠血糖(P ﹤0.01)。黄连解毒汤和桂枝汤均能降低 NF -κB 及Ⅳ型胶原蛋白表达、降低基底膜厚度与 GK 大鼠对照组比较差异有统计学意义(P ﹤0.01,P ﹤0.05)。在抑制 NF -κB阳性表达方面,桂枝汤和黄连解毒汤疗效无统计学意义。在抑制Ⅳ型胶原蛋白表达、降低基底膜厚度方面,桂枝汤作用强于黄连解毒汤(P ﹤0.01,P ﹤0.05)。结论桂枝汤和黄连解毒汤均可以抑制 NF-κB 阳性表达,抑制Ⅳ型胶原蛋白表达,降低基膜厚度,且桂枝汤优于黄连解毒汤,显示了其调和营卫治法的独特作用。%Objective To observe the effect of Guizhi decoction and Huanglianjiedu decoction on the thickness of myocardial basement membrane and the expression of NF - κB and type IV collagen,and to explore the mechanism of inflammatory injury in Spontaneous diabetic rats(GK rats). Methods Ten Wister rats were included in the control group,while forty GK rats were randomly divided into GK control group,met-formin group,Huanglianjiedu decoction group and Guizhi decoction group. Each group was respectively ad-ministrated the corresponding dose of drugs for 12 weeks. Then blood glucose,the degree of positive expres-sion of myocardial NF - κB and type IV collagen and the changes of basement membrane thickness were de

  19. Evaluation of a bioresorbable collagen membrane of fish origin in the treatment of periodontal intrabony defects: A prospective clinical study

    Directory of Open Access Journals (Sweden)

    B B Santosh Kumar


    Full Text Available Background: Recently, there has been interest in non-mammalian collagen sources such as fish collagen in the development of biomatrices and scaffolds for periodontal regeneration. In the present study, a novel collagen barrier membrane of fish origin was assessed in the treatment of periodontal intra-bony defects. Materials and Methods: Ten systemically healthy chronic periodontitis patients having an osseous defect in the mandibular posterior teeth were selected and following the open flap debridement, a collagen membrane was placed over the defect and the flap was sutured with interrupted sutures. Clinical parameters such as Plaque Index, Gingival Bleeding Index, probing pocket depth (PPD, relative attachment level (RAL, and recession (R were recorded at baseline, 6 and 9 months, whereas radiographic evaluation was done to assess alveolar crestal bone level and defect depth fill at 6 and 9 months using Auto-computer aided design (ACAD 2007 software. Statistical significance was set at 5% level of significance. Results: There was statistical significant differences with respect to periodontal clinical parameters such as Plaque Index, Gingival Bleeding Index, PPD, RAL, and gingival recession assessed at baseline, at 6 and 9 months respectively ( P < 0.05, and radiographic evaluation showed a defect fill of 58.62 median % at 9 months. Conclusion: This preliminary study has shown predictable results in using fish collagen membrane, for treating periodontal intra-bony defects. Further, long-term clinical trials are needed to validate the effectiveness of this membrane.

  20. Nonlinear optical response of the collagen triple helix and second harmonic microscopy of collagen liquid crystals (United States)

    Deniset-Besseau, A.; De Sa Peixoto, P.; Duboisset, J.; Loison, C.; Hache, F.; Benichou, E.; Brevet, P.-F.; Mosser, G.; Schanne-Klein, M.-C.


    Collagen is characterized by triple helical domains and plays a central role in the formation of fibrillar and microfibrillar networks, basement membranes, as well as other structures of the connective tissue. Remarkably, fibrillar collagen exhibits efficient Second Harmonic Generation (SHG) and SHG microscopy proved to be a sensitive tool to score fibrotic pathologies. However, the nonlinear optical response of fibrillar collagen is not fully characterized yet and quantitative data are required to further process SHG images. We therefore performed Hyper-Rayleigh Scattering (HRS) experiments and measured a second order hyperpolarisability of 1.25 10-27 esu for rat-tail type I collagen. This value is surprisingly large considering that collagen presents no strong harmonophore in its amino-acid sequence. In order to get insight into the physical origin of this nonlinear process, we performed HRS measurements after denaturation of the collagen triple helix and for a collagen-like short model peptide [(Pro-Pro-Gly)10]3. It showed that the collagen large nonlinear response originates in the tight alignment of a large number of weakly efficient harmonophores, presumably the peptide bonds, resulting in a coherent amplification of the nonlinear signal along the triple helix. To illustrate this mechanism, we successfully recorded SHG images in collagen liquid solutions by achieving liquid crystalline ordering of the collagen triple helices.

  1. Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin

    DEFF Research Database (Denmark)

    Sasaki, T; Brakebusch, C; Engel, J


    in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. The protein also mediated adhesion of cell lines at comparable strength with laminin. Adhesion to M2BP was inhibited by antibodies to integrin beta1...

  2. Synthesis of collagen nanotubes with highly regular dimensions through membrane-templated layer-by-layer assembly. (United States)

    Landoulsi, Jessem; Roy, Cécile J; Dupont-Gillain, Christine; Demoustier-Champagne, Sophie


    Nanotubes made from a fibrillar protein, namely, collagen, were fabricated by a template-based method combined with layer-by-layer (LbL) deposition. The ability to incorporate collagen in LbL multilayered film was first demonstrated by in situ quartz crystal microbalance and ex situ ellipsometry on a flat model substrate, using poly(styrene sulfonate) (PSS) as polyanion. Collagen-based nanotubes were then fabricated by alternately immersing a polycarbonate membrane, used as template, in PSS and collagen aqueous solutions. Direct evidence for nanotube formation was obtained by dissolving the membrane and imaging the liberated (PSS/collagen)(n) nanostructures by scanning electron microscopy and by transmission electron microscopy. The proposed strategy constitutes a practical alternative to electrospinning as it allows a very good control over the dimensions (outside and inside diameters and length) of the resulting nanotubes. Besides their fundamental interest, collagen-based nanotubes are useful nano-objects for the creation of new nanostructured biomaterials with numerous potential applications in the biomedical field.

  3. Preparation and characterisation of Punica granatum pericarp aqueous extract loaded chitosan-collagen-starch membrane: role in wound healing process. (United States)

    Amal, B; Veena, B; Jayachandran, V P; Shilpa, Joy


    Engineered scaffolds made from natural biomaterials are crucial elements in tissue engineering strategies. In this study, biological scaffold like chitosan-collagen-starch membrane (CCSM) loaded with the antibacterial agent, Punica granatum pericarp aqueous extract was explored for enhanced regeneration of epithelial tissue during wound healing. Collagen was extracted from Rachycentron canadum fish skin. Membranous scaffold was prepared by mixing collagen, starch and chitosan in a fixed proportion, loaded with aqueous extract of P. granatum and its anti-pseudomonal activity was studied. Morphological characterization by SEM and mechanical property like tensile strength of the membrane were studied. Excision wound of 2 cm(2) size was induced in Guinea pig and the effect of P. granatum extract loaded CCSM in wound healing was studied. The SEM image showed deep pores in the membrane and also possessed good tensile strength. Wound surface area was reduced prominently in the experimental group with P. granatum extract loaded CCSM when compared to the group with unloaded membrane and the one with no membrane. Punica granatum extract loaded CCSM has antipseudomonal property and supported enhanced epithelial cell proliferation without leaving a scar after wound healing. This has significant therapeutic application in membranous scaffold mediated skin repair and regeneration.

  4. Regeneration of chronic tympanic membrane perforation using 3D collagen with topical umbilical cord serum. (United States)

    Jang, Chul Ho; Cho, Yong Beom; Yeo, MyungGu; Lee, Hyeongjin; Min, Eun Jung; Lee, Byung Hhwa; Kim, Geun Hyung


    Chronic tympanic membrane (TM) perforation is one of the most common otology complications. Current surgical management of TM perforation includes myringoplasty and tympanoplasty. The purpose of this study was to evaluate the efficacy and feasibility of three dimensional (3D) porous collagen scaffolds with topically applied human umbilical cord serum (UCS) for the regeneration of chronic TM perforation in guinea pigs. To achieve this goal, we fabricated porous 3D collagen scaffolds (avg. strut diameter of 236 ± 51 μm, avg. pore size of 382 ± 67 μm, and a porosity of 96%) by using a 3 axis robot dispensing and low temperature plate systems. Guinea pigs were used in a model of chronic TM perforation. In the experimental group (n=10), 3D collagen scaffold was placed on the perforation and topically applied of UCS every other day for a period of 8 days. The control group ears (n=10) were treated with paper discs and phosphate buffered saline (PBS) only using the same regimen. Healing time, acoustic-mechanical properties, and morphological analysis were performed by otoendoscopy, auditory brainstem response (ABR), single-point laser Doppler vibrometer (LDV), optical coherence tomography (OCT), and light microscopic evaluation. The closure of the TM perforation was achieved in 100% of the experimental group vs. 43% of the control group, and this difference was statistically significant (p=0.034). The ABR threshold at all frequencies of the experimental group was significantly recovered to the normal level compared to the control group. TM vibration velocity in the experimental group recovered similar to the normal control level. The difference is very small and they are not statistically significant below 1 kHz (p=0.074). By OCT and light microscopic examination, regenerated TM of the experimental group showed thickened fibrous and mucosal layer. In contrast, the control group showed absence of fibrous layer like a dimeric TM.

  5. Chitosan nanoparticles enhance the antibacterial activity of chlorhexidine in collagen membranes used for periapical guided tissue regeneration. (United States)

    Barreras, Uriel Soto; Méndez, Fernando Torres; Martínez, Rita Elizabeth Martínez; Valencia, Carolina Samano; Rodríguez, Panfilo Raymundo Martinez; Rodríguez, Juan Pablo Loyola


    Endodontic failure is mainly associated with the persistence of microbial infection in the root canal system and/or the periradicular area. Microorganisms and their toxins located in the root canal system may trigger apical periodontitis and tissue destruction. Tissue regeneration in periapical surgery by using membrane barriers has shown an improved healing and bone closure. However, bacterial membrane contamination is a main reason of failure. In this in vitro study, different brands of chlorhexidine, a combination of chitosan nanoparticles containing chlorhexidine were tested against Enterococcus faecalis on agar plate's cultures and infected collagen membranes. Our results indicated that chitosan nanoparticles acted synergistically with chlorhexidine, inhibiting and eliminating significantly a greater amount of colony former units in both BHI-agar cultures and infected collagen membranes. These results suggested that chitosan nanoparticles could be used to improve regenerative procedures in periapical surgery.

  6. Chondrocyte-seeded type I/III collagen membrane for autologous chondrocyte transplantation

    DEFF Research Database (Denmark)

    Niemeyer, Philipp; Lenz, Philipp; Kreuz, Peter C;


    PURPOSE: We report the 2-year clinical results and identify prognostic factors in patients treated with autologous chondrocyte transplantation by use of a collagen membrane to seed the chondrocytes (ACT-CS). METHODS: This is a prospective study of 59 patients who were treated with ACT-CS...... Repair Society (ICRS) rating, the percentage of patients rated A (normal) and B (nearly normal) increased from 33.9% preoperatively to 92.5% at 24 months after ACT-CS. IKDC and Lysholm scores increased from 50.1 points (SD, 13.4) and 60.5 points (SD, 9.4), respectively, to 76.1 points (SD, 15.2) (P...... CS as a salvage procedure. The rate of failures in patients with isolated cartilage defects was 5.9%. CONCLUSIONS: ACT-CS...

  7. Immunohistochemical localization of chondroitin sulfate, chondroitin sulfate proteoglycan, heparan sulfate proteoglycan, entactin, and laminin in basement membranes of postnatal developing and adult rat lungs

    DEFF Research Database (Denmark)

    Sannes, P L; Burch, K K; Khosla, J


    alveolar, airway, and vascular BMs, in addition to smooth muscle external laminae (EL), in the adult and developing rat. Immunostaining for CSPG required hyaluronidase digestion, whereas CS staining was lost with the same treatment. A polyclonal antibody to the core protein of HSPG was found...... to be similarly distributed to CSPG by immunoperoxidase staining in adult and developing rat lungs, with the notable exception that little immunoreactivity for HSPG was found in smooth muscle EL. Commercially obtained polyclonal antibodies to entactin and laminin gave immunostaining comparable to that seen......Histologic preparations of lungs from 1-, 5-, 10-, 18-, and 25-day-old postnatal and adult rats were examined immunohistochemically with antibodies specific against chondroitin sulfate (CS), basement membrane chondroitin sulfate proteoglycan (BM-CSPG), heparan sulfate proteoglycan (HSPG), entactin...

  8. In vitro blood-brain barrier models for drug research: state-of-the-art and new perspectives on reconstituting these models on artificial basement membrane platforms. (United States)

    Banerjee, Jayati; Shi, Yejiao; Azevedo, Helena S


    In vitro blood-brain barrier (BBB) models are indispensable screening tools for obtaining early information about the brain-penetrating behaviour of promising drug candidates. Until now, in vitro BBB models have focused on investigating the interplay among cellular components of neurovascular units and the effect of fluidic sheer stress in sustaining normal BBB phenotype and functions. However, an area that has received less recognition is the role of the noncellular basement membrane (BM) in modulating BBB physiology. This review describes the state-of-the-art on in vitro BBB models relevant in drug discovery research and highlights their strengths, weaknesses and the utility potential of some of these models in testing the permeability of nanocarriers as vectors for delivering therapeutics to the brain. Importantly, our review also introduces a new concept of engineering artificial BM platforms for reconstituting BBB models in vitro.

  9. Chondroitin 6-sulfate proteoglycan but not heparan sulfate proteoglycan is abnormally expressed in skin basement membrane from patients with dominant and recessive dystrophic epidermolysis bullosa

    DEFF Research Database (Denmark)

    Fine, J D; Couchman, J R


    Two distinct groups of proteoglycans, chondroitin 6-sulfate (C6-S) proteoglycan and heparan sulfate proteoglycan (HSPG), have been recently shown to reside within the lamina densa of normal human skin basement membrane (BM). To determine whether either or both antigens are normally expressed in one...... junctional EB, and all control skin specimens. We have subsequently extracted a greater than 400 kD C6-S proteoglycan from normal skin BM and have found that the core protein may also contain heparan sulfate side chains. Our findings suggest that 3B3 monoclonal antibody recognizes a hybrid proteoglycan...... in human skin, and that its absent or reduced binding in dystrophic EB skin BM may reflect either absence of associated core protein or posttranslational alterations in the proteoglycan side chains....

  10. Reciprocal interactions between Beta1-integrin and epidermal growth factor in three-dimensional basement membrane breast cultures: A different perspective in epithelial biology

    Energy Technology Data Exchange (ETDEWEB)

    Wang, F.; Weaver, V.M.; Petersen, O.W.; Larabell, C.A.; Dedhar, S.; Briand, P.; Lupu, R.; Bissell, M.J.


    Anchorage and growth factor independence are cardinal features of the transformed phenotype. Although it is logical that the two pathways must be coregulated in normal tissues to maintain homeostasis, this has not been demonstrated directly. We showed previously that down-modulation of {beta}1-integrin signaling reverted the malignant behavior of a human breast tumor cell line (T4-2) derived from phenotypically normal cells (HMT-3522) and led to growth arrest in a threedimensional (3D) basement membrane assay in which the cells formed tissue-like acini (14). Here, we show that there is a bidirectional cross-modulation of {beta}1-integrin and epidermal growth factor receptor (EGFR) signaling via the mitogenactivated protein kinase (MAPK) pathway. The reciprocal modulation does not occur in monolayer (2D) cultures. Antibodymediated inhibition of either of these receptors in the tumor cells, or inhibition of MAPK kinase, induced a concomitant downregulation of both receptors, followed by growth-arrest and restoration of normal breast tissue morphogenesis. Crossmodulation and tissue morphogenesis were associated with attenuation of EGF-induced transient MAPK activation. To specifically test EGFR and {beta}1-integrin interdependency, EGFR was overexpressed in nonmalignant cells, leading to disruption of morphogenesis and a compensatory up-regulation of {beta}1-integrin expression, again only in 3D. Our results indicate that when breast cells are spatially organized as a result of contact with basement membrane, the signaling pathways become coupled and bidirectional. They further explain why breast cells fail to differentiate in monolayer cultures in which these events are mostly uncoupled. Moreover, in a subset of tumor cells in which these pathways are misregulated but functional, the cells could be 'normalized' by manipulating either pathway.

  11. Periodontal Responses to Augmented Corticotomy with Collagen Membrane Application during Orthodontic Buccal Tipping in Dogs

    Directory of Open Access Journals (Sweden)

    Dong-Yeol Lee


    Full Text Available This prospective randomized split-mouth study was performed to examine the effects of absorbable collagen membrane (ACM application in augmented corticotomy using deproteinized bovine bone mineral (DBBM, during orthodontic buccal tipping movement in the dog. After buccal circumscribing corticotomy and DBBM grafting into the decorticated area, flaps were repositioned and sutured on control sides. ACM was overlaid and secured with membrane tacks, on test sides only, and the flaps were repositioned and sutured. Closed coil springs were used to apply 200 g orthodontic force in the buccolingual direction on the second and third premolars, immediately after primary flap closure. The buccal tipping angles were 31.19±14.60° and 28.12±11.48° on the control and test sides, respectively. A mean of 79.5 ± 16.0% of the buccal bone wall was replaced by new bone on the control side, and on the test side 78.9±19.5% was replaced. ACM application promoted an even bone surface. In conclusion, ACM application in augmented corticotomy using DBBM might stimulate periodontal tissue reestablishment, which is useful for rapid orthodontic treatment or guided bone regeneration. In particular, ACM could control the formation of mesenchymal matrix, facilitating an even bone surface.

  12. Characterization of the collagen phenotype of rabbit proximal tubule cells in culture. (United States)

    Gibbs, S R; Goins, R A; Belvin, E L; Dimari, S J; Merriam, A P; Bowling-Brown, S; Harris, R C; Haralson, M A


    Studies were performed to characterize the collagen phenotype of cultured rabbit proximal tubule (RPT) epithelial cells grown on plastic and on the reconstituted basement membrane preparation, Matrigel. When grown on a plastic substratum, RPT cells display a cobblestone appearance characteristic of glomerular epithelial cells. While initially forming an interlocking network of cells after subculture on Matrigel, this pattern of culture morphology rapidly develops into one characterized by isolated, organized groups of cells. Notwithstanding the effects of Matrigel on culture morphology, total cellular proliferation was reduced only 25% when RPT cells were grown on this substrate. Greater than 90% of the collagen synthesized by RPT cells grown on plastic was secreted into the culture medium. Qualitative analysis by SDS-PAGE revealed components exhibiting electrophoretic mobilities corresponding to the chains present in type IV and type I collagens. Quantitative analysis by CM-Trisacryl chromatography established that approximately 2/3 of the total collagen synthesized by RPT cells grown on plastic was type IV and approximately 1/3 type I. Quantitative analysis of the collagens produced by RPT cells grown on Matrigel again indicated the synthesis of only type IV and type I molecules but in a slightly more equal ratio of both collagen types and in the ratio of secreted to cell-associated molecules. However, the total amount of collagen synthesized by RPT cells grown on Matrigel was reduced to approximately 1% of the level synthesized by the cells grown on plastic. On plastic, approximately 3/4 of the type I collagen produced was recovered as the type I homotrimer, but on Matrigel type I homotrimers represented only approximately 55% of the total type I collagen synthesized. On Matrigel, the majority of the type IV collagen was recovered as heterotrimers containing alpha1(IV) and alpha2(IV) chains. In contrast, RTP cells grown on plastic predominantly produced type IV

  13. Breaches of the pial basement membrane and disappearance of the glia limitans during development underlie the cortical lamination defect in the mouse model of muscle-eye-brain disease. (United States)

    Hu, Huaiyu; Yang, Yuan; Eade, Amber; Xiong, Yufang; Qi, Yue


    Neuronal overmigration is the underlying cellular mechanism of cerebral cortical malformations in syndromes of congenital muscular dystrophies caused by defects in O-mannosyl glycosylation. Overmigration involves multiple developmental abnormalities in the brain surface basement membrane, Cajal-Retzius cells, and radial glia. We tested the hypothesis that breaches in basement membrane and the underlying glia limitans are the key initial events of the cellular pathomechanisms by carrying out a detailed developmental study with a mouse model of muscle-eye-brain disease, mice deficient in O-mannose beta1,2-N-acetylglucosaminyltransferase 1 (POMGnT1). The pial basement membrane was normal in the knockout mouse at E11.5. It was breached during rapid cerebral cortical expansion at E13.5. Radial glial endfeet, which comprise glia limitans, grew out of the neural boundary. Neurons moved out of the neural boundary through these breaches. The overgrown radial glia and emigrated neurons disrupted the overlying pia mater. The overmigrated neurons did not participate in cortical plate (CP) development; rather they formed a diffuse cell zone (DCZ) outside the original cortical boundary. Together, the DCZ and the CP formed the knockout cerebral cortex, with disappearance of the basement membrane and the glia limitans. These results suggest that disappearance of the basement membrane and the glia limitans at the cerebral cortical surface during development underlies cortical lamination defects in congenital muscular dystrophies and a cellular mechanism of cortical malformation distinct from that of the reeler mouse, double cortex syndrome, and periventricular heterotopia.

  14. Heparanase expression,degradation of basement membrane and low degree of infiltration by immunocytes correlate with invasion and progression of human gastric cancer

    Institute of Scientific and Technical Information of China (English)

    Zun-Jiang Xie; Ying Liu; Li-Min Jia; Ye-Chun He


    AIM: To disclose the mechanisms that accelerate or limit tumor invasion and metastasis in gastric cancer patients.METHODS: The heparanase expression,continuity of basement,degree of infiltration by dendritic cells and lymphocytes in gastric cancer tissues from 33 the early and late stage patients were examined by immunohistochemistry,in situ hybridization and transmission electron microscopy.RESULTS: Heparanase mRNA expression in the late stage patients with gastric cancer was stronger than that in the early stage gastric cancer patients.In the early stage gastric cancer tissues,basement membrane (BlVl) appeared intact,whereas in the late stage,discontinuous BM was often present.The density of $100 protein positive tumor infiltrating dendritic cells (TIDC) in the early stage gastric cancer tissues was higher than that in the late stage.The infiltrating degree of tumor infiltrating lymphocytes (TIL) in the early stage patients whose tumor tissues contained a high density of TIDC was significantly higher than that in the late stage gastric cancer tissues patients with a low density of TIDC.There were few cancer cells penetrated through the continuous BM of cancer nests in the early stage gastric cancers,but many cancer cells were found outside of the defective BM of cancer nests in the late stage.CONCLUSION: Our results suggest that strong heparanase expression is related with the degradation of BM which allows or accelerates tumor invasion and metastasis.However,high density of TIDC and degree of infiltration by TIL are associated with tumor progression in human gastric cancers.

  15. Guided bone regeneration produced by new mineralized and reticulated collagen membranes in critical-sized rat calvarial defects. (United States)

    Veríssimo, Denusa M; Leitão, Renata F C; Figueiró, Sônia D; Góes, Júlio C; Lima, Vilma; Silveira, Charles O; Brito, Gerly A C


    The aim of this study was to evaluate the bone regenerative effect of glutaraldehyde (GA) cross-linking on mineralized polyanionic collagen membranes in critical-sized defects on rat calvarias. Bone calvarial defects were induced in Wistar rats, which were then divided into five groups: a sham group; a control group, which received a commercial membrane; and GA, 25GA, and 75GA groups, which received one of three different polyanionic collagen membranes mineralized by 0, 25, or 75 hydroxyapatite cycles and then cross-linked by GA. Bone formation was evaluated based on digital radiography and computerized tomography. Histological analyses were performed 4 and 12 weeks after the surgical procedure to observe bone formation, membrane resorption, and fibrous tissue surrounding the membranes. Measurement of myeloperoxidase activity, tumor necrosis factor alpha, and interleukin 1beta production was performed 24 h after surgery. The percentage of new bone formation in the GA, 25GA, and 75GA groups was higher compared with the control and sham groups. In the GA and 25 GA groups, the membranes were still in place and were contained in a thick fibrous capsule after 12 weeks. No significant difference was found among the groups regarding myeloperoxidase activity and interleukin 1beta levels, although the GA, 25GA, and 75GA groups presented decreased levels of tumor necrosis factor alpha compared with the control group. These new GA cross-linked membranes accelerated bone healing of the calvarium defects and did not induce inflammation. In addition, unlike the control membrane, the experimental membranes were not absorbed during the analyzed period, so they may offer advantages in large bone defects where prolonged membrane barrier functions are desirable.

  16. Guided periodontal regeneration using bilayered collagen membranes and bovine bone mineral in fenestration defects in the canine. (United States)

    Tal, Haim; Artzi, Zvi; Moses, Ofer; Nemcovsky, Carlos; Kozlovsky, Avital


    This study was performed to evaluate the effect of deproteinized bovine porous bone mineral (BBM) and BBM-collagen (BBMC) used alone or in combination with a bilayer collagen membrane in guided periodontal regeneration. In 12 dogs, contralateral surgical circular fenestration defects 5 mm in diameter were produced at the midbuccal aspect of the alveolar bone in 24 maxillary canines. Bone, periodontal ligament, and cementum were completely removed. Experimental sites were filled with BBM or BBMC. Bilayered collagen membranes covered half the experimental sites (BBM+M and BBMC+M), and the other half were left uncovered. Control sites remained empty; half were covered with collagen membranes (cont+M) and the underlying space spontaneously filled with blood, and half were left uncovered (cont). Three months postsurgery, undecalcified sections were prepared. Measurements were made using a caliper on a projection microscope, and the surface area of new bone and BBM particles within the healed surgical defect was evaluated using the point-counting method. In the experimental defects, new cementum covered 31% to 67% of the exposed dentin, with a significant difference between defects covered with membranes and defects that were not covered (P tissue in the covered defects than in the uncovered defects (P tissue/bone marrow, and bovine bone particles. New bone area fraction was 23.4% to 25.2% in defects filled with BBMC and BBM, respectively (P = NS). Bone fraction area in membrane-covered defects ranged from 34.4% to 36.8% in experimental defects (P = NS). All membrane-treated defects showed higher values for bone area fraction in comparison to the uncovered control defects. Particle area fraction ranged between 17.4% and 26.2%, with only BBMC and BBM+M defects showing a statistically significant difference (P regeneration than experimental defects filled with BBM or BBMC. Treatment of defects with BBM or BBMC showed similar influences on bone and cementum regeneration in

  17. Polyanionic collagen membranes for guided tissue regeneration: Effect of progressive glutaraldehyde cross-linking on biocompatibility and degradation. (United States)

    Veríssimo, D M; Leitão, R F C; Ribeiro, R A; Figueiró, S D; Sombra, A S B; Góes, J C; Brito, G A C


    The ultimate goal of periodontal therapy is to control periodontal tissue inflammation and to produce predictable regeneration of that part of the periodontium which has been lost as a result of periodontal disease. In guided tissue regeneration membranes function as mechanical barriers, excluding the epithelium and gingival corium from the root surface and allowing regeneration by periodontal ligament cells. This report aims to study the effect of glutaraldehyde (GA) cross-linking on mineralized polyanionic collagen (PAC) membranes by conducting a histological evaluation of the tissue response (biocompatibility) and by assessing the biodegradation of subcutaneous membrane implants in rats. We studied six different samples: a PAC, a PAC mineralized by alternate soaking processes for either 25 or 75 cycles (PAC 25 and PAC 75, respectively) and these films cross-linked by GA. Inflammatory infiltrate, cytokine dosage, fibrosis capsule thickness, metalloproteinase immunohistochemistry and membrane biodegradation after 1, 7, 15 and 30 days were measured. The inflammatory response was found to be more intense in membranes without cross-linking, while the fibrosis capsules became thicker in cross-linked membranes after 30 days. The membranes without cross-linking suffered intense biodegradation, while the membranes with cross-linking remained intact after 30 days. The cross-linking with GA reduced the inflammatory response and prevented degradation of the membranes over the entire course of the observation period. These membranes are thus an attractive option when the production of new bone depends on the prolonged presence of a mechanical barrier.

  18. A three-layered nano-carbonated hydroxyapatite/collagen/PLGA composite membrane for guided tissue regeneration. (United States)

    Liao, Susan; Wang, Wei; Uo, Motohiro; Ohkawa, Shoji; Akasaka, Tsukasa; Tamura, Kazuchika; Cui, Fuzhai; Watari, Fumio


    Functional graded materials (FGM) provided us one new concept for guided tissue regeneration (GTR) membrane design with graded component and graded structure where one face of the membrane is porous thereby allowing cell growth thereon and the opposite face of the membrane is smooth, thereby inhibiting cell adhesion in periodontal therapy. The goal of the present study was to develop a three-layered graded membrane, with one face of 8% nano-carbonated hydroxyapatite/collagen/poly(lactic-co-glycolic acid) (nCHAC/PLGA) porous membrane, the opposite face of pure PLGA non-porous membrane, the middle layer of 4% nCHAC/PLGA as the transition through layer-by-layer casting method. Then the three layers were combined well with each other with flexibility and enough high mechanical strength as membrane because the three layers all contained PLGA polymer that can be easily used for practical medical application. This high biocompatibility and osteoconductivity of this biodegraded composite membrane was enhanced by the nCHAC addition, for the same component and nano-level crystal size with natural bone tissue. The osteoblastic MC3T3-E1 cells were cultured on the three-layered composite membrane, the primary result shows the positive response compared with pure PLGA membrane.

  19. Toward guided tissue and bone regeneration: morphology, attachment, proliferation, and migration of cells cultured on collagen barrier membranes. A systematic review.

    NARCIS (Netherlands)

    Behring, J.; Junker, R.; Walboomers, X.F.; Chessnut, B.; Jansen, J.A.


    Collagen barrier membranes are frequently used in both guided tissue regeneration (GTR) and guided bone regeneration (GBR). Collagen used for these devices is available from different species and is often processed to alter the properties of the final product. This is necessary because unprocessed c

  20. Possible association of elevated serum collagen type IV level with skin sclerosis in systemic sclerosis. (United States)

    Motegi, Sei-Ichiro; Sekiguchi, Akiko; Fujiwara, Chisako; Toki, Sayaka; Ishikawa, Osamu


    Collagen type IV is the primary collagen in the basement membranes around blood vessels and in the dermoepidermal junction in the skin. Perivascular collagen type IV is synthesized by endothelial cells and pericytes, and contributes to the homeostasis and remodeling of blood vessels. It has been well recognized that elevated serum collagen type IV levels are associated with the liver fibrosis. The objective was to examine serum collagen type IV levels and their clinical associations in patients with systemic sclerosis (SSc), and to examine the expression of collagen type IV in the fibrotic skin in SSc. Serum collagen type IV levels in SSc patients and diffuse cutaneous type SSc patients were significantly higher than those in healthy individuals. Serum collagen type IV levels were positively correlated with modified Rodnan total skin score. Serum collagen type IV levels in early stage (disease duration ≤3 years) diffuse cutaneous SSc patients were significantly elevated. Serum collagen type IV levels in SSc patients with digital ulcers (DU) were significantly elevated. In immunohistochemical staining, the expression of collagen type IV around dermal small vessels in the affected skin was reduced compared with those of normal individuals. These results suggest that elevated serum collagen type IV levels may be associated with the skin sclerosis in the early stage of SSc. The measurement of serum collagen type IV levels in SSc patients may be useful as a disease activity marker in skin sclerosis and DU.

  1. [Monoclonal autoantibodies to the epithelial basement membrane cells of human skin and thymus obtained through immunization with Rickettsia prowazekii antigens]. (United States)

    Drobyshevskaia, E I; Spitsyn, S V; Nedialkov, Iu A; Shchekotikhina, Iu A; Tarasevich, I V


    As the result of immunization of BALB/c mice with the commercial preparation of typhus vaccine and R. prowazekii corpuscular antigen, in 29.2% and 40.3% of cases (respectively) the appearance of hybridomas synthesizing monoclonal antibodies (McAb) to different autologous structures (skin and thymic epithelium, cell nuclei, conjunctive tissue structures and vascular endothelium) has been revealed. The McAb under test have proved to be IgM-autoantibodies. McAb M-6, active against the basal membrane of human skin and thymic epithelium, produce quite a definite picture of disturbances in the differentiation of epithelium and can be used for the diagnosis of dyskeratosis.

  2. Cells that emerge from embryonic explants produce fibers of type IV collagen. (United States)

    Chen, J M; Little, C D


    Double immunofluorescence staining experiments designed to examine the synthesis and deposition of collagen types I and IV in cultured explants of embryonic mouse lung revealed the presence of connective tissue-like fibers that were immunoreactive with anti-type IV collagen antibodies. This observation is contrary to the widely accepted belief that type IV collagen is found only in sheet-like arrangements beneath epithelia or as a sheath-like layer enveloping bundles of nerve or muscle cells. The extracellular matrix produced by cells that migrate from embryonic mouse lung rudiments in vitro was examined by double indirect immunofluorescence microscopy. Affinity-purified monospecific polyclonal antibodies were used to examine cells after growth on glass or native collagen substrata. The data show that embryonic mesenchymal cells can produce organized fibers of type IV collagen that are not contained within a basement membrane, and that embryonic epithelial cells deposit fibers and strands of type IV collagen beneath their basal surface when grown on glass; however, when grown on a rat tail collagen substratum the epithelial cells produce a fine meshwork. To our knowledge this work represents the first report that type IV collagen can be organized by cells into a fibrous extracellular matrix that is not a basement membrane.

  3. The collagen turnover profile is altered in patients with inguinal and incisional hernia

    DEFF Research Database (Denmark)

    Henriksen, Nadia A; Mortensen, Joachim H; Sorensen, Lars T;


    BACKGROUND: Disturbed metabolism in the extracellular matrix (ECM) contributes to formation of abdominal wall hernias. The aim of this study was to gain deeper insight into the ECM turnover in hernia patients by analyzing serum biomarkers specifically reflecting collagen synthesis and breakdown...... in the interstitial matrix (types I, III, and V collagens) and in the basement membrane (type IV collagen). MATERIAL AND METHODS: Patients with 3 different types of hernias were included: Primary unilateral inguinal hernia (n = 17), multiple hernias defined as ≥3 hernias (n = 21), and incisional hernia (n = 25......). Patients without hernias scheduled to undergo elective operation for gallstones (n = 18) served as controls. Whole venous blood was collected preoperatively. Biomarkers for synthesis of interstitial matrix (PINP, Pro-C3, P5CP) and basement membrane (P4NP) as well as corresponding degradation (C1M, C3M, C5M...

  4. Discoidin domain receptors regulate the migration of primary human lung fibroblasts through collagen matrices

    Directory of Open Access Journals (Sweden)

    Ruiz Pedro A


    Full Text Available Abstract Background The two discoidin domain receptors (DDRs, DDR1 and DDR2 are receptor tyrosine kinases (RTKs with the unique ability among RTKs to respond to collagen. We have previously shown that collagen I induces DDR1 and matrix metalloproteinase (MMP-10 expression through DDR2 activation and a Janus kinase (JAK2 and extracellular signal-regulated kinase (ERK1/2-mediated mechanism in primary human lung fibroblasts suggesting that these signaling pathways play a role in fibroblast function. Fibroblasts can traverse basement membrane barriers during development, wound healing and pathological conditions such as cancer and fibrosis by activating tissue-invasive programs, the identity of which remain largely undefined. In the present work, we investigated the role of DDRs and DDR-associated signal transduction in these processes. Results Transwell migration experiments showed that normal human lung fibroblast (NHLF transmigration through collagen I-coated inserts is mediated by DDR2 and the DDR2-associated signaling kinases JAK2 and ERK1/2, but not DDR1. Additionally, experiments with specific small interfering (siRNAs revealed that collagen I-induced expression of MMP-10 and MMP-2 is DDR2 but not DDR1 dependent in NHLFs. Our data showed that collagen I increases NHLF migration through collagen IV, the main component of basement membranes. Furthermore, basal and collagen I-induced NHLF migration through collagen IV-coated inserts was both DDR2 and DDR1 dependent. Finally, DDR2, but not DDR1 was shown to be involved in fibroblast proliferation. Conclusions Our results suggest a mechanism by which the presence of collagen I in situations of excessive matrix deposition could induce fibroblast migration through basement membranes through DDR2 activation and subsequent DDR1 and MMP-2 gene expression. This work provides new insights into the role of DDRs in fibroblast function.

  5. Collagen alpha5 and alpha2(IV) chain coexpression: analysis of skin biopsies of Alport patients. (United States)

    Patey-Mariaud de Serre, N; Garfa, M; Bessiéres, B; Noël, L H; Knebelmann, B


    Alport syndrome is a collagen type IV disease caused by mutations in the COL4A5 gene with the X-linked form being most prevalent. The resultant alpha5(IV) collagen chain is a component of the glomerular and skin basement membranes (SBMs). Immunofluorescent determination of the alpha5(IV) chain in skin biopsies is the procedure of choice to identify patients. In 30% of patients, however, the mutant protein is still found in the SBM resulting in a normal staining pattern. In order to minimize or eliminate false results, we compared the distribution of the alpha2(IV) chain (another SBM component) and the alpha5(IV) chain by standard double label immunofluorescence (IF) and by confocal laser scanning microscopy. The study was performed on 55 skin biopsies of patients suspected of Alports and five normal control specimens. In normal skin, IF showed the classical linear pattern for both collagens along the basement membrane. Additionally, decreased alpha5(IV) was found in the bottom of the dermal papillary basement membrane. Confocal analysis confirmed the results and show alpha5(IV) focal interruptions. In suspected patients, both techniques showed the same rate of abnormal alpha5(IV) expression: segmental in women and absent in men. Our results show a physiological variation of alpha5(IV) location with focal interruptions and decreased expression in the bottom of the dermal basement membrane. Comparison of alpha5(IV) with alpha2(IV) expression is simple and eliminates technical artifacts.

  6. Evaluation of 3D printed PCL/PLGA/β-TCP versus collagen membranes for guided bone regeneration in a beagle implant model. (United States)

    Won, J-Y; Park, C-Y; Bae, J-H; Ahn, G; Kim, C; Lim, D-H; Cho, D-W; Yun, W-S; Shim, J-H; Huh, J-B


    Here, we compared 3D-printed polycaprolactone/poly(lactic-co-glycolic acid)/β-tricalcium phosphate (PCL/PLGA/β-TCP) membranes with the widely used collagen membranes for guided bone regeneration (GBR) in beagle implant models. For mechanical property comparison in dry and wet conditions and cytocompatibility determination, we analyzed the rate and pattern of cell proliferation of seeded fibroblasts and preosteoblasts using the cell counting kit-8 assay and scanning electron microscopy. Osteogenic differentiation was verified using alizarin red S staining. At 8 weeks following implantation in vivo using beagle dogs, computed tomography and histological analyses were performed after sacrifice. Cell proliferation rates in vitro indicated that early cell attachment was higher in collagen than in PCL/PLGA/β-TCP membranes; however, the difference subsided by day 7. Similar outcomes were found for osteogenic differentiation, with approximately 2.5 times greater staining in collagen than PCL/PLGA/β-TCP, but without significant difference by day 14. In vivo, bone regeneration in the defect area, represented by new bone formation and bone-to-implant contact, paralleled those associated with collagen membranes. However, tensile testing revealed that whereas the PCL/PLGA/β-TCP membrane mechanical properties were conserved in both wet and dry states, the tensile property of collagen was reduced by 99% under wet conditions. Our results demonstrate in vitro and in vivo that PCL/PLGA/β-TCP membranes have similar levels of biocompatibility and bone regeneration as collagen membranes. In particular, considering that GBR is always applied to a wet environment (e.g. blood, saliva), we demonstrated that PCL/PLGA/β-TCP membranes maintained their form more reliably than collagen membranes in a wet setting, confirming their appropriateness as a GBR membrane.

  7. Concomitant presentation of collagenous sprue and HFE hemochromatosis. (United States)

    Parisian, Keely R; Plesec, Thomas P; Fairbanks, Kyrsten D; Tavill, Anthony S; Shen, Bo


    Collagenous sprue (CS) is a progressive malabsorptive disorder characterized by collagen deposition beneath the basement membrane of small bowel epithelium in refractory celiac sprue. CS is a pathologically distinct entity from celiac disease, despite a similar clinical presentation. The etiology of CS is unclear, although there are speculations that CS and celiac disease may share similar pathogenetic pathways. On the other hand, HFE hemochromatosis (HH) is a distinct disease entity. Celiac disease and HH are common HLA-associated genetic disorders in Northern European populations. There are a few case reports linking celiac disease and HH. We present a patient diagnosed with concurrent CS and HH.

  8. Cell cytoskeletal changes effected by static compressive stress lead to changes in the contractile properties of tissue regenerative collagen membranes

    Directory of Open Access Journals (Sweden)

    K Gellynck


    Full Text Available Static compressive stress can influence the matrix, which subsequently affects cell behaviour and the cell’s ability to further transform the matrix. This study aimed to assess response to static compressive stress at different stages of osteoblast differentiation and assess the cell cytoskeleton’s role as a conduit of matrix-derived stimuli. Mouse bone marrow mesenchymal stem cells (MSCs (D1 ORL UVA, osteoblastic cells (MC3T3-E1 and post-osteoblast/pre-osteocyte-like cells (MLO-A5 were seeded in hydrated and compressed collagen gels. Contraction was quantified macroscopically, and cell morphology, survival, differentiation and mineralisation assessed using confocal microscopy, alamarBlue® assay, real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR and histological stains, respectively. Confocal microscopy demonstrated cell shape changes and favourable microfilament organisation with static compressive stress of the collagen matrix; furthermore, cell survival was greater compared to the hydrated gels. The stage of osteoblast differentiation determined the degree of matrix contraction, with MSCs demonstrating the greatest amount. Introduction of microfilament disrupting inhibitors confirmed that pre-stress and tensegrity forces were under the influence of gel density, and there was increased survival and differentiation of the cells within the compressed collagen compared to the hydrated collagen. There was also relative stiffening and differentiation with time of the compressed cell-seeded collagen, allowing for greater manipulation. In conclusion, the combined collagen chemistry and increased density of the microenvironment can promote upregulation of osteogenic genes and mineralisation; MSCs can facilitate matrix contraction to form an engineered membrane with the potential to serve as a ‘pseudo-periosteum’ in the regeneration of bone defects.

  9. Crucial Role of Mesangial Cell-derived Connective Tissue Growth Factor in a Mouse Model of Anti-Glomerular Basement Membrane Glomerulonephritis (United States)

    Toda, Naohiro; Mori, Kiyoshi; Kasahara, Masato; Ishii, Akira; Koga, Kenichi; Ohno, Shoko; Mori, Keita P.; Kato, Yukiko; Osaki, Keisuke; Kuwabara, Takashige; Kojima, Katsutoshi; Taura, Daisuke; Sone, Masakatsu; Matsusaka, Taiji; Nakao, Kazuwa; Mukoyama, Masashi; Yanagita, Motoko; Yokoi, Hideki


    Connective tissue growth factor (CTGF) coordinates the signaling of growth factors and promotes fibrosis. Neonatal death of systemic CTGF knockout (KO) mice has hampered analysis of CTGF in adult renal diseases. We established 3 types of CTGF conditional KO (cKO) mice to investigate a role and source of CTGF in anti-glomerular basement membrane (GBM) glomerulonephritis. Tamoxifen-inducible systemic CTGF (Rosa-CTGF) cKO mice exhibited reduced proteinuria with ameliorated crescent formation and mesangial expansion in anti-GBM nephritis after induction. Although CTGF is expressed by podocytes at basal levels, podocyte-specific CTGF (pod-CTGF) cKO mice showed no improvement in renal injury. In contrast, PDGFRα promoter-driven CTGF (Pdgfra-CTGF) cKO mice, which predominantly lack CTGF expression by mesangial cells, exhibited reduced proteinuria with ameliorated histological changes. Glomerular macrophage accumulation, expression of Adgre1 and Ccl2, and ratio of M1/M2 macrophages were all reduced both in Rosa-CTGF cKO and Pdgfra-CTGF cKO mice, but not in pod-CTGF cKO mice. TGF-β1-stimulated Ccl2 upregulation in mesangial cells and macrophage adhesion to activated mesangial cells were decreased by reduction of CTGF. These results reveal a novel mechanism of macrophage migration into glomeruli with nephritis mediated by CTGF derived from mesangial cells, implicating the therapeutic potential of CTGF inhibition in glomerulonephritis. PMID:28191821

  10. Collagen binding specificity of the discoidin domain receptors: binding sites on collagens II and III and molecular determinants for collagen IV recognition by DDR1. (United States)

    Xu, Huifang; Raynal, Nicolas; Stathopoulos, Stavros; Myllyharju, Johanna; Farndale, Richard W; Leitinger, Birgit


    The discoidin domain receptors, DDR1 and DDR2 are cell surface receptor tyrosine kinases that are activated by triple-helical collagen. While normal DDR signalling regulates fundamental cellular processes, aberrant DDR signalling is associated with several human diseases. We previously identified GVMGFO (O is hydroxyproline) as a major DDR2 binding site in collagens I-III, and located two additional DDR2 binding sites in collagen II. Here we extend these studies to the homologous DDR1 and the identification of DDR binding sites on collagen III. Using sets of overlapping triple-helical peptides, the Collagen II and Collagen III Toolkits, we located several DDR2 binding sites on both collagens. The interaction of DDR1 with Toolkit peptides was more restricted, with DDR1 mainly binding to peptides containing the GVMGFO motif. Triple-helical peptides containing the GVMGFO motif induced DDR1 transmembrane signalling, and DDR1 binding and receptor activation occurred with the same amino acid requirements as previously defined for DDR2. While both DDRs exhibit the same specificity for binding the GVMGFO motif, which is present only in fibrillar collagens, the two receptors display distinct preferences for certain non-fibrillar collagens, with the basement membrane collagen IV being exclusively recognised by DDR1. Based on our recent crystal structure of a DDR2-collagen complex, we designed mutations to identify the molecular determinants for DDR1 binding to collagen IV. By replacing five amino acids in DDR2 with the corresponding DDR1 residues we were able to create a DDR2 construct that could function as a collagen IV receptor.

  11. A two-dimensional model of the colonic crypt accounting for the role of the basement membrane and pericryptal fibroblast sheath.

    Directory of Open Access Journals (Sweden)

    Sara-Jane Dunn

    Full Text Available The role of the basement membrane is vital in maintaining the integrity and structure of an epithelial layer, acting as both a mechanical support and forming the physical interface between epithelial cells and the surrounding connective tissue. The function of this membrane is explored here in the context of the epithelial monolayer that lines the colonic crypt, test-tube shaped invaginations that punctuate the lining of the intestine and coordinate a regular turnover of cells to replenish the epithelial layer every few days. To investigate the consequence of genetic mutations that perturb the system dynamics and can lead to colorectal cancer, it must be possible to track the emerging tissue level changes that arise in the crypt. To that end, a theoretical crypt model with a realistic, deformable geometry is required. A new discrete crypt model is presented, which focuses on the interaction between cell- and tissue-level behaviour, while incorporating key subcellular components. The model contains a novel description of the role of the surrounding tissue and musculature, based upon experimental observations of the tissue structure of the crypt, which are also reported. A two-dimensional (2D cross-sectional geometry is considered, and the shape of the crypt is allowed to evolve and deform. Simulation results reveal how the shape of the crypt may contribute mechanically to the asymmetric division events typically associated with the stem cells at the base. The model predicts that epithelial cell migration may arise due to feedback between cell loss at the crypt collar and density-dependent cell division, an hypothesis which can be investigated in a wet lab. This work forms the basis for investigation of the deformation of the crypt structure that can occur due to proliferation of cells exhibiting mutant phenotypes, experiments that would not be possible in vivo or in vitro.

  12. Biological Evaluation (In Vitro and In Vivo) of Bilayered Collagenous Coated (Nano Electrospun and Solid Wall) Chitosan Membrane for Periodontal Guided Bone Regeneration. (United States)

    Lotfi, Ghogha; Shokrgozar, Mohammad Ali; Mofid, Rasoul; Abbas, Fatemeh Mashhadi; Ghanavati, Farzin; Baghban, Alireza Akbarzadeh; Yavari, Seyedeh Kimia; Pajoumshariati, Seyedramin


    The application of barrier membranes in guided bone regeneration (GBR) has become a commonly used surgical technique in periodontal research. The objectives of this study were to evaluate the in vitro biocompatibility and osteogenic differentiation of mesenchymal stem cells (MSCs) on two different collagenous coatings (nano electrospun fibrous vs. solid wall) of bilayered collagen/chitosan membrane and their histological evaluation on bone regeneration in rabbit calvarial defects. It was found that chitosan-nano electrospun collagen (CNC) membranes had higher proliferation/metabolic activity compared to the chitosan-collagen (CC) and pristine chitosan membranes. The qRT-PCR analysis demonstrated the CNC membranes induced significant expression of osteogenic genes (Osteocalcin, RUNX2 and Col-α1) in MSCs. Moreover, higher calcium content and alkaline phosphatase activity of MSCs were observed compared to the other groups. Histologic and histomorphometric evaluations were performed on the uncovered (negative control) as well as covered calvarial defects of ten adult white rabbits with different membranes (CNC, CC, BioGide (BG, positive control)) at 1 and 2 months after surgery. More bone formation was detected in the defects covered with CNC and BG membranes than those covered by CC and the negative control. No inflammation and residual biomaterial particles were observed on the membrane surface or in the surrounding tissues in the surgical areas. These results suggest that bilayer CNC membrane can have the potential for use as a GBR membrane material facilitating bone formation.

  13. A comparative evaluation of bovine-derived xenograft (Bio-Oss Collagen) and type I collagen membrane (Bio-Gide) with bovine-derived xenograft (Bio-Oss Collagen) and fibrin fibronectin sealing system (TISSEEL) in the treatment of intrabony defects: A clinico-radiographic study


    Palachur, Deepthi; K V Prabhakara Rao; Murthy, K. Raja V.; Kishore, D. Trinath; Reddy, M. Narendra; Bhupathi, Anitha


    Background and Objectives: The purpose of this study was to compare the efficacy of bovine-derived xenograft (Bio-Oss Collagen) and Type I collagen membrane (Bio-Gide) with bovine-derived xenograft (Bio-Oss Collagen) and fibrin fibronectin sealing system (TISSEEL) in the treatment of periodontal infrabony defects. Materials and Methods: Fourteen healthy patients in the age range of 20 to 60 years, showing bilateral or contralateral infrabony defects were selected. The defects were assigned ra...

  14. Relationship between the changes in ischemia/reperfusion cerebro-microvessel basement membrane injury and gelatinase system in senile rat%老龄大鼠脑缺血/再灌注致脑微血管基底膜损伤的变化及与明胶酶系的关系

    Institute of Scientific and Technical Information of China (English)

    李建生; 刘轲; 刘敬霞; 王明航; 赵跃武; 刘正国


    Objective To study the relationship of cerebro-microvessel basement membrane injury and gelatinase system after cerebral ischemia/reperfusion(I/R)in aged rats.Methods Cerebral I/R injury model was reproduced by intraluminal silk ligature thrombosis of the middle cerebral artery occlusion (MCAO).Rats were divided randomly into sham control and I/R groups in young rats Cischemia 3 hours (13 h)and reperfusion 6 hours(I/R 6 h),12 hours(1/R 12 h),24 hours(I/R 24 h),3 days(1/R 3 d),6 days(I/R 6 d)],and sham control group and I/R group in aged rats(1 3 h and I/R 6 h,I/R 12 h,I/R 24 h,1/R 3 d,1/R 6 d).The change in cerebro-cortex microvessel basement membrane structure,basement membrane type Ⅳ collagen(Col Ⅳ)and laminin(LN)contents,matrix metalloproteinases(MMPs)and tissue inhibitor of metalloproteinases(TIMPs)expression in every group were determined with immunohis tochemical method and zymogram analysis.Results With the inerease in age,Col Ⅳ and LN contents of the microvessel basement membrane were increased,and MMP-2 and MMP-9 expressions were stronger.With prolongation of I/R.the degradation of microvessel basement membrane components(Col Ⅳ and LN)was positively correlated with the duration of cerebral I/R.MMP-2 expression was increased gradually,and MMP-9 and TIMP-1 expression increased at the beginning and decreased subsequently.Col Ⅳ(I 3 h,I/R 6 h,I/R 12 h),LN(1 3 h,I/R 6-24 h),MMP-2(I 3 h,1/R 6 h-6 d)and MMP-9(1 3 h,I/R 6-24 h)expression level in aged rats with I/R injury were higher,and TIMP-1(I/R 24 h)expression was lower than those in young rats(P<0.05 or P<0.01).In addition,changes in MMP-2 and MMP-9 contents as determined by zymogram analysis method coincided with their immunoexpression.Conclusion With the increase of age,alteration in membrane components of eerebro-microvessel basement membrane in rats is related with MMPs and TlMP.Cerebro-mierovesseI basement membrane injury is more serious in aged rats than that of young rats.Changes in

  15. Change in Long-Spacing Collagen in Descemet's Membrane of Diabetic Goto-Kakizaki Rats and Its Suppression by Antidiabetic Agents

    Directory of Open Access Journals (Sweden)

    Yoshihiro Akimoto


    Full Text Available We examined changes in the ultrastructure and localization of major extracellular matrix components, including 5 types of collagen (type I, III, IV, VI, and VIII, laminin, fibronectin, and heparan sulfate proteoglycan in Descemet's membrane of the cornea of diabetic GK rats. In the cornea of diabetic GK rats, more long-spacing collagen fibrils were observed in Descemet's membrane than in the membrane of the nondiabetic Wistar rats. Both GK and Wistar rats showed an age-dependent increase in the density of the long-spacing collagen. Immunoelectron microscopy showed that type VIII collagen was localized in the internodal region of the long-spacing collagen, which was not labelled by any of the other antibodies used. The antidiabetic agents nateglinide and glibenclamide significantly suppressed the formation of the long-spacing collagen in the diabetic rats. Long-spacing collagen would thus be a useful indicator for studying diabetic changes in the cornea and the effect of antidiabetic agents.

  16. Fras1, a basement membrane-associated protein mutated in Fraser syndrome, mediates both the initiation of the mammalian kidney and the integrity of renal glomeruli. (United States)

    Pitera, Jolanta E; Scambler, Peter J; Woolf, Adrian S


    FRAS1 is mutated in some individuals with Fraser syndrome (FS) and the encoded protein is expressed in embryonic epidermal cells, localizing in their basement membrane (BM). Syndactyly and cryptophthalmos in FS are sequelae of skin fragility but the bases for associated kidney malformations are unclear. We demonstrate that Fras1 is expressed in the branching ureteric bud (UB), and that renal agenesis occurs in homozygous Fras1 null mutant blebbed (bl) mice on a C57BL6J background. In vivo, the bl/bl bud fails to invade metanephric mesenchyme which undergoes involution, events replicated in organ culture. The expression of glial cell line-derived neurotrophic factor and growth-differentiation factor 11 was defective in bl/bl renal primordia in vivo, whereas, in culture, the addition of either growth factor restored bud invasion into the mesenchyme. Mutant primordia also showed deficient expression of Hoxd11 and Six2 transcription factors, whereas the activity of bone morphogenetic protein 4, an anti-branching molecule, was upregulated. In wild types, Fras1 was also expressed by nascent nephrons. Foetal glomerular podocytes expressed Fras1 transcripts and Fras1 immunolocalized in a glomerular BM-like pattern. On a mixed background, bl mutants, and also compound mutants for bl and my, another bleb strain, sometimes survive into adulthood. These mice have two kidneys, which contain subsets of glomeruli with perturbed nephrin, podocin, integrin alpha3 and fibronectin expression. Thus, Fras1 protein coats branching UB epithelia and is strikingly upregulated in the nephron lineage after mesenchymal/epithelial transition. Fras1 deficiency causes defective interactions between the bud and mesenchyme, correlating with disturbed expression of key nephrogenic molecules. Furthermore, Fras1 may also be required for the formation of normal glomeruli.

  17. Poliomyelitis in MuLV-infected ICR-SCID mice after injection of basement membrane matrix contaminated with lactate dehydrogenase-elevating virus. (United States)

    Carlson Scholz, Jodi A; Garg, Rohit; Compton, Susan R; Allore, Heather G; Zeiss, Caroline J; Uchio, Edward M


    The arterivirus lactate dehydrogenase-elevating virus (LDV) causes life-long viremia in mice. Although LDV infection generally does not cause disease, infected mice that are homozygous for the Fv1(n) allele are prone to develop poliomyelitis when immunosuppressed, a condition known as age-dependent poliomyelitis. The development of age-dependent poliomyelitis requires coinfection with endogenous murine leukemia virus. Even though LDV is a common contaminant of transplantable tumors, clinical signs of poliomyelitis after inadvertent exposure to LDV have not been described in recent literature. In addition, LDV-induced poliomyelitis has not been reported in SCID or ICR mice. Here we describe the occurrence of poliomyelitis in ICR-SCID mice resulting from injection of LDV-contaminated basement membrane matrix. After exposure to LDV, a subset of mice presented with clinical signs including paresis, which was associated with atrophy of the hindlimb musculature, and tachypnea; in addition, some mice died suddenly with or without premonitory signs. Mice presenting within the first 6 mo after infection had regions of spongiosis, neuronal necrosis and astrocytosis of the ventral spinal cord, and less commonly, brainstem. Axonal degeneration of ventral roots prevailed in more chronically infected mice. LDV was identified by RT-PCR in 12 of 15 mice with typical neuropathology; positive antiLDV immunolabeling was identified in all PCR-positive animals (n = 7) tested. Three of 8 mice with neuropathology but no clinical signs were LDV negative by RT-PCR. RT-PCR yielded murine leukemia virus in spinal cords of all mice tested, regardless of clinical presentation or neuropathology.

  18. Nonautonomous Roles of MAB-5/Hox and the Secreted Basement Membrane Molecule SPON-1/F-Spondin in Caenorhabditis elegans Neuronal Migration. (United States)

    Josephson, Matthew P; Miltner, Adam M; Lundquist, Erik A


    Nervous system development and circuit formation requires neurons to migrate from their birthplaces to specific destinations.Migrating neurons detect extracellular cues that provide guidance information. In Caenorhabditis elegans, the Q right (QR) and Q left (QL) neuroblast descendants migrate long distances in opposite directions. The Hox gene lin-39 cell autonomously promotes anterior QR descendant migration, and mab-5/Hox cell autonomously promotes posterior QL descendant migration. Here we describe a nonautonomous role of mab-5 in regulating both QR and QL descendant migrations, a role masked by redundancy with lin-39 A third Hox gene, egl-5/Abdominal-B, also likely nonautonomously regulates Q descendant migrations. In the lin-39 mab-5 egl-5 triple mutant, little if any QR and QL descendant migration occurs. In addition to well-described roles of lin-39 and mab-5 in the Q descendants, our results suggest that lin-39, mab-5, and egl-5 might also pattern the posterior region of the animal for Q descendant migration. Previous studies showed that the spon-1 gene might be a target of MAB-5 in Q descendant migration. spon-1 encodes a secreted basement membrane molecule similar to vertebrate F-spondin. Here we show that spon-1 acts nonautonomously to control Q descendant migration, and might function as a permissive rather than instructive signal for cell migration. We find that increased levels of MAB-5 in body wall muscle (BWM) can drive the spon-1 promoter adjacent to the Q cells, and loss of spon-1 suppresses mab-5 gain of function. Thus, MAB-5 might nonautonomously control Q descendant migrations by patterning the posterior region of the animal to which Q cells respond. spon-1 expression from BWMs might be part of the posterior patterning necessary for directed Q descendant migration.

  19. Comparative analysis of the noncollagenous NC1 domain of type IV collagen: identification of structural features important for assembly, function, and pathogenesis.


    Netzer, K. O.; Suzuki, K; Itoh, Y.; Hudson, B.G.; Khalifah, R. G.


    Type IV collagen alpha1-alpha6 chains have important roles in the assembly of basement membranes and are implicated in the pathogenesis of Goodpasture syndrome, an autoimmune disorder, and Alport syndrome, a hereditary renal disease. We report comparative sequence analyses and structural predictions of the noncollagenous C-terminal globular NC1 domain (28 sequences). The inferred tree verified that type IV collagen sequences fall into two groups, alpha1-like and alpha2-like, and suggested tha...

  20. Mechanical properties of collagen membranes modified with pores--are they still sufficient for orbital floor reconstruction? (United States)

    Birkenfeld, F; Flörke, C; Behrens, E; Rohnen, M; Kern, M; Gassling, V; Wiltfang, J


    Adequate mechanical strength is essential for materials used to reconstruct the orbital floor, and collagen membranes have recently been suggested for the repair of isolated fractures of the orbital floor. However, their mechanical properties after modification with pores for increased drainage of blood into the sinus have not been sufficiently investigated. We have tested the mechanical resistance of polydioxanone foils (PDS) to distortion and compared it with that of 3 resorbable collagen membranes (Smartbrane(®), Bio-Gide(®), and Creos(®)) in mint condition and when artificially aged (3 weeks, 6 weeks, and 8 weeks) after modification with pores (diameter 2mm) in a standard configuration (n=12 in each group). PDS and Creos(®) had comparable initial values for mechanical resistance of about 2.3N/mm(2), and Bio-Gide(®) and Smartbrane(®) had about 20% and 80% lower initial mechanical resistance, respectively. All materials tested had lower values after artificial ageing. After eight weeks of ageing, PDS lost about 99% of its initial mechanical resistance, Creos(®) about 66%, Bio-Gide(®) about 30%, and Smartbrane(®) about 95%. After 3 weeks the mechanical resistance in all groups was significantly less than the initial values (p=0.05), but there was no difference between samples aged artificially for 6 compared with 8 weeks. The mechanical resistance of the tested materials was not influenced by the presence of pores in a standard configuration and was in the appropriate range for moderate fractures of the orbital floor. We recommend further clinical investigations of collagen membranes modified with pores.

  1. Induction of rat yolk sac carcinomas with consistent pattern of laminin, entactin, and type IV collagen biosynthesis

    DEFF Research Database (Denmark)

    Wewer, U


    Twenty-four yolk sac carcinomas in Lewis rats were experimentally induced by puncturing the pregnant uterine wall with a hypodermic needle at day 9-13 of gestation. Morphologically, the tumours were composed of parietal- and visceral yolk sac carcinoma and to a less degree of trophoblastic giant ...... to their biosynthesis of the basement membrane components laminin, entactin, and type IV collagen. This model system offers a simple approach to inducing rat yolk sac carcinomas for further morphological and biochemical characterization of the basement membrane....

  2. Structure and properties of collagen vitrigel membranes for ocular repair and regeneration applications. (United States)

    Calderón-Colón, Xiomara; Xia, Zhiyong; Breidenich, Jennifer L; Mulreany, Daniel G; Guo, Qiongyu; Uy, Oscar M; Tiffany, Jason E; Freund, David E; McCally, Russell L; Schein, Oliver D; Elisseeff, Jennifer H; Trexler, Morgana M


    The frequency of ocular injuries on the battlefield has been steadily increasing during recent conflicts. Combat-related eye injuries are difficult to treat and solutions requiring donor tissue are not ideal and are often not readily available. Collagen vitrigels have previously been developed for corneal reconstruction, but increased transparency and mechanical strength are desired for improved vision and ease of handling. In this study, by systematically varying vitrification temperature, relative humidity and time, the collagen vitrigel synthesis conditions were optimized to yield the best combination of high transparency and high mechanical strength. Optical, mechanical, and thermal properties were characterized for each set of conditions to evaluate the effects of the vitrification parameters on material properties. Changes in denaturing temperature and collagen fibril morphology were evaluated to correlate properties with structure. Collagen vitrigels with transmittance up to 90%, tensile strength up to 12 MPa, and denaturing temperatures that significantly exceed the eye/body temperature have been synthesized at 40 °C and 40% relative humidity for one week. This optimal set of conditions enabled improvements of 100% in tensile strength and 11% in transmittance, compared to the previously developed collagen vitrigels.

  3. Bone density of defects treated with lyophilized amniotic membrane versus collagen membrane: a tomographic and histomorfogenic study in rabbit´s femur

    Directory of Open Access Journals (Sweden)

    Liz Katty Ríos


    Full Text Available ABSTRACT The aim of this study was to compare the bone density of bone defects treated with lyophilizated amniotic membrane (LAM and collagen Membrane (CM, at 3 and 5 weeks. Two bone defects of 4 mm in diameter and 6 mm deep were created in left distal femoral diaphysis of New Zealand rabbits (n = 12. The animals were randomly divided into 2 groups. One of the defects was covered with lyophilized amniotic membrane (Rosa Chambergo Tissue Bank/National Institute of Child Health-IPEN, Lima, Peru or collagen Membrane (Dentium Co, Seoul, Korea. The second was left uncovered (NC. The rabbits were killed after 3 and 5 weeks (3 rabbits/period. The results showed a high bone density and repair of the defect by new bone. The tomographic study revealed that the bone density of the defects treated with LAM at 3 weeks was equivalent to the density obtained with CM and higher density compared with NC (p 0.05. The results show that lyophilizated amniotic membrane provides bone density equal or higher to the collagen membrane. RESUMEN El propósito de este estudio fue comparar la densidad ósea (DO de defectos óseos tratados con membrana amniótica liofilizada (MAL y membrana de colágeno (MC, a las 3 y 5 semanas. Se crearon dos defectos óseos, de 4 mm de diámetro y 6 mm de profundidad, en la diáfisis femoral distal izquierda de conejos Nueva Zelanda (n=12. Los animales fueron divididos aleatoriamente en 2 grupos. Uno de los defectos fue cubierto con membrana amniótica liofilizada (Banco de tejidos Rosa Chambergo/INSN-IPEN, Lima, Perú o membrana de colágeno (Dentium Co, Seoul, Korea. El segundo se dejó sin cubrir (NC. Los conejos fueron sacrificados después de 3 y 5 semanas (3 conejos/periodo. Los resultados mostraron una alta DO y reparación del defecto por hueso neoformado. El estudio tomográfico reveló que la DO de los defectos tratados con MAL a las 3 semanas fue comparable a la densidad obtenida con MC y mayor comparado con la densidad de NC (p

  4. Healing of intra-bony defects following treatment with a composite bovine-derived xenograft (Bio-Oss Collagen) in combination with a collagen membrane (Bio-Gide PERIO).

    NARCIS (Netherlands)

    Sculean, A.; Chiantella, G.C.; Windisch, P.; Arweiler, N.B.; Brecx, M.; Gera, I.


    AIM: The purpose of the present study was to compare clinically the treatment of deep intra-bony defects with a combination of a composite bovine-derived xenograft (BDX Coll) and a bioresorbable collagen membrane [guided tissue regeneration (GTR)] to access flap surgery only. METHODS: Thirty-two pat

  5. Collagen-Coated Polytetrafluoroethane Membrane Inserts Enhances Chondrogenic Differentiation of Human Cord Blood Multi-Lineage Progenitor Cells

    DEFF Research Database (Denmark)

    Munir, Samir; Søballe, Kjeld; Ulrich-Vinther, Michael;

    Background: Articular chondrocytes and bone marrow-derived multipotent mesenchymal stromal cells (MSCs) are the favoured cells for cartilage tissue engineering. Umbilical cord blood has proven an alternative source of MSCs and moreover they may be more potent chondroprogenitor cells than bonemarrow...... MSCs. Purpose / Aim of Study: Multilineage progenitor cells (MLPCs) are clonal cord blood-derived MSCs and may therefore provide a cell source with more reproducible outcomes compared to heterogeneous primary MSC cultures. Materials and Methods: We evaluated the chondrogenic potency of MLPCs...... in standard micromass pellet system, layered on calcium polyphosphate (CPP), and on semi-permeable polytetrafluoroethane membranes with and without collagen type I, II or IV pre-coating. Findings / Results: The MPLC cell line used in this study possessed poor chondrogenic potency overall, but membrane...

  6. How to Study Basement Membrane Stiffness as a Biophysical Trigger in Prostate Cancer and Other Age-related Pathologies or Metabolic Diseases (United States)

    Rodriguez-Teja, Mercedes; Breit, Claudia; Clarke, Mitchell; Talar, Kamil; Wang, Kai; Mohammad, Mohammad A.; Pickwell, Sage; Etchandy, Guillermina; Stasiuk, Graeme J.; Sturge, Justin


    Here we describe a protocol that can be used to study the biophysical microenvironment related to increased thickness and stiffness of the basement membrane (BM) during age-related pathologies and metabolic disorders (e.g. cancer, diabetes, microvascular disease, retinopathy, nephropathy and neuropathy). The premise of the model is non-enzymatic crosslinking of reconstituted BM (rBM) matrix by treatment with glycolaldehyde (GLA) to promote advanced glycation endproduct (AGE) generation via the Maillard reaction. Examples of laboratory techniques that can be used to confirm AGE generation, non-enzymatic crosslinking and increased stiffness in GLA treated rBM are outlined. These include preparation of native rBM (treated with phosphate-buffered saline, PBS) and stiff rBM (treated with GLA) for determination of: its AGE content by photometric analysis and immunofluorescent microscopy, its non-enzymatic crosslinking by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) as well as confocal microscopy, and its increased stiffness using rheometry. The procedure described here can be used to increase the rigidity (elastic moduli, E) of rBM up to 3.2-fold, consistent with measurements made in healthy versus diseased human prostate tissue. To recreate the biophysical microenvironment associated with the aging and diseased prostate gland three prostate cell types were introduced on to native rBM and stiff rBM: RWPE-1, prostate epithelial cells (PECs) derived from a normal prostate gland; BPH-1, PECs derived from a prostate gland affected by benign prostatic hyperplasia (BPH); and PC3, metastatic cells derived from a secondary bone tumor originating from prostate cancer. Multiple parameters can be measured, including the size, shape and invasive characteristics of the 3D glandular acini formed by RWPE-1 and BPH-1 on native versus stiff rBM, and average cell length, migratory velocity and persistence of cell movement of 3D spheroids formed by PC3 cells under

  7. Structure-function analysis of peroxidasin provides insight into the mechanism of collagen IV crosslinking. (United States)

    Lázár, Enikő; Péterfi, Zalán; Sirokmány, Gábor; Kovács, Hajnal A; Klement, Eva; Medzihradszky, Katalin F; Geiszt, Miklós


    Basement membranes provide structural support and convey regulatory signals to cells in diverse tissues. Assembly of collagen IV into a sheet-like network is a fundamental mechanism during the formation of basement membranes. Peroxidasin (PXDN) was recently described to catalyze crosslinking of collagen IV through the formation of sulfilimine bonds. Despite the significance of this pathway in tissue genesis, our understanding of PXDN function is far from complete. In this work we demonstrate that collagen IV crosslinking is a physiological function of mammalian PXDN. Moreover, we carried out structure-function analysis of PXDN to gain a better insight into its role in collagen IV synthesis. We identify conserved cysteines in PXDN that mediate the oligomerization of the protein into a trimeric complex. We also demonstrate that oligomerization is not an absolute requirement for enzymatic activity, but optimal collagen IV coupling is only catalyzed by the PXDN trimers. Localization experiments of different PXDN mutants in two different cell models revealed that PXDN oligomers, but not monomers, adhere on the cell surface in "hot spots," which represent previously unknown locations of collagen IV crosslinking.

  8. Cell-collagen interactions: the use of peptide Toolkits to investigate collagen-receptor interactions. (United States)

    Farndale, Richard W; Lisman, Ton; Bihan, Dominique; Hamaia, Samir; Smerling, Christiane S; Pugh, Nicholas; Konitsiotis, Antonios; Leitinger, Birgit; de Groot, Philip G; Jarvis, Gavin E; Raynal, Nicolas


    Fibrillar collagens provide the most fundamental platform in the vertebrate organism for the attachment of cells and matrix molecules. We have identified specific sites in collagens to which cells can attach, either directly or through protein intermediaries. Using Toolkits of triple-helical peptides, each peptide comprising 27 residues of collagen primary sequence and overlapping with its neighbours by nine amino acids, we have mapped the binding of receptors and other proteins on to collagens II or III. Integrin alpha2beta1 binds to several GXX'GER motifs within the collagens, the affinities of which differ sufficiently to control cell adhesion and migration independently of the cellular regulation of the integrin. The platelet receptor, Gp (glycoprotein) VI binds well to GPO (where O is hydroxyproline)-containing model peptides, but to very few Toolkit peptides, suggesting that sequence in addition to GPO triplets is important in defining GpVI binding. The Toolkits have been applied to the plasma protein vWF (von Willebrand factor), which binds to only a single sequence, identified by truncation and amino acid substitution within Toolkit peptides, as GXRGQOGVMGFO in collagens II and III. Intriguingly, the receptor tyrosine kinase, DDR2 (discoidin domain receptor 2) recognizes three sites in collagen II, including its vWF-binding site, although the amino acids that support the interaction differ slightly within this motif. Furthermore, the secreted protein BM-40 (basement membrane protein 40) also binds well to this same region. Thus the availability of extracellular collagen-binding proteins may be important in regulating and facilitating direct collagen-receptor interaction.

  9. Guided Bone Regeneration in Long-Bone Defects with a Structural Hydroxyapatite Graft and Collagen Membrane (United States)


    is currently limited by high costs and associated complications, including life-threatening cervical swelling4 and ectopic bone formation.5...fibrous ver- sus collagen zones. Biomechanical evaluation Immediately after euthanasia, eight excised radii and ul- nae per group (seven for the wrap group...serve as controls for the biomechanical evaluation. The specimens were tested to flexural failure in a 4-point bending configuration with 10- mm spacing

  10. Comparative study of chitosan/fibroin-hydroxyapatite and collagen membranes for guided bone regeneration in rat calvarial defects: micro-computed tomography analysis. (United States)

    Song, Jae Min; Shin, Sang Hun; Kim, Yong Deok; Lee, Jae Yeol; Baek, Young Jae; Yoon, Sang Yong; Kim, Hong Sung


    This study aimed to utilize micro-computed tomography (micro-CT) analysis to compare new bone formation in rat calvarial defects using chitosan/fibroin-hydroxyapatite (CFB-HAP) or collagen (Bio-Gide) membranes. Fifty-four (54) rats were studied. A circular bony defect (8 mm diameter) was formed in the centre of the calvaria using a trephine bur. The CFB-HAP membrane was prepared by thermally induced phase separation. In the experimental group (n=18), the CFB-HAP membrane was used to cover the bony defect, and in the control group (n=18), a resorbable collagen membrane (Bio-Gide) was used. In the negative control group (n=18), no membrane was used. In each group, six animals were euthanized at 2, 4 and 8 weeks after surgery. The specimens were then analysed using micro-CT. There were significant differences in bone volume (BV) and bone mineral density (BMD) (Pmembrane groups. However, there were no significant differences between the CFB-HAP group and the collagen group. We concluded that the CFB-HAP membrane has significant potential as a guided bone regeneration (GBR) membrane.

  11. Measurement of the quadratic hyperpolarizability of the collagen triple helix and application to second harmonic imaging of natural and biomimetic collagenous tissues (United States)

    Deniset-Besseau, A.; Strupler, M.; Duboisset, J.; De Sa Peixoto, P.; Benichou, E.; Fligny, C.; Tharaux, P.-L.; Mosser, G.; Brevet, P.-F.; Schanne-Klein, M.-C.


    Collagen is a major protein of the extracellular matrix that is characterized by triple helical domains. It plays a central role in the formation of fibrillar and microfibrillar networks, basement membranes, as well as other structures of the connective tissue. Remarkably, fibrillar collagen exhibits efficient Second Harmonic Generation (SHG) so that SHG microscopy proved to be a sensitive tool to probe the three-dimensional architecture of fibrillar collagen and to assess the progression of fibrotic pathologies. We obtained sensitive and reproducible measurements of the fibrosis extent, but we needed quantitative data at the molecular level to further process SHG images. We therefore performed Hyper- Rayleigh Scattering (HRS) experiments and measured a second order hyperpolarisability of 1.25 10-27 esu for rat-tail type I collagen. This value is surprisingly large considering that collagen presents no strong harmonophore in its aminoacid sequence. In order to get insight into the physical origin of this nonlinear process, we performed HRS measurements after denaturation of the collagen triple helix and for a collagen-like short model peptide [(Pro-Pro- Gly)10]3. It showed that the collagen large nonlinear response originates in the tight alignment of a large number of weakly efficient harmonophores, presumably the peptide bonds, resulting in a coherent amplification of the nonlinear signal along the triple helix. To illustrate this mechanism, we successfully recorded SHG images in collagenous biomimetic matrices.

  12. Socket preservation using bovine bone mineral and collagen membrane: a randomized controlled clinical trial with histologic analysis. (United States)

    Cardaropoli, Daniele; Tamagnone, Lorenzo; Roffredo, Alessandro; Gaveglio, Lorena; Cardaropoli, Giuseppe


    After tooth extraction, varying amounts of bone resorption occur because of qualitative and quantitative changes at the edentulous site of the alveolar process. The aims of this randomized controlled clinical trial were (1) to compare the postextraction changes in residual ridge dimensions during spontaneous healing with those during socket preservation, (2) to analyze the histologic and histomorphometric aspects of the grafted sockets, and (3) to compare probing procket depth (PPD) and clinical attachment level (CAL) changes at teeth adjacent to extraction sites. Forty-eight teeth were extracted from 41 patients referred for extraction of 1 or more maxillary or mandibular premolars or molars. The edentulous sites were randomly assigned to the control (EXT, extraction alone) or experimental groups (SP, extraction and socket preservation). In the SP group, the sockets were filled with bovine bone mineral and covered with porcine collagen membrane. At baseline and after 4 months, PPD, gingival recession (REC), and CAL were measured at teeth adjacent to the edentulous sites. The changes in ridge dimensions from baseline to 4 months were assessed on dental casts. At 4 months, bone was harvested from the grafted areas in the SP group and the edentulous areas in the EXT group. PPD, REC, and CAL were comparable between groups. However, from baseline to 4 months, the SP group showed significantly less reduction in ridge width (1.04 ± 1.08 mm vs 4.48 ± 0.65 mm, P collagen membrane considerably limits the amount of horizontal and vertical bone resorption when compared with extraction alone.

  13. Seismic basement in Poland (United States)

    Grad, Marek; Polkowski, Marcin


    The area of contact between Precambrian and Phanerozoic Europe in Poland has complicated structure of sedimentary cover and basement. The thinnest sedimentary cover in the Mazury-Belarus anteclize is only 0.3-1 km thick, increases to 7-8 km along the East European Craton margin, and 9-12 km in the Trans-European Suture Zone (TESZ). The Variscan domain is characterized by a 1- to 2-km-thick sedimentary cover, while the Carpathians are characterized by very thick sediments, up to c. 20 km. The map of the basement depth is created by combining data from geological boreholes with a set of regional seismic refraction profiles. These maps do not provide data about the basement depth in the central part of the TESZ and in the Carpathians. Therefore, the data set is supplemented by 32 models from deep seismic sounding profiles and a map of a high-resistivity (low-conductivity) layer from magnetotelluric soundings, identified as a basement. All of these data provide knowledge about the basement depth and of P-wave seismic velocities of the crystalline and consolidated type of basement for the whole area of Poland. Finally, the differentiation of the basement depth and velocity is discussed with respect to geophysical fields and the tectonic division of the area.

  14. Peptide- and collagen-based hydrogel substrates for in vitro culture of chick cochleae. (United States)

    Spencer, Nathaniel J; Cotanche, Douglas A; Klapperich, Catherine M


    The overall goal of this work is to improve the culture of the auditory organ of birds for the dual use of developing a hair cell regeneration model and charting a pathway to the eventual replacement of the hearing organ. In doing so, we develop a protocol for removing the auditory organ from its basement membrane in the inner ear, attach the organ to a series of artificial basement membranes, and conduct qualitative and quantitative analysis of how cell morphology, viability and function change with time. Native matrix cultures, where the epithelium was floating in media with the basement membrane and accessory structures attached, were used as a basis of comparison. PuraMatrix, collagen I, collagen I/chondroitin-sulfate and Matrigel were chosen to encompass a diverse range of mechanical properties and macromolecule moieties. Surprisingly, we find that PuraMatrix outperformed the other matrices as a scaffold for sensory organ culture. PuraMatrix a self-assembled peptide hydrogel, is a biochemically specific culture substrate that contains none of the extracellular matrix (ECM) molecules and growth factors contained in the inner ear's basement membrane. Rheological measurements reveal that PuraMatrix may be a closer approximation to the stiffness of the soft tissue supporting the auditory organ. Cell density on the PuraMatrix substrate is comparable to that of the native matrix cultures, despite the absence of the basement membrane and accessory structures. Further studies show that PuraMatrix supports the culture of functional hair cells over a 72 h period, with a significant increase in the number of functional hair cells in comparison to the organ cultured without a matrix. This is the first example of adhesion of the adult auditory epithelium to a biomaterial for an extended period of time. With further optimization, this system will enable the performance of many novel biophysical and pharmacological studies involving hair cells and supporting cells.

  15. Enhancement of tendon–bone healing via the combination of biodegradable collagen-loaded nanofibrous membranes and a three-dimensional printed bone-anchoring bolt (United States)

    Chou, Ying-Chao; Yeh, Wen-Lin; Chao, Chien-Lin; Hsu, Yung-Heng; Yu, Yi-Hsun; Chen, Jan-Kan; Liu, Shih-Jung


    A composite biodegradable polymeric model was developed to enhance tendon graft healing. This model included a biodegradable polylactide (PLA) bolt as the bone anchor and a poly(D,L-lactide-co-glycolide) (PLGA) nanofibrous membrane embedded with collagen as a biomimic patch to promote tendon–bone interface integration. Degradation rate and compressive strength of the PLA bolt were measured after immersion in a buffer solution for 3 months. In vitro biochemical characteristics and the nanofibrous matrix were assessed using a water contact angle analyzer, pH meter, and tetrazolium reduction assay. In vivo efficacies of PLGA/collagen nanofibers and PLA bolts for tendon–bone healing were investigated on a rabbit bone tunnel model with histological and tendon pullout tests. The PLGA/collagen-blended nanofibrous membrane was a hydrophilic, stable, and biocompatible scaffold. The PLA bolt was durable for tendon–bone anchoring. Histology showed adequate biocompatibility of the PLA bolt on a medial cortex with progressive bone ingrowth and without tissue overreaction. PLGA nanofibers within the bone tunnel also decreased the tunnel enlargement phenomenon and enhanced tendon–bone integration. Composite polymers of the PLA bolt and PLGA/collagen nanofibrous membrane can effectively promote outcomes of tendon reconstruction in a rabbit model. The composite biodegradable polymeric system may be useful in humans for tendon reconstruction. PMID:27601901

  16. Comparative study of chitosan/fibroin–hydroxyapatite and collagen membranes for guided bone regeneration in rat calvarial defects: micro-computed tomography analysis


    Song, Jae Min; Shin, Sang Hun; Kim, Yong Deok; Lee, Jae Yeol; Baek, Young Jae; Yoon, Sang Yong; Kim, Hong Sung


    This study aimed to utilize micro-computed tomography (micro-CT) analysis to compare new bone formation in rat calvarial defects using chitosan/fibroin–hydroxyapatite (CFB–HAP) or collagen (Bio-Gide) membranes. Fifty-four (54) rats were studied. A circular bony defect (8 mm diameter) was formed in the centre of the calvaria using a trephine bur. The CFB–HAP membrane was prepared by thermally induced phase separation. In the experimental group (n=18), the CFB–HAP membrane was used to cover the...

  17. Abcc6 deficiency in the mouse leads to calcification of collagen fibers in Bruch's membrane

    NARCIS (Netherlands)

    Gorgels, T.G.; Teeling, P.; Meeldijk, J.D.; Nillesen, S.T.M.; Wal, A.C. van der; Kuppevelt, A.H.M.S.M. van; Bergen, A.A.B.


    Pseudoxanthoma elasticum (PXE) is a heritable disorder characterized by mineralization of connective tissue, which leads to pathology in eye, skin and blood vessels. The disease is caused by mutations in ABCC6. To learn more about PXE eye pathology, we analyzed Bruch's membrane (BM) of the eye of an

  18. Rapid Upregulation of Orai1 Abundance in the Plasma Membrane of Platelets Following Activation with Thrombin and Collagen Related Peptide

    Directory of Open Access Journals (Sweden)

    Guilai Liu


    Full Text Available Background: Blood platelets accomplish primary hemostasis following vascular injury and contribute to the orchestration of occlusive vascular disease. Platelets are activated by an increase of cytosolic Ca2+-activity ([Ca2+]i, which is accomplished by Ca2+-release from intracellular stores and subsequent store operated Ca2+ entry (SOCE through Ca2+ release activated Ca2+ channel moiety Orai1. Powerful activators of platelets include thrombin and collagen related peptide (CRP, which are in part effective by activation of small G- protein Rac1. The present study explored the influence of thrombin and CRP on Orai1 protein abundance and cytosolic Ca2+-activity ([Ca2+]i in platelets drawn from wild type mice. Methods: Orai1 protein surface abundance was quantified utilizing CF™488A conjugated antibodies, and [Ca2+]i was determined with Fluo3-fluorescence. Results: In resting platelets, Orai1 protein abundance and [Ca2+]i were low. Thrombin (0.02 U/ml and CRP (5ug/ml within 2 min increased [Ca2+]i and Orai1 protein abundance at the platelet surface. [Ca2+]i was further increased by Ca2+ ionophore ionomycin (1 µM and by store depletion with the sarcoendoplasmatic Ca2+ ATPase inhibitor thapsigargin (1 µM. However, Orai1 protein abundance at the platelet surface was not significantly affected by ionomycin and only slightly increased by thapsigargin. The effect of thrombin and CRP on Orai1 abundance and [Ca2+]i was significantly blunted by Rac1 inhibitor NSC23766 (50 µM. Conclusion: The increase of [Ca2+]i following stimulation of platelets with thrombin and collagen related peptide is potentiated by ultrarapid Rac1 sensitive translocation of Orai1 into the cell membrane.

  19. Biological role of prolyl 3-hydroxylation in type IV collagen. (United States)

    Pokidysheva, Elena; Boudko, Sergei; Vranka, Janice; Zientek, Keith; Maddox, Kerry; Moser, Markus; Fässler, Reinhard; Ware, Jerry; Bächinger, Hans Peter


    Collagens constitute nearly 30% of all proteins in our body. Type IV collagen is a major and crucial component of basement membranes. Collagen chains undergo several posttranslational modifications that are indispensable for proper collagen function. One of these modifications, prolyl 3-hydroxylation, is accomplished by a family of prolyl 3-hydroxylases (P3H1, P3H2, and P3H3). The present study shows that P3H2-null mice are embryonic-lethal by embryonic day 8.5. The mechanism of the unexpectedly early lethality involves the interaction of non-3-hydroxylated embryonic type IV collagen with the maternal platelet-specific glycoprotein VI (GPVI). This interaction results in maternal platelet aggregation, thrombosis of the maternal blood, and death of the embryo. The phenotype is completely rescued by producing double KOs of P3H2 and GPVI. Double nulls are viable and fertile. Under normal conditions, subendothelial collagens bear the GPVI-binding sites that initiate platelet aggregation upon blood exposure during injuries. In type IV collagen, these sites are normally 3-hydroxylated. Thus, prolyl 3-hydroxylation of type IV collagen has an important function preventing maternal platelet aggregation in response to the early developing embryo. A unique link between blood coagulation and the ECM is established. The newly described mechanism may elucidate some unexplained fetal losses in humans, where thrombosis is often observed at the maternal/fetal interface. Moreover, epigenetic silencing of P3H2 in breast cancers implies that the interaction between GPVI and non-3-hydroxylated type IV collagen might also play a role in the progression of malignant tumors and metastasis.

  20. Effects of collagen membranes enriched with in vitro-differentiated N1E-115 cells on rat sciatic nerve regeneration after end-to-end repair

    Directory of Open Access Journals (Sweden)

    Fornaro Michele


    Full Text Available Abstract Peripheral nerves possess the capacity of self-regeneration after traumatic injury but the extent of regeneration is often poor and may benefit from exogenous factors that enhance growth. The use of cellular systems is a rational approach for delivering neurotrophic factors at the nerve lesion site, and in the present study we investigated the effects of enwrapping the site of end-to-end rat sciatic nerve repair with an equine type III collagen membrane enriched or not with N1E-115 pre-differentiated neural cells. After neurotmesis, the sciatic nerve was repaired by end-to-end suture (End-to-End group, end-to-end suture enwrapped with an equine collagen type III membrane (End-to-EndMemb group; and end-to-end suture enwrapped with an equine collagen type III membrane previously covered with neural cells pre-differentiated in vitro from N1E-115 cells (End-to-EndMembCell group. Along the postoperative, motor and sensory functional recovery was evaluated using extensor postural thrust (EPT, withdrawal reflex latency (WRL and ankle kinematics. After 20 weeks animals were sacrificed and the repaired sciatic nerves were processed for histological and stereological analysis. Results showed that enwrapment of the rapair site with a collagen membrane, with or without neural cell enrichment, did not lead to any significant improvement in most of functional and stereological predictors of nerve regeneration that we have assessed, with the exception of EPT which recovered significantly better after neural cell enriched membrane employment. It can thus be concluded that this particular type of nerve tissue engineering approach has very limited effects on nerve regeneration after sciatic end-to-end nerve reconstruction in the rat.

  1. Morphometric Changes of the Socket after Site Preservation Using Nanobone and Collagen Membrane or Stypro Versus Extraction Alone

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    Salahi S


    Full Text Available Statement of Problem: The long-term success of a dental implant relies on implant osseointegration into native and viable bone, implant placement in an ideal position, and optimal hard and soft tissue contour. This requires the presence of sufficient alveolar bone volume, good alveolar ridge (Practically with no sign of atrophy and good surgical technique. Objectives: The aim of this randomized controlled clinical study was to evaluate morphometric changes after different alveolar ridge preservation procedures. Materials and Methods: In this study, 33 patients who had single-rooted premolar, which needed to be extracted, were recruited. Patients were randomly divided into 3 groups and after tooth extraction the following treatments were administered: in group A: NanoBone and a collagen membrane; in group B: NanoBone and Stypro; and in group C: natural healing. The following clinical parameters were evaluated at baseline and 6 months after the extraction: buccolingual width, midbuccal height (with the use of a custom made stent and width of keratinized gingiva. For data analysis, Paired t-test,one-way ANOVA and Tukey’s tests were used. Results: The average reduction in the buccolingual width, midbuccal height and keratinized gingiva was as follows: group A: 1.18±0.6, 0.64±0.92 and 3.45±1.75 mm; group B: 2.18±0.75, 0.73±0.78 and 4.73±0.9 mm; and group C: 1±0.89, 2.36±1.21 and 5±0.63 mm, respectively. Moreover, a significantly reduced resorption was found in both the buccolingual width and the width of keratinized gingiva in group A as compared to groups B and C (p<0.05. Conclusions: This study showed that the use of collagen membrane+Nano bone (group A can significantly reduce the horizontal resorption of the alveolar ridge and keratinized tissue more effectively than stypro+Nano bone (group B and blood clot alone and natural healing (group C.

  2. The membrane bound LRR lipoprotein Slr, and the cell wall-anchored M1 protein from Streptococcus pyogenes both interact with type I collagen.

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    Marta Bober

    Full Text Available Streptococcus pyogenes is an important human pathogen and surface structures allow it to adhere to, colonize and invade the human host. Proteins containing leucine rich repeats (LRR have been identified in mammals, viruses, archaea and several bacterial species. The LRRs are often involved in protein-protein interaction, are typically 20-30 amino acids long and the defining feature of the LRR motif is an 11-residue sequence LxxLxLxxNxL (x being any amino acid. The streptococcal leucine rich (Slr protein is a hypothetical lipoprotein that has been shown to be involved in virulence, but at present no ligands for Slr have been identified. We could establish that Slr is a membrane attached horseshoe shaped lipoprotein by homology modeling, signal peptidase II inhibition, electron microscopy (of bacteria and purified protein and immunoblotting. Based on our previous knowledge of LRR proteins we hypothesized that Slr could mediate binding to collagen. We could show by surface plasmon resonance that recombinant Slr and purified M1 protein bind with high affinity to collagen I. Isogenic slr mutant strain (MB1 and emm1 mutant strain (MC25 had reduced binding to collagen type I as shown by slot blot and surface plasmon resonance. Electron microscopy using gold labeled Slr showed multiple binding sites to collagen I, both to the monomeric and the fibrillar structure, and most binding occurred in the overlap region of the collagen I fibril. In conclusion, we show that Slr is an abundant membrane bound lipoprotein that is co-expressed on the surface with M1, and that both these proteins are involved in recruiting collagen type I to the bacterial surface. This underlines the importance of S. pyogenes interaction with extracellular matrix molecules, especially since both Slr and M1 have been shown to be virulence factors.

  3. Clinical and radiographical evaluation of a bioresorbable collagen membrane of fish origin in the treatment of periodontal intrabony defects: A preliminary study

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    B B Santosh Kumar


    Full Text Available Background: Recently, there has been interest in non-mammalian collagen sources such as fish collagen in periodontal regeneration. In the present study, collagen barrier membrane of fish origin was assessed in the treatment of periodontal intrabony defects. Materials and Methods: Ten systemically healthy chronic periodontitis patients having a paired osseous defect in the mandibular posterior teeth were selected and randomly assigned to receive a collagen membrane (test or open flap debridement (control in a split mouth design. Clinical parameters such as Plaque index, Gingival bleeding index, Probing pocket depth, Relative attachment level, and Recession were recorded at baseline, 3, 6, and at 9 months, while radiographic evaluation was done to assess alveolar crestal bone level and percentage of defect fill at 6 and 9 months using autoCAD 2007 software. Student′s t test (two-tailed, dependent was used to find the significance of study parameters on continuous scale. Significance was set at 5% level of significance. Wilcoxon signed rank test was used to find the significance of percentage change of defect fill. Results: The comparison between the two groups did not show any statistically significant differences in the parameters assessed (P > 0.05 but, within each group, clinical parameters showed statistically significant differences from baseline to 9 months (P < 0.05. Conclusion: Within the limits of the study, it can be inferred that no significant differences were found either by using collagen membrane of fish origin or open flap debridement in the treatment of periodontal intrabony defects.

  4. Enhancement of tendon–bone healing via the combination of biodegradable collagen-loaded nanofibrous membranes and a three-dimensional printed bone-anchoring bolt

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    Chou YC


    Full Text Available Ying-Chao Chou,1,2 Wen-Lin Yeh,2 Chien-Lin Chao,1 Yung-Heng Hsu,1,2 Yi-Hsun Yu,1,2 Jan-Kan Chen,3 Shih-Jung Liu1,2 1Department of Mechanical Engineering, Chang Gung University, 2Department of Orthopedic Surgery, Chang Gung Memorial Hospital, 3Department of Physiology and Pharmacology, Chang Gung University, Taoyuan, Taiwan Abstract: A composite biodegradable polymeric model was developed to enhance tendon graft healing. This model included a biodegradable polylactide (PLA bolt as the bone anchor and a poly(D,L-lactide-co-glycolide (PLGA nanofibrous membrane embedded with collagen as a biomimic patch to promote tendon–bone interface integration. Degradation rate and compressive strength of the PLA bolt were measured after immersion in a buffer solution for 3 months. In vitro biochemical characteristics and the nanofibrous matrix were assessed using a water contact angle analyzer, pH meter, and tetrazolium reduction assay. In vivo efficacies of PLGA/collagen nanofibers and PLA bolts for tendon–bone healing were investigated on a rabbit bone tunnel model with histological and tendon pullout tests. The PLGA/collagen-blended nanofibrous membrane was a hydrophilic, stable, and biocompatible scaffold. The PLA bolt was durable for tendon–bone anchoring. Histology showed adequate biocompatibility of the PLA bolt on a medial cortex with progressive bone ingrowth and without tissue overreaction. PLGA nanofibers within the bone tunnel also decreased the tunnel enlargement phenomenon and enhanced tendon–bone integration. Composite polymers of the PLA bolt and PLGA/collagen nanofibrous membrane can effectively promote outcomes of tendon reconstruction in a rabbit model. The composite biodegradable polymeric system may be useful in humans for tendon reconstruction. Keywords: polylactide–polyglycolide nanofibers, PLGA, collagen, 3D printing, polylactide, PLA, bone-anchoring bolts, tendon healing

  5. Collagen I but not Matrigel matrices provide an MMP-dependent barrier to ovarian cancer cell penetration

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    Brown Theodore J


    Full Text Available Abstract Background The invasive potential of cancer cells is usually assessed in vitro using Matrigel as a surrogate basement membrane. Yet cancer cell interaction with collagen I matrices is critical, particularly for the peritoneal metastatic route undertaken by several cancer types including ovarian. Matrix metalloprotease (MMP activity is important to enable cells to overcome the barrier constraints imposed by basement membranes and stromal matrices in vivo. Our objective was to compare matrices reconstituted from collagen I and Matrigel as representative barriers for ovarian cancer cell invasion. Methods The requirement of MMP activity for ovarian cancer cell penetration of Matrigel and collagen matrices was assessed in 2D transwell and 3D spheroid culture systems. Results The broad range MMP inhibitor GM6001 completely prevented cell perforation of polymerised collagen I-coated transwell membranes. In contrast, GM6001 decreased ES-2 cell penetration of Matrigel by only ~30% and had no effect on HEY cell Matrigel penetration. In 3D culture, ovarian cancer cells grown as spheroids also migrated into surrounding Matrigel matrices despite MMP blockade. In contrast, MMP activity was required for invasion into 3D matrices of collagen I reconstituted from acid-soluble rat-tail collagen I, but not from pepsin-extracted collagen I (Vitrogen/Purecol, which lacks telopeptide regions. Conclusion Matrigel does not form representative barriers to ovarian cancer cells in either 2D or 3D culture systems. Our findings support the use of collagen I rather than Matrigel as a matrix barrier for invasion studies to better approximate critical interactions and events associated with peritoneal metastasis.

  6. Collagen-based mechanical anisotropy of the tectorial membrane: implications for inter-row coupling of outer hair cell bundles.

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    Núria Gavara

    Full Text Available BACKGROUND: The tectorial membrane (TM in the mammalian cochlea displays anisotropy, where mechanical or structural properties differ along varying directions. The anisotropy arises from the presence of collagen fibrils organized in fibers of approximately 1 microm diameter that run radially across the TM. Mechanical coupling between the TM and the sensory epithelia is required for normal hearing. However, the lack of a suitable technique to measure mechanical anisotropy at the microscale level has hindered understanding of the TM's precise role. METHODOLOGY/PRINCIPAL FINDINGS: Here we report values of the three elastic moduli that characterize the anisotropic mechanical properties of the TM. Our novel technique combined Atomic Force Microscopy (AFM, modeling, and optical tracking of microspheres to determine the elastic moduli. We found that the TM's large mechanical anisotropy results in a marked transmission of deformations along the direction that maximizes sensory cell excitation, whereas in the perpendicular direction the transmission is greatly reduced. CONCLUSIONS/SIGNIFICANCE: Computational results, based on our values of elastic moduli, suggest that the TM facilitates the directional cooperativity of sensory cells in the cochlea, and that mechanical properties of the TM are tuned to guarantee that the magnitude of sound-induced tip-link stretching remains similar along the length of the cochlea. Furthermore, we anticipate our assay to be a starting point for other studies of biological tissues that require directional functionality.

  7. Clinical comparison of guided tissue regeneration, with collagen membrane and bone graft, versus connective tissue graft in the treatment of gingival recessions


    Haghighati F; Akbari S


    Background and Aim: Increasing patient demands for esthetic, put the root coverage procedures in particular attention. Periodontal regeneration with GTR based root coverage methods is the most common treatment used. The purpose of this study was to compare guided tissue regeneration (GTR) with collagen membrane and a bone graft, with sub-epithelial connective tissue graft (SCTG), in treatment of gingival recession. Materials and Methods: In this randomized clinical trial study, eleven healthy...

  8. Collagenous gastritis. (United States)

    Jin, Xiaoyi; Koike, Tomoyuki; Chiba, Takashi; Kondo, Yutaka; Ara, Nobuyuki; Uno, Kaname; Asano, Naoki; Iijima, Katsunori; Imatani, Akira; Watanabe, Mika; Shirane, Akio; Shimosegawa, Tooru


    In the present paper, we report a case of rare collagenous gastritis. The patient was a 25-year-old man who had experienced nausea, abdominal distention and epigastralgia since 2005. Esophagogastroduodenoscopy (EGD) carried out at initial examination by the patient's local doctor revealed an extensively discolored depression from the upper gastric body to the lower gastric body, mainly including the greater curvature, accompanied by residual mucosa with multiple islands and nodularity with a cobblestone appearance. Initial biopsies sampled from the nodules and accompanying atrophic mucosa were diagnosed as chronic gastritis. In August, 2011, the patient was referred to Tohoku University Hospital for observation and treatment. EGD at our hospital showed the same findings as those by the patient's local doctor. Pathological findings included a membranous collagen band in the superficial layer area of the gastric mucosa, which led to a diagnosis of collagenous gastritis. Collagenous gastritis is an extremely rare disease, but it is important to recognize its characteristic endoscopic findings to make a diagnosis.

  9. Neural stem cell transplantation in a double-layer collagen membrane with unequal pore sizes for spinal cord injur y repair

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    Ning Yuan; Wei Tian; Lei Sun; Runying Yuan; Jianfeng Tao; Dafu Chen


    A novel double-layer collagen membrane with unequal pore sizes in each layer was designed and tested in this study. The inner, loose layer has about 100-μm-diameter pores, while the outer, compact layer has about 10-μm-diameter pores. In a rat model of incomplete spinal cord injury, a large number of neural stem cells were seeded into the loose layer, which was then adhered to the injured side, and the compact layer was placed against the lateral side. The results showed that the transplantation of neural stem cells in a double-layer collagen membrane with unequal pore sizes promoted the differentiation of neural stem cells, attenuated the pathological lesion, and signiifcantly improved the motor function of the rats with incomplete spinal cord injuries. These experimental ifndings suggest that the transplantation of neural stem cells in a double-lay-er collagen membrane with unequal pore sizes is an effective therapeutic strategy to repair an injured spinal cord.

  10. Influence of basement membrane proteins and endothelial cell-derived factors on the morphology of human fetal-derived astrocytes in 2D.

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    Amanda F Levy

    Full Text Available Astrocytes are the most prevalent type of glial cell in the brain, participating in a variety of diverse functions from regulating cerebral blood flow to controlling synapse formation. Astrocytes and astrocyte-conditioned media are widely used in models of the blood-brain barrier (BBB, however, very little is known about astrocyte culture in 2D. To test the hypothesis that surface coating and soluble factors influence astrocyte morphology in 2D, we quantitatively analyzed the morphology of human fetal derived astrocytes on glass, matrigel, fibronectin, collagen IV, and collagen I, and after the addition soluble factors including platelet-derived growth factor (PDGF, laminin, basic fibroblast growth factor (bFGF, and leukemia inhibitory factor (LIF. Matrigel surface coatings, as well as addition of leukemia inhibitory factor (LIF to the media, were found to have the strongest effects on 2D astrocyte morphology, and may be important in improving existing BBB models. In addition, the novel set of quantitative parameters proposed in this paper provide a test for determining the influence of compounds on astrocyte morphology, both to screen for new endothelial cell-secreted factors that influence astrocytes, and to determine in a high-throughput way which factors are important for translation to more complex, 3D BBB models.

  11. Type IV collagen is an activating ligand for the adhesion G protein-coupled receptor GPR126. (United States)

    Paavola, Kevin J; Sidik, Harwin; Zuchero, J Bradley; Eckart, Michael; Talbot, William S


    GPR126 is an orphan heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptor (GPCR) that is essential for the development of diverse organs. We found that type IV collagen, a major constituent of the basement membrane, binds to Gpr126 and activates its signaling function. Type IV collagen stimulated the production of cyclic adenosine monophosphate in rodent Schwann cells, which require Gpr126 activity to differentiate, and in human embryonic kidney (HEK) 293 cells expressing exogenous Gpr126. Type IV collagen specifically bound to the extracellular amino-terminal region of Gpr126 containing the CUB (complement, Uegf, Bmp1) and pentraxin domains. Gpr126 derivatives lacking the entire amino-terminal region were constitutively active, suggesting that this region inhibits signaling and that ligand binding relieves this inhibition to stimulate receptor activity. A new zebrafish mutation that truncates Gpr126 after the CUB and pentraxin domains disrupted development of peripheral nerves and the inner ear. Thus, our findings identify type IV collagen as an activating ligand for GPR126, define its mechanism of activation, and highlight a previously unrecognized signaling function of type IV collagen in basement membranes.

  12. Expression of Haemophilus ducreyi collagen binding outer membrane protein NcaA is required for virulence in swine and human challenge models of chancroid. (United States)

    Fulcher, Robert A; Cole, Leah E; Janowicz, Diane M; Toffer, Kristen L; Fortney, Kate R; Katz, Barry P; Orndorff, Paul E; Spinola, Stanley M; Kawula, Thomas H


    Haemophilus ducreyi, the etiologic agent of the sexually transmitted genital ulcer disease chancroid, has been shown to associate with dermal collagen fibers within infected skin lesions. Here we describe NcaA, a previously uncharacterized outer membrane protein that is important for H. ducreyi collagen binding and host colonization. An H. ducreyi strain lacking the ncaA gene was impaired in adherence to type I collagen but not fibronectin (plasma or cellular form) or heparin. The mutation had no effect on serum resistance or binding to HaCaT keratinocytes or human foreskin fibroblasts in vitro. Escherichia coli expressing H. ducreyi NcaA bound to type I collagen, demonstrating that NcaA is sufficient to confer collagen attachment. The importance of NcaA in H. ducreyi pathogenesis was assessed using both swine and human experimental models of chancroid. In the swine model, 20% of lesions from sites inoculated with the ncaA mutant were culture positive for H. ducreyi 7 days after inoculation, compared to 73% of wild-type-inoculated sites. The average number of CFU recovered from mutant-inoculated lesions was also significantly reduced compared to that recovered from wild-type-inoculated sites at both 2 and 7 days after inoculation. In the human challenge model, 8 of 30 sites inoculated with wild-type H. ducreyi progressed to the pustular stage, compared to 0 of 30 sites inoculated with the ncaA mutant. Together these results demonstrate that the collagen binding protein NcaA is required for H. ducreyi infection.

  13. Complex collagen fiber and membrane morphologies of the whole porcine aortic valve.

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    Christopher A Rock

    Full Text Available OBJECTIVES: Replacement aortic valves endeavor to mimic native valve function at the organ, tissue, and in the case of bioprosthetic valves, the cellular levels. There is a wealth of information about valve macro and micro structure; however, there presently is limited information on the morphology of the whole valve fiber architecture. The objective of this study was to provide qualitative and quantitative analyses of whole valve and leaflet fiber bundle branching patterns using a novel imaging system. METHODS: We developed a custom automated microscope system with motor and imaging control. Whole leaflets (n = 25 were imaged at high resolution (e.g., 30,000×20,000 pixels using elliptically polarized light to enhance contrast between structures without the need for staining or other methods. Key morphologies such as fiber bundle size and branching were measured for analyses. RESULTS: The left coronary leaflet displayed large asymmetry in fiber bundle organization relative to the right coronary and non-coronary leaflets. We observed and analyzed three main patterns of fiber branching; tree-like, fan-like, and pinnate structures. High resolution images and quantitative metrics are presented such as fiber bundle sizes, positions, and branching morphological parameters. SIGNIFICANCE: To our knowledge there are currently no high resolution images of whole fresh leaflets available in the literature. The images of fiber/membrane structures and analyses presented here could be highly valuable for improving the design and development of more advanced bioprosthetic and/or bio-mimetic synthetic valve replacements.

  14. Collagen XVIII/endostatin expression in experimental endotoxemic acute renal failure

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    M.C. Cichy


    Full Text Available Acute renal failure (ARF is a frequent complication of Gram-negative sepsis, with a high risk of mortality. Lipopolysaccharide (LPS-induced ARF is associated with hemodynamic changes that are strongly influenced by the overproduction of nitric oxide (NO through the cytokine-mediated up-regulation of inducible NO synthase. LPS-induced reductions in systemic vascular resistance paradoxically culminate in renal vasoconstriction. Collagen XVIII is an important component of the extracellular matrix expressed in basement membranes. Its degradation by matrix metalloproteases, cathepsins and elastases results in the formation of endostatin, claimed to have antiangiogenic activity and to be a prominent vasorelaxing agent. We evaluated the expression of endostatin/collagen XVIII in an endotoxemic ARF model. ARF was induced in C57BL/6 mice by intraperitoneal injection of LPS (10 mg/kg followed by sacrifice 4 and 12 h later. Kidney tissue was the source of RNA and protein and the subject of histological analysis. As early as 4 h after LPS administration, blood urea, creatinine and NO levels were significantly increased compared to control. Endostatin/collagen XVIII mRNA levels were 0.71 times lower than sham-inoculated mice 4 h after LPS inoculation, returning to normal levels 12 h after LPS inoculation. Immunohistological examination revealed that acute injury caused by LPS leads to an increase of endostatin basement membrane staining in association with the decrease of CD31 endothelial basement membrane staining. These results indicate that in the early phase of endotoxemic ARF the endostatin levels were not regulated by gene expression, but by the metabolism of collagen XVIII.

  15. Type IV collagen is a novel DEJ biomarker that is reduced by radiotherapy. (United States)

    McGuire, J D; Gorski, J P; Dusevich, V; Wang, Y; Walker, M P


    The dental basement membrane (BM) is composed of collagen types IV, VI, VII, and XVII, fibronectin, and laminin and plays an inductive role in epithelial-mesenchymal interactions during tooth development. The BM is degraded and removed during later-stage tooth morphogenesis; however, its original position defines the location of the dentin-enamel junction (DEJ) in mature teeth. We recently demonstrated that type VII collagen is a novel component of the inner enamel organic matrix layer contiguous with the DEJ. Since it is frequently co-expressed with and forms functional complexes with type VII collagen, we hypothesized that type IV collagen should also be localized to the DEJ in mature human teeth. To identify collagen IV, we first evaluated defect-free erupted teeth from various donors. To investigate a possible stabilizing role, we also evaluated extracted teeth exposed to high-dose radiotherapy--teeth that manifest post-radiotherapy DEJ instability. We now show that type IV collagen is a component within the morphological DEJ of posterior and anterior teeth from individuals aged 18 to 80 yr. Confocal microscopy revealed that immunostained type IV collagen was restricted to the 5- to 10-µm-wide optical DEJ, while collagenase treatment or previous in vivo tooth-level exposure to > 60 Gray irradiation severely reduced immunoreactivity. This assignment was confirmed by Western blotting with whole-tooth crown and enamel extracts. Without reduction, type IV collagen contained macromolecular α-chains of 225 and 250 kDa. Compositionally, our results identify type IV collagen as the first macromolecular biomarker of the morphological DEJ of mature teeth. Given its network structure and propensity to stabilize the dermal-epidermal junction, we propose that a collagen-IV-enriched DEJ may, in part, explain its well-known fracture toughness, crack propagation resistance, and stability. In contrast, loss of type IV collagen may represent a biochemical rationale for the DEJ

  16. In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration. (United States)

    Wodewotzky, T I; Lima-Neto, J F; Pereira-Júnior, O C M; Sudano, M J; Lima, S A F; Bersano, P R O; Yoshioka, S A; Landim-Alvarenga, F C


    Support structures for dermal regeneration are composed of biodegradable and bioresorbable polymers, animal skin or tendons, or are bacteria products. The use of such materials is controversial due to their low efficiency. An important area within tissue engineering is the application of multipotent mesenchymal stromal cells (MSCs) to reparative surgery. The combined use of biodegradable membranes with stem cell therapy may lead to promising results for patients undergoing unsuccessful conventional treatments. Thus, the aim of this study was to test the efficacy of using membranes composed of anionic collagen with or without the addition of hyaluronic acid (HA) as a substrate for adhesion and in vitro differentiation of bone marrow-derived canine MSCs. The benefit of basic fibroblast growth factor (bFGF) on the differentiation of cells in culture was also tested. MSCs were collected from dog bone marrow, isolated and grown on collagen scaffolds with or without HA. Cell viability, proliferation rate, and cellular toxicity were analyzed after 7 days. The cultured cells showed uniform growth and morphological characteristics of undifferentiated MSCs, which demonstrated that MSCs successfully adapted to the culture conditions established by collagen scaffolds with or without HA. This demonstrates that such scaffolds are promising for applications to tissue regeneration. bFGF significantly increased the proliferative rate of MSCs by 63% when compared to groups without the addition of the growth factor. However, the addition of bFGF becomes limiting, since it has an inhibitory effect at high concentrations in culture medium.

  17. In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration

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    T.I. Wodewotzky


    Full Text Available Support structures for dermal regeneration are composed of biodegradable and bioresorbable polymers, animal skin or tendons, or are bacteria products. The use of such materials is controversial due to their low efficiency. An important area within tissue engineering is the application of multipotent mesenchymal stromal cells (MSCs to reparative surgery. The combined use of biodegradable membranes with stem cell therapy may lead to promising results for patients undergoing unsuccessful conventional treatments. Thus, the aim of this study was to test the efficacy of using membranes composed of anionic collagen with or without the addition of hyaluronic acid (HA as a substrate for adhesion and in vitro differentiation of bone marrow-derived canine MSCs. The benefit of basic fibroblast growth factor (bFGF on the differentiation of cells in culture was also tested. MSCs were collected from dog bone marrow, isolated and grown on collagen scaffolds with or without HA. Cell viability, proliferation rate, and cellular toxicity were analyzed after 7 days. The cultured cells showed uniform growth and morphological characteristics of undifferentiated MSCs, which demonstrated that MSCs successfully adapted to the culture conditions established by collagen scaffolds with or without HA. This demonstrates that such scaffolds are promising for applications to tissue regeneration. bFGF significantly increased the proliferative rate of MSCs by 63% when compared to groups without the addition of the growth factor. However, the addition of bFGF becomes limiting, since it has an inhibitory effect at high concentrations in culture medium.

  18. In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration

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    Wodewotzky, T.I.; Lima-Neto, J.F. [Departamento de Reprodução Animal e Radiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Pereira-Júnior, O.C.M. [Departamento de Reprodução Animal e Radiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Departamento de Cirurgia e Anestesiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Sudano, M.J.; Lima, S.A.F. [Departamento de Reprodução Animal e Radiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Bersano, P.R.O. [Departamento de Patologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Yoshioka, S.A. [Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos, SP (Brazil); Landim-Alvarenga, F.C. [Departamento de Reprodução Animal e Radiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil)


    Support structures for dermal regeneration are composed of biodegradable and bioresorbable polymers, animal skin or tendons, or are bacteria products. The use of such materials is controversial due to their low efficiency. An important area within tissue engineering is the application of multipotent mesenchymal stromal cells (MSCs) to reparative surgery. The combined use of biodegradable membranes with stem cell therapy may lead to promising results for patients undergoing unsuccessful conventional treatments. Thus, the aim of this study was to test the efficacy of using membranes composed of anionic collagen with or without the addition of hyaluronic acid (HA) as a substrate for adhesion and in vitro differentiation of bone marrow-derived canine MSCs. The benefit of basic fibroblast growth factor (bFGF) on the differentiation of cells in culture was also tested. MSCs were collected from dog bone marrow, isolated and grown on collagen scaffolds with or without HA. Cell viability, proliferation rate, and cellular toxicity were analyzed after 7 days. The cultured cells showed uniform growth and morphological characteristics of undifferentiated MSCs, which demonstrated that MSCs successfully adapted to the culture conditions established by collagen scaffolds with or without HA. This demonstrates that such scaffolds are promising for applications to tissue regeneration. bFGF significantly increased the proliferative rate of MSCs by 63% when compared to groups without the addition of the growth factor. However, the addition of bFGF becomes limiting, since it has an inhibitory effect at high concentrations in culture medium.

  19. Two-layer membranes of calcium phosphate/collagen/PLGA nanofibres: in vitro biomineralisation and osteogenic differentiation of human mesenchymal stem cells (United States)

    Hild, Nora; Schneider, Oliver D.; Mohn, Dirk; Luechinger, Norman A.; Koehler, Fabian M.; Hofmann, Sandra; Vetsch, Jolanda R.; Thimm, Benjamin W.; Müller, Ralph; Stark, Wendelin J.


    The present study evaluates the in vitro biomedical performance of an electrospun, flexible, anisotropic bilayer with one layer containing a collagen to mineral ratio similar to that in bone. The double membrane consists of a poly(lactide-co-glycolide) (PLGA) layer and an amorphous calcium phosphate (a-CaP)/collagen (Col)/PLGA layer. In vitro biomineralisation and a cell culture study with human mesenchymal stem cells (hMSC) were conducted to characterise such membranes for possible application as biomaterials. Nanofibres with different a-CaP/Col/PLGA compositions were synthesised by electrospinning to mimic the actual composition of bone tissue. Immersion in simulated body fluid and in cell culture medium resulted in the deposition of a hydroxyapatite layer. Incubation of hMSC for 4 weeks allowed for assessment of the proliferation and osteogenic differentiation of the cells on both sides of the double membrane. Confocal laser scanning microscopy was used to observe the proper adhesion of the cells. Calcium and collagen content was proven by Alizarin red S and Sirius red assays. Acute cytotoxic effects of the nanoparticles or the chemicals used in the scaffold preparation could be excluded based on viability assays (alamarBlue and alkaline phosphatase activity). The findings suggest possible application of such double membranes is in treatment of bone defects with complex geometries as wound dressing material.The present study evaluates the in vitro biomedical performance of an electrospun, flexible, anisotropic bilayer with one layer containing a collagen to mineral ratio similar to that in bone. The double membrane consists of a poly(lactide-co-glycolide) (PLGA) layer and an amorphous calcium phosphate (a-CaP)/collagen (Col)/PLGA layer. In vitro biomineralisation and a cell culture study with human mesenchymal stem cells (hMSC) were conducted to characterise such membranes for possible application as biomaterials. Nanofibres with different a

  20. Effect of Supercoiling on the Mechanical and Permeability Properties of Model Collagen IV Networks. (United States)

    Gyoneva, Lazarina; Segal, Yoav; Dorfman, Kevin D; Barocas, Victor H


    Collagen IV networks in the glomerular basement membrane (GBM) are essential for the maintenance and regulation of blood filtration in the kidneys. The GBM contains two different types of collagen IV networks: [α1(IV)]2α2(IV) and α3(IV)α4(IV)α5(IV), the latter of which has a higher number of supercoils (two or more collagens coiling around each other). To investigate the effects of supercoiling on the mechanical and permeability properties of collagen IV networks, we generated model collagen IV networks in the GBM and reconnected them to create different levels of supercoiling. We found that supercoiling greatly increases the stiffness of collagen IV networks but only minimally decreases the permeability. Also, doubling the amount of supercoils in a network had a bigger effect than doubling the stiffness of the supercoils. Our results suggest that the formation of supercoils is a specialized mechanism by the GBM that provides with a network stiff and strong enough to withstand the high hydrostatic pressures of filtration, yet porous enough that filtration is not hindered. Clinically, understanding the effects of supercoiling gives us insight into the mechanisms of GBM failure in some disease states where the normal collagen IV structure is disrupted.

  1. Quaternary epitopes of α345(IV) collagen initiate Alport post-transplant anti-GBM nephritis

    DEFF Research Database (Denmark)

    Olaru, Florina; Luo, Wentian; Wang, Xu-Ping


    Alport post-transplant nephritis (APTN) is an aggressive form of anti-glomerular basement membrane disease that targets the allograft in transplanted patients with X-linked Alport syndrome. Alloantibodies develop against the NC1 domain of α5(IV) collagen, which occurs in normal kidneys, including...... of alloantibodies against allogeneic collagen IV. Some alloantibodies targeted alloepitopes within α5NC1 monomers, shared by α345NC1 and α1256NC1 hexamers. Other alloantibodies specifically targeted alloepitopes that depended on the quaternary structure of α345NC1 hexamers. In Col4a5-null mice, immunization...... with native forms of allogeneic collagen IV exclusively elicited antibodies to quaternary α345NC1 alloepitopes, whereas alloimmunogens lacking native quaternary structure elicited antibodies to shared α5NC1 alloepitopes. These results imply that quaternary epitopes within α345NC1 hexamers may initiate...

  2. [The enlarged diagnosis of the fatal penicillin accident. Immunehistologic demonstration of antigen-antibody complexes and of antibodies against the tubular basement membrane after administraiton of depot penicillin]. (United States)

    Dirnhofer, R; Sonnabend, W; Sigrist, T


    In a case of fatal penicillin allergy it proved possible at autopsy to demonstrate (by immunohistological examination of basal membranes of proximal renal tubuli) antigen-antibody complexes belonging to the penicillin (BPO) group and to an anti-penicilloyl antibody of the IgG type. In addition, complement C3 was detected. Antibodies against the basal membranes or renal tubuli were also demonstrated in material eluted from the kidney, although an inflammatory reaction ot the immunoligical changes had not yet been observed in light microscopy. It is undecided whether this discrepancy is due to the low dose of penicillin administered or the relatively short time lag between first injection and time of fatality. It is assumed that, pathogenetically, a reaction of the serum sickness type is probably involved. For etiological clarification the use of immunohistological methods in addition to serological procedures provides further indices for an antecedent sensitization to penicillin, because assay effectiveness does not decrease even after a lengthy postmortal time-lapse. On the other hand, tissues and serum for examination should be frozen at low temperatures immediately after autopsy.

  3. Clinical comparison of guided tissue regeneration, with collagen membrane and bone graft, versus connective tissue graft in the treatment of gingival recessions

    Directory of Open Access Journals (Sweden)

    Haghighati F


    Full Text Available Background and Aim: Increasing patient demands for esthetic, put the root coverage procedures in particular attention. Periodontal regeneration with GTR based root coverage methods is the most common treatment used. The purpose of this study was to compare guided tissue regeneration (GTR with collagen membrane and a bone graft, with sub-epithelial connective tissue graft (SCTG, in treatment of gingival recession. Materials and Methods: In this randomized clinical trial study, eleven healthy patients with no systemic diseases who had miller’s class I or II recession defects (gingival recession  2mm were treated with SCTG or GTR using a collagen membrane and a bone graft. Clinical measurements were obtained at baseline and 6 months after surgery. These clinical measurements included recession depth (RD, recession width (RW, probing depth (PD, and clinical attachment level (CAL. Data were analyzed using independent t test with p<0.05 as the limit of significance. Results: Both treatment methods resulted in a statistically significant reduction of recession depth (SCTG=2.3mm, GTR=2.1mm; P<0.0001. CAL gain after 6 months was also improved in both groups (SCG= 2.5mm, GTR=2.1mm, compared to baseline (P<0.0001. No statistical differences were observed in RD, RW, CAL between test and control groups. Root coverage was similar in both methods (SCTG= 74.2%, GTR= 62.6%, P=0.87. Conclusion: Based on the results of this study, the two techniques are clinically comparable. Therefore the use of collagen membrane and a bovine derived xenograft may alleviate the need for connective tissue graft.

  4. Comparative evaluation of a bioabsorbable collagen membrane and connective tissue graft in the treatment of localized gingival recession: A clinical study

    Directory of Open Access Journals (Sweden)

    Harsha Mysore Babu


    Full Text Available Background: Gingival recession (GR can result in root sensitivity, esthetic concern to the patient, and predilection to root caries. The purpose of this randomized clinical study was to evaluate (1 the effect of guided tissue regeneration (GTR procedure using a bioabsorbable collagen membrane, in comparison to autogenous subepithelial connective tissue graft (SCTG for root coverage in localized gingival recession defects; and (2 the change in width of keratinized gingiva following these two procedures. Materials and Methods: A total of 10 cases, showing at least two localized Miller′s Class I or Class II gingival recession, participated in this study. In a split mouth design, the pairs of defects were randomly assigned for treatment with either SCTG (SCTG Group or GTR-based collagen membrane (GTRC Group. Both the grafts were covered with coronally advanced flap. Recession depth (RD, recession width (RW, width of keratinized gingiva (KG, probing depth (PD, relative attachment level (RAL, plaque index (PI, and gingival index (GI were recorded at baseline, 3 and 6 months postoperatively. Results: Six months following root coverage procedures, the mean root coverage was found to be 84.84% ± 16.81% and 84.0% ± 15.19% in SCTG Group and GTRC Group, respectively. The mean keratinized gingival width increase was 1.50 ± 0.70 mm and 2.30 ± 0.67 mm in the SCTG and GTRC group, respectively, which was not statistically significant. Conclusion: It may be concluded that resorbable collagen membrane can be a reliable alternative to autogenous connective tissue graft in the treatment of gingival recession.

  5. An Immunohistochemical Study of Retinal Collagen IV Expression during Pre- and Postnatal Postnatal Periods in Balb/c Mice

    Directory of Open Access Journals (Sweden)

    Mehdi Jalali


    Full Text Available Objective: Basement membranes are specialized extracellular matrices which playimportant roles such as cell regulation, proliferation and migration. Collagen fibers,especially type IV, are the most important basement membrane constituents. As retinais one of the target organs in diabetes mellitus, and nephropathy is a major cause ofend stage renal and retinal diseases resulting in increased morbidity and mortality,early diagnosis leads to better treatment. Hence, in this investigation, the appearanceand distribution of collagen IV during gestational days and early postnatal periodswere observed.Materials and Methods: 24 intact female Balb/c mice were kept under normal conditions.After mating, appearance of a vaginal plug was assumed as day zero of the pregnancy.From days 13-18 of gestation, the pregnant mice were euthanised and theirembryos as well as pups from days 1 to 5 were collected. For histochemichal studies,heads of the specimens were fixed, serially sectioned and immunohistochemicallstudies were performed by using monoclonal antibodies for tracing of collagentype IV.Results: Our findings revealed that the amount of collagen IV in the internal limiting membrane(ILM and extra cellular matrix (ECM of the retina, as well as vessels of the vitreusbody appear on embryonic day 16. Also, a patchy distribution was observed in the pigmentedepithelium which continued to further develop until the end stage of embryoniclife. Strong labeling was observed until postnatal day 3 but did not increase significantlythereafter.Conclusion: These findings establish the importance of collagen IV during the critical periodof retinal development. In addition, this study indicates that high levels of collagen IVare present in the basal membrane (BM of the inner limiting membranes and pigmentedepithelium (3rd post natal on the 3rd postnatal day.

  6. [An experience of treatment of double positive myeloperoxidase-antineutrophil cytoplasmic antibodies (MPO-ANCA) and anti-glomerular basement membrane antibodies in Goodpasture's syndrome onset of crescentic glomerulonephritis]. (United States)

    Takeda, T; Takeda, T; Naiki, Y; Yonekawa, S; Sakaguchi, M; Iwamoto, I; Tanaka, H; Hasegawa, H; Imada, A; Kanamaru, A; Hiruma, S; Maekura, S; Hashimoto, S; Yamazumi, T


    A 68-year-old woman was admitted to Kinki University Hospital because of progressive renal failure. She had been well until two months before admission. Laboratory data were as follows: serum creatinine 4.1 mg/dl, BUN 69 mg/dl, MPO-ANCA 33 EU, anti-glomerular basement membrane antibodies (AGBMA) 118 U. Histological findings showed cellular and fibrocellular crescents in many glomeruli. Therefore, we diagnosed rapidly progressive glomerulonephritis (RPGN) due to MPO-ANCA and anti-GBM associated renal disease. The patient was started on prednisolone and double filtration plasmapheresis (DFPP) therapy. Subsequently, the values of MPO-ANCA and AGBMA decreased. However, the patient's condition suddenly worsened and she died of interstitial pneumonia. Autopsy examination revealed crescentic glomerulonephritis and alveolar hemorrhage with linear deposition of IgG along the glomerular and alveolar capillary walls by immunofluorescence studies. We considered this to be a rare case of Goodpasture's syndrome associated with not only anti-GBM antibodies, but also MPO-ANCA.

  7. Postnatal development of epididymis and ductus deferens in the rat. A correlation between the ultrastructure of the epithelium and tubule wall, and the fluorescence-microscopic distribution of actin, myosin, fibronectin, and basement membrane. (United States)

    Francavilla, S; Moscardelli, S; Properzi, G; De Matteis, M A; Scorza Barcellona, P; Natali, P G; De Martino, C


    The postnatal maturation of regions of the epididymis and intragonadal segment of the deferens duct was studied in the rat by light- and transmission electron microscopy. Maturation of the genital duct starts in the distal cauda epididymidis and ductus deferens after one week of life, and one week later, in the more cranial segments of the epididymis. Epithelial principal cells and peritubular contractile cells are structurally mature 35 days after birth. The synchronous changes of these cells indicate that the same factors control their postnatal maturation. The epithelial principal cells obtain an endocytotic apparatus and long stereocilia, whereas peritubular cells acquire contractile features. These changes are associated with a progressive increase in the immunoreaction for smooth muscle actin in both cell types. Smooth muscle myosin is detected in the apical region of the epithelial cells and the peritubular cell cytoplasm by day one of postnatal development. The differentiation of contractile cells in the wall is accompanied by progressive organization of the pericellular matrix into a continuous basement membrane. Although fibronectin is visible at birth, it is gradually removed from the tubule wall.

  8. Tissue engineering of composite grafts: Cocultivation of human oral keratinocytes and human osteoblast-like cells on laminin-coated polycarbonate membranes and equine collagen membranes under different culture conditions. (United States)

    Glaum, R; Wiedmann-Al-Ahmad, M; Huebner, U; Schmelzeisen, R


    In complex craniomaxillofacial defects, the simultaneous reconstruction of hard and soft tissue is often necessary. Until now, oral keratinocytes and osteoblast-like cells have not been cocultivated on the same carrier. For the first time, the cocultivation of human oral keratinocytes and human osteoblast-like cells has been investigated in this study. Different carriers (laminin-coated polycarbonate and equine collagen membranes) and various culture conditions were examined. Human oral keratinocytes and human osteoblast-like cells from five patients were isolated from tissue samples, seeded on the opposite sides of the carriers and cultivated for 1 and 2 weeks under static conditions in an incubator and in a perfusion chamber. Proliferation and morphology of the cells were analyzed by EZ4U-tests, light microscopy, and scanning electron microscopy. Cocultivation of both cell-types seeded on one carrier was possible. Quantitative and qualitative growth was significantly better on collagen membranes when compared with laminin-coated polycarbonate membranes independent of the culture conditions. Using perfusion culture in comparison to static culture, the increase of cell proliferation after 2 weeks of cultivation when compared with the proliferation after 1 week was significantly lower, independent of the carriers used. In conclusion, the contemporaneous cultivation of human oral keratinocytes and human osteoblast-like cells on the same carrier is possible, a prerequisite for planned in vivo studies. As carrier collagen is superior to laminin-coated polycarbonate membranes. Regarding the development over time, the increase of proliferation rate is lower in perfusion culture. Examinations of cellular differentiation over time under various culture conditions will be subject of further investigations.

  9. The collagen receptor uPARAP/Endo180 in tissue degradation and cancer (Review). (United States)

    Melander, Maria C; Jürgensen, Henrik J; Madsen, Daniel H; Engelholm, Lars H; Behrendt, Niels


    The collagen receptor uPARAP/Endo180, the product of the MRC2 gene, is a central component in the collagen turnover process governed by various mesenchymal cells. Through the endocytosis of collagen or large collagen fragments, this recycling receptor serves to direct basement membrane collagen as well as interstitial collagen to lysosomal degradation. This capacity, shared only with the mannose receptor from the same protein family, endows uPARAP/Endo180 with a critical role in development and homeostasis, as well as in pathological disruptions of the extracellular matrix structure. Important pathological functions of uPARAP/Endo180 have been identified in various cancers and in several fibrotic conditions. With a particular focus on matrix turnover in cancer, this review presents the necessary background for understanding the function of uPARAP/Endo180 at the molecular and cellular level, followed by an in-depth survey of the available knowledge of the expression and role of this receptor in various types of cancer and other degenerative diseases.

  10. Animal Experiment of Periodontal Defect Reconstruction Using Collagen-hydroxyapatite Artificial Bone Plus Collagen Membrane%胶原—羟基磷灰石人工骨与胶原膜引导牙周组织再生的动物实验研究

    Institute of Scientific and Technical Information of China (English)

    吴文蕾; 葛久禹; 李升; 黄晓峰; 陈湘华


    目的:将胶原—羟基磷灰石人工骨与胶原膜联合应用于修复牙周缺损的动物实验,探讨其用于引导牙周组织再生的可行性.方法:人工构建4只成年Beagle犬下颌后牙区牙周缺损模型,分别随机采用:胶原—羟基磷灰石人工骨/胶原膜、胶原—羟基磷灰石人工骨、空白对照治疗,每组8颗牙,12周后处死动物,进行组织学观察并测量新生组织高度.结果:与单纯植入胶原—羟基磷灰石人工骨组相比,胶原—羟基磷灰石人工骨/胶原膜组获得了更多的新附着,表现为有较多的新生牙槽骨、新生牙周膜和新生牙骨质样组织生长,2组之间新生组织差异有显著性(P<0.05).结论:胶原—羟基磷灰石人工骨与胶原膜联合运用修复牙周牙槽骨缺损引导牙周组织再生的效果优于单纯植入人工骨.%Objective: To investigate the possibility of using collagen-hydroxyapatite artificial bone plus collagen membrane in the reconstruction of periodontal defect. Methods: Four beagle dogs were constructed periodontal defect in mandibular premolars or molars. 24 teeth were divided into three groups randomly. They were collagen - hydroxyapatite artificial bone plus collagen membrane, collagen-hydroxyapatite artificial bone and blank control. The dogs were sacrificed 12 weeks post-surgery, and healing was evaluated histologically. Results: Periodontal defects implanted with collagen - hydroxyapatite artificial bone plus collagen membrane got more neonatal alveolar bone (P<0. 05) , periodontal membrane (P<0. 05) and cementum- like tissue (P<0. 05) than the group with collagen-hydroxyapatite artificial bone only. Conclusion: Collagen-hydroxyapatite artificial bone plus collagen membrane is better than artificial bone only in periodontal regeneration.

  11. Laminin peptide YIGSR induces collagen synthesis in Hs27 human dermal fibroblasts

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Jong Hyuk; Kim, Jaeyoon; Lee, Hyeongjoo [NovaCell Technology Inc., Pohang, Kyungbuk 790-784 (Korea, Republic of); Kim, So Young [Department of Dermatology, Chung-Ang University College of Medicine, Seoul 156-756 (Korea, Republic of); Department of Convergence Medicine and Pharmaceutical Biosciences, Graduate School, Chung-Ang University, Seoul 156-756 (Korea, Republic of); Jang, Hwan-Hee [Functional Food and Nutrition Division, Department of Agrofood Resources, Rural Development Administration, Suwon 441-853 (Korea, Republic of); Ryu, Sung Ho [Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Pohang, Kyungbuk 790-784 (Korea, Republic of); Kim, Beom Joon [Department of Dermatology, Chung-Ang University College of Medicine, Seoul 156-756 (Korea, Republic of); Department of Convergence Medicine and Pharmaceutical Biosciences, Graduate School, Chung-Ang University, Seoul 156-756 (Korea, Republic of); Lee, Taehoon G., E-mail: [NovaCell Technology Inc., Pohang, Kyungbuk 790-784 (Korea, Republic of)


    Highlights: Black-Right-Pointing-Pointer We identify a function of the YIGSR peptide to enhance collagen synthesis in Hs27. Black-Right-Pointing-Pointer YIGSR peptide enhanced collagen type 1 synthesis both of gene and protein levels. Black-Right-Pointing-Pointer There were no changes in cell proliferation and MMP-1 level in YIGSR treatment. Black-Right-Pointing-Pointer The YIGSR effect on collagen synthesis mediated activation of FAK, pyk2 and ERK. Black-Right-Pointing-Pointer The YIGSR-induced FAK and ERK activation was modulated by FAK and MEK inhibitors. -- Abstract: The dermal ECM is synthesized from fibroblasts and is primarily compromised of fibrillar collagen and elastic fibers, which support the mechanical strength and resiliency of skin, respectively. Laminin, a major glycoprotein located in the basement membrane, promotes cell adhesion, cell growth, differentiation, and migration. The laminin tyrosine-isoleucine-glycine-serine-arginine (YIGSR) peptide, corresponding to the 929-933 sequence of the {beta}1 chain, is known to be a functional motif with effects on the inhibition of tumor metastasis, the regulation of sensory axonal response and the inhibition of angiogenesis through high affinity to the 67 kDa laminin receptor. In this study, we identified a novel function of the YIGSR peptide to enhance collagen synthesis in human dermal fibroblasts. To elucidate this novel function regarding collagen synthesis, we treated human dermal fibroblasts with YIGSR peptide in both a time- and dose-dependent manner. According to subsequent experiments, we found that the YIGSR peptide strongly enhanced collagen type 1 synthesis without changing cell proliferation or cellular MMP-1 level. This YIGSR peptide-mediated collagen type 1 synthesis was modulated by FAK inhibitor and MEK inhibitor. This study clearly reveals that YIGSR peptide plays a novel function on the collagen type 1 synthesis of dermal fibroblasts and also suggests that YIGSR is a strong candidate

  12. 高脂高糖家兔脑梗死模型基底膜损伤机制的探讨%Mechanism of basement membrane injury in cerebral infarction model in rabbit with hyperglycemia and hyperlipidemia

    Institute of Scientific and Technical Information of China (English)

    徐娉; 庄强


    Objective To discuss the mechanism of microvascular endothelial basement membrane injury following cerebral ischemla. Methods Twenty-four New Zealand rabbits were randomly divided into two groups (n=12): normal control group and model group. The latter were fed high-fat, high-sucrose diet, but the former was fed with normal diet. They all were made into the models of cerebral infarction. The change of Laminin (LN) in blood and the expression of LN in brain tissue were observed simultaneously. All groups received pathological examination. Results The levels of blood lipids and blood glucose of rabbit and the content of LN were significantly increased after fed with high fat and high sucrose about 1-2 times in 10 d. The plasma concentration of LN was decreased significantly after the operation of cerebral infarction (P<0.05). Pathological observation revealed that LN was mainly located in cytoplasm of microvascular matrix, and its positive reaction was yellow to brown. Conclusions Injury of the microvascular endothelial basement membrane exists in the early phase of rabbit cerebral ischemia, leading to decrease of LN in extracellular matrix. The early intervention of blood lipids and blood glucose, accompanied by the brain protection with agents and the application of inhibitors of MMPs, will be able to effectively improve the cerebral infarction.%目的 探讨脑缺血后微血管内皮细胞基底膜的损伤机制.方法 24只新西兰家兔采用随机数字表法分为非高脂高糖脑梗死模型对照组(简称空白对照组),高脂高糖脑梗死模型对照组(简称模型对照组),每组12只.空白对照组给予普通饲料,模型对照组给予高脂高糖饲料,均制作成脑梗死模型.监测造模前后血浆中层粘连蛋白(LN)的表达,并观察腩组织中LN表达情况.结果模型对照组高脂高糖模型制作后,家兔血脂血糖增高,与造模前比较差异有统计学意义(P<0.05).脑梗死模型制作后两组血浆中LN表达

  13. Application of membrane separation in the desalination process of pilot fish scale collagen extraction%膜分离在鱼鳞胶原蛋白中试提取液脱盐过程中的应用

    Institute of Scientific and Technical Information of China (English)

    陈晖; 易瑞灶; 陈俊德; 陈思谨; 张怡评; 谢全灵


    Membrane separation was applied for the desalination of pilot collagen extraction from fish scale.The effect of ultrafiltration membranes with different MWCO or different material on the desalination rate and collagen recovery rate were researched.The result showed that the optimal membrane was spiral ultrafiltration membrane with MWCO 8ku, the waste water could be recycled for desalination process after treated by nanofiltration membrane.The results of three pilot batch tests of the collagen extraction desalination showed that when the collagen extraction was four times concentrated ,the collagen recovery rate was more than 75% ,the removal rate of inorganic salt related to ash in the extraction was more than 85% ,the ash content in the final collagen product was less than 1 %.%采用膜分离技术进行鱼鳞胶原蛋白中试提取液的脱盐实验。研究了不同分子量、不同类型的超滤膜对料液脱盐效果及胶原蛋白回收率的影响。结果表明,最适用膜为截留分子量8ku的卷式超滤膜,脱盐废水经纳滤膜处理后可循环用于脱盐过程,从而减少了用水量。经过三批次胶原蛋白中试规模脱盐实验验证,当提取料液浓缩倍数为4倍时,胶原蛋白总回收率〉75%,胶原蛋白提取液中与灰分相关的无机盐脱除率〉85%,最终产品灰分〈1%。

  14. 酶法提取鸡蛋壳膜胶原蛋白工艺%Enzymatic Extraction of Collagen from Egg Shell Membrane

    Institute of Scientific and Technical Information of China (English)

    任萌; 宫新统; 赵颂宁; 张元元; 孙研博; 殷建伟; 刘静波


    In order to establish an enzymatic method for collagen extraction from egg shell membrane, the influence of enzyme type, pH, enzyme dose, material-to-water ratio and extraction time on extraction efficiency was investigated and these extraction parameters were optimized one-factor-at-a-time experiments combined with an L9(3a) orthogonal array design. The optimal extraction condition was hydrolysis for 5 h with 6% papain at normal temperature and a material-to-water ratio of 1:100 (g/mL); resulting in an extraction rate of 0.91%. As a result, an efficient extraction method was obtained. Collagen extraction from egg shell membrane by this method not only avoids environmental pollution caused by abandoned egg shell membrane, but also has great practical value in the collagen production industry.%采用酶解法从鸡蛋膜中提取胶原蛋白,研究酶的种类、pH值、酶用量、料水比和提取时间对胶原蛋白提取的影响,采用单因素试验并结合L9(34)正交试验优化,得出酶法提取蛋壳膜中的胶原蛋白的最佳工艺条件为常温条件下质量分数6%木瓜蛋白酶在pH5、料水比1:100(g/mL)条件下、酶解5h,胶原蛋白的提取率达到0.91%。得到高效提取胶原蛋白的工艺,不仅解决了废弃鸡蛋壳膜的环境污染问题,而且在胶原蛋白生产工业有较大的实际利用价值。

  15. A clinical stydy on the effectiveness of slow - resorbing collagen membrane barrier therapy to guide regeneration in mandibular class II furcations in human

    Directory of Open Access Journals (Sweden)

    Abolfazli N


    Full Text Available The present clinical trial was designed to evaluate the regenerative potential of periodontal tissues in degree II"nfurcation defects at mandibular molars of human using a slow-resorbing collagen membrane and a surgical treatment"ntechnique based on the principles of guided tissue regeneration."nThe patient sampleinclude 8 subjects who had periodontal lessions in right and left mandibular molars regions, including moderate to advance periodonal destruction within the radicular area. Following a baseline examination including recording the clinical measurements (PD, Al, HC, F.G.M , the furcation- involved molars were randomly assigned in each patient to either a test or a control treatment procedure. Included the evevation of mucoperiosteal flaps, recording measurement from the cemento enamel junction (C.E.J directly coronal to the furcation area to the alveolar crest and to the base of the defect-Horizontal furcation measurements were also made using a William's probe, finally a collagen membrrane placed on the involved area to cover the entrance of the furcation and adjucent root surfaces as well as a portion of the alveolar bone apical to the crest. The flaps were repositioned and secured with interdental sutures. A procedure identical to the one used at the test teeth was Performed at the control teeth region with the exception of the placement of the collagen membrance. Following surgery all patients were placed on a plaque control regimen. All Patients received normal postsurgical care and at 6 month post-surgery were scheduled for re-entry surgery. Before re-entry surgery all clinical parameters recorded again. The re-entry mucoperiosteal flaps were designed to expose the furcation area for measurements, as describedabove. There was clinical improvement in all measurements made in both the test and control patients (especially in test group over the 6 month period. The horizontal and vertical furcation measurements did yield a

  16. Collagenous gastroduodenitis. (United States)

    Rustagi, Tarun; Rai, Mridula; Scholes, John V


    Collagenous gastroduodenitis is a rare histopathologic entity characterized by marked subepithelial collagen deposition with associated mucosal inflammatory infiltrate. Only 4 cases have been reported, of which 3 had associated collagenous colitis. Collagenous gastroduodenitis without colonic involvement is exceptionally rare with only 1 case reported so far in the literature. We present a case of a 68-year-old woman with dyspepsia and mild anemia, who was found to have nodular gastric and duodenal mucosa on endoscopic examination. Histopathology showed collagenous gastroduodenitis. To the best of our knowledge, this is the second (and first in English literature) reported case of isolated collagenous gastroduodenitis.

  17. Modeling pulmonary fibrosis by abnormal expression of telomerase/apoptosis/collagen V in experimental usual interstitial pneumonia

    Energy Technology Data Exchange (ETDEWEB)

    Parra, E.R.; Pincelli, M.S. [Departamento de Patologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP (Brazil); Teodoro, W.R.; Velosa, A.P.P. [Disciplina de Reumatologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP (Brazil); Martins, V.; Rangel, M.P.; Barbas-Filho, J.V.; Capelozzi, V.L. [Departamento de Patologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP (Brazil)


    Limitations on tissue proliferation capacity determined by telomerase/apoptosis balance have been implicated in pathogenesis of idiopathic pulmonary fibrosis. In addition, collagen V shows promise as an inductor of apoptosis. We evaluated the quantitative relationship between the telomerase/apoptosis index, collagen V synthesis, and epithelial/fibroblast replication in mice exposed to butylated hydroxytoluene (BHT) at high oxygen concentration. Two groups of mice were analyzed: 20 mice received BHT, and 10 control mice received corn oil. Telomerase expression, apoptosis, collagen I, III, and V fibers, and hydroxyproline were evaluated by immunohistochemistry, in situ detection of apoptosis, electron microscopy, immunofluorescence, and histomorphometry. Electron microscopy confirmed the presence of increased alveolar epithelial cells type 1 (AEC1) in apoptosis. Immunostaining showed increased nuclear expression of telomerase in AEC type 2 (AEC2) between normal and chronic scarring areas of usual interstitial pneumonia (UIP). Control lungs and normal areas from UIP lungs showed weak green birefringence of type I and III collagens in the alveolar wall and type V collagen in the basement membrane of alveolar capillaries. The increase in collagen V was greater than collagens I and III in scarring areas of UIP. A significant direct association was found between collagen V and AEC2 apoptosis. We concluded that telomerase, collagen V fiber density, and apoptosis evaluation in experimental UIP offers the potential to control reepithelization of alveolar septa and fibroblast proliferation. Strategies aimed at preventing high rates of collagen V synthesis, or local responses to high rates of cell apoptosis, may have a significant impact in pulmonary fibrosis.

  18. Collagenous sprue

    DEFF Research Database (Denmark)

    Soendergaard, Christoffer; Riis, Lene Buhl; Nielsen, Ole Haagen


    Collagenous sprue is a rare clinicopathological condition of the small bowel. It is characterised by abnormal subepithelial collagen deposition and is typically associated with malabsorption, diarrhoea and weight loss. The clinical features of collagenous sprue often resemble those of coeliac dis...

  19. Histological and histomorphometric analysis of animal experimental dehiscence defect treated with three bio absorbable GTR collagen membrane

    Directory of Open Access Journals (Sweden)

    Parichehr Behfarnia


    Conclusion: The membrane-treated groups had a statistically significant increase in bone formation and connective tissue attachment compared to control groups. However, there are some differences among experimental groups, which should be considered in GTR treatments.

  20. Angiogenesis and collagen type IV expression in different endothelial cell culture systems. (United States)

    Bahramsoltani, M; Slosarek, I; De Spiegelaere, W; Plendl, J


    In vitro angiogenesis assays constitute an important tool for studying the mechanisms of angiogenesis and for identification of pro- and anti-angiogenic substances. Therefore, endothelial cell and media systems used for in vitro angiogenesis assays are required to mimic the angiogenic process in vivo including endothelial capability to express collagen type IV as a component of the basement membrane. In this study, the expression of collagen type IV and its α chains (α1-6) was investigated in different endothelial cell culture systems in vitro qualitatively and quantitatively. These systems included four different batches of microvascular endothelial cells derived from the human skin, heart and lung, from which only two batches were found to be angiogenic and two batches were classified as non-angiogenic. Distribution of the transcripts of the α chains of collagen type IV was similar in all cell and media systems investigated. However, secretion and deposition of a stable extracellular network of collagen type IV could only be observed in the angiogenic cultures. In conclusion, the consecutive steps of the angiogenic cascade in vivo as well as in vitro depend on an increasing secretion and subsequent extracellular deposition of collagen type IV.

  1. The Role of Type IV Collagen in Developing Lens in Mouse Fetuses

    Directory of Open Access Journals (Sweden)

    Mehdi Jalali


    Full Text Available Objective(sExtracellular matrix (ECM and basement membrane (BM play important roles in many developmental processes during development and after birth. Among the components of the BM, collagen fibers specially type IV are the most important parts. The aim of this study was to determine the time when collagen type IV appears in the BM of lens structure during mouse embryonic development.Materials and MethodsIn this experimental study, 22 female Balb/C mice were randomly selected and were kept under normal condition, finding vaginal plug was assumed as day zero of pregnancy. From embryonic day 10 to 20, all specimens were sacrificed by cervical dislocation and their heads were fixed, serially sectioned and immunohistochemistry study for tracing collagen type IV in lens were carried out.ResultsOur data revealed that collagen type IV appeared at the early stage of gestation day 12 in BM of anterior epithelial lens cells and the amount of this protein gradually increased until days 15-17 in ECM and posterior capsule epithelium. After this period, severe reaction was not observed in any part of the lens.ConclusionThese findings establish the important role of collagen IV in developing optic cup and any changes during critical period of pregnancy may be result in severe visual system defect

  2. Measure Guideline: Basement Insulation Basics

    Energy Technology Data Exchange (ETDEWEB)

    Aldrich, R.; Mantha, P.; Puttagunta, S.


    This guideline is intended to describe good practices for insulating basements in new and existing homes, and is intended to be a practical resources for building contractors, designers, and also to homeowners.

  3. Immunohistochmical Study of Glomerular Mesengial Collagen IV Expression in Diabetic Balb/c Mice

    Directory of Open Access Journals (Sweden)

    M. Jalali


    Full Text Available Introduction & Objective: Extra-cellular matrix and basement membrane play important roles in many developmental phenamenon during development and after birth. Among the components of the basement membrane, collagen fibers specially type IV, are the most important part of this area. As kidney is one of the target organs in diabetes mellitus and diabetic nephropathy is a major cause of end stage-renal disease and result an increase in morbidity and mortality of effected individuals, therefore early diagnosis leads to better treatment. The aim of this investigation was to study the primary diagnostic parameters by special regards to collagen IV fibers.Materials & Methods: In this study, 24 male balb/c mice were divided into experimental and control groups. In experimental group, the beta cells of Langerhance were chemically destroyed by an injection of 160 mg/kg alloxan and subdivided in experimental groups 1 and 2. Controls were kept untreated. Experimental group 1and 2 were sacrified 8 and 16 weeks after treatment with alloxan respectively. The same procedure was performed for control group. Immunohistochmical studies were carried out using monocolonal antibody against collagen type IV in Glomeruli. In addition, using morphometrical and stereological methods the volume of the glumerles was compared in all groups.Results: Our finding showed that in experimental groups with special regards in 16 weeks diabetic mice, the rate of collagen type IV in basement membrane around the parietal layer of Bowman capsule, mesangial cells and endothelium of capillary in glomerules increased significantly compared to controls and experimental group 1 (p<0.05, while there was not a significant difference among experimental group 2 and controls. Our data also revealed that the number of mesangial cells as well as glomerular volume increased significantly in experimental 2 (4.635±0.289×106µm3 compared to experimental 1 (3.504±0.189×106µm3 and controls (3.422

  4. Preparation of Propanedioic Acid Derivative and its Modification of Collagen Membrane%一种丙二酸衍生物的制备及其对胶原膜的改性应用

    Institute of Scientific and Technical Information of China (English)

    谢树忠; 李国英


    Propanedioic acid-NHS ester was prepared by modifying propanedioic acid with N-hydroxysuccinimide(NHS) in the presence of dicyclohexylcarbodiimide(DCC). Propanedioic acid-NHS ester was used as a crosslinking agent for collagen membrane, and the effect of crosslinking agent dosage on the physicochemical properties of collagen membrane was investigated. The physicochemical properties of the collagen membrane, such as denaturation temperature, water absorption, pancreatin resistance and mechanical strength were tested before and after crosslinking. The results demonstrate that after crosslinking, the thermal stability, morphological stability, enzymatic resistance and mechanical strength of collagen membrane are all improved. It is suggested that propanedioic acid-NHS ester could improve the physicochemical properties of collagen membrane as a crosslinking agent.%将丙二酸与N-羟基琥珀酰亚胺(NHS)在二环己基碳二亚胺(DCC)催化下反应得到丙二酸NHS酯。以丙二酸NHS酯为交联剂对胶原膜进行交联改性,考察不同交联剂浓度对胶原膜物理化学性能的影响,通过测试其变性温度、吸水率、酶解率和机械强度等,来表征交联前后膜性能的改变。结果表明:经过丙二酸NHS酯交联改性后的胶原膜热稳定性、形态稳定性增强,抵抗酶解的能力增加,机械强度有所提高。说明丙二酸NHS酯作为一种交联剂,可有效改善胶原膜的物理化学性能。

  5. Collagenous spherulosis: An interesting cytological finding in breast lesion

    Directory of Open Access Journals (Sweden)

    Shailja Puri


    Full Text Available Collagenous spherulosis (CS is a rare and interesting entity associated with benign breast lesions. CS is an incidental finding picked up only on 0.2% of cytology specimen. Typically cytologically of CS of breast consists of central spherical hyaline spherule surrounded by myoepithelial cells. The central hyaline spherule has been found to be basement membrane material histochemically and immunohistochemically. The importance of recognizing CS of breast lies in the fact that similar hyaline globules can occur in adenoid cystic carcinoma of the breast (ACCB. The two lesions need to be differentiated on cytology failing which inappropriate treatment can be given for either of the two lesions. We present here a case of CS associated with benign breast lesion and a case of ACCB to describe their cytological features and key points to differentiate them cytologically.

  6. Increased angiogenic response in aortic explants of collagen XVIII/endostatin-null mice. (United States)

    Li, Qing; Olsen, Bjorn R


    Endostatin, a proteolytic fragment of basement membrane-associated collagen XVIII, has been shown to be a potent angiogenesis inhibitor both in vivo and in vitro when given at high concentrations. The precise molecular mechanisms by which it functions and whether or not it plays a role in physiological regulation of angiogenesis are not clear. In mice with targeted null alleles of Col18a1, there appears to be no major abnormality in vascular patterns or capillary density in most organs. Furthermore, the growth of experimental tumors is not increased. However, a detailed analysis of induced angiogenesis in these mice has not been performed. Therefore, we compared the angiogenic responses induced by in vitro culture of aortic explants from collagen XVIII/endostatin-null mice (ko) to wild-type (wt) littermates. We found a twofold increase in microvessel outgrowth in explants from ko mice, relative to wt explants. This increased angiogenesis was reduced to the wt level by the addition of low levels (0.1 microg/ml) of recombinant mouse or human endostatin during the culture period. To address cellular/molecular mechanisms underlying this difference in angiogenic response between ko and wt mice, we isolated endothelial cells from both strains and compared their biological behavior. Proliferation assays showed no difference between the two types of endothelial cells. In contrast, adhesion assays showed a striking difference in their ability to adhere to fibronectin suggesting that collagen XVIII/endostatin may regulate interactions between endothelial cells and underlying basement membrane-associated components, including fibronectin, such that in the absence of collagen XVIII/endostatin, endothelial cells are more adhesive to fibronectin. In the aortic explant assay, characterized by dynamic processes of microvessel elongation and regression, this may result in stabilization of newly formed vessels, reduced regression, and a net increase in microvessel outgrowth in

  7. AGE-breakers cleave model compounds, but do not break Maillard crosslinks in skin and tail collagen from diabetic rats. (United States)

    Yang, Shengzu; Litchfield, John E; Baynes, John W


    Advanced glycation end products (AGE), formed by nonenzymatic Maillard reactions between carbohydrate and protein, contribute to the increase in chemical modification and crosslinking of tissue proteins with age. Acceleration of AGE formation in collagen during hyperglycemia, with resultant effects on vascular elasticity and basement membrane permeability, is implicated in the pathogenesis of diabetic complications. AGE-breakers, such as N-phenacylthiazolium (PTB) and N-phenacyl-4,5-dimethylthiazolium (PMT) halides, have been proposed as therapeutic agents for reversing the increase in protein crosslinking in aging and diabetes. We have confirmed that these compounds, as well as the AGE-inhibitor pyridoxamine (PM), cleave the model AGE crosslink, phenylpropanedione, and have studied the effects of these compounds in reversing the increased crosslinking of skin and tail collagen isolated from diabetic rats. Crosslinking of skin collagen, measured as the half-time for solubilization of collagen by pepsin in 0.5M acetic acid, was increased approximately 5-fold in diabetic, compared to nondiabetic rats. Crosslinking of tail tendon collagen, measured as insolubility in 0.05 N acetic acid, was increased approximately 10-fold. Collagen preparations were incubated in the presence or absence of AGE-breakers or PM in phosphate buffer, pH 7.4, for 24h at 37 degrees C. These treatments did not decrease the half-time for solubilization of diabetic skin collagen by pepsin or increase the acid solubility of diabetic tail tendon collagen. We conclude that, although AGE-breakers and PM cleave model crosslinks, they do not significantly cleave AGE crosslinks formed in vivo in skin collagen of diabetic rats.

  8. 胶原蛋白/壳聚糖复合纳米纤维膜在生物医学工程中的应用%Application of collagen/chitosan compound nanofiber membrane in biomedical engineering

    Institute of Scientific and Technical Information of China (English)

    余丕军; 陈炜


    背景:胶原蛋白-壳聚糖复合纳米纤维膜以其优异的力学性能和良好的组织细胞相容性而成为近年来科学研究的热点.目的:总结胶原蛋白/壳聚糖复合纳米纤维膜在生物医学工程中的应用进展.方法:以"胶原蛋白、壳聚糖、复合纳米纤维膜、胶原蛋白/壳聚糖复合纳米纤维膜、collagen/chitosan、compound Nanofiber membrane、collagen/chitosan compound nanofiber membrane、development of research" 为检索词,应用计算机检索Pubmed数据库、Elsevier数据库、万方数据库1993-01/2010-05关于胶原蛋白/壳聚糖复合纳米纤维膜研究的相关文章,对53篇文献进行分析.结果与结论:研究表明,将胶原蛋白/壳聚糖共混,在不同条件下交联,其共混复合物在力学性能方面较单一的胶原蛋白有一定的改善,其共混膜可以作为较小软骨缺损的修复的支架材料.研究证实胶原蛋白/壳聚糖复合纳米纤维膜有着优异的力学性能、很好的组织细胞相容性和生物可降解性.文章从胶原蛋白和壳聚糖单一生物材料的缺陷性、复合纤维膜的优势及其在生物医药工程中的应用方面进行了探讨.%BACKGROUND: Coil agen/chitosan compound nanofiber membrane has become a research hotspot in recent years in respect of excellent mechanical properties and good biocompatibilrty.OBJECTIVE: To review the research progress of collagen/chitosan compound nanofiber membrane in biomedica I engineering. METHODS: Literatures related to collogen/chitosan compound nanofiber membranefrom PubMed database, Elsevier database and Wanfang database (1993-01/2010-05) were retrieved by computer with the key words of "collagen, chitosan, compound nanofiber membrane, collagen/chitosan compound nanofiber membrane" in Chinese and Eng is h According to indus ion criteria, 53 literatures were selected in this study.RESULTS AND CONCLUSION: The studies have demonstrated that collagen combined with chitosan

  9. Viruses in the Oceanic Basement (United States)

    Jungbluth, Sean P.; Lin, Huei-Ting; Hsieh, Chih-Chiang; Miranda, Jaclyn A.; Schvarcz, Christopher R.; Rappé, Michael S.


    ABSTRACT Microbial life has been detected well into the igneous crust of the seafloor (i.e., the oceanic basement), but there have been no reports confirming the presence of viruses in this habitat. To detect and characterize an ocean basement virome, geothermally heated fluid samples (ca. 60 to 65°C) were collected from 117 to 292 m deep into the ocean basement using seafloor observatories installed in two boreholes (Integrated Ocean Drilling Program [IODP] U1362A and U1362B) drilled in the eastern sediment-covered flank of the Juan de Fuca Ridge. Concentrations of virus-like particles in the fluid samples were on the order of 0.2 × 105 to 2 × 105 ml−1 (n = 8), higher than prokaryote-like cells in the same samples by a factor of 9 on average (range, 1.5 to 27). Electron microscopy revealed diverse viral morphotypes similar to those of viruses known to infect bacteria and thermophilic archaea. An analysis of virus-like sequences in basement microbial metagenomes suggests that those from archaeon-infecting viruses were the most common (63 to 80%). Complete genomes of a putative archaeon-infecting virus and a prophage within an archaeal scaffold were identified among the assembled sequences, and sequence analysis suggests that they represent lineages divergent from known thermophilic viruses. Of the clustered regularly interspaced short palindromic repeat (CRISPR)-containing scaffolds in the metagenomes for which a taxonomy could be inferred (163 out of 737), 51 to 55% appeared to be archaeal and 45 to 49% appeared to be bacterial. These results imply that the warmed, highly altered fluids in deeply buried ocean basement harbor a distinct assemblage of novel viruses, including many that infect archaea, and that these viruses are active participants in the ecology of the basement microbiome. PMID:28270584

  10. Role of α1 and α2 chains of type IV collagen in early fibrotic lesions of idiopathic interstitial pneumonias and migration of lung fibroblasts. (United States)

    Urushiyama, Hirokazu; Terasaki, Yasuhiro; Nagasaka, Shinya; Terasaki, Mika; Kunugi, Shinobu; Nagase, Takahide; Fukuda, Yuh; Shimizu, Akira


    Early fibrotic lesions are thought to be the initial findings of fibrogenesis in idiopathic interstitial pneumonias, but little is known about their properties. Type IV collagen comprises six gene products, α1-α6, and although it is known as a major basement membrane component, its abnormal deposition is seen in fibrotic lesions of certain organs. We studied the expression of type I and III collagen and all α chains of type IV collagen in lung specimens from patients with usual interstitial pneumonia (UIP) or organizing pneumonia (OP) via immunohistochemistry. With cultured lung fibroblasts, we analyzed the expression and function of all α chains of type IV collagen via immunohistochemistry, western blotting, real-time quantitative PCR, and a Boyden chamber migration assay after the knockdown of α1 and α2 chains. Although we observed type I and III collagens in early fibrotic lesions of both UIP and OP, we found type IV collagen, especially α1 and α2 chains, in early fibrotic lesions of UIP but not OP. Fibroblasts enhanced the expression of α1 and α2 chains of type IV collagen after transforming growth factor-β1 stimulation. Small interfering RNA against α1 and α2 chains increased fibroblast migration, with upregulated phosphorylation of focal adhesion kinase (FAK), and adding medium containing fibroblast-produced α1 and α2 chains reduced the increased levels of fibroblast migration and phosphorylation of FAK. Fibroblasts in OP were positive for phosphorylated FAK but fibroblasts in UIP were not. These results suggest that fibroblasts in UIP with type IV collagen deposition, especially α1 and α2 chains, have less ability to migrate from early fibrotic lesions than fibroblasts in OP without type IV collagen deposition. Thus, type IV collagen deposition in early fibrotic lesions of UIP may be implicated in refractory pathophysiology including migration of lesion fibroblasts via a FAK pathway.

  11. Screening and identification of neutralizated single-chain antibody of anti-glomerular basement membrane antibody%中和抗肾小球基底膜抗体的单链抗体的筛选与鉴定

    Institute of Scientific and Technical Information of China (English)

    肖静; 刘章锁; 王沛; 黄留玉; 宋宏彬; 赵明辉


    Objective To screen a human single-chain variable fragments(scFv)against antiGBM antibody.Methods Using phage display technique,the phage antibody library was panned by antiglomerular basement membrane(GBM)antibody which was coated in a micro-titer plate,one clone was found to have high affinity to anti-GBM antibody.The DNA sequence of the positive clone was determined.Results Along with the increase of rounds anti-GBM antibody specific phage antibody was highly enriched and screening efficiency was increased 137 folds than the firest round.ELISA and competition inhibition assay showed that the scFv had a specific combination character with anti-GBM antibody.DNA sequencing confirmed that the whole gene of scFv was 750 bp,and in accordance with humanized single-chain variable region antibody sequence structure.Conclusion The results suggested that the scFv fragment to anti-GBM antibody could be successfully selected by recombinant phage antibody technique,which will laid an experimental foundation for further research of the therapy of Goodpasture syndrome.%目的 制备人抗肾小球基底膜(GBM)抗体的特异性人源化单链可变区抗体.方法 采用噬菌体表面展示技术,获得一个与人抗GBM抗体结合活性较强的单链可变区抗体片段的阳性克隆,并对该克隆进行DNA序列测定分析.结果 对噬菌体单链可变区抗体库经过3轮筛选后,与第1轮相比富集了137倍.噬菌体抗体与人抗GBM抗体的结合活性其中有35株克隆ELISA的吸光度较高.对这些噬菌体抗体进行交叉反应后,确定其中有10株交叉反应较弱.确定1株(C31)阳性克隆提取质粒,进行DNA序列测定,大小为750 bp,并符合人源化单链可变区抗体的序列结构.结论 应用噬菌体展示技术成功获得人-抗GBM抗体的单链可变区抗体基因,为临床上治疗Goodpasture综合征奠定实验基础.

  12. Characterization of collagen-chitosan/hydroxyapatite composite membrane for guided tissue regeneration%胶原蛋白-壳聚糖/羟基磷灰石复合组织引导再生膜的性能表征

    Institute of Scientific and Technical Information of China (English)

    李志宏; 武继民; 关静; 黄姝杰; 李瑞欣; 郭勇; 马军; 张西正


    A novel kind of guided tissue regeneration barrier membrane has been fabricated using an alternate soaking technique on collagen and chitosan composite membranes. The prepared guided tissue regeneration membranes were characterized by infrared spectroscopy, X-ray diffraction, end scanning electron microscopy, and the mechanical property was tested by INSTRON-5865. The results showed that collagen-chitosan/hydroxyapatite membrane were similar to natural bone structure in crystal phase, chemical composition, and crystal size; the morphology and content of hydroxyapatite gradually transformed with increasing immersion cycles in calcium and phosphate solutions, which lead to the tensile strength of collagen-chitosan/hydroxyapatite reduced to 23.67 MPa after five immersion cycles. The results also indicated that collagen-chitosan/hydroxyapatite membrane had better mechanical property than pure collagen membrane; therefore, it is a potential candidate as guided tissue regeneration membrane.%采用生物矿化交替沉积法制备了胶原蛋白-壳聚糖/羟基磷灰石复合组织引导再生膜材料,用红外光谱、X射线衍射、扫描电子显微镜及INSTRON-5865力学实验机等方法对膜材料性能进行了分析表征.结果表明:交替沉积可制备在晶相组成、化学成分、羟基磷灰石尺寸上具有类骨结构的羟基磷灰石涂层;随沉积次数增加,羟基磷灰石形态发生改变,组织引导再生膜力学性能呈下降趋势,沉积5次后拉伸强度为23.67 MPa,仍优于纯胶原膜,适作组织引导再生膜.

  13. Bioengineered collagens (United States)

    Ramshaw, John AM; Werkmeister, Jerome A; Dumsday, Geoff J


    Mammalian collagen has been widely used as a biomedical material. Nevertheless, there are still concerns about the variability between preparations, particularly with the possibility that the products may transmit animal-based diseases. Many groups have examined the possible application of bioengineered mammalian collagens. However, translating laboratory studies into large-scale manufacturing has often proved difficult, although certain yeast and plant systems seem effective. Production of full-length mammalian collagens, with the required secondary modification to give proline hydroxylation, has proved difficult in E. coli. However, recently, a new group of collagens, which have the characteristic triple helical structure of collagen, has been identified in bacteria. These proteins are stable without the need for hydroxyproline and are able to be produced and purified from E. coli in high yield. Initial studies indicate that they would be suitable for biomedical applications. PMID:24717980

  14. Novel feline autoimmune blistering disease resembling bullous pemphigoid in humans: IgG autoantibodies target the NC16A ectodomain of type XVII collagen (BP180/BPAG2). (United States)

    Olivry, T; Chan, L S; Xu, L; Chace, P; Dunston, S M; Fahey, M; Marinkovich, M P


    In humans and dogs, bullous pemphigoid (BP) is an autoimmune blistering disease associated with the production of basement membrane autoantibodies that target the 180-kd type XVII collagen (BP180, BPAG2) and/or the 230-kd plakin epidermal isoform BPAG1e (BP230). In two adult cats, an acquired dermatosis and stomatitis was diagnosed as BP subsequent to the fulfillment of the following criteria: 1) presence of cutaneous vesicles, erosions, and ulcers; 2) histologic demonstration of subepidermal vesiculation with inflammatory cells, including eosinophils; 3) in vivo deposition of IgG autoantibodies at the epidermal basement membrane zone; and 4) serum IgG autoantibodies targeting a 180-kd epidermal protein identified as type XVII collagen. In both cats, the antigenic epitopes targeted by IgG autoantibodies were shown to be situated in the NC16A ectodomain of type XVII collagen, a situation similar to that of humans and dogs with BP. Feline BP therefore can be considered a clinical, histopathologic, and immunologic homologue of BP in humans and dogs.

  15. Clinical evaluation of regenerative potential of type I collagen membrane along with xenogenic bone graft in the treatment of periodontal intrabony defects assessed with surgical re-entry and radiographic linear and densitometric analysis

    Directory of Open Access Journals (Sweden)

    Sowmya N


    Full Text Available Background and Objectives: The primary goal of periodontal therapy is to restore the tooth supporting tissues lost due to periodontal disease. The aim of the present study was to compare the efficacy of combination of type I collagen (GTR membrane and xenogenic bone graft with open flap debridement (OFD in treatment of periodontal intrabony defects. Materials and Methods: Twenty paired intrabony defects were surgically treated using split mouth design. The defects were randomly assigned to treatment with OFD + collagen membrane + bone graft (Test or OFD alone (Control. The clinical efficacy of two treatment modalities was evaluated at 9 month postoperatively by clinical, radiographical, and intrasurgical (re-entry parameters. The measurements included probing pocket depth (PD, clinical attachment level (CAL, gingival recession (GR, bone fill (BF, bone density (BD and intra bony component (INTRA. Results: The mean reduction in PD at 0-9 month was 3.3±0.82 mm and CAL gain of 3.40±1.51 mm occurred in the collagen membrane + bone graft (Test group; corresponding values for OFD (Control were 2.20±0.63 mm and 1.90±0.57 mm. Similar pattern of improvement was observed when radiographical and intra-surgical (re-entry post operative evaluation was made. All improvement in different parameters was statistically significant (P< 0.01. Interpretation and Conclusion: Treatment with a combination of collagen membrane and bone graft led to a significantly more favorable clinical outcome in intrabony defects as compared to OFD alone.

  16. A comparative evaluation of the effectiveness of guided tissue regeneration by using a collagen membrane with or without decalcified freeze-dried bone allograft in the treatment of infrabony defects: A clinical and radiographic study


    Kher, Vishal Kiran; Manohar L. Bhongade; Shori, Tony D.; Kolte, Abhay P.; Dharamthok, Swarup B.; Shrirao, Tushar S.


    Background: The present, randomized, controlled clinical and radiographic study was undertaken to compare the effectiveness of guided tissue regeneration (GTR) by using a collagen membrane barrier with or without decalcified freeze-dried bone allograft (DFDBA) in the treatment of periodontal infrabony defects characterized by unfavorable architecture. Materials and Methods: Sixteen systemically healthy patients with 20 periodontal infrabony defects were selected for the study. Each patient ha...

  17. Perlecan antagonizes collagen IV and ADAMTS9/GON-1 in restricting the growth of presynaptic boutons. (United States)

    Qin, Jianzhen; Liang, Jingjing; Ding, Mei


    In the mature nervous system, a significant fraction of synapses are structurally stable over a long time scale. However, the mechanisms that restrict synaptic growth within a confined region are poorly understood. Here, we identified that in the C. elegans neuromuscular junction, collagens Type IV and XVIII, and the secreted metalloprotease ADAMTS/GON-1 are critical for growth restriction of presynaptic boutons. Without these components, ectopic boutons progressively invade into the nonsynaptic region. Perlecan/UNC-52 promotes the growth of ectopic boutons and functions antagonistically to collagen Type IV and GON-1 but not to collagen XVIII. The growth constraint of presynaptic boutons correlates with the integrity of the extracellular matrix basal lamina or basement membrane (BM), which surrounds chemical synapses. Fragmented BM appears in the region where ectopic boutons emerge. Further removal of UNC-52 improves the BM integrity and the tight association between BM and presynaptic boutons. Together, our results unravel the complex role of the BM in restricting the growth of presynaptic boutons and reveal the antagonistic function of perlecan on Type IV collagen and ADAMTS protein.

  18. In vivo evidence for a bridging role of a collagen V subtype at the epidermis-dermis interface. (United States)

    Bonod-Bidaud, Christelle; Roulet, Muriel; Hansen, Uwe; Elsheikh, Ahmed; Malbouyres, Marilyne; Ricard-Blum, Sylvie; Faye, Clément; Vaganay, Elisabeth; Rousselle, Patricia; Ruggiero, Florence


    Collagen V is the defective product in most cases of classical Ehlers-Danlos syndrome (EDS), a connective tissue disorder typically characterized by skin fragility and abnormal wound healing. Collagen V assembles into diverse molecular forms. The predominant α1(V)(2)α2(V) heterotrimer controls fibrillogenesis in skin and other tissues. The α1(V)(3) minor form is thought to occur in skin, but its function is unknown. To elucidate its role, we generated transgenic mice that overexpress the human α1(V)(3) homotrimer in the epidermis. The transgene-derived product is deposited as thin unstriated fibrillar material in the basement membrane zone of embryonic and perinatal epidermis and hair follicles. Accumulation of α1(V)(3)-containing fibrils leads to ultrastructural modifications at the epidermis-dermis interface and provokes changes in biomechanical properties, although not statistically significant. Using superparamagnetic immunobeads to isolate authentic suprastructures and protein-binding assays, we demonstrate that the homotrimer is part of a protein network containing collagen IV, laminin-111, and the dermal collagen VI. Our data show that the homotrimer serves as a bridging molecule that contributes to the stabilization of the epidermal-dermal interface. This finding strongly suggests that collagen V may be expressed in skin as different subtypes with important but distinct roles in matrix organization and stability.

  19. Cutaneous collagenous vasculopathy: a new case series with clinicopathologic and ultrastructural correlation, literature review, and insight into the pathogenesis. (United States)

    Salama, Samih S


    Cutaneous collagenous vasculopathy (CCV) is a rare distinct idiopathic microangiopathy of the superficial cutaneous vasculature. Seven new cases are reported (6 females and 1 male) ranging in age from 42 to 85 years, with some showing unusual clinical and histopathological findings. All presented with macular telangiectases starting on the lower extremities and spreading progressively in 5 cases and were suspected to have generalized essential telangiectasia. Two cases had a history of over 20 years. One case had lesions in the abdominal striae, and 1 was markedly ecchymotic. All skin biopsies showed the characteristic features of CCV with dilatation and marked thickening of the walls of superficial dermal blood vessels displaying reduplication of the basement membrane on periodic acid-Schiff-diastase stain and deposition of hyaline collagenous material immunostaining as collagen type IV, and showing decreased or absent actin staining. However, the changes were subtle and only seen focally in some biopsies. Few lymphoid cells were present around occasional vessels. Electron microscopy showed increased basement membrane lamellae with marked deposition of normal and some abnormal collagen (Luse-like bodies) and focal endothelial damage, suggesting reparative perivascular fibrosis resulting from repeated endothelial injury. These cases (and all 18 previously reported ones) are of a wide age range and no gender predilection. This disorder is underdiagnosed, and it is likely that some cases clinically suspected to be generalized essential telangiectasia may actually represent CCV. Better recognition by dermatologists may lead to more biopsies from patients with generalized telangiectasia and a further understanding of the pathogenesis of CCV and its relationship to other cutaneous vascular disorders.

  20. [Collagenous colitis]. (United States)

    Lindström, C G


    Collagenous colitis is now regarded by an overwhelming majority of authors as a clinicopathological entity and has been taken up as a such in many text-books and diagnostic atlases (Morson & Dawson, 1990, Fenoglio-Preiser et al., 1989, Whitehead 1985, Whitehead 1989). A good, detailed review of cases of collagenous colitis published up to 1988 was performed by Perri et al. Collagenous colitis was also presented to a wider medical public through a clinicopathological conference case at Massachusetts General Hospital (Case 29-1988). Finally it may be added that collagenous colitis has been included in the new fourth edition of Robbins Pathologic Basis of Disease (Cotran, Kumar, Robbins, 1989), where the possibility of an autoimmune disease is stressed.

  1. Hybrid Membranes of PLLA/Collagen for Bone Tissue Engineering: A Comparative Study of Scaffold Production Techniques for Optimal Mechanical Properties and Osteoinduction Ability

    Directory of Open Access Journals (Sweden)

    Flávia Gonçalves


    Full Text Available Synthetic and natural polymer association is a promising tool in tissue engineering. The aim of this study was to compare five methodologies for producing hybrid scaffolds for cell culture using poly-l-lactide (PLLA and collagen: functionalization of PLLA electrospun by (1 dialkylamine and collagen immobilization with glutaraldehyde and by (2 hydrolysis and collagen immobilization with carbodiimide chemistry; (3 co-electrospinning of PLLA/chloroform and collagen/hexafluoropropanol (HFP solutions; (4 co-electrospinning of PLLA/chloroform and collagen/acetic acid solutions and (5 electrospinning of a co-solution of PLLA and collagen using HFP. These materials were evaluated based on their morphology, mechanical properties, ability to induce cell proliferation and alkaline phosphatase activity upon submission of mesenchymal stem cells to basal or osteoblastic differentiation medium (ODM. Methods (1 and (2 resulted in a decrease in mechanical properties, whereas methods (3, (4 and (5 resulted in materials of higher tensile strength and osteogenic differentiation. Materials yielded by methods (2, (3 and (5 promoted osteoinduction even in the absence of ODM. The results indicate that the scaffold based on the PLLA/collagen blend exhibited optimal mechanical properties and the highest capacity for osteodifferentiation and was the best choice for collagen incorporation into PLLA in bone repair applications.

  2. 纳滤膜对鱼鳞小分子胶原肽提取液脱盐性能的研究%Study on desalination performance of small-molecule collagen peptide extraction from fish scale with nanofiltration membranes

    Institute of Scientific and Technical Information of China (English)

    林谢凤; 郭洪辉


    采用纳滤技术进行鱼鳞小分子胶原肽提取液的脱盐小试实验,研究了不同分子量的纳滤膜对料液脱盐效果及胶原肽回收率的影响。实验结果表明,截留分子量为700 u的卷式纳滤膜能够有效地除去鱼鳞胶原肽提取液中的无机盐类。脱盐后,鱼鳞胶原肽提取液的电导率从26.9 ms/cm下降到80.3μs/cm,无机盐浓度从13.9 g/L下降到0.04 g/L ,胶原肽的回收率为72.7%,最终产品的灰分为0.79%。%Nanofiltration technology was applied to the desalination of small-molecule collagen peptide ex-traction from fish scale. The impact of nanofiltration membranes with different molecular weight cut off (MW-CO)on the desalination effect and collagen recovery was researched. The result showed that the optimal mem-brane was the nanofiltration membrane with MWCO 700 u. After desalination,the conductivity of collagen peptide extraction decreased from 26.9 ms/cm to 80.3 μs/cm and the inorganic salt concentration decreased from 13.9 g/L to 0.04 g/L. The recovery rate of collagen peptide was 72.7%. The ash content in the final product was 0.79%.

  3. Collagen scaffolds loaded with collagen-binding NGF-beta accelerate ulcer healing. (United States)

    Sun, Wenjie; Lin, Hang; Chen, Bing; Zhao, Wenxue; Zhao, Yannan; Xiao, Zhifeng; Dai, Jianwu


    Studies have shown that exogenous nerve growth factor (NGF) accelerates ulcer healing, but the inefficient growth factor delivery system limits its clinical application. In this report, we found that the native human NGF-beta fused with a collagen-binding domain (CBD) could form a collagen-based NGF targeting delivery system, and the CBD-fused NGF-beta could bind to collagen membranes efficiently. Using the rabbit dermal ischemic ulcer model, we have found that this targeting delivery system maintains a higher concentration and stronger bioactivity of NGF-beta on the collagen membranes by promoting peripheral nerve growth. Furthermore, it enhances the rate of ulcer healing through accelerating the re-epithelialization of dermal ulcer wounds and the formation of capillary lumens within the newly formed tissue area. Thus, collagen membranes loaded with collagen-targeting human NGF-beta accelerate ulcer healing efficiently.

  4. Membraner

    DEFF Research Database (Denmark)

    Bach, Finn


    Notatet giver en kort introduktion til den statiske virkemåde af membraner og membrankonstruktioner......Notatet giver en kort introduktion til den statiske virkemåde af membraner og membrankonstruktioner...

  5. NMR studies demonstrate a unique AAB composition and chain register for a heterotrimeric type IV collagen model peptide containing a natural interruption site. (United States)

    Xiao, Jianxi; Sun, Xiuxia; Madhan, Balaraman; Brodsky, Barbara; Baum, Jean


    All non-fibrillar collagens contain interruptions in the (Gly-X-Y)n repeating sequence, such as the more than 20 interruptions found in chains of basement membrane type IV collagen. Two selectively doubly labeled peptides are designed to model a site in type IV collagen with a GVG interruption in the α1(IV) and a corresponding GISLK sequence within the α2(IV) chain. CD and NMR studies on a 2:1 mixture of these two peptides support the formation of a single-component heterotrimer that maintains the one-residue staggering in the triple-helix, has a unique chain register, and contains hydrogen bonds at the interruption site. Formation of hydrogen bonds at interruption sites may provide a driving force for self-assembly and chain register in type IV and other non-fibrillar collagens. This study illustrates the potential role of interruptions in the structure, dynamics, and folding of natural collagen heterotrimers and forms a basis for understanding their biological role.

  6. Closure of 1.5-cm alveolar oral antral fistula with intra-alveolar sinus membrane elevation and bone morphogenetic protein-2/collagen graft followed by dental implant restoration: case report. (United States)

    Cottam, Jared R; Jensen, Ole T; Beatty, Lucas; Ringeman, Jason


    Closure of a 1.5-cm oral antral fistula was done in combination with sinus floor and extraction socket grafting using recombinant human bone morphogenetic protein-2 within a collagen sponge matrix. The approach to the sinus was transalveolar, with elevation of the sinus membrane done through a molar extraction socket. Following graft placement, soft tissue repair was done with a buccal advancement flap. A dental implant was subsequently placed and restored. Peri-implant bone and implant stability were well maintained at the 1-year follow up examination.

  7. Persistence of collagen type II-specific T-cell clones in the synovial membrane of a patient with rheumatoid arthritis

    Energy Technology Data Exchange (ETDEWEB)

    Londei, M.; Savill, C.M.; Verhoef, A.; Brennan, F.; Leech, Z.A.; Feldmann, M. (Charing Cross Sunley Research Centre, London (England)); Duance, V. (Bristol Laboratory (England)); Maini, R.N. (Kennedy Institute of Rheumatology, London (England))


    Rheumatoid arthritis is an autoimmune disease characterized by T-cell infiltration of the synovium of joints. Analysis of the phenotype and antigen specificity of the infiltrating cells may thus provide insight into the pathogenesis of rheumatoid arthritis. T cells were cloned with interleukin 2, a procedure that selects for in vivo-activated cells. All clones had the CD4 CDW29 phenotype. Their antigen specificity was tested by using a panel of candidate joint autoantigens. Four of 17 reacted against autologous blood mononuclear cells. Two clones proliferated in response to collagen type II. After 21 months, another set of clones was derived from synovial tissue of the same joint. One of eight clones tested showed a strong proliferative response against collagen type II. The uncloned synovial T cells of a third operation from another joint also responded to collagen type II. The persistence of collagen type II-specific T cells in active rheumatoid joints over a period of 3 years suggests that collagen type II could be one of the autoantigens involved in perpetuating the inflammatory process in rheumatoid arthritis.

  8. Long-term results of dual anti-collagen membrane in guided tissue regeneration%双抗胶原膜应用于引导组织再生的长期疗效观察

    Institute of Scientific and Technical Information of China (English)

    尚姝环; 李成章; 樊明文


    目的 观察双抗胶原膜应用于引导组织再生(guided tissue regeneration,GTR)的长期疗效,以期为临床提供参考.方法 对24例重度牙周炎患者的26颗患牙按单双数法随机分组,分别应用双抗胶原膜(试验组)和牛腱胶原膜(对照组)对3l处垂直型骨吸收和根分叉病变进行GTB治疗,试验组共13例患者,14颗患牙,16处骨缺损;对照组共11例患者,12颗患牙,15处骨缺损.分别于术前、术后6个月、1、3和6年进行临床附着水平、计算机辅助密度影像分析(computer assisted densitometry image analysis,CADIA)等指标的临床检查,并对结果进行统计学分析.结果 GTR术后1年,试验组临床附着获得是(3.93±1.74)mm,对照组是(2.25±1.90)mm(t=2.146,P=0.044),CADIA值分别增加(53.14±21.35)和(32.96±17.97)(t=2.319,P=0.031).GTR术后6年与术后1年临床附着水平相比,试验组和对照组仅分别平均有0.35 mm和0.34 mm的附着丧失.结论 在6年随访的病例中,双抗胶原膜应用于引导组织再生所获得的再生牙周组织可以长期保持稳定.%Objective To evaluate the long-term clinical effect of dual anti-collagen membranes in guided tissue regeneration(GTR). Methods This randomized clinical trial included 26 teeth in 24 patients, presenting a total of 31 lesions consisting of intrabony defects and furcation defects. Twenty-six teeth were divided into two groups and treated by GTR with dual anti-collagen membranes and atelocollagen membranes, respectively. At baseline, 6 months, 1, 3 and 6 years, the following parameters were recorded; clinical attachment level, probing depth, gingival recession and the quantity of alveolar bone analyzed by computer assisted densitometry image analysis (CADIA). Results At 1 year after GTR surgery, the gain of clinical attachment in dual anti-collagen membranes group was (3. 93 ± 1. 74) mm,compared with (2. 25 ± 1. 90) mm in atelocollagen group(P =0. 044). The increasing of the value of CADIA in dual

  9. Type I collagen as an extracellular matrix for the in vitro growth of human small intestinal epithelium.

    Directory of Open Access Journals (Sweden)

    Ziyad Jabaji

    Full Text Available We previously reported in vitro maintenance and proliferation of human small intestinal epithelium using Matrigel, a proprietary basement membrane product. There are concerns over the applicability of Matrigel-based methods for future human therapies. We investigated type I collagen as an alternative for the culture of human intestinal epithelial cells.Human small intestine was procured from fresh surgical pathology specimens. Small intestinal crypts were isolated using EDTA chelation. Intestinal subepithelial myofibroblasts were isolated from a pediatric sample and expanded in vitro. After suspension in Matrigel or type I collagen gel, crypts were co-cultured above a confluent layer of myofibroblasts. Crypts were also grown in monoculture with exposure to myofibroblast conditioned media; these were subsequently sub-cultured in vitro and expanded with a 1∶2 split ratio. Cultures were assessed with light microscopy, RT-PCR, histology, and immunohistochemistry.Collagen supported viable human epithelium in vitro for at least one month in primary culture. Sub-cultured epithelium expanded through 12 passages over 60 days. Histologic sections revealed polarized columnar cells, with apical brush borders and basolaterally located nuclei. Collagen-based cultures gave rise to monolayer epithelial sheets at the gel-liquid interface, which were not observed with Matrigel. Immunohistochemical staining identified markers of differentiated intestinal epithelium and myofibroblasts. RT-PCR demonstrated expression of α-smooth muscle actin and vimentin in myofibroblasts and E-Cadherin, CDX2, villin 1, intestinal alkaline phosphatase, chromogranin A, lysozyme, and Lgr5 in epithelial cells. These markers were maintained through several passages.Type I collagen gel supports long-term in vitro maintenance and expansion of fully elaborated human intestinal epithelium. Collagen-based methods yield familiar enteroid structures as well as a new pattern of sheet

  10. Blockade of Ets-1 attenuates epidermal growth factor-dependent collagen loss in human carotid plaque smooth muscle cells. (United States)

    Rao, Velidi H; Rai, Vikrant; Stoupa, Samantha; Agrawal, Devendra K


    Although degradation of extracellular matrix by matrix metalloproteinases (MMPs) is thought to be involved in symptomatic (S) carotid plaques in atherosclerosis, the mechanisms of MMP expression are poorly understood. Here, we demonstrate that collagen loss in vascular smooth vessel cells (VSMCs) isolated from S plaques was induced by epidermal growth factor (EGF) through the activation of p38-MAPK and JNK-MAPK pathways. Inhibitors of p38-MAPK and JNK-MAPK signaling pathways downregulated the expression of MMP-1 and MMP-9. In addition, we examined whether v-ets erythroblastosis virus E26 oncogene homologue 1 (Ets-1), an important regulator of different genes, is involved in destabilizing S plaques in patients with carotid stenosis. We demonstrate that EGF induces Ets-1 expression and decreases interstitial and basement membrane collagen in vascular smooth muscle cells (VSMCs) from patients with carotid stenosis. Increased expression of MMP-1 and -9 and decreased collagen mRNA transcripts were also found in Ets-1-overexpressed VSMCs. Transfection with both dominant-negative form of Ets-1 and small interfering RNA blocked EGF-induced MMP-1 and -9 expressions and increased the mRNA transcripts for collagen I (α1) and collagen III (α1) in S compared with asymptomatic (AS) carotid plaques. Inhibitors of p38-MAPK (SB202190) and JNK-MAPK (SP600125) signaling pathways decreased the expression of Ets-1, MMP-1, and MMP-9 and increased collagen type I and III expression in EGF-treated VSMCs. This study provides a mechanistic insight into the role of Ets-1 in the plaque destabilization in patients with carotid stenosis involving p38-MAPK and JNK signaling pathways.

  11. Sequence dependence of kinetics and morphology of collagen model peptide self-assembly into higher order structures (United States)

    Kar, Karunakar; Wang, Yuh-Hwa; Brodsky, Barbara


    The process of self-assembly of the triple-helical peptide (Pro-Hyp-Gly)10 into higher order structure resembles the nucleation-growth mechanism of collagen fibril formation in many features, but the irregular morphology of the self-assembled peptide contrasts with the ordered fibers and networks formed by collagen in vivo. The amino acid sequence in the central region of the (Pro-Hyp-Gly)10 peptide was varied and found to affect the kinetics of self-assembly and nature of the higher order structure formed. Single amino acid changes in the central triplet produced irregular higher order structures similar to (Pro-Hyp-Gly)10, but the rate of self-association was markedly delayed by a single change in one Pro to Ala or Leu. The introduction of a Hyp-rich hydrophobic sequence from type IV collagen resulted in a more regular suprastructure of extended fibers that sometimes showed supercoiling and branching features similar to those seen for type IV collagen in the basement membrane network. Several peptides, where central Pro-Hyp sequences were replaced by charged residues or a nine-residue hydrophobic region from type III collagen, lost the ability to self-associate under standard conditions. The inability to self-assemble likely results from loss of imino acids, and lack of an appropriate distribution of hydrophobic/electrostatic residues. The effect of replacement of a single Gly residue was also examined, as a model for collagen diseases such as osteogenesis imperfecta and Alport syndrome. Unexpectedly, the Gly to Ala replacement interfered with self-assembly of (Pro-Hyp-Gly)10, while the peptide with a Gly to Ser substitution self-associated to form a fibrillar structure. PMID:18441232

  12. Guided bone regeneration at a dehiscence-type defect using chitosan/collagen membranes in dogs%壳聚糖-胶原膜引导种植体颊侧骨缺损再生的实验

    Institute of Scientific and Technical Information of China (English)

    李晓静; 王新木; 苗滪汶; 杨国利; 高波; 董研


    Objective To compare a developed absorbable chitosan/collagen membrane (CCM) with a standard biodegradable collagen membrane for the treatment of implant dehiscence-type defect in dog model.Methods The right four mandibular premolars and the first molar were extracted in each of 10 beagle dogs.Four months later,acute buccal dehiscence-type defects were surgically created following implant site preparation in each dog.Using self-control,defects were randomly assigned to four different groups:CCM-1 (with the ratio of chitosan and collagen of 40∶1),CCM-2 (with the ratio of chitosan and collagen of 20∶1),Bio-Gide collagen membrane (BG collagen),control.The animals were sacrificed after 4 (3 animals),8 (3 animals) and 12 (4 animals) weeks of healing interval for histological observation and histomorphometrical analysis including defect length(DL),new bone height (NBH),bone-to-implant contact (BIC) and area of new bone fill (BA).Results Newly formed bone was observed in all the groups and became mature with time.At 8 weeks,increased mean NBH and BIC values were obtained for all the groups,the mean NBH values of the CCM-1,CCM-2 and BG groups[(1.1 0 ±0.11) ~ (1.48 ±0.07) mm] were significantly higher than that of the control [(0.74 ± 0.12)mm] (P < 0.05).At 12 weeks,the membranes treated groups obtained more mean NBH,BIC and BA values compared with the control.The CCM-1 groups showed the highest mean NBH value [(1.91 ± 0.25)mm],which was significantly higher than the control [(1.20 ± 0.34) mm] (P < 0.05).However,no statistically significant differences in BIC and BA were found between membrane groups and control and among the membranes treated groups.Conclusions The results of this study demonstrated that the developed CCM can enhance bone regeneration and obtaine similar amounts of newly formed bone compared with defects regenerated with a standard collagen membrane.%目的 研究用壳聚糖-胶原膜(chitosan-collagen membrane,CCM)引导种植体颊侧裂

  13. Human collagen produced in plants (United States)

    Shoseyov, Oded; Posen, Yehudit; Grynspan, Frida


    Consequential to its essential role as a mechanical support and affinity regulator in extracellular matrices, collagen constitutes a highly sought after scaffolding material for regeneration and healing applications. However, substantiated concerns have been raised with regard to quality and safety of animal tissue-extracted collagen, particularly in relation to its immunogenicity, risk of disease transmission and overall quality and consistency. In parallel, contamination with undesirable cellular factors can significantly impair its bioactivity, vis-a-vis its impact on cell recruitment, proliferation and differentiation. High-scale production of recombinant human collagen Type I (rhCOL1) in the tobacco plant provides a source of an homogenic, heterotrimeric, thermally stable “virgin” collagen which self assembles to fine homogenous fibrils displaying intact binding sites and has been applied to form numerous functional scaffolds for tissue engineering and regenerative medicine. In addition, rhCOL1 can form liquid crystal structures, yielding a well-organized and mechanically strong membrane, two properties indispensable to extracellular matrix (ECM) mimicry. Overall, the shortcomings of animal- and cadaver-derived collagens arising from their source diversity and recycled nature are fully overcome in the plant setting, constituting a collagen source ideal for tissue engineering and regenerative medicine applications. PMID:23941988

  14. Type XI Collagen

    DEFF Research Database (Denmark)

    Luo, Yunyun


    Type XI collagen is a fibrillary collagen. Type XI collagen is broadly distributed in articular cartilage, testis, trachea, tendons, trabecular bone, skeletal muscle, placenta, lung, and the neoepithelium of the brain. Type XI collagen is able to regulate fibrillogenesis by maintaining the spacing...... and diameter of type II collagen fibrils, and a nucleator for the fibrillogenesis of collagen types I and II. Mutations in type XI collagen are associated with Stickler syndrome, Marshall syndrome, fibrochondrogenesis, otospondylomegaepiphyseal dysplasia deafness, and Weissenbacher–Zweymüller syndrome. Type XI...... collagen binds heparin, heparan sulfate, and dermatan sulfate. Currently there are no biomarkers for type XI collagen....


    Directory of Open Access Journals (Sweden)



    Full Text Available INTRODUCTION: Amniotic membrane is the innermost layer of the f etal membranes. It has a stromal matrix, a collagen layer, and an overlying basement membrane with a single layer of epithelium. (1 Amniotic membrane has unique properties including an ti-adhesive effects, bacterio-static properties, wound protection, pain redu ction, and epithelialisation effects. Another characteristic of amniotic membrane is the lack of imunogenicity. (2 Amniotic membranes have been used as a dressing to promote he aling of chronic ulcers of the leg and as a biological dressing for burned skin and skin woun ds. (3, 4 It has also been used in surgical reconstruction of artificial vagina, for repairing o mphaloceles, and to prevent tissue adhesion in surgeries of the abdomen, head, or pelvis. (5, 6 Amniotic membrane has been successfully used in ocular conditions like persistent epithelial defects (7, pterygium, (8 Symblepharon (9 and for ocular surface reconstruction. (10, 11 The purpose of this study was to evaluate the use of cryo- preserved Human amniotic membrane graft( HAMT, with or without limbal autograft transplantation (LAT in patients with previous and fr esh chemical eye injuries respectively. Institutional ethical committee approval was obtained .

  16. Chemical chaperone treatment reduces intracellular accumulation of mutant collagen IV and ameliorates the cellular phenotype of a COL4A2 mutation that causes haemorrhagic stroke. (United States)

    Murray, Lydia S; Lu, Yinhui; Taggart, Aislynn; Van Regemorter, Nicole; Vilain, Catheline; Abramowicz, Marc; Kadler, Karl E; Van Agtmael, Tom


    Haemorrhagic stroke accounts for ∼20% of stroke cases and porencephaly is a clinical consequence of perinatal cerebral haemorrhaging. Here, we report the identification of a novel dominant G702D mutation in the collagen domain of COL4A2 (collagen IV alpha chain 2) in a family displaying porencephaly with reduced penetrance. COL4A2 is the obligatory protein partner of COL4A1 but in contrast to most COL4A1 mutations, the COL4A2 mutation does not lead to eye or kidney disease. Analysis of dermal biopsies from a patient and his unaffected father, who also carries the mutation, revealed that both display basement membrane (BM) defects. Intriguingly, defective collagen IV incorporation into the dermal BM was observed in the patient only and was associated with endoplasmic reticulum (ER) retention of COL4A2 in primary dermal fibroblasts. This intracellular accumulation led to ER stress, unfolded protein response activation, reduced cell proliferation and increased apoptosis. Interestingly, the absence of ER retention of COL4A2 and ER stress in cells from the unaffected father indicate that accumulation and/or clearance of mutant COL4A2 from the ER may be a critical modifier for disease development. Our analysis also revealed that mutant collagen IV is degraded via the proteasome. Importantly, treatment of patient cells with a chemical chaperone decreased intracellular COL4A2 levels, ER stress and apoptosis, demonstrating that reducing intracellular collagen accumulation can ameliorate the cellular phenotype of COL4A2 mutations. Importantly, these data highlight that manipulation of chaperone levels, intracellular collagen accumulation and ER stress are potential therapeutic options for collagen IV diseases including haemorrhagic stroke.

  17. Marine-derived collagen biomaterials from echinoderm connective tissues

    KAUST Repository

    Ferrario, Cinzia


    The use of marine collagens is a hot topic in the field of tissue engineering. Echinoderms possess unique connective tissues (Mutable Collagenous Tissues, MCTs) which can represent an innovative source of collagen to develop collagen barrier-membranes for Guided Tissue Regeneration (GTR). In the present work we used MCTs from different echinoderm models (sea urchin, starfish and sea cucumber) to produce echinoderm-derived collagen membranes (EDCMs). Commercial membranes for GTR or soluble/reassembled (fibrillar) bovine collagen substrates were used as controls. The three EDCMs were similar among each other in terms of structure and mechanical performances and were much thinner and mechanically more resistant than the commercial membranes. Number of fibroblasts seeded on sea-urchin membranes were comparable to the bovine collagen substrates. Cell morphology on all EDCMs was similar to that of structurally comparable (reassembled) bovine collagen substrates. Overall, echinoderms, and sea urchins particularly, are alternative collagen sources to produce efficient GTR membranes. Sea urchins display a further advantage in terms of eco-sustainability by recycling tissues from food wastes.

  18. Irradiation Alters MMP-2/TIMP-2 System and Collagen Type IV Degradation in Brain

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Won Hee [School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, Blacksburg, Virginia (United States); Warrington, Junie P.; Sonntag, William E. [Reynolds Oklahoma Center on Aging, Department of Geriatric Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma (United States); Lee, Yong Woo, E-mail: [School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, Blacksburg, Virginia (United States); Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia (United States)


    Purpose: Blood-brain barrier (BBB) disruption is one of the major consequences of radiation-induced normal tissue injury in the central nervous system. We examined the effects of whole-brain irradiation on matrix metalloproteinases (MMPs)/tissue inhibitors of metalloproteinases (TIMPs) and extracellular matrix (ECM) degradation in the brain. Methods and Materials: Animals received either whole-brain irradiation (a single dose of 10 Gy {gamma}-rays or a fractionated dose of 40 Gy {gamma}-rays, total) or sham-irradiation and were maintained for 4, 8, and 24 h following irradiation. mRNA expression levels of MMPs and TIMPs in the brain were analyzed by real-time reverse transcriptase-polymerase chain reaction (PCR). The functional activity of MMPs was measured by in situ zymography, and degradation of ECM was visualized by collagen type IV immunofluorescent staining. Results: A significant increase in mRNA expression levels of MMP-2, MMP-9, and TIMP-1 was observed in irradiated brains compared to that in sham-irradiated controls. In situ zymography revealed a strong gelatinolytic activity in the brain 24 h postirradiation, and the enhanced gelatinolytic activity mediated by irradiation was significantly attenuated in the presence of anti-MMP-2 antibody. A significant reduction in collagen type IV immunoreactivity was also detected in the brain at 24 h after irradiation. In contrast, the levels of collagen type IV were not significantly changed at 4 and 8 h after irradiation compared with the sham-irradiated controls. Conclusions: The present study demonstrates for the first time that radiation induces an imbalance between MMP-2 and TIMP-2 levels and suggests that degradation of collagen type IV, a major ECM component of BBB basement membrane, may have a role in the pathogenesis of brain injury.

  19. Molecular characterization of collagen IV evidences early transcription expression related to the immune response against bacterial infection in the red abalone (Haliotis rufescens). (United States)

    Chovar-Vera, Ornella; Valenzuela-Muñoz, Valentina; Gallardo-Escárate, Cristian


    Collagen IV has been described as a structural protein of the basement membrane, which as a whole forms a specialized extracellular matrix. Recent studies have indicated a possible relationship between collagen IV and the innate immune response of invertebrate organisms. The present study characterized the alpha-1 chain of collagen IV in the red abalone Haliotis rufescens (Hr-ColIV) and evaluated its association with the innate immune response against Vibrio anguillarum. To further evidence the immune response, the matrix metalloproteinase-1 (Hr-MMP-1) and C-type lectin (Hr-CLEC) genes were also assessed. The complete sequence of Hr-ColIV was composed of 6658 bp, with a 5'UTR of 154 bp, a 3'UTR of 1177 bp, and an ORF of 5327 bp that coded for 1776 amino acids. The innate immune response generated against V. anguillarum resulted in a significant increase in the transcript levels of Hr-ColIV between 3 and 6 hpi, whereas Hr-MMP-1 and Hr-CLEC had the highest transcript activity 6 and 12 hpi, respectively. The results obtained in this study propose a putative biological function for collagen IV involved in the early innate immune response of the red abalone H. rufescens.

  20. Subcellular localization of transglutaminase. Effect of collagen. (United States)

    Juprelle-Soret, M; Wattiaux-De Coninck, S; Wattiaux, R


    1. The subcellular distribution of transglutaminase was investigated by using the analytical approach of differential and isopycnic centrifugation as applied to three organs of the rat: liver, kidney and lung. After differential centrifugation by the method of de Duve, Pressman, Gianetto, Wattiaux & Appelmans [(1955) Biochem. J. 63, 604-617], transglutaminase is mostly recovered in the unsedimentable fraction S and the nuclear fraction N. After isopycnic centrifugation of the N fraction in a sucrose density gradient, a high proportion of the enzyme remains at the top of the gradient; a second but minor peak of activity is present in high-density regions, where a small proportion of 5'-nucleotidase, a plasma-membrane marker, is present together with a large proportion of collagen recovered in that fraction. 2. Fractions where a peak of transglutaminase was apparent in the sucrose gradient were examined by electron microscopy. The main components are large membrane sheets with extracellular matrix and free collagen fibers. 3. As these results seem to indicate that some correlation exists between particulate transglutaminase distribution and those of collagen and plasma membranes, the possible binding of transglutaminase by collagen (type I) and by purified rat liver plasma membrane was investigated. 4. The binding studies indicated that collagen is able to bind transglutaminase and to make complexes with plasma-membrane fragments whose density is higher than that of plasma-membrane fragments alone. Transglutaminase cannot be removed from such complexes by 1% Triton X-100, but can be to a relatively large extent by 0.5 M-KCl and by 50% (w/v) glycerol. 5. Such results suggest that the apparent association of transglutaminase with plasma membrane originates from binding in vitro of the cytosolic enzyme to plasma membrane bound to collagen, which takes place during homogenization of the tissue, when the soluble enzyme and extracellular components are brought together

  1. Altered spatiotemporal expression of collagen types I, III, IV, and VI in Lpar3-deficient peri-implantation mouse uterus. (United States)

    Diao, Honglu; Aplin, John D; Xiao, Shuo; Chun, Jerold; Li, Zuguo; Chen, Shiyou; Ye, Xiaoqin


    Lpar3 is upregulated in the preimplantation uterus, and deletion of Lpar3 leads to delayed uterine receptivity in mice. Microarray analysis revealed that there was higher expression of Col3a1 and Col6a3 in the Preimplantation Day 3.5 Lpar3(-/-) uterus compared to Day 3.5 wild-type (WT) uterus. Since extracellular matrix (ECM) remodeling is indispensable during embryo implantation, and dynamic spatiotemporal alteration of specific collagen types is part of this process, this study aimed to characterize the expression of four main uterine collagen types: fibril-forming collagen (COL) I and COL III, basement membrane COL IV, and microfibrillar COL VI in the peri-implantation WT and Lpar3(-/-) uterus. An observed delay of COL III and COL VI clearance in the Lpar3(-/-) uterus may be associated with higher preimplantation expression of Col3a1 and Col6a3. There was also delayed clearance of COL I and delayed deposition of COL IV in the decidual zone in the Lpar3(-/-) uterus. These changes were different from the effects of 17beta-estradiol and progesterone on uterine collagen expression in ovariectomized WT uterus, indicating that the altered collagen expression in Lpar3(-/-) uterus is unlikely to be a result of alterations in ovarian hormones. Decreased expression of several genes encoding matrix-degrading metallo- and serine proteinases was observed in the Lpar3(-/-) uterus. These results demonstrate that pathways downstream of LPA3 are involved in the dynamic remodeling of ECM in the peri-implantation uterus.

  2. Novel collagen VI mutations identified in Chinese patients with Ullrich congenital muscular dystrophy

    Institute of Scientific and Technical Information of China (English)

    Yan-Zhi Zhang; Hui Xiong; Dan-Hua Zhao; Hai-Po Yang; Ai-Jie Liu; Xing-Zhi Chang; Dao-Jun Hong; Carsten Bonnemann; Yun Yuan; Xi-Ru Wu


    Background: We determined the clinical and molecular genetic characteristics of 8 Chinese patients with Ullrich congenital muscular dystrophy (UCMD). Methods: Clinical data of probands were collected and muscle biopsies of patients were analyzed. Exons of COL6A1, COL6A2 and COL6A3 were analyzed by direct sequencing. Mutations in COL6A1, COL6A2 and COL6A3 were identifi ed in 8 patients. Results: Among these mutations, 5 were novel [three in the triple helical domain (THD) and 2 in the second C-terminal (C2) domain]. We also identified five known missense or in-frame deletion mutations in THD and C domains. Immunohistochemical studies on muscle biopsies from patients showed reduced level of collagen VI at the muscle basement membrane and mis-localization of the protein in interstitial and perivascular regions. Conclusions: The novel mutations we identified underscore the importance of THD and C2 domains in the assembly and function of collagen VI, thereby providing useful information for the genetic counseling of UCMD patients.

  3. Reconstruction of a full-thickness collagen-based human oral mucosal equivalent. (United States)

    Kinikoglu, Beste; Auxenfans, Céline; Pierrillas, Pascal; Justin, Virginie; Breton, Pierre; Burillon, Carole; Hasirci, Vasif; Damour, Odile


    Tissue engineered human oral mucosa has the potential to be applied to the closure of surgical wounds after tissue deficits due to facial trauma, malignant lesion surgery or preposthetic procedure. It can also be used to elucidate the biology and pathology of oral mucosa and as a model alternative to animals for safety testing of oral care products. Using the technology previously developed in our laboratory for the production of a skin equivalent, we were able to reconstruct a nonkeratinized full-thickness human oral mucosal equivalent closely mimicking human native oral mucosa. The successive coculture of human lamina propria fibroblasts and human oral epithelial cells isolated from the nonkeratinized region of oral cavity in a porous collagen-glycosaminoglycan (GAG)-chitosan scaffold gave rise to a lamina propria equivalent (LPE) and then to an oral mucosa equivalent (OME). The results of the histology, immunohistology and transmission electron microscopy of this OME demonstrated the presence of a nonkeratinized pluristratified and differentiated epithelium as in native nonkeratinized human oral mucosa expressing both K13 and K3/76. This epithelium was firmly anchored to the LPE by a continuous and ultrastructurally well-organized basement membrane. In the LPE, fibroblasts synthesized new extracellular matrix where the average collagen fibre diameter was 28.4 nm, close to that of native oral mucosa. The proliferative capacity of the basal cells was demonstrated by the expression of Ki67.

  4. Clinical evaluation of porous hydroxyapatite bone graft (Periobone G with and without collagen membrane (Periocol in the treatment of bilateral grade II furcation defects in mandibular first permanent molars

    Directory of Open Access Journals (Sweden)

    Sruthy Prathap


    Full Text Available Background: Furcation invasions represent one of the most demanding therapeutic challenges in periodontics. This investigation assessed and compared the clinical efficacy of hydroxyapatite bone graft material when used alone and with collagen membrane in the treatment of grade II furcation defects. Materials and Methods: Ten patients with comparable bilateral furcation defects in relation to mandibular first molars were selected and treated in a split-mouth design. After the hygiene phase of therapy was completed, the groups were selected randomly either for treatment with hydroxyapatite bone graft (Periobone G alone or with a combination of bone graft and guided tissue regeneration (GTR membrane (Periocol. Clinical parameters like plaque index, gingival index, vertical probing depth, horizontal probing depth, clinical attachment level, position of marginal gingiva, and the amount of bone fill were used at baseline and at 3 and 6 months postoperatively. Results: At 6 months, both surgical procedures resulted in statistically significant reduction in vertical and horizontal probing depths and gain in the clinical attachment level. Conclusion: The use of combination technique yielded superior results compared to sites treated with bone graft alone. However, the difference was not statistically significant.

  5. The effects of acellular amniotic membrane matrix on osteogenic differentiation and ERK1/2 signaling in human dental apical papilla cells. (United States)

    Chen, Yi-Jane; Chung, Min-Chun; Jane Yao, Chung-Chen; Huang, Chien-Hsun; Chang, Hao-Hueng; Jeng, Jiiang-Huei; Young, Tai-Horng


    The amniotic membrane (AM) has been widely used in the field of tissue engineering because of the favorable biological properties for scaffolding material. However, little is known about the effects of an acellular AM matrix on the osteogenic differentiation of mesenchymal stem cells. In this study, it was found that both basement membrane side and collagenous stroma side of the acellular AM matrix were capable of providing a preferential environment for driving the osteogenic differentiation of human dental apical papilla cells (APCs) with proven stem cell characteristics. Acellular AM matrix potentiated the induction effect of osteogenic supplements (OS) such as ascorbic acid, β-glycerophosphate, and dexamethasone and enhanced the osteogenic differentiation of APCs, as seen by increased core-binding factor alpha 1 (Cbfa-1) phosphorylation, alkaline phosphatase activity, mRNA expression of osteogenic marker genes, and mineralized matrix deposition. Even in the absence of soluble OS, acellular AM matrix also could exert the substrate-induced effect on initiating APCs' differentiation. Especially, the collagenous stroma side was more effective than the basement membrane side. Moreover, the AM-induced effect was significantly inhibited by U0126, an inhibitor of extracellular signaling-regulated kinase 1/2 (ERK1/2) signaling. Taken together, the osteogenic differentiation promoting effect on APCs is AM-specific, which provides potential applications of acellular AM matrix in bone/tooth tissue engineering.

  6. Biochemical and biophysical characterization of collagens of marine sponge, Ircinia fusca (Porifera: Demospongiae: Irciniidae). (United States)

    Pallela, Ramjee; Bojja, Sreedhar; Janapala, Venkateswara Rao


    Collagens were isolated and partially characterized from the marine demosponge, Ircinia fusca from Gulf of Mannar (GoM), India, with an aim to develop potentially applicable collagens from unused and under-used resources. The yield of insoluble, salt soluble and acid soluble forms of collagens was 31.71 ± 1.59, 20.69 ± 1.03, and 17.38 ± 0.87 mg/g dry weight, respectively. Trichrome staining, Scanning & Transmission Electron microscopic (SEM & TEM) studies confirmed the presence of collagen in the isolated, terminally globular irciniid filaments. The partially purified (gel filtration chromatography), non-fibrillar collagens appeared as basement type collagenous sheets under light microscopy whereas the purified fibrillar collagens appeared as fibrils with a repeated band periodicity of 67 nm under Atomic Force Microscope (AFM). The non-fibrillar and fibrillar collagens were seen to have affinity for anti-collagen type IV and type I antibodies raised against human collagens, respectively. The macromolecules, i.e., total protein, carbohydrate and lipid contents within the tissues were also quantified. The present information on the three characteristic irciniid collagens (filamentous, fibrillar and non-fibrillar) could assist the future attempts to unravel the therapeutically important, safer collagens from marine sponges for their use in pharmaceutical and cosmeceutical industries.

  7. 可吸收胶原膜在即刻种植中引导骨组织再生作用的研究%Guided Bone Regeneration of Bioabsorbable Collagen Membrane in Immediate Dental Implants

    Institute of Scientific and Technical Information of China (English)

    刘向辉; 张磊; 孙卫革; 杨建; 黄辉; 张林


    Objective: This study was proposed to evaluate the effect of bioabsorbable collagen membrane in guided bone regeneration in cases of bone defects with immediate dental implants. Methods: 8 beagle dogs were sedated with ketamine and 2, 3, and 4 premolars in both sides of mandible were extracted. Immediate implants was inserted into the extraction wounds. Each extraction socket was made one implantation. A half circumferential bone defect measured 3 mmx4 mm was prepared around the neck of implant. 6 bone defects in one dog were randomly divided into 3 groups. Group A, coralline hydroxyapatite (CHA) covered with bioabsorbable collagen membrane were used as restorative material in the defects. Group B, CHA only. Group C, blank control. All the dogs were sacrificed at the end of 3 months. The bone specimens including the single implant were collected, which were removed and studied by Gross examination, histological observation, and biomechanical tests. Result: New bone formation in bone defects were observed in all 3 groups. Group A: great amount of matured new bone with no soft tissue growth. Group B: there was scanty new bone and some soft tissue formation. Group C was the worst of three groups. Conclusion: Bioabsorbable collagen membrane with CHA might act as a autograft and stimulate peri-implant bone healing.%目的;通过动物实验研究可吸收胶原膜在即刻种植骨缺损区引导骨组织再生的作用.方法:8只实验用犬,拔除双侧下颌第二、三、四前磨牙,每个拔牙窝植入1枚种植体,并在种植体颈部对应牙槽骨上制备半环状骨缺损,将每只实验犬口内的6处骨缺损随机分为3组,每组两处,并给予不同处理.A组:骨缺损中植入珊瑚羟基磷灰石,并用可吸收胶原膜覆盖;B组:骨缺损中单纯植入珊瑚羟基磷灰石;C组:骨缺损中不植入任何材料,作为空白对照组.3个月后处死所有实验动物,制作含单个种植体的骨标本,进行大体观察、生物力学测

  8. Biomedical applications of collagens. (United States)

    Ramshaw, John A M


    Collagen-based biomedical materials have developed into important, clinically effective materials used in a range of devices that have gained wide acceptance. These devices come with collagen in various formats, including those based on stabilized natural tissues, those that are based on extracted and purified collagens, and designed composite, biosynthetic materials. Further knowledge on the structure and function of collagens has led to on-going developments and improvements. Among these developments has been the production of recombinant collagen materials that are well defined and are disease free. Most recently, a group of bacterial, non-animal collagens has emerged that may provide an excellent, novel source of collagen for use in biomaterials and other applications. These newer collagens are discussed in detail. They can be modified to direct their function, and they can be fabricated into various formats, including films and sponges, while solutions can also be adapted for use in surface coating technologies.

  9. Preparation of collagen-based materials for wound dressing

    Institute of Scientific and Technical Information of China (English)

    吴志谷; 盛志勇; 孙同柱; 耿淼; 黎君友; 姚咏明; 黄祖琇


    Objective To describe the methods which were used to develop collagen-based materials for wound dressing.Methods Fresh frozen bovine tendon was treated with 0.05 mol/L acetic acid at pH 3.2 for 48-72 hours, homogenized, filtered, mixed with 8% chondroitin sulphate, for creating a deaerated 1.5%-2.5% collagen solution. The solution was lyophilized in either a pre-frozen or non-pre-frozen mould. The collagen sponge was then cross-linked with 0.25% glutaraldehyde for 24 hours. Three other types of wound dressings were developed using a similar method: collagen membrane with a polyurethane membrane onlay, polyurethane-coated collagen membrane and collagen membrane on gauze.Results It was demonstrated that the use of frozen bovine tendon was stable, and that the prepared collagen sponge contained pores of 50-400 μm in diameter. Conclusions Collagen could be used as wound dressing.

  10. Mouse endometrial stromal cells produce basement-membrane components

    DEFF Research Database (Denmark)

    Wewer, U M; Damjanov, A; Weiss, J;


    During mouse pregnancy, uterine stromal cells transform into morphologically distinct decidual cells under the influence of the implanting embryo and a proper hormonal environment. Mechanical stimulation of hormonally primed uterine stromal cells leads to the same morphologic alterations. The dec...

  11. Deformation and fracture of echinoderm collagen networks

    CERN Document Server

    Ovaska, Markus; Miksic, Amandine; Sugni, Michela; Di Benedetto, Cristiano; Ferrario, Cinzia; Leggio, Livio; Guidetti, Luca; Alava, Mikko J; La Porta, Caterina A M; Zapperi, Stefano


    Collagen networks provide the main structural component of most tissues and represent an important ingredient for bio-mimetic materials for bio-medical applications. Here we study the mechanical properties of stiff collagen networks derived from three different echinoderms and show that they exhibit non-linear stiffening followed by brittle fracture. The disordered nature of the network leads to strong sample-to-sample fluctuations in elasticity and fracture strength. We perform numerical simulations of a three dimensional model for the deformation of a cross-linked elastic fibril network which is able to reproduce the macroscopic features of the experimental results and provide insights into the internal mechanics of stiff collagen networks. Our numerical model provides an avenue for the design of collagen membranes with tunable mechanical properties.

  12. Proximal collagenous gastroenteritides:

    DEFF Research Database (Denmark)

    Nielsen, Ole Haagen; Riis, Lene Buhl; Danese, Silvio


    AIM: While collagenous colitis represents the most common form of the collagenous gastroenteritides, the collagenous entities affecting the proximal part of the gastrointestinal tract are much less recognized and possibly overlooked. The aim was to summarize the latest information through a syste...

  13. Induction of initial steps of angiogenic differentiation and maturation of endothelial cells by pericytes in vitro and the role of collagen IV. (United States)

    Zhou, Zhigang; Pausch, Friederike; Schlötzer-Schrehardt, Ursula; Brachvogel, Bent; Pöschl, Ernst


    Activation of endothelial cells and recruitment of mural cells define critical steps during the formation of stable vascular elements. Both events are reflected by cocultures of endothelial cells and isolated murine pericyte-like cells and define a versatile platform for the analysis of distinct steps during the angiogenic process in vitro. Isolated pericyte-like cells promote the survival of endothelial cells, induce the assembly of endothelial cells as well as establish direct contacts with forming endothelial alignments. More importantly, they also induce characteristic steps of maturation including the assembly of stable cell-cell junctions, deposition of basement membrane-like matrices and local formation of a central lumen. The presence of pericyte-like cells induces the secretion of extracellular matrices enriched in collagen IV by endothelial cells, which improves endothelial tube formation and provides the adhesive substrate for mural cell recruitment. Collagen-binding integrins contribute differentially to the process, with α1β1 involved in the adhesion of pericyte-like cells to collagen IV and α2β1 mainly involved in endothelial cord formation. These data indicate that pericyte-like cells are essential for the survival of endothelial cells, the efficient formation of endothelial alignments as well as initial steps of maturation of capillary-like structures.

  14. Autoantibodies to Multiple Epitopes on the Non-Collagenous-1 Domain of Type VII Collagen Induce Blisters

    NARCIS (Netherlands)

    Vorobyev, Artem; Ujiie, Hideyuki; Recke, Andreas; Buijsrogge, Jacqueline J. A.; Jonkman, Marcel F.; Pas, Hendrikus; Iwata, Hiroaki; Hashimoto, Takashi; Kim, Soo-Chan; Kim, Jong Hoon; Groves, Richard; Samavedam, Unni; Gupta, Yask; Schmidt, Enno; Zillikens, Detlef; Shimizu, Hiroshi; Ludwig, Ralf J.


    Epidermolysis bullosa acquisita (EBA) is an autoimmune blistering disease of the skin and mucous membranes, characterized by autoantibodies against type VII collagen (COL7), a major component of anchoring fibrils. Different clinical EBA phenotypes are described, including mechanobullous and inflamma

  15. Combined effects of moderately elevated blood glucose and locally produced TGF-beta1 on glomerular morphology and renal collagen production

    DEFF Research Database (Denmark)

    Krag, Søren; Nyengaard, Jens R; Wogensen, Lise


    chain reaction (PCR) and phosphorylated extracellular signal-regulated kinase 1/2 (pERK) expression by immunohistochemistry. RESULTS: Mild hyperglycaemia alone had no effect on glomerular structure or ECM deposition. Over-expression of TGF-beta1 increased basement membrane thickness (BMT......) and the mesangial volume fraction. Furthermore, it augmented ECM, Matrix metalloproteinase-2 (MMP), MMP-9, plasminogen activator inhibitor-1 (PAI) and tissue inhibitor of metalloproteinase-1 (TIMP) gene expression and pERK1/2 immunostaining. Elevated BG in combination with TGF-beta1 resulted in enlargement...... of glomerular volume, total mesangial volume and renal collagen content. Moreover, high BG exaggerated TGF-beta1-induced changes in the BMT, MMP-2 and TIMP-1 expression and pERK1/2 staining. CONCLUSION: Even moderate elevations in BG accelerate the progression of those kidney diseases in which TGF-beta1...

  16. Endocytic collagen degradation

    DEFF Research Database (Denmark)

    Madsen, Daniel H.; Jürgensen, Henrik J.; Ingvarsen, Signe;


    it crucially important to understand both the collagen synthesis and turnover mechanisms in this condition. Here we show that the endocytic collagen receptor, uPARAP/Endo180, is a major determinant in governing the balance between collagen deposition and degradation. Cirrhotic human livers displayed a marked......Fibrosis of the liver and its end-stage, cirrhosis, represent major health problems worldwide. In these fibrotic conditions, activated fibroblasts and hepatic stellate cells display a net deposition of collagen. This collagen deposition is a major factor leading to liver dysfunction, thus making...... up-regulation of uPARAP/Endo180 in activated fibroblasts and hepatic stellate cells located close to the collagen deposits. In a hepatic stellate cell line, uPARAP/Endo180 was shown to be active in, and required for, the uptake and intracellular degradation of collagen. To evaluate the functional...

  17. Complications of collagenous colitis

    Institute of Scientific and Technical Information of China (English)

    Hugh James Freeman


    Microscopic forms of colitis have been described, including collagenous colitis. This disorder generally has an apparently benign clinical course. However, a number of gastric and intestinal complications, possibly coincidental, may develop with collagenous colitis. Distinctive inflammatory disorders of the gastric mucosa have been described, including lymphocytic gastritis and collagenous gastritis. Celiac disease and collagenous sprue (or collagenous enteritis) may occur. Colonic ulceration has been associated with use of nonsteroidal anti-inflammatory drugs, while other forms of inflammatory bowel disease, including ulcerative colitis and Crohn's disease, may evolve from collagenous colitis. Submucosal "dissection", colonic fractures or mucosal tears and perforation from air insufflation during colonoscopy may occur and has been hypothesized to be due to compromise of the colonic wall from submucosal collagen deposition. Similar changes may result from increased intraluminal pressure during barium enema contrast studies. Finally, malignant disorders have also been reported, including carcinoma and lymphoproliferative disease.

  18. The collagen receptor uPARAP/Endo180

    DEFF Research Database (Denmark)

    Engelholm, Lars H; Ingvarsen, Signe; Jürgensen, Henrik J


    The uPAR-associated protein (uPARAP/Endo180), a type-1 membrane protein belonging to the mannose receptor family, is an endocytic receptor for collagen. Through this endocytic function, the protein takes part in a previously unrecognized mechanism of collagen turnover. uPARAP/Endo180 can bind...... and internalize both intact and partially degraded collagens. In some turnover pathways, the function of the receptor probably involves an interplay with certain matrix-degrading proteases whereas, in other physiological processes, redundant mechanisms involving both endocytic and pericellular collagenolysis seem...... in collagen breakdown seems to be involved in invasive tumor growth....

  19. The collagen receptor uPARAP/Endo180

    DEFF Research Database (Denmark)

    Engelholm, Lars H; Ingvarsen, Signe; Jürgensen, Henrik J


    The uPAR-associated protein (uPARAP/Endo180), a type-1 membrane protein belonging to the mannose receptor family, is an endocytic receptor for collagen. Through this endocytic function, the protein takes part in a previously unrecognized mechanism of collagen turnover. uPARAP/Endo180 can bind...... and internalize both intact and partially degraded collagens. In some turnover pathways, the function of the receptor probably involves an interplay with certain matrix-degrading proteases whereas, in other physiological processes, redundant mechanisms involving both endocytic and pericellular collagenolysis seem...... in collagen breakdown seems to be involved in invasive tumor growth Udgivelsesdato: 2009...

  20. Pseudomembranous collagenous colitis. (United States)

    Yuan, Shan; Reyes, Victoria; Bronner, Mary P


    The classic clinical and histologic features of collagenous colitis are well characterized; however, the acute or neutrophilic inflammatory changes that may accompany this entity are less well established. In this report of 10 patients, we describe the first series of pseudomembranous collagenous colitis. Because superimposed Clostridium difficile infection was only demonstrated in one patient and no other causes of pseudomembranous colitis were evident in the remaining nine patients, we conclude that pseudomembranes are part of the spectrum of collagenous colitis itself. This case series illustrates the importance of searching for collagenous colitis in the evaluation of pseudomembranous colitis. At the same time, superimposed infectious or ischemic etiologies need to be excluded clinically in any patient with superimposed pseudomembranes. The existence of pseudomembranes in collagenous colitis also lends support to the hypothesis that toxin- and/or ischemia-mediated injury may be involved in the pathogenesis of collagenous colitis.

  1. Enigmatic insight into collagen

    Directory of Open Access Journals (Sweden)

    Shrutal Narendra Deshmukh


    Full Text Available Collagen is a unique, triple helical molecule which forms the major part of extracellular matrix. It is the most abundant protein in the human body, representing 30% of its dry weight. It is the fibrous structural protein that makes up the white fibers (collagen fibers of skin, tendons, bones, cartilage and all other connective tissues. Collagens are not only essential for the mechanical resistance and resilience of multicellular organisms, but are also signaling molecules defining cellular shape and behavior. The human body has at least 16 types of collagen, but the most prominent types are I, II and III. Collagens are produced by several cell types and are distinguishable by their molecular compositions, morphologic characteristics, distribution, functions and pathogenesis. This is the major fibrous glycoprotein present in the extracellular matrix and in connective tissue and helps in maintaining the structural integrity of these tissues. It has a triple helical structure. Various studies have proved that mutations that modify folding of the triple helix result in identifiable genetic disorders. Collagen diseases share certain similarities with autoimmune diseases, because autoantibodies specific to each collagen disease are produced. Therefore, this review highlights the role of collagen in normal health and also the disorders associated with structural and functional defects in collagen.

  2. Real-time-guided bone regeneration around standardized critical size calvarial defects using bone marrow-derived mesenchymal stem cells and collagen membrane with and without using tricalcium phosphate: an in vivo microcomputed tomographic and histologic experiment in rats

    Institute of Scientific and Technical Information of China (English)

    Khalid Al-Hezaimi; Sundar Ramalingam; Mansour Al-Askar; Aws S ArRejaie; Nasser Nooh; Fawad Jawad; Abdullah Aldahmash; Muhammad Atteya; Cun-Yu Wang


    The aimof the present real time in vivo micro-computed tomography (mCT) and histologic experiment was to assess the efficacy of guided bone regeneration (GBR) around standardized calvarial critical size defects (CSD) using bone marrow-derived mesenchymal stem cells (BMSCs), and collagen membrane (CM) with and without tricalcium phosphate (TCP) graft material. In the calvaria of nine female Sprague-Dawley rats, full-thickness CSD (diameter 4.6 mm) were created under general anesthesia. Treatment-wise, rats were divided into three groups. In group 1, CSD was covered with a resorbable CM; in group 2, BMSCs were filled in CSD and covered with CM; and in group 3, TCP soaked in BMSCs was placed in CSD and covered with CM. All defects were closed using resorbable sutures. Bone volume and bone mineral density of newly formed bone (NFB) and remaining TCP particles and rate of new bone formation was determined at baseline, 2, 4, 6, and 10 weeks using in vivo mCT. At the 10th week, the rats were killed and calvarial segments were assessed histologically. The results showed that the hardness ofNFBwas similar to that of the native bone in groups 1 and 2 as compared to theNFB in group 3. Likewise, values for the modulus of elasticity were also significantly higher in group 3 compared to groups 1 and 2. This suggests that TCP when used in combination with BMSCs and without CM was unable to form bone of significant strength that could possibly provide mechanical “lock” between the natural bone and NFB. The use of BMSCs as adjuncts to conventional GBR initiated new bone formation as early as 2 weeks of treatment compared to when GBR is attempted without adjunct BMSC therapy.

  3. The efficacy of collagen-hydroxyapatite composite membrane on bone regeneration%胶原-羟磷灰石复合膜引导骨组织再生的动物实验研究

    Institute of Scientific and Technical Information of China (English)

    金琼; 王晓敏; 王晓飞; 李旭东; 麻健丰


    目的 探讨自制胶原-羟磷灰石(COL-HA)复合膜修复SD大鼠颅骨缺损的效果.方法在24只SD大鼠的颅顶骨上各制备4个缺损,其中左、右侧缺损区分别覆盖COL-HA复合单层致密膜(第2组)和COL-HA复合双层膜(第3组),额骨缺损区保留作为空白对照(第1组),枕骨缺损区覆盖Bio-Gide膜作为对比(第4组).术后2、4、8、12周各处死6只大鼠取材,进行肉眼观察、X线检查、组织切片观察和新生骨量测定,对结果进行广义线性模型,析因设计方差分析和KSD-t检验.结果 从术后2周开始除第1组外,其余3组均可见少量新生骨质,12周时3组缺损区由不透明硬组织封闭,并可见分解的部分膜碎片.X线显示,术后12周时第3组和第4组缺损区修复骨密度与原骨质接近,第2组稍低.新生骨量分析显示,术后初期第4组新牛骨量大于其余3组,术后12周第4组与第3组和第2组的成骨量无统计学筹异.结论 COL-HA复合膜能引导大鼠颅骨组织再生,且复合双层膜成骨效果要优于复合单层致密膜,其结构更有利于成骨细胞的附着和生长.%Objective To investigate the efficacy of a new collagen-hydroxyapatite (COL-HA) composite membrane on bone regeneration of SD rat cranial defects.Methods Four defects were produced in the calvaria of 24 SD rats.The animals were divided into four groups: Empty defects without membrane (group 1); defects covered by COL-HA single-layer dense membranes (group 2); defects covered by COL-HA double-layer membranes (group 3);defects covered by Bio-Gide membranes(group 4).At 2, 4, 8 and 12 weeks after surgery, 6 rats were sacrificed and the following parameters were analyzed: Macroscopic observation, X-ray examination, descriptive histology, regenerate bone quantitative histology.Statistical analysis consisted of generalized linear models/factorial design analysis of variance and LSD-t test was performed.Results Since two weeks after surgery, there were a small amount

  4. The expression of collagen type Ⅳ in the temporomandibular joint disorders%Ⅳ型胶原在颞下颌关节紊乱病中的表达

    Institute of Scientific and Technical Information of China (English)

    熊卉; 龙星; 邓末宏; 蔡恒星; 孟庆功; 李金荣


    目的 探讨颞下颌关节紊乱病病变与Ⅳ型胶原之间的关系.方法 应用链菌素亲生物素-过氧化酶连结法检测19例颞下颌关节病变关节盘和盘后组织中Ⅳ型胶原的表达.结果 19例颞下颌关节紊乱病患者中,不可复性盘前移位3例,盘前移位伴关节盘穿孔5例,骨关节病伴关节盘穿孔11例.Ⅳ型胶原主要存在于病变关节盘和双板区的血管基底膜.其中骨关节病伴关节盘穿孔的阳性反应最为明显.6例在软骨细胞周围有弱阳性反应.结论 Ⅳ型胶原在病变颞下颌关节盘和盘后组织的血管基底膜以及软骨细胞周围可出现表达,Ⅳ型胶原免疫组织化学的染色强度与病变的严重程度有一定关系.%Objective To discuss the relationship between collagen type Ⅳ and the pathologic changes of temporomandibular joint disorders. Methods The distribution of collagen type Ⅳ in 19 human disc of temporomandibular joint disorders were studied by inmmunohistochemical Streptavidium Peroxdase Conjugated method. Results There were 3 cases with anterior disc displacement without reduction,5 cases ADD with perforation and 11 eases esteoarthrosis with perforation. Collagen type Ⅳ existed in the basement membrane of blood vesseLs in the disc and retrodisc tissues of temporomandibular joint disorders. There were weak immunoreactivities around the chondrocytes in 6 cases. Conclusion Collagen type Ⅳ can be detected in the basement membrane of blood vessels in the disc and retrodisc tissues of temperomandibular joint disorders. It is suggested that the stronger immunoreactivities of collagen type Ⅳ, the more severe arthritic change in the disc and retrodisc tissues of temperomandibular joint disorders.

  5. Characterization of three-dimensional cancer cell migration in mixed collagen-Matrigel scaffolds using microfluidics and image analysis (United States)

    Maška, Martin; Ederra, Cristina; Peláez, Rafael; Morales, Xabier; Muñoz-Arrieta, Gorka; Mujika, Maite; Kozubek, Michal; Muñoz-Barrutia, Arrate; Rouzaut, Ana; Arana, Sergio; Garcia-Aznar, José Manuel; Ortiz-de-Solorzano, Carlos


    Microfluidic devices are becoming mainstream tools to recapitulate in vitro the behavior of cells and tissues. In this study, we use microfluidic devices filled with hydrogels of mixed collagen-Matrigel composition to study the migration of lung cancer cells under different cancer invasion microenvironments. We present the design of the microfluidic device, characterize the hydrogels morphologically and mechanically and use quantitative image analysis to measure the migration of H1299 lung adenocarcinoma cancer cells in different experimental conditions. Our results show the plasticity of lung cancer cell migration, which turns from mesenchymal in collagen only matrices, to lobopodial in collagen-Matrigel matrices that approximate the interface between a disrupted basement membrane and the underlying connective tissue. Our quantification of migration speed confirms a biphasic role of Matrigel. At low concentration, Matrigel facilitates migration, most probably by providing a supportive and growth factor retaining environment. At high concentration, Matrigel slows down migration, possibly due excessive attachment. Finally, we show that antibody-based integrin blockade promotes a change in migration phenotype from mesenchymal or lobopodial to amoeboid and analyze the effect of this change in migration dynamics, in regards to the structure of the matrix. In summary, we describe and characterize a robust microfluidic platform and a set of software tools that can be used to study lung cancer cell migration under different microenvironments and experimental conditions. This platform could be used in future studies, thus benefitting from the advantages introduced by microfluidic devices: precise control of the environment, excellent optical properties, parallelization for high throughput studies and efficient use of therapeutic drugs. PMID:28166248

  6. De Novo and Inherited Mutations in COL4A2, Encoding the Type IV Collagen α2 Chain Cause Porencephaly (United States)

    Yoneda, Yuriko; Haginoya, Kazuhiro; Arai, Hiroshi; Yamaoka, Shigeo; Tsurusaki, Yoshinori; Doi, Hiroshi; Miyake, Noriko; Yokochi, Kenji; Osaka, Hitoshi; Kato, Mitsuhiro; Matsumoto, Naomichi; Saitsu, Hirotomo


    Porencephaly is a neurological disorder characterized by fluid-filled cysts or cavities in the brain that often cause hemiplegia. It has been suggested that porencephalic cavities result from focal cerebral degeneration involving hemorrhages. De novo or inherited heterozygous mutations in COL4A1, which encodes the type IV α1 collagen chain that is essential for structural integrity for vascular basement membranes, have been reported in individuals with porencephaly. Most mutations occurred at conserved Gly residues in the Gly-Xaa-Yaa repeats of the triple-helical domain, leading to alterations of the α1α1α2 heterotrimers. Here we report on two individuals with porencephaly caused by a heterozygous missense mutation in COL4A2, which encodes the type IV α2 collagen chain. Mutations c.3455G>A and c.3110G>A, one in each of the individuals, cause Gly residues in the Gly-Xaa-Yaa repeat to be substituted as p.Gly1152Asp and p.Gly1037Glu, respectively, probably resulting in alterations of the α1α1α2 heterotrimers. The c.3455G>A mutation was found in the proband's mother, who showed very mild monoparesis of the left upper extremity, and the maternal elder uncle, who had congenital hemiplegia. The maternal grandfather harboring the mutation is asymptomatic. The c.3110G>A mutation occurred de novo. Our study confirmed that abnormalities of the α1α1α2 heterotrimers of type IV collagen cause porencephaly and stresses the importance of screening for COL4A2 as well as for COL4A1. PMID:22209246

  7. Comparative analysis of the noncollagenous NC1 domain of type IV collagen: identification of structural features important for assembly, function, and pathogenesis. (United States)

    Netzer, K O; Suzuki, K; Itoh, Y; Hudson, B G; Khalifah, R G


    Type IV collagen alpha1-alpha6 chains have important roles in the assembly of basement membranes and are implicated in the pathogenesis of Goodpasture syndrome, an autoimmune disorder, and Alport syndrome, a hereditary renal disease. We report comparative sequence analyses and structural predictions of the noncollagenous C-terminal globular NC1 domain (28 sequences). The inferred tree verified that type IV collagen sequences fall into two groups, alpha1-like and alpha2-like, and suggested that vertebrate alpha3/alpha4 sequences evolved before alpha1/alpha2 and alpha5/alpha6. About one fifth of NC1 residues were identified to confer either the alpha1 or alpha2 group-specificity. These residues accumulate opposite charge in subdomain B of alpha1 (positive) and alpha2 (negative) sequences and may play a role in the stoichiometric chain selection upon type IV collagen assembly. Neural network secondary structure prediction on multiple aligned sequences revealed a subdomain core structure consisting of six hydrophobic beta-strands and one short alpha-helix with a significant hydrophobic moment. The existence of opposite charges in the alpha-helices may carry implications for intersubdomain interactions. The results provide a rationale for defining the epitope that binds Goodpasture autoantibodies and a framework for understanding how certain NC1 mutations may lead to Alport syndrome. A search algorithm, based entirely on amino acid properties, yielded a possible similarity of NC1 to tissue inhibitor of metalloproteinases (TIMP) and prompted an investigation of a possible functional relationship. The results indicate that NC1 preparations decrease the activity of matrix metalloproteinases 2 and 3 (MMP-2, MMP-3) toward a peptide substrate, though not to [14C]-gelatin. We suggest that an ancestral NC1 may have been incorporated into type IV collagen as an evolutionarily mobile domain carrying proteinase inhibitor function.

  8. Gangliosides regulate tumor cell adhesion to collagen. (United States)

    Kazarian, Tamara; Jabbar, Adnan A; Wen, Fei-Qui; Patel, Dharmesh A; Valentino, Leonard A


    The ability of tumor cells to adhere to extracellular matrix proteins is critical for migration and invasion. The factors that regulate tumor cell adhesion are poorly characterized. Gangliosides promote platelet adhesion and may also play a role in the adhesion of other cell types. We hypothesized that pharmacological depletion of membrane gangliosides from adherent cells would abrogate adhesion to collagen and promote migration and invasion. To test these hypotheses, LA-N1 neuroblastoma cells, which avidly adhere to collagen and are rich with membrane gangliosides (43.69 nmol/10(8) cells), were cultured in the presence of D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol-HCl. Endogenous gangliosides were reduced by 98% (0.76 nmol/10(8) cells) and adhesion to collagen decreased by 67%. There were no changes in cell morphology, viability, proliferation rate or apoptosis. Pre-incubation of ganglioside-depleted cells in conditioned medium from control cells restored adhesion to collagen (0.45 +/- 0.002), comparable to that of control cells (0.49 +/- 0.035). Similarly, pre-incubation of ganglioside-depleted cells with purified GD2 completely restored adhesion in a concentration-dependent manner. When LA-N1 cells were cultured with retinoic acid, a biological response modifier known to increase endogenous gangliosides, adhesion to collagen increased. Next, we questioned whether changes in adhesion would be reflected as changes in migration and invasion. Cells depleted of endogenous cellular gangliosides migrated more than control cells. Finally, control cells replete with their endogenous gangliosides demonstrated less invasive potential than control cells. The data demonstrate that endogenous tumor gangliosides increase neuroblastoma cell adhesion to collagen and reduce migration and invasion in vitro.

  9. Design and synthesis of collagen mimetic peptide derivatives for studying triple helix assembly and collagen mimetic peptide-collagen binding interaction (United States)

    Mo, Xiao


    region, one of the thermally unstable domains in the collagen molecule. The binding results at various temperatures were in good agreement with our hypothesis that CMP-collagen binding occurs through strand invasion reaction of the thermally unstable collagen domain. In addition, purple membrane (PM), a protein crystal patch from halobacteria cell membrane, was genetically engineered to display cysteine or histidine groups on the membrane surface with defined nanoscale symmetry. X-ray diffraction results showed that the engineered PM formed a stable 2D PM-like hexagonal crystal lattice (unit cell: 6.2 mm). We utilized the genetically engineered PM as a bio-template for inorganic nanoparticle nucleation and assembly. The specific functional groups (cysteine or histidine) on the surfaces of genetically engineered PMs exhibited designed reactivity for inorganic nanoparticles while promoting the formation of nanoscale 2D particle assembly.

  10. Arrangement of type IV collagen on NH₂ and COOH functionalized surfaces. (United States)

    Coelho, Nuno Miranda; González-García, Cristina; Salmerón-Sánchez, Manuel; Altankov, George


    Apart from the paradigm that cell-biomaterials interaction depends on the adsorption of soluble adhesive proteins we anticipate that upon distinct conditions also other, less soluble ECM proteins such as collagens, associate with the biomaterials interface with consequences for cellular response that might be of significant bioengineering interest. Using atomic force microscopy (AFM) we seek to follow the nanoscale behavior of adsorbed type IV collagen (Col IV)--a unique multifunctional matrix protein involved in the organization of basement membranes (BMs) including vascular ones. We have previously shown that substratum wettability significantly affects Col IV adsorption pattern, and in turn alters endothelial cells interaction. Here we introduce two new model surfaces based on self-assembled monolayers (SAMs), a positively charged -NH(2) , and negatively charged -COOH surface, to learn more about their particular effect on Col IV behavior. AFM studies revealed distinct pattern of Col IV assembly onto the two SAMs resembling different aspects of network-like structure or aggregates (suggesting altered protein conformation). Moreover, the amount of adsorbed FITC-labeled Col IV was quantified and showed about twice more protein on NH(2) substrata. Human umbilical vein endothelial cells attached less efficiently to Col IV adsorbed on negatively charged COOH surface judged by altered cell spreading, focal adhesions formation, and actin cytoskeleton development. Immunofluorescence studies also revealed better Col IV recognition by both α(1) and α(2) integrins on positively charged NH(2) substrata resulting in higher phosphorylated focal adhesion kinase recruitment in the focal adhesion complexes. On COOH surface, no integrin clustering was observed. Taken altogether these results, point to the possibility that combined NH(2) and Col IV functionalization may support endothelization of cardiovascular implants.

  11. Altered stress fibers and integrin expression in the Malpighian epithelium of Drosophila type IV collagen mutants

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    András A. Kiss


    Full Text Available Basement membranes (BMs are highly specialized extracellular matrices (ECMs that provide support and polarization cues for epithelial cells. Proper adhesion to the BM is pivotal in epithelial cell function and survival. Type IV collagens are the predominant components of all types of BMs, that form an irregular, polygonal lattice and serve as a scaffold for numerous other BM components and BM-associated cells. Mutations in the ubiquitous human BM components COL4A1 and COL4A2 cause a multisystem disorder involving nephropathy. Affected patients develop renal dysfunction and chronic kidney failure with or without hematuria. Mouse Col4a1 and Col4a2 mutants recapitulate the human symptoms. In vertebrates, excretion is accomplished by the kidneys and by the Malpighian tubules in insects, including the fruit fly Drosophila. Our present results with dominant, temperature-sensitive mutation of the Drosophila col4a1 gene demonstrate altered integrin expression and amplified effects of mechanical stress on the Malpighian epithelial cytoskeleton.

  12. Different assembly of type IV collagen on hydrophilic and hydrophobic substrata alters endothelial cells interaction. (United States)

    Coelho, N Miranda; González-García, C; Planell, J A; Salmerón-Sánchez, M; Altankov, G


    Considering the structural role of type IV collagen (Col IV) in the assembly of the basement membrane (BM) and the perspective of mimicking its organization for vascular tissue engineering purposes, we studied the adsorption pattern of this protein on model hydrophilic (clean glass) and hydrophobic trichloro(octadecyl)silane (ODS) surfaces known to strongly affect the behavior of other matrix proteins. The amount of fluorescently labeled Col IV was quantified showing saturation of the surface for concentration of the adsorbing solution of about 50microg/ml, but with approximately twice more adsorbed protein on ODS. AFM studies revealed a fine - nearly single molecular size - network arrangement of Col IV on hydrophilic glass, which turns into a prominent and growing polygonal network consisting of molecular aggregates on hydrophobic ODS. The protein layer forms within minutes in a concentration-dependent manner. We further found that human umbilical vein endothelial cells (HUVEC) attach less efficiently to the aggregated Col IV (on ODS), as judged by the significantly altered cell spreading, focal adhesions formation and the development of actin cytoskeleton. Conversely, the immunofluorescence studies for integrins revealed that the fine Col IV network formed on hydrophilic substrata is better recognized by the cells via both alpha1 and alpha2 heterodimers which support cellular interaction, apart from these on hydrophobic ODS where almost no clustering of integrins was observed.

  13. Different assembly of type IV collagen on hydrophilic and hydrophobic substrata alters endothelial cells interaction

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    NM Coelho


    Full Text Available Considering the structural role of type IV collagen (Col IV in the assembly of the basement membrane (BM and the perspective of mimicking its organization for vascular tissue engineering purposes, we studied the adsorption pattern of this protein on model hydrophilic (clean glass and hydrophobic trichloro(octadecylsilane (ODS surfaces known to strongly affect the behavior of other matrix proteins. The amount of fluorescently labeled Col IV was quantified showing saturation of the surface for concentration of the adsorbing solution of about 50μg/ml, but with approximately twice more adsorbed protein on ODS. AFM studies revealed a fine – nearly single molecular size – network arrangement of Col IV on hydrophilic glass, which turns into a prominent and growing polygonal network consisting of molecular aggregates on hydrophobic ODS. The protein layer forms within minutes in a concentration-dependent manner. We further found that human umbilical vein endothelial cells (HUVEC attach less efficiently to the aggregated Col IV (on ODS, as judged by the significantly altered cell spreading, focal adhesions formation and the development of actin cytoskeleton. Conversely, the immunofluorescence studies for integrins revealed that the fine Col IV network formed on hydrophilic substrata is better recognized by the cells via both α1 and α2 heterodimers which support cellular interaction, apart from these on hydrophobic ODS where almost no clustering of integrins was observed.

  14. Study and progress of collagen Ⅳ gene COL4A1 mutations%Ⅳ型胶原基因COL4A1突变的研究进展

    Institute of Scientific and Technical Information of China (English)

    安丽梅; 夏欣一


    Ⅳ型胶原蛋白基因α1 (COL4A1)编码Ⅳ型胶原的α1链,是各种组织基底膜的主要结构成分.基底膜由胶原、层粘连蛋白、蛋白多糖和巢蛋白组成,在所有胶原成分中,最重要的是Ⅳ型胶原.COL4A1基因突变能引起多种疾病,在一定的环境压力下会引发HANAC综合征、肾病、孔洞脑、白内障等疾病.本文主要从COL4A1的结构、组成及在基底膜中的作用等方面,对COI4A1基因突变引发的人类疾病及其发病机制进行综述.%α1 type IV collagen (COL4A1) encodes the α1 chain of type Ⅳ collagen. Collagen type Ⅳ is the most abundant structural BM component in a variety of tissues. In general, Basement membranes ( BMs) are composed of collagens, laminins, perle-can, and nidogens. Collagen IV is important to BM. Mutations in COL4A1 gene have been linked to a spectrum of diverse diseases caused by abnormalities of these molecules in the kidney, brain and eyes. In this review, we will cover, after a short overview of the molecular basis of the COL4A1 and the main BM components, the current knowledge of diseases caused by COL4A1 mutations and summarize the possible mechanisms of mutations and human diseases.

  15. Blood flow interplays with elastin: collagen and MMP: TIMP ratios to maintain healthy vascular structure and function

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    Poulami Basu


    Full Text Available Poulami Basu, Utpal Sen, Neetu Tyagi, Suresh C TyagiDepartment of Physiology and Biophysics, University of Louisville School of Medicine, Louisville, Kentucky, USAAbstract: Differential vascular remodeling is one of the major mechanisms of heterogeneity in atherosclerosis. The structural and functional heterogeneity between arteries and veins determines the degree of vascular remodeling. Matrix metalloproteases (MMPs and their tissue inhibitors (TIMPs play key roles in vascular structural and functional remodeling. We hypothesized that the level of blood flow in different arteries and veins caused structural and functional heterogeneity that ultimately determined potential vascular remodeling. To test this hypothesis, in vivo blood flow and blood pressure in the aorta, carotid, femoral artery, and femoral vein was measured in male Sprague-Dawley rats (weight 380–400 gm. Arterial and venous pressures were measured by PE-50 catheter cannulation. Blood flow was measured by a transonic ultrasound system. The aortic arch, femoral and carotid arteries, and abdominal vena cava were isolated to determine the expression of MMP-2, -9, -12, and -13 and TIMP-1, -3, and -4 by Western blot and in gelatin gel zymography. Masson trichrome and van Gieson stains were used to stain the histologic tissue sections. The results revealed that blood flow was higher in the aorta and carotid artery than the femoral artery and vein. MMP-9 and MMP-13 were higher in the carotid artery in comparison with the other blood vessels, while TIMP-3 showed higher expression in the aorta than the arteries. Further, the MMP-9 activity was significantly higher in the carotid artery than in the aorta and femoral artery. There was a higher degree of basement membrane collagen in the femoral artery and therefore a low elastin: collagen ratio, while in the carotid artery a higher level of elastin and, therefore, a high elastin: collagen ratio was found. The results suggested that medial

  16. Role of collagens and perlecan in microvascular stability: exploring the mechanism of capillary vessel damage by snake venom metalloproteinases.

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    Teresa Escalante

    Full Text Available Hemorrhage is a clinically important manifestation of viperid snakebite envenomings, and is induced by snake venom metalloproteinases (SVMPs. Hemorrhagic and non-hemorrhagic SVMPs hydrolyze some basement membrane (BM and associated extracellular matrix (ECM proteins. Nevertheless, only hemorrhagic SVMPs are able to disrupt microvessels; the mechanisms behind this functional difference remain largely unknown. We compared the proteolytic activity of the hemorrhagic P-I SVMP BaP1, from the venom of Bothrops asper, and the non-hemorrhagic P-I SVMP leucurolysin-a (leuc-a, from the venom of Bothrops leucurus, on several substrates in vitro and in vivo, focusing on BM proteins. When incubated with Matrigel, a soluble extract of BM, both enzymes hydrolyzed laminin, nidogen and perlecan, albeit BaP1 did it at a faster rate. Type IV collagen was readily digested by BaP1 while leuc-a only induced a slight hydrolysis. Degradation of BM proteins in vivo was studied in mouse gastrocnemius muscle. Western blot analysis of muscle tissue homogenates showed a similar degradation of laminin chains by both enzymes, whereas nidogen was cleaved to a higher extent by BaP1, and perlecan and type IV collagen were readily digested by BaP1 but not by leuc-a. Immunohistochemistry of muscle tissue samples showed a decrease in the immunostaining of type IV collagen after injection of BaP1, but not by leuc-a. Proteomic analysis by LC/MS/MS of exudates collected from injected muscle revealed higher amounts of perlecan, and types VI and XV collagens, in exudates from BaP1-injected tissue. The differences in the hemorrhagic activity of these SVMPs could be explained by their variable ability to degrade key BM and associated ECM substrates in vivo, particularly perlecan and several non-fibrillar collagens, which play a mechanical stabilizing role in microvessel structure. These results underscore the key role played by these ECM components in the mechanical stability of

  17. Real-Time Assessment of Guided Bone Regeneration in Standardized Calvarial Defects in Rats Using Bio-Oss With and Without Collagen Membrane: An In Vivo Microcomputed Tomographic and Histologic Experiment. (United States)

    Nooh, Nasser; Ramalingam, Sundar; Al-Kindi, Mohammed; Al-Rasheed, Abdulaziz; Al-Hamdan, Khalid S; Al-Hezaimi, Khalid


    In vivo microcomputed tomography (μCT) enables real-time assessment of bone regeneration. The aim of this μCT and histologic experiment was to assess guided bone regeneration (GBR) around standardized calvarial defects in rats using particulate graft material (Bio-Oss) with and without collagen membranes (CMs). Eighteen female Sprague-Dawley rats aged 6 weeks and weighing 300 g were used. With the rats under general anesthesia, calvaria were exposed and a full-thickness standardized defect was created on the parietal bone. For treatment, rats were randomly assigned to the following three groups: (1) CM group; (2) Bio-Oss group; and (3) Bio-Oss + CM group. Bone volume and bone mineral density (BMD) of newly formed bone (NFB) and remnant bone particles were measured at baseline and 2, 4, 6, and 10 weeks after the operations using real-time in vivo μCT. At 10 weeks, all animals were sacrificed and calvarial tissues were assessed histologically. In the CM group, a significant increase in mean ± standard deviation (SD) BMD of NFB was observed at 6 weeks (0.32 ± 0.02 g/mm(3)) (P Bio-Oss group, mean ± SD volume (3.03 ± 0.14 mm(3)) (P Bio-Oss + CM group, mean ± SD volume (0.98 ± 0.19 mm(3)) and BMD (0.13 ± 0.01 g/mm(3)) of NFB significantly increased at 4 weeks compared with baseline (P Bio-Oss + CM group, mean ± SD volume (3.5 ± 0.7 mm(3)) and BMD (0.44 ± 0.03 g/mm(3)) of remnant bone particles were significantly reduced at 10 weeks compared with baseline values (5.8 ± 0.96 mm(3) and 1.3 ± 0.02 g/mm(3)) (P Bio-Oss + CM group exhibited the most. The results of this study revealed that, in real time, new bone formation starts as early as 4 weeks in standardized calvarial defects undergoing GBR with Bio-Oss + CM, compared with new bone formation at 6 weeks in defects undergoing GBR with Bio-Oss alone.

  18. Avaliação do potencial osteogênico do periósteo em associação com uma membrana de colágeno Evaluation of osteogenic capacity of the periosteum in combination with collagen membrane

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    Wagner Costa Rossi Junior


    Full Text Available OBJETIVOS: Este trabalho avaliou o potencial osteogênico de enxertos de periósteo livre associado a uma membrana de colágeno. MÉTODOS: Vinte ratos albinos Wistar com idade média de 100 dias foram submetidos à cirurgia para criação de um defeito ósseo de 2,5 a 3,0 mm de comprimento na diáfise das fíbulas. Após 30 dias os animais foram então divididos em dois grupos: Grupo I recebeu o implante de periósteo associado à membrana de colágeno e Grupo II, somente a membrana de colágeno. Os animais foram radiografados antes do implante de periósteo e 15 ou 30 dias após o mesmo. RESULTADOS: Os resultados mostraram que o enxerto de periósteo livre associado à membrana de colágeno não foi eficiente no processo de reparo do defeito ósseo. CONCLUSÃO: Sugere-se que enxertos periosteais não vascularizados não apresentam potencial para formar novo osso. O fato de o enxerto ter sido implantado 30 dias após a criação do defeito ósseo interferiu negativamente no processo de osteogênese.OBJECTIVE: The present study evaluated the osteogenic potential of free periosteum graft in combination with collagen membranes. METHODS: Twenty white Wistar rats aged 100 days underwent surgery to create a bone defect measuring 2.5-3.0 mm in length in the diaphysis of the fibula. After thirty days, the animals were divided into two groups. Group I received periosteum along with a collagen membrane, while Group II received only a collagen membrane. The animals were X-rayed before the implant surgery and 15 or 30 days post-operation. RESULTS: The results demonstrated that free periosteum graft in combination with collagen membranes was not efficient in the repair of bone defects. CONCLUSION: We suggest that nonvascularized periosteal grafts do not show potential to form new bone, and that making the implant 30 days after the creation of the bone defect may have interfered negatively in osteogenic process.

  19. Mechanical properties of collagen fibrils


    Wenger, M. P. E.; Bozec, L.; Horton, M. A.; Mesquida, P


    The formation of collagen fibers from staggered subfibrils still lacks a universally accepted model. Determining the mechanical properties of single collagen fibrils ( diameter 50 - 200 nm) provides new insights into collagen structure. In this work, the reduced modulus of collagen was measured by nanoindentation using atomic force microscopy. For individual type 1 collagen fibrils from rat tail, the modulus was found to be in the range from 5 GPa to 11.5 GPa ( in air and at room temperature)...

  20. Collagenous gastritis: Review

    Institute of Scientific and Technical Information of China (English)

    Kenya Kamimura; Masaaki Kobayashi; Yuichi Sato; Yutaka Aoyagi; Shuji Terai


    Collagenous gastritis is a rare disease characterizedby the subepithelial deposition of collagen bandsthicker than 10 μm and the infiltration of inflammatorymononuclear cells in the lamina propria. Collagenouscolitis and collagenous sprue have similar histologicalcharacteristics to collagenous gastritis and are thoughtto be part of the same disease entity. However, whilecollagenous colitis has become more common inthe field of gastroenterology, presenting with clinicalsymptoms of chronic diarrhea in older patients,collagenous gastritis is rare. Since the disease was firstreported in 1989, only 60 cases have been documentedin the English literature. No safe and effective treatmentshave been identified from randomized, controlled trials.Therefore, better understanding of the disease and thereporting of more cases will help to establish diagnosticcriteria and to develop therapeutic strategies. Therefore,here we review the clinical characteristics, endoscopicand histological findings, treatment, and clinical outcomesfrom case reports and case series published to date,and provide a summary of the latest information on thedisease. This information will contribute to improvedknowledge of collagenous gastritis so physicians canrecognize and correctly diagnose the disease, and willhelp to develop a standard therapeutic strategy forfuture clinical trials.

  1. A biomimetic strategy to form calcium phosphate crystals on type I collagen substrate

    Energy Technology Data Exchange (ETDEWEB)

    Xu Zhang [Department of Restorative Dentistry, Faculty of Dentistry, National University of Singapore, 5 Lower Kent Ridge Road 119074, Singapore (Singapore); Neoh, Koon Gee [Department of Chemical and Biomolecular Engineering, National University of Singapore, Kent Ridge 119260, Singapore (Singapore); Kishen, Anil, E-mail: [Discipline of Endodontics, Faculty of Dentistry, University of Toronto, 124 Edward Street, Toronto, ON (Canada)


    Objective: The aim of this study is to induce mineralization of collagen by introducing phosphate groups onto type I collagen from eggshell membrane (ESM) by treating with sodium trimetaphosphate (STMP). This strategy is based on the hypothesis that phosphate groups introduced on collagen can mimic the nucleating role of phosphorylated non-collagenous proteins bound to collagen for inducing mineralization in natural hard tissue. Method: The collagen membrane was phosphorylated by treating it with a solution of STMP and saturated calcium hydroxide. The phosphorylated collagen was subsequently exposed to a mineralization solution and the pattern of mineralization on the surface of phosphorylated collagen substrate was analyzed. Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), field emission electron microscopy (FESEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD) and microhardness test were used to characterize the collagen substrate and the pattern of minerals formed on the collagen surface. Results: The FTIR and EDX results indicated that the phosphate groups were incorporated onto the collagen surface by treatment with STMP. During the mineralization process, the plate-like mineral, octacalcium phosphate (OCP), which was initially formed on the surface of ESM, was later transformed into needle-like hydroxyapatite (HAP) as indicated by the SEM, FESEM, EDX and XRD findings. The microhardness test displayed significant increase in the Knoop hardness number of the mineralized collagen. Conclusions: Phosphate groups can be introduced onto type I collagen surface by treating it with STMP and such phosphorylated collagen can induce the mineralization of type I collagen.

  2. Corneal collagen crosslinking for keratoconus. A review

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    M. M. Bikbov


    Full Text Available Photochemical crosslinking is widely applied in ophthalmology. Its biochemical effect is due to the release of singlet oxygen that promotes anaerobic photochemical reaction. Keratoconus is one of the most common corneal ectasia affecting 1 in 250 to 250 000 persons. Currently, the rate of iatrogenic ectasia following eximer laser refractive surgery increases due to biomechanical weakening of the cornea. Morphologically and biochemically, ectasia is characterized by corneal layers thinning, contact between the stroma and epithelium resulting from Bowman’s membrane rupture, chromatin fragmentation in keratocyte nuclei, phagocytosis, abnormal staining and arrangement of collagen fibers, enzyme system disorders, and keratocyte apoptosis. In corneal ectasia, altered enzymatic processes result in the synthesis of abnormal collagen. Collagen packing is determined by the activity of various extracellular matrix enzymes which bind amines and aldehydes of collagen fiber amino acids. In the late stage, morphological changes of Descemet’s membrane (i.e., rupture and detachment develop. Abnormal hexagonal-shaped keratocytes and their apoptosis are the signs of endothelial dystrophy. The lack of analogs in domestic ophthalmology encouraged the scientists of Ufa Eye Research Institute to develop a device for corneal collagen crosslinking. The parameters of ultraviolet (i.e., wavelength, exposure time, power to achieve the desired effect were identified. The specifics of some photosensitizers in the course of the procedure were studied. UFalink, a device for UV irradiation of cornea, and photosensitizer Dextralink were developed and adopted. Due to the high risk of endothelial damage, this treatment is contraindicated in severe keratoconus (CCT less than 400 microns. Major effects of corneal collagen crosslinking are the following: Young’s modulus (modulus of elasticity increase by 328.9 % (on average, temperature tolerance increase by 5

  3. Corneal collagen crosslinking for keratoconus. A review

    Directory of Open Access Journals (Sweden)

    M. M. Bikbov


    Full Text Available Photochemical crosslinking is widely applied in ophthalmology. Its biochemical effect is due to the release of singlet oxygen that promotes anaerobic photochemical reaction. Keratoconus is one of the most common corneal ectasia affecting 1 in 250 to 250 000 persons. Currently, the rate of iatrogenic ectasia following eximer laser refractive surgery increases due to biomechanical weakening of the cornea. Morphologically and biochemically, ectasia is characterized by corneal layers thinning, contact between the stroma and epithelium resulting from Bowman’s membrane rupture, chromatin fragmentation in keratocyte nuclei, phagocytosis, abnormal staining and arrangement of collagen fibers, enzyme system disorders, and keratocyte apoptosis. In corneal ectasia, altered enzymatic processes result in the synthesis of abnormal collagen. Collagen packing is determined by the activity of various extracellular matrix enzymes which bind amines and aldehydes of collagen fiber amino acids. In the late stage, morphological changes of Descemet’s membrane (i.e., rupture and detachment develop. Abnormal hexagonal-shaped keratocytes and their apoptosis are the signs of endothelial dystrophy. The lack of analogs in domestic ophthalmology encouraged the scientists of Ufa Eye Research Institute to develop a device for corneal collagen crosslinking. The parameters of ultraviolet (i.e., wavelength, exposure time, power to achieve the desired effect were identified. The specifics of some photosensitizers in the course of the procedure were studied. UFalink, a device for UV irradiation of cornea, and photosensitizer Dextralink were developed and adopted. Due to the high risk of endothelial damage, this treatment is contraindicated in severe keratoconus (CCT less than 400 microns. Major effects of corneal collagen crosslinking are the following: Young’s modulus (modulus of elasticity increase by 328.9 % (on average, temperature tolerance increase by 5

  4. Viability of corneal epithelial cells and cleavage of corneal basement after ethanol effect%乙醇对角膜上皮瓣活性及基底膜分离定位研究

    Institute of Scientific and Technical Information of China (English)

    陈颖欣; 蓝平; 高明宏; 范忠义; 宋福林; 徐旭; 于静


    Objective To evaluate the viability of corneal epithelial cells and to determine the anatomic cleavage on the epithelial basement membrane after various exposure times to 20% ethanol during epithelial flap preparation in laser-assisted subepithelial keratectomy(LASEK)in cadaver eyes.Methods Six human cadaver eyes were exposed to 20% ethanol for 20,30 and 40 seconds(2 eyes for each group),and another one eye was used as the control.PCNA staining was performed to determine the viability of corneal epithelial cells.Immunofluorescence staining using monoclonal antibodies against collagen Ⅶ,and immunohistological staining using monoclonal antibodies against laminin were performed to detect the anatomic location of the cleavage plane on the corneal epithelial flaps created by 20 seconds exposure to 20% ethanol in cadaver eyes.Results Hematoxylin and eosin staining of epithelial flaps revealed a coherent stratified epithelium.The PCNA positive rates of the epithelial cells in the flap decreased in the 20-second group,30-seconcl group and 40-second group successively.Immunohistological staining to laminin was patchy in the lifted flap and the remaining corneal basement membrane.Immunofluorescence to collagen Ⅶ,the main component of anchoring fibrils remained exclusively in the corneal bed.Conclusions Viability of the epithelial flap decreased with longer time exposure to ethanol.The cleavage plane of the ethanol-treated corneal epithelial flap is located between the lamina lucida and the lamina densa of the basement membrane where laminin forms hemidesmesome.%目的 探讨不同乙醇作用时间对角膜上皮瓣活性的影响,并确定角膜上皮瓣的解剖分离层面.方法 按标准准分子激光角膜上皮瓣下磨镶术(LASEK)方法制备7只尸体眼角膜上皮瓣,其中对照组1只眼,其余6只分为A、B、C 3个组,每组2只眼,乙醇浸润时间分别为20、30和40 S.对7只眼进行HE、增殖细胞核抗原(PCNA)、层黏连蛋白免疫组化

  5. Stabilization of collagen through bioconversion: An insight in protein-protein interaction. (United States)

    Usharani, Nagarajan; Jayakumar, Gladstone Christopher; Kanth, Swarna Vinodh; Rao, Jonnalagadda Raghava


    Collagen is a natural protein, which is used as a vital biomaterial in tissue engineering. The major concern about native collagen is lack of its thermal stability and weak resistance to proteolytic degradation. In this scenario, the crosslinking compounds used for stabilization of collagen are mostly of chemical nature and exhibit toxicity. The enzyme mediated crosslinking of collagen provides a novel alternative, nontoxic method for stabilization. In this study, aldehyde forming enzyme (AFE) is used in the bioconversion of hydroxylmethyl groups of collagen to formyl groups that results in the formation of peptidyl aldehyde. The resulted peptidyl aldehyde interacts with bipolar ions of basic amino acid residues of collagen. Further interaction leads to the formation of conjugated double bonds (aldol condensation involving the aldehyde group of peptidyl aldehyde) within the collagen. The enzyme modified collagen matrices have shown an increase in the denaturation temperature, when compared with native collagen. Enzyme modified collagen membranes exhibit resistance toward collagenolytic activity. Moreover, they exhibited a nontoxic nature. The catalytic activity of AFE on collagen as a substrate establishes an efficient modification, which enhances the structural stability of collagen. This finds new avenues in the context of protein-protein stabilization and discovers paramount application in tissue engineering.

  6. Deforming Etna's Basement: Implications for Edifice stability. (United States)

    Bakker, Richard; Benson, Philip; Vinciguerra, Sergio


    At over 3 kilometers in height, Mt. Etna (Italy) is the largest volcano of continental Europe. The volcano formed on top of the alpine fold and thrust belt, with basaltic outflows lying unconformably on top of an alternation between sandstones, limestones and clays. Presently Etna's eastern flank is moving with speeds up to 2cm/yr to the east [Tibaldi and Groppelli, 2002]. It is the sequence of layers below the volcano that is thought to provide a complex, structurally controlled, mechanism to the volcano deformation as a whole. This is due to the interplay of gravitational forces, volcanic pressurization, and regional tectonics, which combine to play a complex role that remains poorly understood, especially when the physical and mechanical properties of the rocks are considered. In this study, we concentrate on the rock mechanical component, and in particular the formation known as Comiso Limestone. This limestone forms of one of the key lithologies of Etna's basement. The formation has been suggested to be affected by thermal weakening [Heap et al., 2013]. Previous work on Comiso Limestone suggests brittle behavior for the range of temperatures (up to 760 ˚C) and a significant reduction in strength with higher temperatures. [Mollo et al., 2011]. Chiodini et al [2011], speculate carbonate assimilation. This implies that the Carbondioxide created by decarbonatization, is able to escape. Using an internally heated "Paterson" type pressure vessel, we recreated conditions at 2-4 km depth (50-100 MPa) and using an anomalously high geotherm, as expected in volcanic settings (ranging from room to 600 ˚C). With the addition of confining pressure, we show a brittle to ductile transition occurs at a relatively low temperature of 300 ˚C. A significant decrease in strength occurs when the rock is exposed to temperatures exceeding 400 ˚C. In addition, we observe a significant difference in mechanical behavior between vented and unvented situations when decarbonatization is

  7. Collagen metabolism in obesity

    DEFF Research Database (Denmark)

    Rasmussen, M H; Jensen, L T; Andersen, T


    OBJECTIVE: To investigate the impact of obesity, fat distribution and weight loss on collagen turnover using serum concentrations of the carboxyterminal propeptide of type I procollagen (S-PICP) and the aminoterminal propeptide of type III pro-collagen (S-PIIINP) as markers for collagen turnover...... (r = 0.37; P = 0.004), height (r = 0.27; P = 0.04), waist circumference (r = 0.35; P = 0.007), as well as with WHR (r = 0.33; P = 0.01) and was inversely correlated to age (r = -0.40; P = 0.002). Compared with randomly selected controls from a large pool of healthy volunteers, the obese patients had...... restriction (P obesity and associated with body fat distribution, suggesting...

  8. Collagen Homeostasis and Metabolism

    DEFF Research Database (Denmark)

    Magnusson, S Peter; Heinemeier, Katja M; Kjaer, Michael


    The musculoskeletal system and its collagen rich tissue is important for ensuring architecture of skeletal muscle, energy storage in tendon and ligaments, joint surface protection, and for ensuring the transfer of muscular forces into resulting limb movement. Structure of tendon is stable...... inactivity or immobilization of the human body will conversely result in a dramatic loss in tendon stiffness and collagen synthesis. This illustrates the importance of regular mechanical load in order to preserve the stabilizing role of the connective tissue for the overall function of the musculoskeletal...

  9. Collagen membrane guided tissue regeneration and planting bone surgery in the combined application of the periodontal therapy%胶原膜引导组织再生与植骨术在牙周病治疗中的联合应用

    Institute of Scientific and Technical Information of China (English)

    谢红帼; 严加林; 吕敏; 彭澜


    Objective:to study the collagen membrane guided tissue regeneration and planting bone surgery in the role of periodontal disease. Method:To gather 112 cases of patients with periodontal disease were divided radomly into two groups,the control group only with collagen membrane guided tissue regeneration treatments ,the observation group in the control group plus the basis of surgical treatment of grafts are compared between the two groups in postoperative situation. Results:in membrane exposure,1 week and 2 weeks is no significant difference(P>0.05),and in 4 weeks have obvious dif-ference (P0.05), two groups in new bone mass,the vertical bone height average increase on the imaging examination also have obvious differ-ence (P <0.05). Conclusion:collagen membrane guided tissue regeneration for bone graft and surgical treatment of peri-odontal disease clinical effect is remarkable.%目的:探讨胶原膜引导组织再生与植骨术在牙周病治疗中的作用。方法:选取112例牙周病患者为研究对象,随机平均分成2组,对照组单纯采用胶原膜引导组织再生治疗,治疗组在对照组的基础上加用植骨术治疗,比较两组临床效果。结果:在膜暴露上,第1周和第2周比较两组间无明显差异(P>0.05),第4周则有明显差异(P<0.05);两组术前和术后PD、AL、GR以及在术后第6个月、12个月时比较有明显差异(P<0.05),术后第3个月两组间比较无明显差异(P>0.05),两组病例在新生骨量、垂直向骨高度平均增加等影像学检查上也有明显差异(P<0.05)。结论:胶原膜引导组织再生与植骨术联合应用治疗牙周病临床效果显著。

  10. Building America Top Innovations 2012: Basement Insulation Systems

    Energy Technology Data Exchange (ETDEWEB)



    This Building America Top Innovations profile describes research on basement insulation, which identifies the wall installation methods and materials that perform best in terms of insulation and water resistance.

  11. Basement configuration of KG offshore basin from magnetic anomalies (United States)

    Subrahmanyam, V.; Swamy, K. V.; Raj, Neetha


    Marine magnetic anomalies along three representative profiles falling between shelf break and continent-ocean boundary in the offshore Krishna-Godavari basin were quantitatively interpreted for understanding the nature and structure of the magnetic basement using inversion technique. The interpretation of the anomalies shows that the magnetic basement lies deeper than the base of the sediments, i.e., acoustic basement identified by the seismic studies. This interpretation also shows that the magnetic basement is faulted along the NW-SE direction with the upthrown side lying to the north of the anomaly trend of this region. The coincidence of magnetizations observed through the present interpretation with that of charnockites of neighbouring EGMB and onshore K-G basin areas indicates that EGMB geology (charnockites, granitic gneiss, etc.) extends up to COB in the offshore K-G basin.

  12. [The genetics of collagen diseases]. (United States)

    Kaplan, J; Maroteaux, P; Frezal, J


    Heritable disorders of collagen include Ehler-Danlos syndromes (11 types are actually known), Larsen syndrome and osteogenesis imperfecta. Their clinical, genetic and biochemical features are reviewed. Marfan syndrome is closely related to heritable disorders of collagen.

  13. Basement domain map of the conterminous United States and Alaska (United States)

    Lund, Karen; Box, Stephen E.; Holm-Denoma, Christopher S.; San Juan, Carma A.; Blakely, Richard J.; Saltus, Richard W.; Anderson, Eric D.; DeWitt, Ed


    The basement-domain map is a compilation of basement domains in the conterminous United States and Alaska designed to be used at 1:5,000,000-scale, particularly as a base layer for national-scale mineral resource assessments. Seventy-seven basement domains are represented as eighty-three polygons on the map. The domains are based on interpretations of basement composition, origin, and architecture and developed from a variety of sources. Analysis of previously published basement, lithotectonic, and terrane maps as well as models of planetary development were used to formulate the concept of basement and the methodology of defining domains that spanned the ages of Archean to present but formed through different processes. The preliminary compilations for the study areas utilized these maps, national-scale gravity and aeromagnetic data, published and limited new age and isotopic data, limited new field investigations, and conventional geologic maps. Citation of the relevant source data for compilations and the source and types of original interpretation, as derived from different types of data, are provided in supporting descriptive text and tables.

  14. Collagen hydrolysate based collagen/hydroxyapatite composite materials (United States)

    Ficai, Anton; Albu, Madalina Georgiana; Birsan, Mihaela; Sonmez, Maria; Ficai, Denisa; Trandafir, Viorica; Andronescu, Ecaterina


    The aim of this study was to study the influence of collagen hydrolysate (HAS) on the formation of ternary collagen-hydrolysate/hydroxyapatite composite materials (COLL-HAS/HA). During the precipitation process of HA, a large amount of brushite is resulted at pH = 7 but, practically pure HA is obtained at pH ⩾ 8. The FTIR data reveal the duplication of the most important collagen absorption bands due to the presence of the collagen hydrolysate. The presence of collagen hydrolysate is beneficial for the management of bone and joint disorders such as osteoarthritis and osteoporosis.

  15. Prognostic Value of Glomerular Collagen IV Immunofluorescence Studies in Male Patients with X-Linked Alport Syndrome (United States)

    Gangemi, Concetta; Giannakakis, Kostas; Crisafi, Antonella; Faraggiana, Tullio; Fallerini, Chiara; Renieri, Alessandra; Muda, Andrea Onetti; Emma, Francesco


    Summary Background and objectives X-linked Alport syndrome (X-AS) is caused by mutations of the COL4A5 gene, which encodes for the collagen IV α5 chain (α5[COLIV]), resulting in structural and functional abnormalities of the glomerular basement membrane (GBM) and leading to CKD. The aim of the present study was to evaluate the prognostic value of residual collagen IV chain expression in the GBM of patients with X-AS. Design, setting, participants, & measurements The medical records of 22 patients with X-AS from 21 unrelated families collected between 1987 and 2009 were reviewed (median age at last follow-up, 19.9 years; range, 5.4–35.1 years); GBM expression of α1, α3, and α5(COLIV) chains was assessed by immunofluorescence microscopy. Results GBM distribution of the α5(COLIV) chain was diffuse in 1 and segmental or absent in 21 of the 22 patients; the expression of the α3(COLIV) chain was diffuse in 5 of 22 patients and segmental or absent in 17 of 22 patients. Patients with diffuse staining for the α3(COLIV) chain presented with proteinuria significantly later (median age, 16.9 versus 6.1 years; P=0.02) and reached an estimated GFR < 90 ml/min per 1.73 m2 at an older age (median age, 27.0 versus 14.9 years; P=0.01) compared with patients with segmental or absent staining. Two thirds of patients with abnormal α3(COLIV) expression by immunofluorescence studies had null or truncating COL4A5 mutations, as opposed to none of the 4 tested patients with diffuse α3(COLIV) chain glomerular distribution. Conclusions These results indicate that maintained expression of the α3(COLIV) chain is an early positive prognostic marker in patients with X-linked Alport symdrome. PMID:23371956

  16. Construction and characterization of a tissue-engineered oral mucosa equivalent based on a chitosan-fish scale collagen composite. (United States)

    Terada, Michiko; Izumi, Kenji; Ohnuki, Hisashi; Saito, Taro; Kato, Hiroko; Yamamoto, Marie; Kawano, Yoshiro; Nozawa-Inoue, Kayoko; Kashiwazaki, Haruhiko; Ikoma, Toshiyuki; Tanaka, Junzo; Maeda, Takeyasu


    This study was designed to (1) assess the in vitro biocompatibility of a chitosan-collagen composite scaffold (C3) constructed by blending commercial chitosan and tilapia scale collagen with oral mucosa keratinocytes, (2) histologically and immunohistochemically characterize an ex vivo-produced oral mucosa equivalent constructed using the C3 (EVPOME-C), and (3) compare EVPOME-C with oral mucosa constructs utilizing AlloDerm® (EVPOME-A), BioMend® Extend™ (EVPOME-B), and native oral mucosa. C3 scaffold had a well-developed fibril network and a sufficiently small porosity to prevent keratinocytes from growing inside the scaffold after cell-seeding. The EVPOME oral mucosa constructs were fabricated in a chemically defined culture system. After culture at an air-liquid interface, EVPOME-C and EVPOME-B had multilayered epithelium with keratinization, while EVPOME-A had a more organized stratified epithelium. Ki-67 and p63 immunolabeled cells in the basal layer of all EVPOMEs suggested a regenerative ability. Compared with native oral mucosa, the keratin 15 and 10/13 expression patterns in all EVPOMEs showed a less-organized differentiation pattern. In contrast to the β1-integrin and laminin distribution in EVPOME-A and native oral mucosa, the subcellular deposition in EVPOME-C and EVPOME-B indicated that complete basement membrane formation failed. These findings demonstrated that C3 has a potential application for epithelial tissue engineering and provides a new potential therapeutic device for oral mucosa regenerative medicine.

  17. Histomorphometric analysis of collagen and elastic fibres in the cranial and caudal fold of the porcine glottis. (United States)

    Lang, A; Koch, R; Rohn, K; Gasse, H


    The porcine glottis differs from the human glottis in its cranial and caudal vocal folds (CraF, CauF). The fibre apparatus of these folds was studied histomorphometrically in adult minipigs. For object definition and quantification, the colour-selection tools of the Adobe-Photoshop program were used. Another key feature was the subdivision of the cross-sections of the folds into proportional subunits. This allowed a statistical analysis irrespective of differences in thickness of the folds. Both folds had a distinct, dense subepithelial layer equivalent to the basement membrane zone in humans. The subsequent, loose layer was interpreted - in principle - as being equivalent to Reinke's space of the human vocal fold. The next two layers were not clearly separated. Due to this, the concept of a true vocal ligament did not appear applicable to neither CauF nor CraF. Instead, the body-cover model was emphasized by our findings. The missing vocalis muscle in the CraF is substituted by large collagen fibre bundles in a proportional depth corresponding to the position of the muscle of the CauF. The distribution of elastic fibres made the CraF rather than the CauF more similar to the human vocal fold. We suggest that these data are useful for those wishing to use the porcine glottis as a model for studying oscillatory properties during phonation.

  18. Comparative immunolocalisation of perlecan with collagen II and aggrecan in human foetal, newborn and adult ovine joint tissues demonstrates perlecan as an early developmental chondrogenic marker. (United States)

    Smith, Susan M; Shu, Cindy; Melrose, James


    We undertook a comparative immunolocalisation study on type II collagen, aggrecan and perlecan in a number of 12- to 14-week-old human foetal and postnatal (7-19 months) ovine joints including finger, toe, knee, elbow, hip and shoulder. This demonstrated that perlecan followed a virtually identical immunolocalisation pattern to that of type II collagen in the foetal tissues, but a slightly divergent localisation pattern in adult tissues. Aggrecan was also localised in the cartilaginous joint tissues, which were clearly delineated by toluidine blue staining and the type II collagen immunolocalisations. It was also present in the capsular joint tissues and in ligaments and tendons in the joint, which stained poorly or not at all with toluidine blue. In higher power microscopic views, antibodies to perlecan also stained small blood vessels in the synovial lining tissues of the joint capsule; however, this was not discernable in low power macroscopic views where the immunolocalisation of perlecan to pericellular regions of cells within the cartilaginous rudiments was a predominant feature. Perlecan was also evident in small blood vessels in stromal connective tissues associated with the cartilage rudiments and with occasional nerves in the vicinity of the joint tissues. Perlecan was expressed by rounded cells in the enthesis attachment points of tendons to bone and in rounded cells in the inner third of the meniscus, which stained prominently with type II collagen and aggrecan identifying the chondrogenic background of these cells and local compressive loads. Flattened cells within the tendon and in the surface laminas of articular cartilages and the meniscus did not express perlecan. Collected evidence presented herein, therefore, indicates that besides being a basement membrane component, perlecan is also a marker of chondrogenic cells in prenatal cartilages. In postnatal cartilages, perlecan displayed a pericellular localisation pattern rather than the territorial

  19. Collagen turnover after tibial fractures

    DEFF Research Database (Denmark)

    Joerring, S; Krogsgaard, M; Wilbek, H;


    collagen. A group comparison showed characteristic sequential changes in the turnover of types I and III collagen in fractures of the tibial diaphysis and tibial condyles. The turnover of type III collagen reached a maximum after 2 weeks in both groups. The synthesis of type I collagen reached a maximum......Collagen turnover after tibial fractures was examined in 16 patients with fracture of the tibial diaphysis and in 8 patients with fracture in the tibial condyle area by measuring sequential changes in serological markers of turnover of types I and III collagen for up to 26 weeks after fracture....... The markers were the carboxy-terminal extension peptide of type I procollagen (PICP), the amino-terminal extension peptide of type III procollagen (PIIINP), and the pyridinoline cross-linked carboxy-terminal telopeptide of type I collagen (ICTP). The latter is a new serum marker of degradation of type I...

  20. MT1-MMP and type II collagen specify skeletal stem cells and their bone and cartilage progeny

    DEFF Research Database (Denmark)

    Szabova, L.; Yamada, S.S.; Wimer, H.;


    Skeletal formation is dependent on timely recruitment of skeletal stem cells and their ensuing synthesis and remodeling of the major fibrillar collagens, type I collagen and type II collagen, in bone and cartilage tissues during development and postnatal growth. Loss of the major collagenolytic...... activity associated with the membrane-type 1 matrix metalloproteinase (MT1-MMP) results in disrupted skeletal development and growth in both cartilage and bone, where MT1-MMP is required for pericellular collagen dissolution. We show here that reconstitution of MT1-MMP activity in the type II collagen...

  1. Examination of the Basement of Historic Buildings in Investment Activity

    Directory of Open Access Journals (Sweden)

    Ulybin Aleksey


    Full Text Available The process and methodology of the survey of basements rarely mentioned in the various construction rules and regulations. Basically describes the procedure of conducting a detailed survey of some of the individual elements. These surveys are fundamental in nature, include a large number of estimates and require significant financial and time costs. Usually the purpose of these surveys is to check the state of the building as a whole, it’s safe operation or before starting of reconstruction. In the process of selecting areas of investment activity such large-scale survey is not possible. Needed a quick and inexpensive method intended for decision about investment in a particular object. At the same time, the survey should cover all the elements of the basement significantly affect the cost of reconstruction of the basement associated with his penetration. The article presents the general conception of conducting a rapid survey. The described methods and technologies applicable to the examination for the purpose of making decisions about investments in reconstruction of a basement level rooms. The composition of the works and their sequence. A comparison of the advantages and disadvantages of different methods. The practical examples. Scheme of conducting a rapid survey of the basement. The article analyzes the materials used in the construction of historic buildings in St. Petersburg.

  2. Effect of papain-based gel on type I collagen--spectroscopy applied for microstructural analysis. (United States)

    Silva Júnior, Zenildo Santos; Botta, Sergio Brossi; Ana, Patricia Aparecida; França, Cristiane Miranda; Fernandes, Kristianne Porta Santos; Mesquita-Ferrari, Raquel Agnelli; Deana, Alessandro; Bussadori, Sandra Kalil


    Considering the improvement of biomaterials that facilitate atraumatic restorative techniques in dentistry, a papain-based gel can be used in the chemomechanical removal of decayed dental tissue. However, there is no information regarding the influence of this gel on the structure of sound collagen. The aim of the present study was to investigate the adsorption of a papain-based gel (Papacarie(TM)) to collagen and determine collagen integrity after treatment. A pilot study was first performed with 10 samples of type I collagen membrane obtained from bovine Achilles deep tendon to compare the influence of hydration (Milli-Q water) on infrared bands of collagen. In a further experiment, 10 samples of type I collagen membrane were used to evaluate the effects of Papacarie(TM) on the collagen microstructure. All analyses were performed using the attenuated total reflectance technique of Fourier transform infrared (ATR-FTIR). The results demonstrated that the application of Papacarie(TM) does not lead to the degradation of collagen and this product can be safely used in minimally invasive dentistry. As the integrity of sound collagen is preserved after the application of the papain-based gel, this product is indicated for the selective removal of infected dentin, leaving the affected dentin intact and capable of re-mineralization.

  3. Regulation of collagen fibrillogenesis by cell-surface expression of kinase dead DDR2. (United States)

    Blissett, Angela R; Garbellini, Derek; Calomeni, Edward P; Mihai, Cosmin; Elton, Terry S; Agarwal, Gunjan


    The assembly of collagen fibers, the major component of the extracellular matrix (ECM), governs a variety of physiological processes. Collagen fibrillogenesis is a tightly controlled process in which several factors, including collagen binding proteins, have a crucial role. Discoidin domain receptors (DDR1 and DDR2) are receptor tyrosine kinases that bind to and are phosphorylated upon collagen binding. The phosphorylation of DDRs is known to activate matrix metalloproteases, which in turn cleave the ECM. In our earlier studies, we established a novel mechanism of collagen regulation by DDRs; that is, the extracellular domain (ECD) of DDR2, when used as a purified, soluble protein, inhibits collagen fibrillogenesis in-vitro. To extend this novel observation, the current study investigates how the DDR2-ECD, when expressed as a membrane-anchored, cell-surface protein, affects collagen fibrillogenesis by cells. We generated a mouse osteoblast cell line that stably expresses a kinase-deficient form of DDR2, termed DDR2/-KD, on its cell surface. Transmission electron microscopy, fluorescence microscopy, and hydroxyproline assays demonstrated that the expression of DDR2/-KD reduced the rate and abundance of collagen deposition and induced significant morphological changes in the resulting fibers. Taken together, our observations extend the functional roles that DDR2 and possibly other membrane-anchored, collagen-binding proteins can play in the regulation of cell adhesion, migration, proliferation and in the remodeling of the extracellular matrix.

  4. Inhibition of collagen fibrillogenesis by cells expressing soluble extracellular domains of DDR1 and DDR2. (United States)

    Flynn, Lisa A; Blissett, Angela R; Calomeni, Edward P; Agarwal, Gunjan


    Collagen fiber assembly affects many physiological processes and is tightly controlled by collagen-binding proteins. However, to what extent membrane-bound versus cell-secreted collagen-binding proteins affect collagen fibrillogenesis is not well understood. In our previous studies, we had demonstrated that the membrane-anchored extracellular domain (ECD) of the collagen receptor discoidin domain receptor 2 (DDR2) inhibits fibrillogenesis of collagen endogenously secreted by the cells. These results led to a novel functional role of the DDR2 ECD. However, since soluble forms of DDR1 and DDR2 containing its ECD are known to naturally exist in the extracellular matrix, in this work we investigated if these soluble DDR ECDs may have a functional role in modulating collagen fibrillogenesis. For this purpose, we created mouse osteoblast cell lines stably secreting DDR1 or DDR2 ECD as soluble proteins. Transmission electron microscopy, fluorescence microscopy, and hydroxyproline assays were used to demonstrate that DDR ECD expression reduced the rate and quantity of collagen deposition and induced significant changes in fiber morphology and matrix mineralization. Collectively, our studies advance our understanding of DDR receptors as powerful regulators of collagen deposition in the ECM and elucidate their multifaceted role in ECM remodeling.

  5. 引导骨再生膜技术对较大根尖囊肿术后骨腔修复的临床研究%Clinical analysis of guided bone regeneration and collagen membrane in the treatment of larger radicular cyst

    Institute of Scientific and Technical Information of China (English)

    李志萍; 孟箭; 张杰; 孟庆飞; 顾倩平; 刘颖


    Objective To evaluate the clinical effects of the guided bone regeneration ( Bio-oss) and collagen membrane in the treatment of larger radicular cyst. Methods 63 patients suffering radicular cyst were enrolled. All these cases were randomly divided into 3 groups. 21 cases were treated with Bio-Gide and Bio-oss,and 21 were treated with collagen membrane in fossa only. The last 21 eases who received cyst extraction were the control group. Clinical parameters evaluated included changes in tooth mobility and the bone loss of the alveolar process according to radiograms preoperatively and at the 3rd,6th and 12th months after operation. Results At the 3rd,6th and 12th months postoperatively,two observation groups showed significantly more bone density increased according to the radiographs than the control group (P <0.05). The 1 st group with Bio-oss and collagen membrane had the best efficacy. The teeth with radicular cyst remained well. Conclusions Bio-oss and absorbable collagen membranes show excellent bio-compatibility, which can accelerate the bone repair progress and can be successfully used for reconstruction of alveolar bone to obtain satisfactory clinical effects.%目的 探讨Bio-oss骨粉复合胶原膜在较大根尖囊肿术后骨腔修复中的临床效果.方法 将63例根尖囊肿的患者随机分成实验组和对照组.第1组21例行根尖囊肿摘除后给予Bio - oss骨粉复合胶原膜填塞;第2组21例仅在术后骨腔外放置胶原膜;选择21例术后囊腔内不放置充填物作为对照组.进行第3、6、12个月的临床观察,分析2组X线下成骨变化.结果 术后第3、6、12个月骨缺损区骨密度改变中骨粉复合胶原膜组均显著优于对照组(P<0.05),且以植骨复合胶原膜组骨密度增高明显,有较多的新骨小梁修复,实验组患牙未见瘘管形成,患牙保存较好.结论 Bio -oss复合胶原膜具有良好的生物相容性、骨传导和骨诱导作用,有利于骨组织长入及囊

  6. Genetics Home Reference: multicentric osteolysis, nodulosis, and arthropathy (United States)

    ... to cut (cleave) a protein called type IV collagen. Type IV collagen is a major structural component of basement membranes, ... 2 enzyme, preventing the normal cleavage of type IV collagen. It is unclear how a loss of enzyme ...

  7. Basement structure of the Granada basin, Betic Cordilleras, southern Spain (United States)

    Morales, J.; Vidal, F.; De Miguel, F.; Alguacil, G.; Posadas, A. M.; Ibañez, J. M.; Guzmán, A.; Guirao, J. M.


    The analysis and interpretation of geophysical data (gravity and seismic reflection) has facilitated the definition of the Granada basin structure. The reflector showing the contact between the Betic-Subbetic basement and the Neogene-Quaternary sedimentary filling has been identified. Mapping of the basement in two and three dimensions is presented. The presence of four important depocenters (Genil, Chimeneas, Cubillas and Granada) has been determined. These troughs are limited by ridge areas through important sets of fractures. In some cases the accumulation of Neogene-Quaternary sediments reaches a thickness exceeding 3 km as in the Genii and Cubillas depocenters. The mapping of the most important fractures affecting the basement has been achieved, defining four systems that have influenced and conditioned the genesis and late evolution of the Granada basin. The directions of the most important groups of fractures are: NE-SW, N70W to E-W, N45W and N10-30E.

  8. Primary hepatocyte culture in collagen gel mixture and collagen sandwich

    Institute of Scientific and Technical Information of China (English)

    Ying-Jie Wang; Hong-Ling Liu; Hai-Tao Guo; Hong-Wei Wen; Jun Liu


    AIM: To explore the methods of hepatocytes culture in a collagen gel mixture or between double layers of collagen sandwich configuration and to examine the functional and cytomorphological characteristics of cultured hepatocytes.METHODS: A two-step collagenase perfusion technique was used to isolate the hepatocytes from Wistar rats or newborn Chinese experimental piglets. The isolated hepatocytes were cultured in a collagen gel mixture or between double layers of collagen sandwich configuration respectively. The former was that rat hepatocytes were mixed with type I rat tail collagen solution till gelled, and the medium was added onto the gel. The latter was that swine hepatocytes were seeded on a plate precoated with collagen gel for 24 h, then another layer of collagen gel was overlaid, resulting in a sandwich configuration. The cytomorphological characteristics, albumin secretion, and LDH-release of the hepatocytes cultured in these two models were examined.RESULTS: Freshly isolated rat hepatocytes were successfully mixed and fixed in collagen gel, and cultured in the gel condition. During the culture period, the urea synthesized and secreted by rat hepatocytes was detected throughout the period. Likewise, newborn experimental piglet hepatocytes were successfully fixed between the double layers of collagen gel, forming a sandwich configuration.Within a week of culture, the albumin secreted by swine hepatocytes was detected by SDS/PAGE analysis. The typical cytomorphological characteristics of the hepatocytes cultured by the above two culture models were found under a phasecontrast microscope. There was little LDH-release during the culture period.CONCLUSION: Both collagen gel mixture and double layers of collagen sandwich configuration can provide cultural conditions much closer to in vivoenvironment, and are helpful for maintaining specific hepatic fiJnctions and cytomorphological characteristics. A collagen gel mixture culture may be more eligible for the

  9. Shining light on collagen: expressing collagen in plants. (United States)

    Brodsky, Barbara; Kaplan, David L


    Collagens are a remarkable group of proteins that are critical from a physiological perspective due to their diverse and versatile functions in vivo. However, collagens are challenging to generate ex vivo for biomaterials or regenerative medicine due to their complex processing and assembly into functional materials. Therefore, collagen availability remains a major unmet need for biomaterials, as relatively limited supplies of the protein in pure form are available mainly through harvesting bovine tissues. This animal source, subsequent to purification, remains associated with significant safety concerns due to the potential carryover of animal-derived diseases. Other more limited sources of animal collagens are also commercially available, as well as collagens generated in heterologous hosts; however, the challenge to these sources remains both economic and structural. The need for new safe sources of collagens remains high, with a significant potential impact in areas of medicine when considering the opportunity to mimic native collagen features. The articles in this issue of the journal focus on plant-derived collagens to address some of these needs. Progress toward plant production of collagens, the ability to self-assemble these recombinant proteins into higher-order structures, and the utility of these materials in various medical applications suggest an important path forward for the field.

  10. UV damage of collagen: insights from model collagen peptides. (United States)

    Jariashvili, Ketevan; Madhan, Balaraman; Brodsky, Barbara; Kuchava, Ana; Namicheishvili, Louisa; Metreveli, Nunu


    Fibrils of Type I collagen in the skin are exposed to ultraviolet (UV) light and there have been claims that collagen photo-degradation leads to wrinkles and may contribute to skin cancers. To understand the effects of UV radiation on collagen, Type I collagen solutions were exposed to the UV-C wavelength of 254 nm for defined lengths of time at 4°C. Circular dichroism (CD) experiments show that irradiation of collagen leads to high loss of triple helical content with a new lower thermal stability peak and SDS-gel electrophoresis indicates breakdown of collagen chains. To better define the effects of UV radiation on the collagen triple-helix, the studies were extended to peptides which model the collagen sequence and conformation. CD studies showed irradiation for days led to lower magnitudes of the triple-helix maximum at 225 nm and lower thermal stabilities for two peptides containing multiple Gly-Pro-Hyp triplets. In contrast, the highest radiation exposure led to little change in the T(m) values of (Gly-Pro-Pro)(10) and (Ala-Hyp-Gly)(10) , although (Gly-Pro-Pro)(10) did show a significant decrease in triple helix intensity. Mass spectroscopy indicated preferential cleavage sites within the peptides, and identification of some of the most susceptible sites of cleavage. The effect of radiation on these well defined peptides gives insight into the sequence and conformational specificity of photo-degradation of collagen.

  11. Discoidin domain receptor 2 regulates the adhesion of fibroblasts to 3D collagen matrices. (United States)

    Kim, Daehwan; You, Eunae; Min, Na Young; Lee, Kwang-Ho; Kim, Hyoung Kyu; Rhee, Sangmyung


    The collagen matrix constitutes the primary extracellular matrix (ECM) portion of mammalian connective tissues in which the interaction of the cell and the surrounding collagen fibers has a significant impact on cell and tissue physiology, including morphogenesis, development and motility. Discoidin domain receptors (DDR1 and DDR2) have been identified as the receptor tyrosine kinases that are activated upon collagen binding. However, there is a lack of evidence regarding the effect of DDRs on the mechanical interaction between fibroblasts and ECM. In this study, we demonstrated that one of the major phosphotyrosine proteins in human fibroblasts during 3D collagen matrix polymerization is DDR2. Treatment of fibroblasts in 3D collagen matrices with platelet-derived growth factor (PDFG) has been shown to increase DDR2 phosphorylation. Silencing of DDR2 with siRNA in fibroblasts significantly reduced the number of dendritic extensions regardless of whether cells were cultured in the collagen or fibronectin 3D matrices. Decreasing dendritic extensions in DDR2-silenced cells has also been shown to decrease the ability of fibroblast entanglement to collagen fibrils in 3D collagen matrices. Finally, we also showed that the silencing of DDR2 decreased the cell migration in 3D nested collagen matrices but had no effect on 3D floating matrix contraction. Collectively, these results suggest that DDR2 functioning is required for the membrane dynamics to control the mechanical attachment of fibroblasts to the 3D collagen matrices in an integrin-independent manner.

  12. Arterial calcification: Conscripted by collagen (United States)

    Miller, Jordan D.


    In atherosclerotic plaques, patterns of calcification -- which have profound implications for plaque stability and vulnerability to rupture -- are determined by the collagen's content and patterning throughout the plaque.

  13. Magnetic basement in the central Bay of Bengal

    Digital Repository Service at National Institute of Oceanography (India)

    Sarma, K.V.L.N; Ramana, M.V.; Ramprasad, T.; Desa, M.; Subrahmanyam, V.; Krishna, K.S.; Rao, M.M.M.

    . The N10-12 degrees W trending subsurface 85 degrees E Ridge buried under 2 to 3 km thick sediments is a prominent tectonic feature. Offshore basins characterised by deeper magnetic basement (approx. 9 km) and 100-200 km wide are present on either sides...

  14. Production, characterization and biocompatibility of marine collagen matrices from an alternative and sustainable source: the sea urchin Paracentrotus lividus. (United States)

    Benedetto, Cristiano Di; Barbaglio, Alice; Martinello, Tiziana; Alongi, Valentina; Fassini, Dario; Cullorà, Emanuele; Patruno, Marco; Bonasoro, Francesco; Barbosa, Mario Adolfo; Carnevali, Maria Daniela Candia; Sugni, Michela


    Collagen has become a key-molecule in cell culture studies and in the tissue engineering field. Industrially, the principal sources of collagen are calf skin and bones which, however, could be associated to risks of serious disease transmission. In fact, collagen derived from alternative and riskless sources is required, and marine organisms are among the safest and recently exploited ones. Sea urchins possess a circular area of soft tissue surrounding the mouth, the peristomial membrane (PM), mainly composed by mammalian-like collagen. The PM of the edible sea urchin Paracentrotus lividus therefore represents a potential unexploited collagen source, easily obtainable as a food industry waste product. Our results demonstrate that it is possible to extract native collagen fibrils from the PM and produce suitable substrates for in vitro system. The obtained matrices appear as a homogeneous fibrillar network (mean fibril diameter 30-400 nm and mesh collagen source for mechanically resistant biomedical devices.

  15. 术前正畸联合可吸收胶原生物膜用于牙槽突裂植骨修复30例分析%Analysis of 30 patients with pre-surgical orthodontics and application of absorbable collagen bio-membrane on alveolar clefe bone grafting

    Institute of Scientific and Technical Information of China (English)

    李志强; 黄诺蓓; 艾伟健; 刘曙光


    目的 探讨术前正畸联合术中应用可吸收胶原生物膜对单侧牙槽突裂植骨效果的影响.方法 选择牙弓狭窄、上颌前牙舌倾或扭转、牙槽突裂隙不规则、难以进行牙槽突裂檀骨术的单侧完全性牙槽突裂患者30例,年龄9 ~13岁,先进行植骨前正畸治疗,再应用髂骨松质骨加可吸收胶原生物膜覆盖行植骨修复,术后定期拍X线片检查,观察植骨效果.牙槽骨高度评价标准采用Bergland标准进行,术后观察期为1~3年.结果 30例患者术后成骨情况Ⅰ型11例,Ⅱ型17例,植骨成功率达93.3%.结论 对于上颌牙弓狭窄、牙槽突裂隙不规则、牙颌畸形严重的牙槽突裂患者,建议先行植骨前正畸治疗,植骨术中联合应用可吸收胶原生物膜可有效提高植骨成功率.%Objective To discuss the effect of bone grafting in patients with unilateral alveolar cleft,treated by presurgical orthodontics and secondary alveolar bone grafting surgery with absorbable collagen bio-membrane,in order to improve the success rate of graft.Methods Thirty complete unilateral alveolar cleft patients,aged 9-13 years,with collapsed upper arch or severe malpositioned upper incisors were selected.The patients received pre-surgical orthodontics before secondary alveolar bone grafting used ilium and absorbable bio-membrane.The observation period was 1-3 years and X-ray was taken regularly.Bergland criteria were used to evaluate the alveolar bone height.Results Pre-surgical orthodontics expanded the collapsed upper arch to benefit the secondary alveolar bone grafting with absorbable collagen bio-membrane,and the success rate of graft was improved to 93.3%.Conclusion It is worth popularizing that patients with collapsed upper arch or severe mal-positioned upper incisors should receive pre-surgical orthodontics and then secondary alveolar bone graft surgery with absorbable collagen bio-membrane.

  16. Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells. (United States)

    Matsumoto, Rena; Uemura, Toshimasa; Xu, Zhefeng; Yamaguchi, Isamu; Ikoma, Toshiyuki; Tanaka, Junzo


    We studied the effect of fibril formation of fish scale collagen on the osteoblastic differentiation of human mesenchymal stem cells (hMSCs). We found that hMSCs adhered easily to tilapia scale collagen, which remarkably accelerated the early stage of osteoblastic differentiation in hMSCs during in vitro cell culture. Osteoblastic markers such as ALP activity, osteopontin, and bone morphogenetic protein 2 were markedly upregulated when the hMSCs were cultured on a tilapia collagen surface, especially in the early osteoblastic differentiation stage. We hypothesized that this phenomenon occurs due to specific fibril formation of tilapia collagen. Thus, we examined the time course of collagen fibril formation using high-speed atomic force microscopy. Moreover, to elucidate the effect of the orientation of fibril formation on the differentiation of hMSCs, we measured ALP activity of hMSCs cultured on two types of tilapia scale collagen membranes with different degrees of fibril formation. The ALP activity in hMSCs cultured on a fibrous collagen membrane was significantly higher than on a non-fibrous collagen membrane even before adding osteoblastic differentiation medium. These results showed that the degree of the fibril formation of tilapia collagen was essential for the osteoblastic differentiation of hMSCs.

  17. Collagen for bone tissue regeneration. (United States)

    Ferreira, Ana Marina; Gentile, Piergiorgio; Chiono, Valeria; Ciardelli, Gianluca


    In the last decades, increased knowledge about the organization, structure and properties of collagen (particularly concerning interactions between cells and collagen-based materials) has inspired scientists and engineers to design innovative collagen-based biomaterials and to develop novel tissue-engineering products. The design of resorbable collagen-based medical implants requires understanding the tissue/organ anatomy and biological function as well as the role of collagen's physicochemical properties and structure in tissue/organ regeneration. Bone is a complex tissue that plays a critical role in diverse metabolic processes mediated by calcium delivery as well as in hematopoiesis whilst maintaining skeleton strength. A wide variety of collagen-based scaffolds have been proposed for different tissue engineering applications. These scaffolds are designed to promote a biological response, such as cell interaction, and to work as artificial biomimetic extracellular matrices that guide tissue regeneration. This paper critically reviews the current understanding of the complex hierarchical structure and properties of native collagen molecules, and describes the scientific challenge of manufacturing collagen-based materials with suitable properties and shapes for specific biomedical applications, with special emphasis on bone tissue engineering. The analysis of the state of the art in the field reveals the presence of innovative techniques for scaffold and material manufacturing that are currently opening the way to the preparation of biomimetic substrates that modulate cell interaction for improved substitution, restoration, retention or enhancement of bone tissue function.

  18. Stimulation of MMP-11 (stromelysin-3 expression in mouse fibroblasts by cytokines, collagen and co-culture with human breast cancer cell lines

    Directory of Open Access Journals (Sweden)

    Matthaei Klaus I


    Full Text Available Abstract Background Matrix metalloproteinases (MMPs are central to degradation of the extracellular matrix and basement membrane during both normal and carcinogenic tissue remodeling. MT1-MMP (MMP-14 and stromelysin-3 (MMP-11 are two members of the MMP family of proteolytic enzymes that have been specifically implicated in breast cancer progression. Expressed in stromal fibroblasts adjacent to epithelial tumour cells, the mechanism of MT1-MMP and MMP-11 induction remains unknown. Methods To investigate possible mechanisms of induction, we examined the effects of a number of plausible regulatory agents and treatments that may physiologically influence MMP expression during tumour progression. Thus NIH3T3 and primary mouse embryonic fibroblasts (MEFs were: a treated with the cytokines IL-1β, IL-2, IL-6, IL-8 and TGF-β for 3, 6, 12, 24, and 48 hours; b grown on collagens I, IV and V; c treated with fibronectin, con-A and matrigel; and d co-cultured with a range of HBC (human breast cancer cell lines of varied invasive and metastatic potential. Results Competitive quantitative RT-PCR indicated that MMP-11 expression was stimulated to a level greater than 100%, by 48 hour treatments of IL-1β, IL-2, TGF-β, fibronectin and collagen V. No other substantial changes in expression of MMP-11 or MT1-MMP in either tested fibroblast culture, under any treatment conditions, were observed. Conclusion We have demonstrated significant MMP-11 stimulation in mouse fibroblasts using cytokines, matrix constituents and HBC cell lines, and also some inhibition of MT1-MMP. Our data suggest that the regulation of these genes in the complex stromal-epithelial interactions that occur in human breast carcinoma, is influenced by several mechanisms.

  19. COOH-terminal collagen Q (COLQ) mutants causing human deficiency of endplate acetylcholinesterase impair the interaction of ColQ with proteins of the basal lamina. (United States)

    Arredondo, Juan; Lara, Marian; Ng, Fiona; Gochez, Danielle A; Lee, Diana C; Logia, Stephanie P; Nguyen, Joanna; Maselli, Ricardo A


    Collagen Q (ColQ) is a key multidomain functional protein of the neuromuscular junction (NMJ), crucial for anchoring acetylcholinesterase (AChE) to the basal lamina (BL) and accumulating AChE at the NMJ. The attachment of AChE to the BL is primarily accomplished by the binding of the ColQ collagen domain to the heparan sulfate proteoglycan perlecan and the COOH-terminus to the muscle-specific receptor tyrosine kinase (MuSK), which in turn plays a fundamental role in the development and maintenance of the NMJ. Yet, the precise mechanism by which ColQ anchors AChE at the NMJ remains unknown. We identified five novel mutations at the COOH-terminus of ColQ in seven patients from five families affected with endplate (EP) AChE deficiency. We found that the mutations do not affect the assembly of ColQ with AChE to form asymmetric forms of AChE or impair the interaction of ColQ with perlecan. By contrast, all mutations impair in varied degree the interaction of ColQ with MuSK as well as basement membrane extract (BME) that have no detectable MuSK. Our data confirm that the interaction of ColQ to perlecan and MuSK is crucial for anchoring AChE to the NMJ. In addition, the identified COOH-terminal mutants not only reduce the interaction of ColQ with MuSK, but also diminish the interaction of ColQ with BME. These findings suggest that the impaired attachment of COOH-terminal mutants causing EP AChE deficiency is in part independent of MuSK, and that the COOH-terminus of ColQ may interact with other proteins at the BL.

  20. Efficacy of Combined Therapy of Q-switched Alexandrite Laser and Collagen Protein Membrane on Naevus Fuscocaeruleus Zygomaticus%调Q翠绿宝石激光联合胶原蛋白膜治疗褐青色痣的疗效观察

    Institute of Scientific and Technical Information of China (English)

    杨超; 刘刚; 陈小霞; 严蕾; 王芳; 谢军; 袁晓慧


    目的 探讨调Q翠绿宝石激光联合胶原蛋白膜治疗褐青色痣的疗效.方法 褐青色痣女性患者267例,按就诊年度,分为2组,实验组138例,对照组129例.两组患者均先以调Q翠绿宝石激光(波长755 nm,光斑直径3mm,频率2或5 Hz,脉宽100 ns,能量密度6~7 J/cm2)垂直照射患处.照射后即刻实验组患者以胶原蛋白膜冷敷患处20 min,对照组患者以水袋冷敷患处20 min.每日治疗1次,共5次.治疗后6个月比较两组的治愈率、总有效率和色素沉着发生率.结果 试验组有效率为92%,痊愈率为64.5%,色素沉着率为13.8%;对照组有效率为86%,痊愈率为59.7%,色素沉着率为28.7%%.两组痊愈率比较,差异有非常显著意义(P<0.01);两组总有效率比较,差异无显著意义(P>0.05);两组色素沉着发生率比较,差异有显著意义.结论 调Q翠绿宝石激光联合胶原蛋白膜治疗褐青色斑痣效果好,可减少色素沉着的发生.%Objective To observe the effect of the therapy combining Q-switched alexandrite laser and collagen protein membrane on naevus fuscocaeruleus zygomaticus ( NFZ). Methods Altogether 129 patients with NEZ were assigned to a control group, who were given Q-switched alexandrite laser and applied with ice pack for 20 minutes after the laser treatment. A test group was set up with 138 patients of NEZ, who were treated with the same laser but given collagen protein membrane for 20 minutes after the treatment. Each patient received five treatments in total, once per day. All the patients were required to use the same antibiotics ointment for 7 days and avoid water and the sun. The pictures of the focus before and after the treatment were taken, and the adverse reactions of the treatment were observed. Results The efficacy, recovery and pigmentation rates of the test group are 92% , 64. 5% and 13. 8 % respectively while those of the control group being 86% , 59.7% , and 28.7% respectively. According to the

  1. Collagenous Gastritis: A Rare Entity

    Directory of Open Access Journals (Sweden)

    Faten Limaiem


    Full Text Available Collagenous gastritis is a rare entity of unknown etiology characterized histologically by the presence of a thick subepithelial collagen band associated with an inflammatory infiltrate of gastric mucosa. A 40-year-old male presented with a history of chronic intermittent abdominal pain for about 6 months. Physical examination was unremarkable, and biological tests were within normal range. The patient underwent esophagogastroduodenoscopy and colonoscopy which showed a nodular mucosa of the stomach. Biopsies of the duodenum and colon were unremarkable. However, biopsies of the gastric fundus revealed a mild chronic gastritis characterized by lymphocytic and plasma cell infiltration of deep mucosa, without lymphoid follicle formation or active inflammation. No microorganisms were identified on routine hematoxylin and eosin or Giemsa-stained sections. Subepithelial collagen in the gastric biopsies was thickened and showed entrapped capillaries. Subepithelial collagen was highlighted by Masson's trichrome staining and was negative for amyloid by Congo Red. In the areas containing thickened collagen, there were no intraepithelial lymphocytes. The final pathological diagnosis was collagenous gastritis. Collagenous gastritis is an extremely rare disease, but it is important to recognize its characteristic endoscopic and pathologic findings to make a correct diagnosis. Specific therapy for this rare entity has not yet been established. [J Interdiscipl Histopathol 2015; 3(2.000: 68-70

  2. Basement configuration of KG offshore basin from magnetic anomalies

    Indian Academy of Sciences (India)

    V Subrahmanyam; K V Swamy; Neetha Raj


    Marine magnetic anomalies along three representative profiles falling between shelf break and continent–ocean boundary in the offshore Krishna–Godavari basin were quantitatively interpreted for understandingthe nature and structure of the magnetic basement using inversion technique. The interpretation of theanomalies shows that the magnetic basement lies deeper than the base of the sediments, i.e., acousticbasement identified by the seismic studies. This interpretation also shows that the magnetic basementis faulted along the NW–SE direction with the upthrown side lying to the north of the anomaly trendof this region. The coincidence of magnetizations observed through the present interpretation with thatof charnockites of neighbouring EGMB and onshore K–G basin areas indicates that EGMB geology(charnockites, granitic gneiss, etc.) extends up to COB in the offshore K–G basin.

  3. Complementary roles of intracellular and pericellular collagen degradation pathways in vivo

    DEFF Research Database (Denmark)

    Wagenaar-Miller, Rebecca A; Engelholm, Lars H; Gavard, Julie


    these two pathways is unclear and even controversial. Here we show that intracellular and pericellular collagen turnover pathways have complementary roles in vivo. Individual deficits in intracellular collagen degradation (urokinase plasminogen activator receptor-associated protein/Endo180 ablation......Collagen degradation is essential for cell migration, proliferation, and differentiation. Two key turnover pathways have been described for collagen: intracellular cathepsin-mediated degradation and pericellular collagenase-mediated degradation. However, the functional relationship between......) or pericellular collagen degradation (membrane type 1-matrix metalloproteinase ablation) were compatible with development and survival. Their combined deficits, however, synergized to cause postnatal death by severely impairing bone formation. Interestingly, this was mechanistically linked to the proliferative...

  4. Electrostatic effects in collagen fibrillization (United States)

    Morozova, Svetlana; Muthukumar, Murugappan


    Using light scattering and AFM techniques, we have measured the kinetics of fibrillization of collagen (pertinent to the vitreous of human eye) as a function of pH and ionic strength. At higher and lower pH, collagen triple-peptides remain stable in solution without fibrillization. At neutral pH, the fibrillization occurs and its growth kinetics is slowed upon either an increase in ionic strength or a decrease in temperature. We present a model, based on polymer crystallization theory, to describe the observed electrostatic nature of collagen assembly.

  5. Sterile Keratitis following Collagen Crosslinking (United States)

    Javadi, Mohammad-Ali; Feizi, Sepehr


    Purpose: To report a keratoconic eye that developed severe sterile keratitis and corneal scar after collagen crosslinking necessitating corneal transplantation. Case Report: A 26-year-old man with progressive keratoconus underwent collagen crosslinking and presented with severe keratitis 72 hours after the procedure. The initial impression was infectious corneal ulcer and a fortified antibiotic regimen was administered. However, the clinical course and confocal microscopy results prompted a diagnosis of sterile keratitis. The eye developed severe corneal scars leading to reduced visual acuity and necessitating corneal transplantation. Conclusion: Sterile keratitis may develop after collagen crosslinking resulting in profound visual loss leading to corneal transplantation. PMID:25709779

  6. Stability of collagen during denaturation. (United States)

    Penkova, R; Goshev, I; Gorinstein, S; Nedkov, P


    The stability of calf skin collagen (CSC) type I during thermal and chemical denaturation in the presence of glycerol was investigated. Thermal denaturation of type I collagen was performed in the presence of glycerol or in combination with urea and sodium chloride. The denaturation curves obtained in the presence of urea or sodium chloride retained their original shape without glycerol. These curves were shifted upward proportionally to the glycerol concentration in the reaction medium. This means that glycerol and the denaturants act independently. The explanation is based on the difference in the mechanism of their action on the collagen molecule.

  7. The Grenville-age basement of the Andes (United States)

    Ramos, Victor A.


    The analysis of the basement of the Andes shows the strong Grenville affinities of most of the inliers exposed in the different terranes from Colombia to Patagonia. The terranes have different histories, but most of them participated in the Rodinia supercontinent amalgamation during the Mesoproterozoic between 1200 and 1000 Ma. After Rodinia break-up some terranes were left in the Laurentian side such as Cuyania and Chilenia, while others stayed in the Gondwanan side. Some of the terranes once collided with the Amazon craton remained attached, experiencing diverse rifting episodes all along the Phanerozoic, as the Arequipa and Pampia terranes. Some other basement inliers were detached in the Neoproterozoic and amalgamated again to Gondwana in the Early Cambrian, Middle Ordovician or Permian times. A few basement inliers with Permian metamorphic ages were transferred to Gondwana after Pangea break-up from the Laurentian side. Some of them were part of the present Middle America terrane. An exceptional case is the Oaxaquia terrane that was detached from the Gondwana margin after the Early Ordovician and is now one of the main Mexican terranes that collided with Laurentia. These displacements, detachments, and amalgamations indicate a complex terrane transfer between Laurentia and Gondwana during Paleozoic times, following plate reorganizations and changes in the absolute motion of Gondwana.

  8. 矿化胶原复合物引导组织再生膜体外性能测试%In vitro fest of mineralized collagen-based composite as guided tissue regeneration membrane

    Institute of Scientific and Technical Information of China (English)

    潘韶霞; 冯海兰


    目的:研究一种新型可吸收性引导组织再生(Guided Tissue Regeneration,GTR)膜的机械力学性能,以及其体外降解过程.方法:通过在聚乳酸(Polylactide,PLA)中加入矿化胶原复合物(nano-hydroxyapatite/collagen,nHAC)获得复合GTR膜,矿化胶原中的无机物为纳米相的羟基磷灰石.测量不同nHAC含量的复合膜的弹性模量和最大拉伸强度,通过热失重分析评估模拟体液浸泡后样本无机相的变化,并通过扫描电镜观察样本表面形态的变化.结果:当nHAC/PLA比例为0.4:1,各项力学性能指标达到最高值.复合膜表面可以观察到比较活跃的晶体溶解和再沉积过程.结论:在本实验条件下,复合GTR膜nHAC/PLA比例为0.4:1力学性能最佳.体外情况下复合膜表面有较活跃的晶体溶解和再沉积过程.

  9. Modern Collagen Wound Dressings: Function and Purpose


    Fleck, Cynthia Ann; Simman, Richard


    Collagen, which is produced by fibroblasts, is the most abundant protein in the human body. A natural structural protein, collagen is involved in all 3 phases of the wound-healing cascade. It stimulates cellular migration and contributes to new tissue development. Because of their chemotactic properties on wound fibroblasts, collagen dressings encourage the deposition and organization of newly formed collagen, creating an environment that fosters healing. Collagen-based biomaterials stimulate...

  10. MT1-MMP and type II collagen specify skeletal stem cells and their bone and cartilage progeny

    DEFF Research Database (Denmark)

    Szabova, Ludmila; Yamada, Susan S; Wimer, Helen;


    Skeletal formation is dependent on timely recruitment of skeletal stem cells and their ensuing synthesis and remodeling of the major fibrillar collagens, type I collagen and type II collagen, in bone and cartilage tissues during development and postnatal growth. Loss of the major collagenolytic...... activity associated with the membrane-type 1 matrix metalloproteinase (MT1-MMP) results in disrupted skeletal development and growth in both cartilage and bone, where MT1-MMP is required for pericellular collagen dissolution. We show here that reconstitution of MT1-MMP activity in the type II collagen......-expressing cells of the skeleton rescues not only diminished chondrocyte proliferation, but surprisingly, also results in amelioration of the severe skeletal dysplasia associated with MT1-MMP deficiency through enhanced bone formation. Consistent with this increased bone formation, type II collagen was identified...

  11. Human Amnion Membrane Serves as a Substratum for Growing Axons in vitro and in vivo (United States)

    Davis, George E.; Blaker, Scott N.; Engvall, Eva; Varon, Silvio; Manthorpe, Marston; Gage, Fred H.


    The epithelial cell layer of human amnion membrane can be removed while the basement membrane and stromal surfaces remain morphologically intact. Such a preparation has been used as a substratum for the in vitro culture of dissociated neurons. Embryonic motor neurons from chick ciliary ganglion attached to both surfaces but grew extensive neurites only on the basement membrane. On cross sections of rolled amnion membranes, regenerating axons of cultured neurons were guided along pathways of basement membrane that were immunoreactive with an antibody to laminin. In addition, when rolled amnion membranes were implanted into a lesion cavity between the rat septum and hippocampus, cholinergic neurons extended axons through the longitudinally oriented implant into the hippocampus. Thus, this amnion preparation can serve as a bridge to promote axonal regeneration in vivo in damaged adult brain.

  12. Increasing extracellular matrix collagen level and MMP activity induces cyst development in polycystic kidney disease

    Directory of Open Access Journals (Sweden)

    Liu Bin


    Full Text Available Abstract Background Polycystic Kidney Disease (PKD kidneys exhibit increased extracellular matrix (ECM collagen expression and metalloproteinases (MMPs activity. We investigated the role of these increases on cystic disease progression in PKD kidneys. Methods We examined the role of type I collagen (collagen I and membrane bound type 1 MMP (MT1-MMP on cyst development using both in vitro 3 dimensional (3D collagen gel culture and in vivo PCK rat model of PKD. Results We found that collagen concentration is critical in controlling the morphogenesis of MDCK cells cultured in 3D gels. MDCK cells did not form 3D structures at collagen I concentrations lower than 1 mg/ml but began forming tubules when the concentration reaches 1 mg/ml. Significantly, these cells began to form cyst when collagen I concentration reached to 1.2 mg/ml, and the ratios of cyst to tubule structures increased as the collagen I concentration increased. These cells exclusively formed cyst structures at a collagen I concentration of 1.8 mg/ml or higher. Overexpression of MT1-MMP in MDCK cells significantly induced cyst growth in 3D collagen gel culture. Conversely, inhibition of MMPs activity with doxycycline, a FDA approved pan-MMPs inhibitor, dramatically slowed cyst growth. More importantly, the treatment of PCK rats with doxycycline significantly decreased renal tubule cell proliferation and markedly inhibited the cystic disease progression. Conclusions Our data suggest that increased collagen expression and MMP activity in PKD kidneys may induce cyst formation and expansion. Our findings also suggest that MMPs may serve as a therapeutic target for the treatment of human PKD.

  13. A simple dot-blot-Sirius red-based assay for collagen quantification. (United States)

    Rodríguez-Rodríguez, Pilar; Arribas, Silvia M; de Pablo, Angel Luis López; González, M Carmen; Abderrahim, Fatima; Condezo-Hoyos, Luis


    The assessment of collagen content in tissues is important in biomedical research, since this protein is altered in numerous diseases. Hydroxyproline and Sirius red based assays are the most common methods for collagen quantification. However, these procedures have some pitfalls, such as the requirement of oxygen-free medium or expensive equipment and large sample size or being unsuitable for hydrolyzed collagen, respectively. Our objective was to develop a specific, versatile, and user-friendly quantitative method applicable to small tissue samples and extracts obtained from elastin purification, therefore, suitable for simultaneous quantification of elastin. This method is based on the binding of Sirius red to collagen present in a sample immobilized on a PVDF membrane, as in the dot-blot technique, and quantified by a scanner and image analysis software. Sample loading, Sirius red concentration, temperature and incubation time, type of standard substance, albumin interference, and quantification time are optimized. The method enabled the quantification of (1) intact collagen in several rat tissue homogenates, including small resistance-sized arteries, (2) partially hydrolyzed collagen obtained from NaOH extracts, compatible with elastin purification, and (3) for the detection of differences in collagen content between hypertensive and normotensive rats. We conclude that the developed technique can be widely used since it is versatile (quantifies intact and hydrolyzed collagen), requires small sample volumes, is user-friendly (low-cost, easy to use, minimum toxic materials, and reduced time of test), and is specific (minimal interference with serum albumin).

  14. Platelet-rich fibrinversus collagen membrane in the repair of gingival defects%富血小板纤维蛋白与胶原膜修复牙龈缺损创面的能力

    Institute of Scientific and Technical Information of China (English)

    董露; 肖琼; 杨琴秋; 孙勇; 陈红亮


    背景:前期实验中发现富血小板纤维蛋白具有良好的诱导牙龈软组织修复再生的能力。  目的:观察富血小板纤维蛋白膜与胶原膜对牙龈软组织愈合的影响,评价富血小板纤维蛋白膜诱导牙龈软组织缺损修复再生的能力。  方法:选因各种原因需要拔除前磨牙或磨牙同期进行位点保存的患者22例(前磨牙2颗,磨牙20颗),随机分为2组,22个软组织缺损区域,拔牙窝内植入Bio-Oss ,表面覆盖富血小板纤维蛋白膜或海奥胶原膜。通过追踪植入Bio-Oss后牙龈创面的愈合时间和植入Bio-Oss后1,2周的创面愈合率,评价富血小板纤维蛋白促进牙龈组织愈合的能力。  结果与结论:①富血小板纤维蛋白组愈合时间为(12.17±2.25) d,胶原膜组愈合时间为(17.30±2.58) d;植入Bio-Oss后1,2周富血小板纤维蛋白组愈合率明显高于胶原膜组。②结果说明,相同时间节点时富血小板纤维蛋白促进牙龈软组织的愈合能力明显优于胶原膜,达到相同愈合状态时富血小板纤维蛋白组所需时间明显短于胶原膜组。%BACKGROUND:Previous studies have found that platelet-rich fibrin has a good ability to induce gingival soft tissue repair and regeneration. OBJECTIVE:To observe the effects of platelet-rich fibrinversus colagen membrane on gingival soft tissue healing, and to evaluate the ability of platelet-rich fibrin to repair gingival defects. METHODS:Twenty-two patients (2 premolar teeth and 20 molars) scheduled for premolar or molar removal or ridge preservation due to various reasons were selected and randomized into two groups. Bio-Oss was implanted into the extraction socket folowed by covering with platelet-rich fibrin or colagen membrane. Healing time and healing rate of gingival defects were detected to evaluate the ability of platelet-rich fibrin to promote gingival tissue healing at 1-2 weeks after Bio-Oss implantation

  15. 汽油机尾气致大鼠慢性肺损伤及对不同类型胶原蛋白表达的影响%Expressions of Collagen Proteins in Lungs of Rats Exposed to Exhaust of Gasoline Engine

    Institute of Scientific and Technical Information of China (English)

    王晔; 程薇波; 张诗珩; 粟小恬; 罗建


    expressed in the basement membrane of rat alveolar epithelial cells and vascular endothelial cells, its positive area and its intensity decreasing with the extending of exposure time. However, around the peribronchial and perivascular, the positive area and intensity of collagen IV increased with the exposure time extending. Conclusion During the process of lung damage induced by gasoline engine exhaust, the expressions of collagen I , collagen III and collagen IV protein can be affected which may be involved in the damage.

  16. NID1 — EDRN Public Portal (United States)

    NID1, a sulfated glycoprotein widely distributed in basement membranes and tightly associated with laminin, is a member of the nidogen family of basement membrane glycoproteins. NID1 interacts with several other components of basement membranes, like collagen IV and perlecan, and may play a role in cell interactions with the extracellular matrix.

  17. Basement faults and volcanic rock distributions in the Ordos Basin

    Institute of Scientific and Technical Information of China (English)


    Volcanic rocks in the Ordos Basin are of mainly two types: one in the basin and the other along the margin of the basin. Besides those along the margin, the marginal volcanic rocks also include the volcanic rocks in the Yinshanian orogenic belt north of the basin. Based on the latest collection of gravitational and aeromagnetic data, here we interpret basement faults in the Ordos Basin and its peripheral region, compare the faults derived from aeromagnetic data with those from seismic data, and identify the geological ages of the fault development. Two aeromagnetic anomaly zones exist in the NE-trending faults of the southern basin, and they are in the volcanic basement formed in pre-Paleozoic. These NE-trending faults are the channel of volcanic material upwelling in the early age (Archean-Neoproterozoic), where igneous rocks and sedimentary rocks stack successively on both sides of the continental nucleus. In the Cambrian, the basin interior is relatively stable, but in the Late Paleozoic and Mesozoic, the basin margin underwent a number of volcanic activities, accompanied by the formation of nearly north-south and east-west basement faults in the basin periphery and resulting in accumulation of great amount of volcanic materials. Volcanic tuff from the basin periphery is discovered in the central basin and volcanic materials are exposed in the margins of the basin. According to the source-reservoir-cap rock configuration, the basin peripheral igneous traps formed in the Indosinian-Early Yanshanian and Late Hercynian are favorable exploration objectives, and the volcanic rocks in the central basin are the future target of exploration.

  18. Formation and Evolution of the Junggar basin basement (United States)

    He, D.


    Junggar Basin is located in the central part of the Central Asian Orogenic Belt (CAOB). Its basement nature is a highly controversial scientific topic, involving the basic style and processes of crustal growth.Based on the borehole data from over 300 wells drilled into the Carboniferous System, together with the high-resolution gravity and magnetic data (in a 1:50,000 scale), we made a detailed analysis of the basement structure, formation timing and process and later evolution on basis of core geochemical and isotopic analysis. Firstly, we defined the Mahu Precambrian micro-continental block in the juvenile crust of Junggar Basin according to the Hf isotopic analysis of the Carboniferous volcanic rocks. Secondly, the results of the tectonic setting and basin analysis suggest that the Junggar area incorporates three approximately E-W trending island arc belts (from north to south: Yemaquan-Wulungu-Chingiz, Jiangjunmiao-Luliang-Darbut and Zhongguai-Mosuowan-Baijiahai-Qitai respectively) and intervened three approximately E-W trending retro-arc or inter-arc basin belts from north to south, such as Santanghu-Suosuoquan-Emin, Wucaiwan-Dongdaohaizi-Mahu (Mahu block sunk as a bathyal basin during this phase) and Fukang-western well Pen1 accordingly. Thirdly, the closure of these retro-arc or inter-arc basins gradually toward the south led to the occurrence of collision and amalgamation of the above-mentioned island arcs during the Carboniferous, constituting the basic framework of the Junggar "block". Fourthly, the emplacement of large-scale mantle-derived magmas occurred in the latest Carboniferous or Early Permian. For instance, the well Mahu 5 penetrate the latest Carboniferous basalts with a thickness of over 20m, and these mantle-derived magmas concreted the above-mentioned island arc-collaged body. Therefore, the Junggar basin basement mainly comprises pre-Carboniferous collaged basement, and its formation is characterized by two-stage growth model, involving the

  19. Comparative study on the clinical effects of two collagen barrier membranes in guided bone regeneration in dental implant%两种胶原膜在牙种植中引导骨再生的对照研究

    Institute of Scientific and Technical Information of China (English)

    石艳; 严宁; 何维兴


    目的:比较在牙种植引导骨再生术中,海奥口腔修复膜和Bio-Gide胶原膜的临床修复效果。方法:选取单颗牙缺失的患者82例,行引导骨再生手术修复种植区骨缺损并同期植入种植体82枚,随机分为实验组和对照组,每组41例。两组患者均采用天博骨粉作骨移植物,实验组采用海奥口腔修复膜行引导骨再生;对照组采用Bio-Gide胶原膜行引导骨再生。观察二期手术时植骨区外形及牙龈状况,比较两组的骨再生效果及不良反应发生率;修复后随访观察1年,比较两组的修复成功率。结果:82枚种植体均与骨组织形成良好的骨结合,骨再生效果及不良反应发生率实验组与对照组相当,差异无统计学意义(P>0.05),种植修复成功率均为100%。修复后随访1年,种植体均成功负载。结论:采用海奥口腔修复膜和Bio-Gide胶原膜均能取得满意的骨再生效果,但采用海奥口腔修复膜更为经济,值得临床推广。%ObjectiveThe aim of this study was to compare clinical effects of Heal-all oral bioiflm and Bio-Gide membrane on guided bone regeneration in dental implant.Methods Eighty-two implants were placed in Eighty-two single tooth missing patients with bone defects in the implantation area and simultaneous guided bone regeneration. All the subjects were randomly divided into experimental group and the control group (n = 41). Bone defects around implants were repaired by guided bone regeneration technique with Heal-all oral bioiflm and Bio-Gide membrane respectively. To observe the shape and gingival status of bone defects when stage II operation was performed,the amount of new-formed bone tissue and adverse reaction rate of the two groups was compared. After 1 year follow-up, the success rate of the two groups was compared. Results Osseointegration was formed well between implants and bone tissue in all Eighty-two patients. The difference of bone

  20. Ordovician Basement Hydrocarbon Reservoirs in the Tarim Basin, China

    Institute of Scientific and Technical Information of China (English)

    YAN Xiangbin; LI Tiejun; ZHANG Tao


    Ordovician marine carbonate basement traps are widely developed in the paleo-highs and paleo-slopes in the Tarim Basin. Reservoirs are mainly altered pore-cavity-fissure reservoirs. Oil sources are marine carbonate rocks of the Lower Paleozoic. Thus, the paleo-highs and paleo-slopes have good reservoiring conditions and they are the main areas to explore giant and large-scale oil reservoirs. The main factors for their reservoiring are: (1) Effective combination of fenestral pore-cavity-fracture reservoirs, resulting from multi-stage, multi-cyclic karstification (paleo-hypergene and deep buried) and fracturing, with effective overlying seals, especially mudstone and gypsum mudstone in the Carboniferous Bachu Formation, is essential to hydrocarbon reservoiring and high and stable production; (2) Long-term inherited large rises and multi-stage fracture systems confine the development range of karst reservoirs and control hydrocarbon migration, accumulation and reservoiring; (3) Long-term multi-source hydrocarbon supply, early reservoiring alteration and late charging adjustment are important reservoiring mechanisms and determine the resource structure and oil and gas properties. Favorable areas for exploration of Ordovician carbonate basement hydrocarbon reservoirs in the Tarim Basin are the Akekule rise, Katahe uplift, Hetianhe paleo-high and Yakela faulted rise.

  1. Provenance of Neoproterozoic sedimentary basement of northern Iran, Kahar Formation (United States)

    Etemad-Saeed, Najmeh; Hosseini-Barzi, Mahboubeh; Adabi, Mohammad Hossein; Sadeghi, Abbas; Houshmandzadeh, Abdolrahim


    This article presents new data to understand the nature of the hidden crystalline basement of northern Iran and the tectonic setting of Iran during late Neoproterozoic time. The siliciclastic-dominated Kahar Formation represents the oldest known exposures of northern Iran and comprises late Ediacaran (ca. 560-550 Ma) compositionally immature sediments including mudrocks, sandstones, and conglomerates. This work focuses on provenance of three well preserved outcrops of this formation in Alborz Mountains: Kahar Mountain, Sarbandan, and Chalus Road, through petrographic and geochemical methods. Mineralogical Index of Alteration (MIA) and Chemical Index of Alteration (CIA-after correction for K-metasomatism) values combined with A-CN-K relations suggest moderate weathering in the source areas. The polymictic nature of Kahar conglomerates indicates a mixed provenance for them. However, modal analysis of Kahar sandstones (volcanic to plagioclase-rich lithic arkose) and whole rock geochemistry of mudrocks suggest that they are largely first-cycle sediments and that their sources were remarkably late Ediacaran, intermediate-felsic igneous rocks from proximal arc settings. Tectonic setting discrimination diagrams also indicate a convergent plate margin and continental arc related basin for Kahar sediments. This interpretation is supported by the phyllo-tectic to tectic composition and geochemistry of mudrocks. These results reveal the presence of a felsic/intermediate subduction-related basement