Showing papers on "Keratan sulfate published in 2010"
TL;DR: The growth factor mediated synthesis ofSeveral different collagen types and the core proteins of several different leucine-rich type proteoglycans are required to produce collagen fibrils with the size and spacing needed for corneal stromal transparency.
444 citations
Journal Article•
TL;DR: In this paper, the authors found that ADSC contained a side population and exhibited differentiation to adipocytes and chondrocytes indicating adult stem-cell potential, and showed that these unique keratocyte products can adopt a Keratocyte phenotype and therefore have potential for use in corneal cell therapy and tissue engineering.
Abstract: Purpose: Adipose-derived stem cells (ADSC) are an abundant population of adult stem cells with the potential to differentiate into several specialized tissue types, including neural and neural crest-derived cells. This study sought to determine if ADSC express keratocyte-specific phenotypic markers when cultured under conditions inducing differentiation of corneal stromal stem cells to keratocytes. Methods: Human subcutaneous adipose tissue was obtained by lipoaspiration. ADSC were isolated by collagenase digestion and differential centrifugation. Side population cells in ADSC were demonstrated using fluorescence-activated cell sorting after staining with Hoechst 33342. Differentiation to keratocyte phenotype was induced in fibrin gels or as pellet cultures with serum-free or reduced-serum media containing ascorbate. Keratocyte-specific gene expression was characterized using western blotting, quantitative RT–PCR, and immunostaining. Results: ADSC contained a side population and exhibited differentiation to adipocytes and chondrocytes indicating adult stem-cell potential. Culture of ADSC in fibrin gels or as pellets in reduced-serum medium with ascorbate and insulin induced expression of keratocan, keratan sulfate, and aldehyde dehydrogenase 3 family, member A1 (ALDH3A1), products highly expressed by differentiated keratocytes. Expression of differentiation markers was quantitatively similar to corneal stromal stem cells and occurred in both serum-free and serum containing media. Conclusions: ADSC cultured under keratocyte-differentiation conditions express corneal-specific matrix components. Expression of these unique keratocyte products suggests that ADSC can adopt a keratocyte phenotype and therefore have potential for use in corneal cell therapy and tissue engineering. The cornea is the outermost tissue of the eye, providing a protective barrier and a clear path for transmission and refraction of light. This organ comprises three distinct cellular layers: epithelium, stroma, and endothelium. The stroma is primarily responsible for the strength and the refractive properties of the cornea [1]. The optical properties of the stroma result from layers of parallel aligned heterotypic collagen fibrils composed of Types I and V collagen. This collagen is associated with several proteoglycans from the small leucine-rich (SLRP) family, including decorin, lumican, keratocan, and mimecan (osteoglycin) [2-4]. The latter three of these proteins are modified with long, highly sulfated keratan sulfate glycosaminoglycan comprising the corneal keratan sulfate proteoglycans (KSPG), a family of molecules unique to corneal stromal extracellular matrix (ECM). After embryonic development, corneal collagen synthesis decreases, but synthesis of the KSPGs is maintained at a high level, suggesting an essential role for these ECM components in homeostasis of stromal transparency. This role has been confirmed by the loss of transparency associated with reduced sulfated keratan sulfate [5,6] or the loss of keratan sulfate core protein, lumican [7,8]. During stromal wound healing,
119 citations
TL;DR: Data suggest that FGF2 and -18 bind to discrete structures on the HS chains attached to chondrocyte-derived perlecan which modulate the growth factor activities, which may be important in the turnover of HS and subsequent signaling required for the establishment and maintenance of functional osteo-chondral junctions in long bone growth.
Abstract: Perlecan is a large multidomain proteoglycan that is essential for normal cartilage development. In this study, perlecan was localized in the pericellular matrix of hypertrophic chondrocytes in developing human cartilage rudiments. Perlecan immunopurified from medium conditioned by cultured human fetal chondrocytes was found to be substituted with heparan sulfate (HS), chondroitin sulfate (CS), and keratan sulfate (KS). Ligand and carbohydrate engagement (LACE) assays demonstrated that immunopurified chondrocyte-derived perlecan formed HS-dependent ternary complexes with fibroblast growth factor (FGF) 2 and either FGF receptors (FGFRs) 1 or 3; however, these complexes were not biologically active in the BaF32 cell system. Chondrocyte-derived perlecan also formed HS-dependent ternary complexes with FGF18 and FGFR3. The proliferation of BaF32 cells expressing FGFR3 was promoted by chondrocyte-derived perlecan in the presence of FGF18, and this activity was reduced by digestion of the HS with either heparinase III or mammalian heparanase. These data suggest that FGF2 and -18 bind to discrete structures on the HS chains attached to chondrocyte-derived perlecan which modulate the growth factor activities. The presence and activity of mammalian heparanase may be important in the turnover of HS and subsequent signaling required for the establishment and maintenance of functional osteo-chondral junctions in long bone growth.
91 citations
TL;DR: Observations provide strong evidence that Langerin mediates diverse functions on Langerhans cells through dual recognition of sulfated as well as mannosylated glycans by its uniquely evolved C-type carbohydrate-recognition domain.
82 citations
TL;DR: The results of this research support the effort to understand the role of GLN and CS in preserving articular cartilage in athletic horses and inhibit the synthesis of several mediators of cartilage degradation.
Abstract: Glucosamine and chondroitin sulphate in many animal and human trials has improved joint health. In vitro studies are beginning to clarify their mode of action. The objective of this research was to: 1) determine at what concentrations glucosamine-HCl (GLN) and/or chondroitin sulphate (CS) would inhibit the cytokine-induced catabolic response in equine articular cartilage explants and 2) to determine if a combination of the 2 was more effective at inhibiting the catabolic response than the individual compounds. Articular cartilage was obtained from carpal joints of horses (age 1-4 years). Cartilage discs (3.5 mm) were biopsied and cultured. Explants were incubated with lipopolysaccharide (LPS) in the presence of varying concentrations of GLN, CS, or both. Control treatments included explants with no LPS and LPS without GLN or CS. Media were analysed for nitric oxide (NO), prostaglandin E2 (PGE2) and keratan sulphate. Cartilage was extracted for analysis of metalloproteinases (MMP). Four experiments were conducted. In all experiments, GLN at concentrations as low as 1 mg/ml decreased NO production relative to LPS stimulated cartilage without GLN over the 4 day period. In general, CS at either 0.25 or 0.5 mg/ml did not inhibit NO production. The addition of CS to GLN containing media did not further inhibit NO production. GLN at concentrations as low as 0.5 mg/ml decreased PGE2 production, whereas CS did not effect on PGE2. The combination of GLN/CS decreased MMP-9 gelatinolytic activity but had no effect on MMP-2 activity. The combination in 2 experiments tended to decrease MMP-13 protein concentrations and decreased keratan sulphate levels in media. Overall, the combination of GLN (1 mg/ml) and CS (0.25 mg/ml) inhibited the synthesis of several mediators of cartilage degradation. These results further support the effort to understand the role of GLN and CS in preserving articular cartilage in athletic horses.
77 citations
TL;DR: In vitro assays using primary cerebellar granule neurons demonstrated that keratan sulfate proteoglycans were required for the proteoglycan-mediated inhibition of neurite outgrowth, and data collectively indicate that keraten sulfate expression is closely associated with functional disturbance after spinal cord injury.
Abstract: Neurons in the adult CNS do not spontaneously regenerate after injuries. The glycosaminoglycan keratan sulfate is induced after spinal cord injury, but its biological significance is not well understood. Here we investigated the role of keratan sulfate in functional recovery after spinal cord injury, using mice deficient in N-acetylglucosamine 6-O-sulfotransferase-1 that lack 5D4-reactive keratan sulfate in the CNS. We made contusion injuries at the 10th thoracic level. Expressions of N-acetylglucosamine 6-O-sulfotransferase-1 and keratan sulfate were induced after injury in wild-type mice, but not in the deficient mice. The wild-type and deficient mice showed similar degrees of chondroitin sulfate induction and of CD11b-positive inflammatory cell recruitment. However, motor function recovery, as assessed by the footfall test, footprint test, and Basso mouse scale locomotor scoring, was significantly better in the deficient mice. Moreover, the deficient mice showed a restoration of neuromuscular system function below the lesion after electrical stimulation at the occipito-cervical area. In addition, axonal regrowth of both the corticospinal and raphespinal tracts was promoted in the deficient mice. In vitro assays using primary cerebellar granule neurons demonstrated that keratan sulfate proteoglycans were required for the proteoglycan-mediated inhibition of neurite outgrowth. These data collectively indicate that keratan sulfate expression is closely associated with functional disturbance after spinal cord injury. N-acetylglucosamine 6-O-sulfotransferase-1-deficient mice are a good model to investigate the roles of keratan sulfate in the CNS.
68 citations
TL;DR: Keratan sulphate is the predominant glycosaminoglycan in the cornea of the eye, where it exists in proteoglycan (PG) form, and mutations in enzymes responsible for the sulphation of KS-GAG chains are causative for the inherited human disease, macular corneal dystrophy.
Abstract: Keratan sulphate (KS) is the predominant glycosaminoglycan (GAG) in the cornea of the eye, where it exists in proteoglycan (PG) form. KS-PGs have long been thought to play a pivotal role in the establishment and maintenance of the array of regularly-spaced and uniformly-thin collagen fibrils which make up the corneal stroma. This characteristic arrangement of fibrils allows light to pass through the cornea. Indeed, perturbations to the synthesis of KS-PG core proteins in genetically altered mice lead to structural matrix alterations and corneal opacification. Similarly, mutations in enzymes responsible for the sulphation of KS-GAG chains are causative for the inherited human disease, macular corneal dystrophy, which is manifested clinically by progressive corneal cloudiness starting in young adulthood.
65 citations
TL;DR: The role of lumican is addressed in the process of extravasation of polymorphonuclear leukocytes (PMNs) during the early inflammatory phase present in the healing of the corneal epithelium following debridement, consistent with the notion that in addition to regulating the collagen fibril architecture, lumican acts to aid neutrophil recruitment and invasion following corneAL damage and inflammation.
Abstract: An important aspect of wound healing is the recruitment of neutrophils to the site of infection or tissue injury Lumican, an extracellular matrix component belonging to the small leucine rich proteoglycan (SLRP) family, is one of the major keratan sulfate proteoglycans (KSPGs) within the corneal stroma Increasing evidence indicates that lumican can serve as a regulatory molecule for several cellular processes, including cell proliferation and migration In the present study, we addressed the role of lumican in the process of extravasation of polymorphonuclear leukocytes (PMNs) during the early inflammatory phase present in the healing of the corneal epithelium following debridement We used Lum−/− mice and a novel transgenic mouse, Lum−/−,Kera-Lum, which expresses lumican only in the corneal stroma, to assess the role of lumican in PMN extravasation into injured corneas Our results showed that PMNs did not readily invade injured corneas of Lum−/− mice and this defect was rescued by the expression of lumican in the corneas of Lum−/−,Kera-Lum mice The presence of lumican in situ facilitates PMN infiltration into the peritoneal cavity in casein-induced inflammation Our findings are consistent with the notion that in addition to regulating the collagen fibril architecture, lumican acts to aid neutrophil recruitment and invasion following corneal damage and inflammation
63 citations
TL;DR: It is shown that enzyme replacement therapy with recombinant human GALNS results in clearance of keratan sulfate accumulation, and that such treatment ameliorates aberrant gene expression in human chondrocytes in vitro.
Abstract: Mucopolysaccharidosis IVA (MPS IVA; Morquio A syndrome) is a lysosomal storage disorder caused by deficiency of N-acetylgalactosamine-6-sulfatase (GALNS), an enzyme that degrades keratan sulfate (KS). Currently no therapy for MPS IVA is available. We produced recombinant human (rh)GALNS as a potential enzyme replacement therapy for MPS IVA. Chinese hamster ovary cells stably overexpressing GALNS and sulfatase modifying factor-1 were used to produce active ( approximately 2 U/mg) and pure (>or=97%) rhGALNS. The recombinant enzyme was phosphorylated and was dose-dependently taken up by mannose-6-phosphate receptor (K(uptake) = 2.5 nM), thereby restoring enzyme activity in MPS IVA fibroblasts. In the absence of an animal model with a skeletal phenotype, we established chondrocytes isolated from two MPS IVA patients as a disease model in vitro. MPS IVA chondrocyte GALNS activity was not detectable and the cells exhibited KS storage up to 11-fold higher than unaffected chondrocytes. MPS IVA chondrocytes internalized rhGALNS into lysosomes, resulting in normalization of enzyme activity and decrease in KS storage. rhGALNS treatment also modulated gene expression, increasing expression of chondrogenic genes Collagen II, Collagen X, Aggrecan and Sox9 and decreasing abnormal expression of Collagen I. Intravenous administration of rhGALNS resulted in biodistribution throughout all layers of the heart valve and the entire thickness of the growth plate in wild-type mice. We show that enzyme replacement therapy with recombinant human GALNS results in clearance of keratan sulfate accumulation, and that such treatment ameliorates aberrant gene expression in human chondrocytes in vitro. Penetration of the therapeutic enzyme throughout poorly vascularized, but clinically relevant tissues, including growth plate cartilage and heart valve, as well as macrophages and hepatocytes in wild-type mouse, further supports development of rhGALNS as enzyme replacement therapy for MPS IVA.
61 citations
TL;DR: Corneal stromal GAGs bind, and thus could alter the availability or conformation of, many proteins that may influence keratocyte and nerve growth cone behavior in the cornea.
Abstract: PURPOSE. Corneal stroma extracellular matrix (ECM) glycosaminoglycans (GAGs) include keratan sulfate (KS), chondroitin sulfate A (CSA), and hyaluronic acid (HA). Embryonic corneal keratocytes and sensory nerve fibers grow and differentiate according to chemical cues they receive from the ECM. This study asked which of the proteins that may regulate keratocytes or corneal nerve growth cone immigration interact with corneal GAGs. METHODS. Biotinylated KS (bKS), CSA (bCSA), and HA (bHA) were prepared and used in microarray protocols to assess their interactions with 8268 proteins and a custom microarray of 85 extracellular epitopes of nerve growth-related proteins. Surface plasmon resonance (SPR) was performed with bKS and SLIT2, and their ka, kd, and KD were determined. RESULTS. Highly sulfated KS interacted with 217 microarray proteins, including 75 kinases, several membrane or secreted proteins, many cytoskeletal proteins, and many nerve function proteins. CSA interacted with 24 proteins, including 10 kinases and 2 cell surface proteins. HA interacted with 6 proteins, including several ECM-related structural proteins. Of 85 ECM nerve-related epitopes, KS bound 40 proteins, including SLIT, 2 ROBOs, 9 EPHs, 8 Ephrins (EFNs), 8 semaphorins (SEMAs), and 2 nerve growth factor receptors. CSA bound nine proteins, including ROBO2, 2 EPHs, 1 EFN, two SEMAs, and netrin 4. HA bound no ECM nerve-related epitopes. SPR confirmed that KS binds SLIT2 strongly. The KS core protein mimecan/osteoglycin bound 15 proteins. CONCLUSIONS. Corneal stromal GAGs bind, and thus could alter the availability or conformation of, many proteins that may influence keratocyte and nerve growth cone behavior in the cornea. (Invest Ophthalmol Vis Sci. 2010;51:4500‐4515) DOI:
58 citations
TL;DR: This study demonstrates the presence of immunocompetent cells in tissue overlying the cartilage surface and adjacent to bone in normal human joints and suggests that cells in the periosteum, rather than those in the marginal synovium, may be involved in attempted "repair" mechanisms, such as osteophyte formation.
Abstract: The immunohistology of the synovium-cartilage junction was studied in 8 normal human knees, using monoclonal antibodies. In all joints at the junction with synovium, a vascular, wedge-shaped tongue of tissue was found to cover the cartilage surface. This marginal tissue overlying cartilage was in continuity with and was immunohistochemically similar to the adjacent synovial tissue, and contained cells possessing class II HLA antigens and antigens present on macrophages and type B synoviocytes. Periosteal tissue adjacent to the synovium-cartilage junction contained not only macrophages and other class II-positive cells, but also cells and matrix that stained with monoclonal antibodies specific for articular cartilage (keratan sulfate and type II collagen). This study demonstrates the presence of immunocompetent cells in tissue overlying the cartilage surface and adjacent to bone in normal human joints. It is likely that pannus in chronic inflammatory rheumatic disease develops by the recruitment of inflammatory cells augmenting this normal marginal tissue. Furthermore, overgrowth of tissue onto the cartilage surface may not be necessary in the pathogenesis of joint destruction in rheumatoid arthritis. In addition, our findings suggest that cells in the periosteum, rather than those in the marginal synovium, may be involved in attempted "repair" mechanisms, such as osteophyte formation.
TL;DR: It is demonstrated that a novel iminosugar, DLHex-DGJ, has potent effects as competitive inhibitor of human acid beta-galactosidase in vitro, and its effects on activity, protein expression, maturation and intracellular transport in vivo in 13 fibroblasts lines with GLB1 mutations are described.
TL;DR: Data indicate that fibromodulin plays important roles in the regulation of region‐specific fibrillogenesis required for the integration of the corneal and scleral matrices and sulcus development required for establishment of the visual axis.
Abstract: Fibromodulin regulates collagen fibrillogenesis, but its existence/role(s) in the cornea is controversial. We hypothesize that fibromodulin regulates fibrillogenesis during postnatal development of the anterior eye. Fibromodulin is weakly expressed in the limbus at post-natal day (P) 4, increases and extends into the central cornea at P14, becomes restricted to the limbus at P30, and decreases at P60. This differential spatial and temporal expression of fibromodulin is coordinated with emmetropization; the developmental increase in axial length and globe size. Genetic analysis demonstrated that fibromodulin regulates fibrillogenesis in a region-specific manner. At the limbus, fibromodulin is dominant in regulating fibril growth during postnatal development. In the posterior peripheral cornea, cooperative interactions of fibromodulin and lumican regulate fibrillogenesis. These data indicate that fibromodulin plays important roles in the regulation of region-specific fibrillogenesis required for the integration of the corneal and scleral matrices and sulcus development required for establishment of the visual axis.
TL;DR: Zebrafish is chosen as an animal model to elucidate the role of lumican in the development of axial myopia and the observation suggests that zebrafish can be used as an in vivo model for screening compounds in treating myopia.
01 Jan 2010
TL;DR: The structure analysis of GAGs is provided based on highly efficient isolation/purification techniques, and high-sensitivity, information-rich analytical instruments to improve the understanding of their biological functions and help in the development of structure–activity relationships for these important biopolymers.
Abstract: Publisher Summary
Glycosaminoglycans (GAGs) are a family of highly sulfated, complex, polydisperse linear polysaccharides that display a variety of important biological roles. Based on the difference of repeating disaccharide units comprising GAGs, they can be categorized into four main groups: heparin/heparan sulfate, chondroitin sulfate/dermatan sulfate, keratan sulfate, and hyaluronan. This chapter provides the structure analysis of GAGs based on highly efficient isolation/purification techniques, and high-sensitivity, information-rich analytical instruments. GAGs perform a variety of biological functions and play an important role in a number of different diseases. Their activities are mainly triggered by interactions with a wide range of proteins. The structure analysis of GAGs improves the understanding of their biological functions and helps in the development of structure–activity relationships for these important biopolymers. With modern mass spectrometry (MS) techniques, structural information, such as molecular weight, monosaccharide composition, number and position of sulfo groups, composition of disaccharide blocks, and sequence of highly charged sulfated carbohydrates can be obtained. High sensitivity of MS is available for the microanalysis of carbohydrates derived from biological samples. Tandem MS has the potential to become a robust method to completely sequence complex oligosaccharides. Mechanistic studies of oligosaccharide behavior in MS and tandem MS provide the basis for future methods development.
TL;DR: Analysis of KS by means of HPLC showed that in vitro cultured stromal keratocytes express and secrete more KS than keratinocytes, thus mirroring results observed in vivo, and expression of the CHST6 gene and of KS proteoglycans such as mimecan, lumican is limited toStromal Keratocytes.
TL;DR: Findings indicate that the inflammatory response in the cornea is regulated by proteoglycan/CXCL1 complexes, and their diffusion into the anterior chamber is consistent with release of a chemokine gradient and resolution of inflammation.
Abstract: Keratocan and lumican are small, leucine-rich repeat KSPGs in the extracellular matrix (ECM) of the mammalian cornea, whose primary role is to maintain corneal transparency. In the current study, we examined the role of these proteoglycans in the breakdown of the chemokine gradient and resolution of corneal inflammation. LPS was injected into the corneal stroma of C57BL/6 mice, and corneal extracts were examined by immunoblot analysis. We found reduced expression of the 52-kD keratocan protein after 6 h and conversely, increased expression of 34/37 kD immunoreactive products. Further, appearance of the 34/37-kD proteins was dependent on neutrophil infiltration to the cornea, as the appearance of these products was coincident with neutrophil infiltration, and the 34/37-kD products were not detected in explanted corneas or in CXCR2 / corneas with deficient neutrophil recruitment. Furthermore, the 34/37-kD products and CXCL1/KC were detected in the anterior chamber, into which the corneal stroma drains; and CXCL1/KC was elevated significantly in keratocan / and lumican / mice. Together, these findings indicate that the inflammatory response in the cornea is regulated by proteoglycan/CXCL1 complexes, and their diffusion into the anterior chamber is consistent with release of a chemokine gradient and resolution of inflammation. J. Leukoc. Biol. 88: 517–522; 2010.
TL;DR: The results of the present study indicate that calpains are involved in the C-terminal truncation of aggrecan and might have a minor role in arthritic diseases.
Abstract: Mature aggrecan is generally C-terminally truncated at several sites in the CS (chondroitin sulfate) region. Aggrecanases and MMPs (matrix metalloproteinases) have been suggested to be responsible for this digestion. To identify whether calpain, a common intracellular protease, has a specific role in the proteolysis of aggrecan we developed neoepitope antibodies (anti-PGVA, anti-GDLS and anti-EDLS) against calpain cleavage sites and used Western blot analysis to identify calpain-generated fragments in normal and OA (osteoarthritis) knee cartilage and SF (synovial fluid) samples. Our results showed that human aggrecan contains six calpain cleavage sites: one in the IGD (interglobular domain), one in the KS (keratan sulfate) region, two in the CS1 and two in the CS2 region. Kinetic studies of calpain proteolysis against aggrecan showed that the aggrecan molecule was cleaved in a specific order where cuts in CS1 was the most preferred and cuts in KS region was the second most preferred cleavage. OA and normal cartilage contained low amounts of a calpain-generated G1-PGVA fragment (0.5-2%) compared with aggrecanase-generated G1-TEGE (71-76%) and MMP-generated G1-IPEN (23-29%) fragments. Significant amounts of calpain-generated GDLS and EDLS fragments were found in OA and normal cartilage, and a ARGS-EDLS fragment was detected in arthritic SF samples. The results of the present study indicate that calpains are involved in the C-terminal truncation of aggrecan and might have a minor role in arthritic diseases.
TL;DR: The accumulation of highly sulfated KS during the E12 to E18 timeframe could serve to fine tune local matrix hydration and collagen fibril spacing during corneal growth, as gross dehydration and compaction of the stroma progress through the action of the nascent endothelial pump.
Abstract: Purpose To investigate structural remodeling of the developing corneal stroma concomitant with changing sulfation patterns of keratan sulfate (KS) glycosaminoglycan (GAG) epitopes during embryogenesis and the onset of corneal transparency Methods Developing chick corneas were obtained from embryonic day (E)12 to E18 of incubation Extracellular matrix composition and collagen fibril spacing were evaluated by synchrotron x-ray diffraction, hydroxyproline assay, ELISA (with antibodies against lesser and more highly sulfated KS), and transmission electron microscopy with specific proteoglycan staining Results A significant relative increase in highly sulfated KS epitope labeling occurred with respect to hydroxyproline content in the final week of chick development, as mean collagen interfibrillar distance decreased Small KS PG filaments increased in frequency with development and were predominantly fibril associated Conclusions The accumulation of highly sulfated KS during the E12 to E18 timeframe could serve to fine tune local matrix hydration and collagen fibril spacing during corneal growth, as gross dehydration and compaction of the stroma progress through the action of the nascent endothelial pump
TL;DR: Data suggest that fibromodulin is one of the small PGs in the PDL-matrix and may fulfill construction and maintenance functions in this tissue.
Abstract: Although several proteoglycans (PGs) have been reported in bovine periodontal ligament (PDL), the composition of PGs in PDL has been poorly characterized. In the present study, we isolated and characterized keratan sulfate-substituted PG (fibromodulin) in bovine PDL. Fibromodulin was purified from 4 M guanidine hydrochloride (GdmCl) extracts of bovine PDL tissues using DEAE Sephacel ion-exchange chromatography and preparative electrophoresis. Fibromodulin appeared as a single polydisperse band with an apparent molecular weight (MW) of 80,000 (80 kDa) on SDS-PAGE. Digestion of fibromodulin with keratanase or neuraminidase reduced the apparent molecular size, and N-glycanase treatment produced core protein bands of around 40 kDa. Fibromodulin reacted with keratan sulfate monoclonal antibody (5D4) and fibromodulin polyclonal antibodies (alpha-FM). The keratanase-digested fibromodulin reacted with alpha-FM, but not with 5D4. These data suggest that fibromodulin is one of the small PGs in the PDL-matrix and may fulfill construction and maintenance functions in this tissue.
TL;DR: The data indicate that exogenous KS ameliorated arthritis in vivo and suppressed cartilage degradation ex vivo, suggesting that KS may have important therapeutic potential in the treatment of inflammatory arthritis.
TL;DR: Higher baseline lnCOMP and lnHA levels were associated with incident knee OA, OST, and JSN over an average follow-up period of 6.3 years, but this interpretation is incomplete due to power limitations and the potential misclassification that comes with self-report and long duration since reported injury.
TL;DR: Abbreviations used: ADAMTS, a disintegrin and metalloproteinase with thrombospondin motifs, sGAG, sulfated glycosaminoglycan, Gu-PG, guanidinium extracted proteogly can, IGD, interglobular domain, keratan sulfate region.
TL;DR: The genetic basis of diseases affecting the cornea is reviewed, includingCorneal dystrophies and systemic diseases with corneal manifestations (e.g. the mucopolysaccharidoses), and stem cell therapy as a potential therapy for Corneal injury and disease is discussed.
Abstract: The current paper reviews the genetic basis of diseases affecting the cornea, including corneal dystrophies and systemic diseases with corneal manifestations (e.g. the mucopolysaccharidoses), discusses the mechanisms by which corneal injury and disease can affect corneal transparency and discusses stem cell therapy as a potential therapy for corneal injury and disease. Several diseases of the cornea have been described, many of which are inheritable. The identification of the genetic mutations responsible for these diseases has led to a better understanding of the mechanisms underlying their corneal manifestations. Corneal disease and injury can affect corneal transparency. Stromal keratocytes play an important role in cellular and extracellular transparency of the cornea. They synthesize type I and V collagen and keratan sulfate proteoglycans that together regulate collagen fibril size and spacing important for extracellular matrix transparency. There are strong indications that cellular transparency of keratocytes relies on the presence of corneal crystallins, which provide a uniform density of proteins in the cytoplasm that increase the spatial order necessary for transparency. After injury, activated keratocytes show heterogeneity of cytoplasmic proteins, decreased spatial ordering and increased light scattering. Stem cell transplantation is regarded as a potential new therapy that can replace keratocytes and restore corneal transparency and normal structure after injury or corneal disease.
18 Jun 2010
TL;DR: The chondroitin sulfate with high purity was extracted by papain hydrolysis, isolated and purified by tricloroacetic acid, alcohol precipitation, dialysis and DEAE-Sepharose Fast Flow ion exchange column chromatography and spectral analysis indicated that ChS from pig laryngeal cartilage contains the repeated units of GlcA, which may explain the internal mechanism of its antioxidation.
Abstract: The chondroitin sulfate with high purity was extracted by papain hydrolysis, isolated and purified by tricloroacetic acid, alcohol precipitation, dialysis and DEAE-Sepharose Fast Flow ion exchange column chromatography. The structure was identified by biological enzymolysis and spectral analysis. GC data exhibited that neutral monosaccharide in chondroitin sulfate only included 0.84% of xylose and 1.28% of galactose. IR data showed that chondroitin sulfate included chondroitin-4-sulfate and chondroitin-6-sulfate isomer in a molecular chain. The results of enzyme hydrolysis and spectra analysis, including reverse phase high performance liquid chromatography, mass-spectrum and NMR, indicated that ChS from pig laryngeal cartilage contains the repeated units of GlcA(β1-3)GalNAc4SO3 and GlcA(β1-3)GalNAc6SO3. It may not have the structure characteristics of keratan sulfate and dermatan sulfate such as NeuAc (α2-3) Gal (β1-4) GlcNAc4SO3 units and GlcA2SO3 (β1-3) GalNAc4SO3, GlcA2SO3 (β1-3) GalNAc6SO3 units. This may explain the internal mechanism of its antioxidation.
Journal Article•
TL;DR: Results suggest that HSKS could detect early signs of cartilage metabolic changes and age had no significant effect on concentrations of KS measured with KS5D4 while 1 year-old horses showed significantly higher amounts than 2 year-olds with HSKS.
Abstract: This study aimed to evaluate a system that identifies cartilage turn over and/or degradation through measurement of a new keratan sulfate (KS) epitope concentration in equine sera. Blood samples were collected from 30 horses, 1 (n = 15) and 2 year-olds (n = 15). Serum samples were analyzed for an epitope of keratan sulfate by 1/20/5D4 (KS5D4) and new epitopes of keratan sulfate using high sensitive keratan sulfate (HSKS), measured by two respective enzyme-linked immunosorbant assays (ELISAs). There was no correlation in serum concentration of KS evaluated using 5D4 and HSKS. Age had no significant effect on concentrations of KS measured with KS5D4 while 1 year-old horses showed significantly higher amounts than 2 year-olds with HSKS. Results suggest that HSKS could detect early signs of cartilage metabolic changes.
TL;DR: The presence of large polysulfated proteoglycans and elevated ADAMTS-4 in laminitic tissue suggest parallels between the pathophysiology of laminitis and osteoarthritis andGene expression and product analyses indicate that the equine digital lamellae are functionally specialized to resist compressive forces.
Patent•
18 Nov 2010
TL;DR: In this paper, the problem was overcome by providing a matrix metalloprotease inhibitor or a matrix-metalloproteinase inhibiting method, an elastase activity inhibitor and a therapeutic agent for treating chronic obstructive pulmonary diseases.
Abstract: PROBLEM TO BE SOLVED: To provide a way to treat chronic obstructive pulmonary diseases etc., by providing new means related to glycosaminoglycan oligosaccharides, which inhibits matrix metalloprotease. SOLUTION: It was found that the problem was overcome by providing a matrix metalloprotease inhibitor or a matrix metalloprotease inhibiting method, an elastase activity inhibitor and a therapeutic agent for treating chronic obstructive pulmonary diseases etc., in which one, or two or more kinds selected from the group consisting of hyaluronan oligosaccharides, keratan sulfate oligosaccharides, 6-O-sulfated N-acetyl heparosan and N, 6-O-sulfated heparosan, as well as pharmaceutically acceptable salts of the glycosaminoglycan oligosaccharides are contained as effective ingredients. COPYRIGHT: (C)2011,JPO&INPIT
01 Jan 2010