heparitin-sulfate and Corneal-Diseases

Corneal transparency relies on the precise arrangement and orientation of collagen fibrils, made of mostly Type I and V collagen fibrils and proteoglycans (PGs). PGs are essential for correct collagen fibrillogenesis and maintaining corneal homeostasis. We investigated the spatial and temporal distribution of glycosaminoglycans (GAGs) and PGs after a chemical injury. The chemical composition of chondroitin sulfate (CS)/dermatan sulfate (DS) and heparan sulfate (HS) were characterized in mouse corneas 5 and 14 days after alkali burn (AB), and compared to uninjured corneas. The expression profile and corneal distribution of CS/DSPGs and keratan sulfate (KS) PGs were also analyzed. We found a significant overall increase in CS after AB, with an increase in sulfated forms of CS and a decrease in lesser sulfated forms of CS. Expression of the CSPGs biglycan and versican was increased after AB, while decorin expression was decreased. We also found an increase in KS expression 14 days after AB, with an increase in lumican and mimecan expression, and a decrease in keratocan expression. No significant changes in HS composition were noted after AB. Taken together, our study reveals significant changes in the composition of the extracellular matrix following a corneal chemical injury.

Topics: Alkalies; Animals; Biomarkers; Burns, Chemical; Corneal Diseases; Dermatan Sulfate; Disease Models, Animal; Extracellular Matrix; Eye Burns; Fluorescent Antibody Technique; Gene Expression; Glycosaminoglycans; Heparitin Sulfate; Keratan Sulfate; Mice; Proteoglycans

Heparan sulfate (HS) is a highly modified glycosaminoglycan (GAG) bound to a core protein to form heparan sulfate proteoglycans (HSPGs) that are vital in many cellular processes ranging from development to adult physiology, as well as in disease, through interactions with various protein ligands. This study aimed to elucidate the role of HS in corneal epithelial homeostasis and wound healing.. An inducible quadruple transgenic mouse model was generated to excise Ext1 and Ndst1, which encode the critical HS chain elongation enzyme and N-deacetylase/N-sulfotransferase, respectively, in keratin 14-positive cells upon doxycycline induction.. EXT(Δ/ΔCEpi) mice (deletion of Ext1 in corneal epithelium) induced at P20 presented progressive thinning of the corneal epithelium with a significant loss in the number of epithelial layers by P55. EXT(Δ/ΔCEpi) mice presented tight junction disruption, loss of cell-basement membrane adhesion complexes, and impaired wound healing. Interestingly, EXT(Δ/ΔCEpi) and NDST(Δ/ΔCEpi) mice presented an increase in cell proliferation, which was assayed by both Ki67 staining and 5-ethynyl-2'-deoxyuridine (EdU) incorporation. Moreover, EXT(Δ/ΔCEpi) mice presented compromised epithelial stratification 7 days after a debridement wound. The conditional knockout of HS from keratocytes using the keratocan promoter led to no corneal abnormalities or any disruption in wound healing.. Corneal epithelial cells require HS for maintaining corneal homeostasis, and the loss of epithelial HS leads to both impaired wound healing and impaired corneal stratification.

Topics: Animals; Corneal Diseases; Epithelium, Corneal; Fluorescent Antibody Technique; Heparitin Sulfate; Homeostasis; Immunohistochemistry; Mice; Mice, Knockout; Microscopy, Electron; Wound Healing

Many microbial pathogens subvert cell surface heparan sulfate proteoglycans (HSPGs) to infect host cells in vitro. The significance of HSPG-pathogen interactions in vivo, however, remains to be determined. In this study, we examined the role of syndecan-1, a major cell surface HSPG of epithelial cells, in Staphylococcus aureus corneal infection. We found that syndecan-1 null (Sdc1(-/-)) mice significantly resist S. aureus corneal infection compared with wild type (WT) mice that express abundant syndecan-1 in their corneal epithelium. However, syndecan-1 did not bind to S. aureus, and syndecan-1 was not required for the colonization of cultured corneal epithelial cells by S. aureus, suggesting that syndecan-1 does not mediate S. aureus attachment to corneal tissues in vivo. Instead, S. aureus induced the shedding of syndecan-1 ectodomains from the surface of corneal epithelial cells. Topical administration of purified syndecan-1 ectodomains or heparan sulfate (HS) significantly increased, whereas inhibition of syndecan-1 shedding significantly decreased the bacterial burden in corneal tissues. Furthermore, depletion of neutrophils in the resistant Sdc1(-/-) mice increased the corneal bacterial burden to that of the susceptible WT mice, suggesting that syndecan-1 moderates neutrophils to promote infection. We found that syndecan-1 does not affect the infiltration of neutrophils into the infected cornea but that purified syndecan-1 ectodomain and HS significantly inhibit neutrophil-mediated killing of S. aureus. These data suggest a previously unknown bacterial subversion mechanism where S. aureus exploits the capacity of syndecan-1 ectodomains to inhibit neutrophil-mediated bacterial killing mechanisms in an HS-dependent manner to promote its pathogenesis in the cornea.

Topics: Animals; Corneal Diseases; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Host-Pathogen Interactions; Mice; Mice, Knockout; Neutrophils; Staphylococcal Infections; Staphylococcus aureus; Syndecan-1

Cultivated limbal epithelial transplantation using an amniotic membrane (AM) carrier is now widely used for ocular surface reconstruction. The reasons for the exceptional success of AM as a carrier are not fully understood but are believed to be related to its unique composition. In this project we characterize, at the ultrastructural level, the extracellular matrix (ECM) components present in AM. We also compare the distribution of ECM components of cellular AM with that of denuded AM.. Scanning, transmission, and atomic force microscopy was used to examine the structure of cellular and denuded amniotic membranes. Immunogold labeling with a panel of antibodies against ECM molecules was carried out on cellular and denuded AM.. Heparan sulfate, fibronectin, and laminin were present at high concentration in the lamina densa, Collagen IV was the major component of the basal lamina. Type I collagen was confined to the stroma along with significant amounts of keratan and chondroitin sulfate. Both cellular and denuded AMs had similar distributions of the ECM components.. We were able to determine the distribution of ECM molecules in the lamina densa, basal lamina, and stroma of AM at the ultrastructural level. The removal of amniotic epithelial cells using our protocol does not appear to have any significant effects on the structure of the basal lamina or the distribution of ECM components.

Topics: Amnion; Basement Membrane; Biological Dressings; Collagen Type I; Collagen Type IV; Conjunctival Diseases; Corneal Diseases; Extracellular Matrix Proteins; Fibronectins; Heparitin Sulfate; Humans; Immunohistochemistry; Laminin; Microscopy, Atomic Force; Microscopy, Electron, Scanning; Microscopy, Immunoelectron

Newer biochemical understanding of the mucopolysaccharidoses now allows a better classification of these diseases. The dermatan and keratan sulfate-storing diseases have corneal clouding. The heparan sulfate-storing diseases have retinal changes and usually central nervous system manifestations.

Topics: Child; Corneal Diseases; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Humans; Keratan Sulfate; Mucopolysaccharidoses; Syndrome