chondroitin-sulfates and 6-carboxyfluorescein

To evaluate the usefulness of viscoelastic in protecting the corneal endothelium from desiccation injury associated with fluid-air exchange in a rabbit model.. Experimental study.. Rabbit eyes undergoing pars plana lensectomy and vitrectomy were insufflated with either dry or humidified air for 20 minutes following introduction of either Opegan (sodium hyaluronate 1.0%; Santen, Osaka, Japan) or Viscoat (sodium hyaluronate 3%-chondroitin sulfate 4%; Alcon, Tokyo, Japan) into the anterior chamber. In two other groups of rabbit eyes, the same procedure was performed without using any viscoelastic agent. Corneas obtained from rabbits undergoing surgery were compared with corneas obtained from rabbits not undergoing surgery. Potential alterations in the corneal endothelium were investigated by scanning electron microscopy, by Phalloidin-FITC staining of actin and by in vitro measurements of corneal permeability for carboxyfluorescein using a diffusion chamber.. Scanning electron microscopy displayed less distortion of corneal endothelium with Opegan and Viscoat compared with the dry air-only exposed corneas. Using humidified air in Opegan and Viscoat coated corneas maintained the normal actin cytoskeleton during fluid-air exchange. Paracellular leakage was much less with Opegan and Viscoat use following infusion of dry air comparing to that of dry air-only group (P =.026 and P =.041). The difference was much more striking following humidified air infusion in Opegan or Viscoat coated corneas comparing to dry air-only infused corneas (P <.002 and P <.002).. Coating of rabbit corneal endothelium with Opegan or Viscoat before fluid-air exchange largely prevents dry air damage to the endothelium. Infusion of humidified air further protects corneal endothelium during fluid-air exchange in aphakic rabbit eyes.

Topics: Actins; Air; Animals; Chondroitin; Chondroitin Sulfates; Desiccation; Drug Combinations; Endothelium, Corneal; Eye Injuries; Fluoresceins; Humidity; Hyaluronic Acid; Lens, Crystalline; Microscopy, Electron, Scanning; Permeability; Rabbits; Vitrectomy

The purpose of this study was to compare Optisol to moist chamber storage for maintaining human corneal endothelial barrier function. Human corneas preserved in Optisol were stored for up to 35 days at 4 C. Endothelial carboxyfluorescein permeability (P(ac)) was measured and endothelial ultrastructure was evaluated by electron microscopy. Endothelial P(ac) (x 10(-4) cm/min) of Optisol-stored corneas was 1.7, 2.0, and 3.1 at five, seven, and 14 days, respectively. The P(ac) increased to 6.5 at 35 days of storage. Endothelial P(ac) in moist chamber stored-eyes was 2.6 at two days, and increased to 13.5 14 days of storage. Multiple regressional analysis showed that storage time and donor age affected P(ac); but time from death to enucleation, time from enucleation to storage, or endothelial cell number did not. Electron microscopy showed that endothelial junctions were maintained through two weeks by Optisol. Large areas of cellular destruction were seen after five days of moist chamber storage. These results show that Optisol can preserve endothelial barrier function through 14 days; barrier function is lost by three days of moist chamber storage.

Topics: Adult; Aged; Cell Count; Cell Membrane Permeability; Chondroitin Sulfates; Complex Mixtures; Cryopreservation; Culture Media, Serum-Free; Dextrans; Endothelium, Corneal; Fluoresceins; Gentamicins; Humans; Intercellular Junctions; Middle Aged; Organ Preservation

As a step in the development of a system for assessing growth of human urothelium and transitional cell carcinoma, the growth of two established cell lines on collagen-based membranes has been evaluated. HT1080 (metastatic human fibrosarcoma) and WI38 VA13 (virus-transformed human fibroblasts) were grown on substrates of collagen, collagen/hyaluronic acid or chondroitin sulphate and reconstituted basement membrane (Matrigel). Cell growth was quantified using a new fluorimetric assay utilizing carboxyfluorescein diacetate. There were differences in morphology between cells grown on collagen and those grown on polystyrene. There were, however, no differences in growth of the WI38 VA13 cells on collagen compared with polystyrene, but growth of the HT1080 cells was increased on membranes of collagen/2.5% hyaluronic acid and collagen/5% chondroitin sulphate, and decreased on Matrigel. Adequate growth on collagen substrates is dependent on cell line. The fluorimetric assay used was suitable for quantifying cell growth on such substrates.

Topics: Basement Membrane; Biocompatible Materials; Carcinoma, Transitional Cell; Cell Count; Chondroitin Sulfates; Collagen; Culture Media; Drug Combinations; Fluoresceins; Fluorometry; Humans; Hyaluronic Acid; In Vitro Techniques; Laminin; Proteoglycans; Tumor Cells, Cultured

Endothelial permeability (Pac) to carboxyfluorescein and Na+/K+ ATPase pump site density were determined in human corneas following storage for 4 or 7 days at 4 degrees C in either modified McCarey-Kaufman (mMK) or K-Sol media. Following 4 days of storage, Pac values for mMK- and K-Sol-preserved corneas were not significantly different from those of their prestorage mates. After 7 days of storage, however, corneas stored in K-Sol media showed a significant increase in Pac compared to their prestorage mates, whereas the mMK-stored corneas showed no change in Pac. Na+/K+ ATPase pump site density determined using [3H]ouabain was similar to a control group in the K-Sol-stored tissue but higher in the mMK-stored tissue following 7 days of storage. These studies suggest that mMK medium maintains endothelial barrier function and Na+/K+ ATPase pump site density at least as well as K-Sol medium through 7 days of corneal storage.

Topics: Adult; Aged; Chondroitin Sulfates; Cornea; Culture Media; Endothelium, Corneal; Fluoresceins; HEPES; Humans; Middle Aged; Organic Chemicals; Permeability; Sodium-Potassium-Exchanging ATPase; Time Factors; Tissue Preservation