chondroitin-sulfates and alizarin

To assess the endothelial toxicity and the microbiological efficacy of moxifloxacin (250 microg/mL) as an additive to Optisol-GS.. Five hundred nine donor rims were studied. One half of each donor rim was placed in standard Optisol-GS and the other half of the rim in Optisol-GS fortified with moxifloxacin (250 microg/mL). All rims were refrigerated for 24 hours at 3 degrees C and placed in thioglycolate broth and incubated at 37 degrees C for 7 days. One pair of donor buttons not used in transplantation stored in each solution was examined for endothelial changes by using electron microscopy (EM). A second pair of cornea buttons was examined for toxicity by endothelial staining with 0.3% trypan blue and 0.2% alizarin red. All endothelial cells that stained (nonviable cells) and nonstained cells (viable cells) were counted, and the ratio of nonviable cells was calculated.. The rate of culture-positive donor rims in the Optisol-GS group was 11.9% (61/509) and in the moxifloxacin-fortified Optisol-GS media was 2.5% (13/509). The difference was statistically significant (P < 0.01; chi test). There was no difference in the cellular morphology of the button stored in moxifloxacin-fortified Optisol-GS compared with Optisol-GS using EM. In the bioassay, the rate of nonviable cells in the moxifloxacin-fortified media compared with the control media was nonsignificant (P > 0.05).. Moxifloxacin (250 microg/mL) seems to be safe as an additive agent for cornea storage media. It significantly reduces the rate of positive rim cultures and shows no signs of endothelial cytotoxicity as viewed by EM and by a bioassay of trypan blue and alizarin red.

Topics: Anthraquinones; Anti-Infective Agents; Aza Compounds; Cell Count; Cell Survival; Chondroitin Sulfates; Coloring Agents; Complex Mixtures; Cornea; Culture Media, Serum-Free; Dextrans; Drug Combinations; Endothelium, Corneal; Fluoroquinolones; Gentamicins; Humans; Middle Aged; Moxifloxacin; Organ Preservation; Organ Preservation Solutions; Quinolines; Tissue Donors; Trypan Blue

To systematically investigate the central, paracentral, and peripheral endothelial cell density (ECD) in normal human corneas.. Observational case series and experimental study.. Noncontact specular microscopy was undertaken to determine the ECD of the central, paracentral (2.7 +/- 0.2 mm from center) and peripheral (4.7 +/- 0.2 mm from center) regions of the cornea of 48 normal eyes. The ECDs of central and peripheral regions were also determined with contact specular microscopy in 21 normal eyes and a group of 30 Optisol-GS eye bank corneas were evaluated with alizarin red stain. Histologic ECD of 13 Optisol-GS stored corneas were also determined.. Paracentral and peripheral ECD measured with the noncontact specular microscope were 5.8% (P <.01) and 9.6% (P <.001) increased compared with central ECD. Superior peripheral ECD was increased compared with the other three peripheral quadrants (P <.05) and was 15.9% higher than central ECD. Contact specular microscopy showed an increase of 8.9% in the peripheral ECD from the center. Alizarin red stained corneas confirmed the specular microscopy numbers with a 9.2% increase in the paracentral region, and a 17.2% increase in the peripheral region. Histological cross sections of human corneas also showed a 22.9% increase in peripheral ECD compared with the central region.. The human cornea has an increased ECD in the paracentral and peripheral regions of cornea compared with the central region. The superior peripheral region of the corneal endothelium has the largest increase in ECD. These data on normal endothelial cell distribution in the human cornea are especially significant as they relate to new surgical techniques and endothelial wound repair.

Topics: Adolescent; Adult; Aged; Anthraquinones; Cell Count; Chondroitin Sulfates; Complex Mixtures; Culture Media, Serum-Free; Dextrans; Endothelium, Corneal; Female; Gentamicins; Humans; Image Processing, Computer-Assisted; Male; Microscopy; Middle Aged; Organ Preservation; Reference Values; Staining and Labeling; Tissue Donors

To investigate the effect of shaking on corneal endothelial preservation.. Thirty-six dog corneal buttons were obtained under standard eye bank procedure, after the animals had been sacrificed for cardiovascular experiments in the surgical department. They were equally divided into two groups. In Group I, buttons were put in Dexsol preservative medium and preserved at 4 degrees C. In Group II, buttons were put in Dexsol preservative medium and shaken in a shaking incubator at a speed of 5 rpm. at 4 degrees C for 10 h. After shaking, they were returned to a 4 degrees C refrigerator until examination. Three buttons from each group underwent specular microscopy, scanning electron microscopy (SEM), and alizarin red with trypan blue stain examinations on days 0, 1, 3, 5, 7 and 9, respectively.. In Group I, intercellular borders became blurred under specular microscopy, beginning on day 5. However, endothelial cell count did not change significantly until day 9. Intercellular digitations and wrinkling of intercellular borders were seen under alizarin red with trypan blue stain and SEM examination, beginning on day 3. Pleomorphism and ill-defined intercellular junctions were seen under SEM on day 7. There was no obvious denudement of the endothelial sheet, but a few scattered exfoliations were seen in Group I. In Group II, endothelial cell count did not decrease on day 1, but an endothelial image could not be obtained by specular microscopy 3 days after treatment. Alizarin red with trypan blue stain and SEM examination revealed that the endothelial cells were denuded from the Descemet's membrane three days after shaking. Severe stromal edema was seen under SEM five days after shaking.. The results showed that shaking had a detrimental effect on endothelial cell preservation. Vigorous shaking should be avoided during transportation of corneal buttons. It is advisable to perform penetrating keratoplasty as soon as possible upon receiving transported donor corneas.

Topics: Animals; Anthraquinones; Cell Count; Chondroitin Sulfates; Coloring Agents; Cryopreservation; Culture Media, Serum-Free; Dogs; Endothelium, Corneal; HEPES; Microscopy, Electron, Scanning; Organ Preservation; Organic Chemicals; Transportation; Trypan Blue; Vibration

We compared the endothelial protection offered by 1% hyaluronate sodium (Healon), 3% hyaluronate sodium and 4% chondroitin sulfate (Viscoat), and a nonviscous irrigating solution (BSS Plus) during phacoemulsification with and without traumatic intraocular lens implantation. Vital-dye staining and scanning electron microscopy were used to determine acute damage to rabbit corneal endothelium. Cell damage during phacoemulsification alone was not significantly different from that in unoperated controls (12.5%). Cell damage after traumatic lens insertion was significantly greater in the groups treated with BSS Plus (76.2%) and Healon (41.4%) than in either paired Viscoat-treated group (21.1% and 17.4%, respectively). Viscoat (but not Healon) was noted to be adherent to the cornea at the end of the procedure in one third of the cases, indicating that Viscoat remains in the anterior chamber during surgery. We attribute this to chondroitin sulfate's newtonian characteristics, allowing it to maintain viscosity in the face of high flow rates.

Topics: Animals; Anthraquinones; Bicarbonates; Cataract Extraction; Cell Survival; Chondroitin; Chondroitin Sulfates; Coloring Agents; Drug Combinations; Endothelium, Corneal; Glutathione; Hyaluronic Acid; Lenses, Intraocular; Microscopy, Electron, Scanning; Ophthalmic Solutions; Rabbits; Trypan Blue; Viscosity