chondroitin-sulfates and Corneal-Injuries

To construct a suitable ex vivo model for the research of molecular mechanisms and the pharmacological screening of fungal adherence on the corneal surface.. Mouse eyes were divided into three groups as follows: a control group with normal corneal epithelium, a group with corneal epithelium that was needle-scarified, and a group with corneal epithelium that was completely debrided. All 96 corneas were placed in organ culture and inoculated with 5 μl spore suspensions of Candida albicans at 10⁹, 10⁸, or 10⁷ colony-forming units (CFU)/ml and incubated for 0, 30, 60, or 120 min. The corneas were homogenated and diluted for quantification by counting the CFU. The effects of amphotericin B or chondroitin sulfate on the adherence of the fungal spores were evaluated with the ex vivo organ culture model and were also compared with the human corneal epithelium monolayer model in vitro.. Compared with the normal corneas with intact epithelium, the corneas with scarified and debrided epithelium adhered more spores for above two and four folds. The spore adhesion on the corneal surface was in an inoculation concentration- and incubation time-dependent manner. Moreover, both amphotericin B and chondroitin sulfate inhibited the adhesion of C. albicans spores on the corneal surface, but the inhibitory rates were different between the ex vivo corneal organ culture model and the in vitro corneal epithelium monolayer model.. The corneal organ culture was a suitable ex vivo model for the research of fungal adhesion mechanisms and drug screening.

Topics: Amphotericin B; Animals; Bacterial Adhesion; Candida albicans; Candidiasis; Chondroitin Sulfates; Colony Count, Microbial; Cornea; Corneal Injuries; Corneal Ulcer; Disease Models, Animal; Eye Infections, Fungal; Mice; Mice, Inbred BALB C; Organ Culture Techniques; Time Factors

To compare a modified chondroitin sulfate aldehyde adhesive with standard sutures for sealing corneal incisions.. A keratome knife was used to create non-self-sealing, uniplanar, 3-mm, clear corneal incisions in enucleated rabbit eyes (n = 18). The wounds were sealed with either a chondroitin sulfate-aldehyde adhesive (n = 8), three 10-0 nylon sutures (n = 5), or one 10-0 nylon suture (n = 5). Wound stability was tested by filling the globes with balanced salt solution through an anterior chamber port and slowly increasing the IOP. The pressure changes were monitored with a digital manometer connected to the anterior chamber, and leak pressure was recorded for each eye. Confocal microscopy was performed on the glued eyes, to document the glue distribution along the wound.. The mean leak pressures in the single-suture and three-suture subgroups were 26.4 +/- 6.0 and 44.3 +/- 8.2 mm Hg (SD), respectively. The maximum IOP achieved in eyes that received the glue was 104.7 mm Hg with a mean of 101.4 +/- 3.2 mm Hg. None of the eyes in which glue was used showed leakage. At confocal microscopy, the glue was distributed inside the wound edges as a homogeneous thin layer of a less dense signal than that of the stroma.. A novel chondroitin sulfate-aldehyde adhesive was shown to be effective ex vivo for sealing corneal incisions in rabbit eyes and was superior to sutures for this purpose.

Topics: Aldehydes; Animals; Chondroitin Sulfates; Corneal Injuries; Eye Enucleation; Intraocular Pressure; Microscopy, Confocal; Rabbits; Suture Techniques; Tissue Adhesives; Wound Healing

We investigated the effect of hyaluronan on corneal epithelial wound healing in rats affected by diabetes. Furthermore, because hyaluronan is thought to affect corneal epithelial wound healing through the mechanism of binding of hyaluronan to provisional fibronectin in the wounded area, we compared the localization of fibronectin immunohistochemically during corneal epithelial wound healing in diabetic and non-diabetic rats. Streptozotocin was used to induce diabetes in half the rats. Two weeks after treatment, the whole corneal epithelium of diabetic and untreated rats was debrided. The rats were divided into groups (seven or eight rats per group), and hyaluronan eye drops at concentrations of 0.03, 0.1, or 0.3%, chondroitin sulfate (3%), or phosphate buffered saline (PBS) was given in eye drops 6 times a day for 4 days, starting immediately after debridement. The area of the corneal epithelial wound was measured immediately after debridement and at 12, 18, 24, 30, 48, 72, and 96 hours afterwards. Although the healing process was similar in non-diabetic and diabetic rats, the healing rate in diabetic rats was slower than that in normal controls. In both diabetic and non-diabetic rats, hyaluronan increased the healing rate in a dose-dependent manner; the difference was significant compared with the PBS-treated group, at hyaluronan doses of 0.1% and 0.3%. However, chondroitin sulfate did not affect corneal epithelial wound closure, regardless of whether the rats were diabetic or not; the healing rates were identical to those of PBS-treated diabetic and non-diabetic controls. In both diabetic and non-diabetic corneas, fibronectin was localized in the corneal subepithelial region, and in streaks between collagen fibers of the stroma. One day after debridement, a layer of fibronectin immunofluorescence was clearly visible on the surface of the denuded stroma. As healing progressed staining of fibronectin diminished at the interface between the new epithelium and the stroma. These changes in localization of fibronectin during corneal epithelial wound healing were similar in both diabetic and non-diabetic rats. Our results demonstrate that hyaluronan facilitates corneal epithelial wound healing in diabetic rats, and suggest that one possible mechanism of its stimulatory effect lies in its binding to a provisional fibronectin matrix, in both diabetic and non-diabetic rats.

Topics: Animals; Chondroitin Sulfates; Cornea; Corneal Injuries; Diabetes Mellitus, Experimental; Dose-Response Relationship, Drug; Epithelium; Fibronectins; Hyaluronic Acid; Male; Rats; Rats, Sprague-Dawley; Wound Healing

Sodium hyaluronate (Healon), chondroitin sulfate, and methylcellulose have been used to protect the corneal endothelium from intraocular lens trauma. A study of the efficacy and toxicity of these compounds showed that 1% sodium hyaluronate, 0.4% methylcellulose, and 20% chondroitin sulfate were nontoxic to the corneal endothelium, but that 20% chondroitin sulfate caused a marked decrease in corneal thickness because of its hypertonicity. Anterior chamber injection of these viscous substances resulted in an increase in intraocular pressure. Within one to four hours the maximum intraocular pressure with 1% sodium hyaluronate was 67 +/- 4.1 mm Hg and that with 20% chondroitin sulfate was 55 +/- 3.5 mm Hg. The intraocular pressure did not increase to these high levels with 10% chondroitin sulfate or 0.4% methylcellulose or when the test substances were washed out of the anterior chamber. The corneal endothelium was protected from injury with 1% sodium hyaluronate and 20% chondroitin sulfate, but 10% chondroitin sulfate and 0.4% methylcellulose provided only minimal protection.

Topics: Animals; Chondroitin; Chondroitin Sulfates; Cornea; Corneal Injuries; Endothelium; Hyaluronic Acid; Intraocular Pressure; Lenses, Intraocular; Methylcellulose; Rabbits