We examined the fibronectin (FN) secretion of cultured trabecular meshwork (TM) cells in a normal human eye by indirect immunofluorescent technique using mouse anti-human FN monoclonal antibody and FITC-conjugated goat anti-mouse IgG. To localize FN on frozen sections of normal TM, which were obtained from 7 enucleated eyes owing to traumatic eyeball rupture, the same indirect immunofluorescent method was used. Immunoelectron microscopy was applied to demonstrate the distribution pattern of FN in the normal TM of 2 human eyes using an avidin-biotin-peroxidase complex method. In the tissue culture of TM, the TM cell walls and extracellular matrices showed an intense staining with antibody to FN. Indirect immunofluorescent staining of FN on frozen sections of TM showed strong positive reactions in the subendothelial region. There was no reaction in the central core of the trabecular beam. Immunoelectron microscopy revealed the reaction products to FN in the areas lining the trabecular endothelial cells.
Adolescent ; Adult ; Antibodies, Monoclonal ; Cells, Cultured ; Fibronectins/biosynthesis/*metabolism ; Fluorescent Antibody Technique ; Humans ; Male ; Microscopy, Immunoelectron ; Trabecular Meshwork/*metabolism/ultrastructure

The changes in intraocular pressure (IOP) following peritomy and wet-field coagulation of 10 rabbit eyes and 7 human cataractous eyes were investigated. The IOP difference before and after 180-degree and 360-degree wet-field coagulation on rabbit eyes showed an average increase of 11.0 +/- 5.8 mmHg and 25.8 +/- 7.1 mmHg(mean +/- SD), respectively (p < 0.05, p < 0.01). The IOP of both groups in rabbit eyes were elevated for 2 to 3 days postoperatively and then became normal by the 7th postoperative day. The IOP difference before and after 180-degree peritomy and wet-field coagulation in human cataractous eyes showed an average increase of 8.4 +/- 5.8 mmHg(mean +/- SD)(p < 0.05). The outflow facility was poor immediately following wet-field coagulation but then improved gradually by means of tonography and concentration changes of anterior chamber neutral-red. Histologically, there was endothelial cell damage, congestion, and red blood cell extravasation of the episcleral and intrascleral vessels at one day after wet-field coagulation in the light microscopic findings. Recanalization of the episcleral and intrascleral capillaries was noted 3 days after wet-field coagulation and was completed 3 weeks after wet-field coagulation. There was poor plastic infusion through the trabecular spaces in the one-day postoperative group compared to the control group by scanning electron micrograph. In conclusion, we have to bear in mind the possibility of an IOP rise after wet-field coagulation following peritomy for cataract and retinal surgery.
Animals ; Aqueous Humor/physiology ; Cataract Extraction ; Conjunctiva/*surgery/ultrastructure ; Humans ; *Intraocular Pressure ; *Light Coagulation ; Microscopy, Electron, Scanning ; Rabbits ; Surgical Flaps ; Tonometry, Ocular ; Trabecular Meshwork/ultrastructure

BACKGROUNDTrabecular meshwork (TM) cell volume may be an important determinant of aqueous humor outflow in the eye. This study aimed to evaluate the role of HepII domain peptides V on corneal permeability, corneal endothelial cells, intraocular pressure (IOP) and morphology of trabecular meshwork in rats.

METHODSThe IOP of rat eyes was measured before and 3, 5, 7 and 8 hours after topical delivery of HepII domain peptides V through intracameral injections. The peptide's concentration in aqueous humor was assessed by high performance liquid chromatography (HPLC). The shape and density of endothelial cells were observed by laser confocal microscopy 8 hours, 3 and 14 days after intracameral injections of HepII domain peptides V. The morphological changes in TM of rat eyes were assessed by transmission electron microscopy (TEM).

RESULTSIntracameral injection of HepII domain peptides V significantly (P < 0.001) decreased IOP by (5.71 ± 2.10) mmHg in rats at 5 hours after injection. There were no obvious changes of the shape and the density of corneal endothelial cells. In addition, morphological changes in the TM of rats were observed including the expansion of intercellular spaces in the juxtacanalicular meshwork, removal of extracellular material, cellular relaxation, and cytoskeleton reorganization.

CONCLUSIONSHepII domain peptides V could not penetrate cornea and was safe to corneal endothelial cells. HepII domain peptides V could significantly decrease IOP in rat probably by disorganizing actin cytoskeleton and cell-junction in the TM.

Animals ; Chromatography, High Pressure Liquid ; Cornea ; cytology ; drug effects ; ultrastructure ; Endothelium, Corneal ; drug effects ; ultrastructure ; Female ; Fibronectins ; chemistry ; pharmacology ; Intraocular Pressure ; drug effects ; Male ; Microscopy, Confocal ; Microscopy, Electron, Transmission ; Rats ; Rats, Sprague-Dawley ; Trabecular Meshwork ; drug effects ; ultrastructure