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PubMed1991

Effect of silicone oil on ocular tissues.

The effect of silicone oil on ocular tissues was investigated histopathologically in rabbit eyes by injecting silicone oil into the anterior chamber or the vitreous cavity after vitrectomy. The eyes were enucleated 3, 6, 9, 12 or 18 months after the injection and various tissues were observed by light and electron microscopy. Silicone oil was first observed in the retrocorneal membrane at 18 months after the injection. In the trabecular meshwork silicone oil was seen for the first time at 12 months after injection. Migrating cells engulfing silicone oil attached to the internal limiting membrane of the retina 3 months after the injection. Twelve and 18 months after the injection, silicone oil passed through the internal limiting membrane and was engulfed by Müller cells. These results indicate that attention should be paid to the effects of silicone oil on ocular tissues when it remains in the eye for a long time since, after a certain period, cells of the trabecular meshwork and retina may engulf silicone oil.

Animals↗
PubMed1992

Immunohistochemical characterization of dendritic cells and macrophages in the aqueous outflow pathways of the rat eye.

Immunohistochemical studies were performed to determine the distribution, phenotype and ontogeny of macrophages and dendritic cells (DCs) in the aqueous humour outflow pathways of the rat eye. Optimal fixation and indirect immunoperoxidase techniques were employed in conjunction with a panel of mAbs on tangential frozen sections of ocular tissues from a total of 37 Wistar Furth rats aged 12-13 days (n = 8), 3 weeks (n = 12), 7 weeks (n = 5) and 15 weeks (n = 12). The density of immunopositive cells was scored qualitatively. A moderate to low density of Ia+ cells with a dendritic morphology were observed in the trabecular meshwork. DCs were also identified in the suprachoroidal space and in the connective tissues of nerves and vessels piercing the sclera, i.e. in association with non-conventional aqueous outflow pathways. The phenotypical and morphological characteristics of these cells would indicate that they may potentially act as antigen presenting cells (APCs). Non-dendritic pleomorphic cells with a macrophage phenotype were also identified in the trabecular meshwork, and bipolar or elongated cells with a macrophage phenotype were a noticeable feature in the perivascular region of collector channels and the limbal episcleral veins. Some macrophage and DC-like cells were observed in intimate association with limbal mast cells. Theories on the mechanisms of Anterior Chamber Associated Immune Deviation (ACAID) have assumed APCs are largely absent from the tissues lining the anterior chamber. Our findings of a low but moderate density of putative APCs in the conventional and non-conventional aqueous humour outflow pathways are discussed in relation to the various theories of ACAID.

Aging↗
PubMed1993

Internal ablative sinostomy using a fiber delivered Q-switched CTE: YAG laser (2.69 microns).

Current trends of laser technology towards low-thermal photoablative pulsed mid-infrared lasers open new, more adequate approaches to experimental surgical procedures which have already been evaluated in the past. Transcorneal laser ablation of the trabecular meshwork (internal sinostomy) in human autopsy eyes was performed with a Q-switched CTE:YAG laser (wavelength: 2.69 microns, pulse width: 1 microsecond). Beam delivery was achieved with conventional optical quartz fibers (Low-hydroxy-fused-silica: 0.3 ppm, 50 cm length, 200 microns diameter). Light- and scanning-electron-microscopy were used for histological examination and micromorphological analysis. By applying two laser pulses (6 J/cm2) to the functional trabecular meshwork, a round sinostomy with a diameter corresponding to the diameter of the fiber-tip was achieved. It was possible to set several internal sinostomies into the chamber angle opposite to the entering paracentesis of the laser fiber-tip. Collateral thermal tissue alteration reached up to 50 microns, and since fiber-tip contact was maintained during laser application, thermal tissue alteration was also found around the opposite wall of Schlemm's canal. At higher energy fluences mechanical (disruptive vaporization) effects were significantly enhanced. It can be concluded, that low-thermal pulsed mid-infrared lasers are adequate instruments to perform transcorneal trabecular ablation (abinterno sinostomy). The laser used in this study (CTE:YAG) bears the advantage that its radiation can easily be delivered in conventional optical quartz fibers.

Fiber Optic Technology↗
PubMed1998

[Endoscopically controlled erbium YAG laser goniotomy. Initial preclinical trials].

INTRODUCTION: Endoscopic erbium-YAG laser treatment is a new approach in glaucoma surgery. In contrast to conventional laser systems, the photoablative erbium-YAG laser allows microperforations of the trabecular meshwork without thermal side effects. We report our first preclinical trials using this new system. METHOD: We used the Endognost system (Schwind Co.). The device combines an endoscope and illumination fiber (0.5 mm diameter), laser fiber (0.5 mm) and a irrigation tube in one probe with a 1.1 mm external diameter. The Endognost system was tested in porcine and enucleated human eyes. All eyes were examined histologically. RESULTS: The endoscopic view into the anterior chamber allows for precise allocation of the laser pulses. Using a single pulse mode with 10 mJ a micropuncture of the trabecular meshwork can be achieved without damaging the adjacent tissue. Using multiple pulses or higher energy levels leads to damage of the posterior wall of Schlemm's canal and to thermal side effects. CONCLUSIONS: Endoscope guided erbium-YAG laser effects on trabecular tissue are comparable to those produced by a 308 nm excimer laser. Therefore, a similar reduction in intraocular pressure can be expected.

Animals↗
PubMed2000

Altered secretion of a TIGR/MYOC mutant lacking the olfactomedin domain.

TIGR/MYOC, a novel 504 amino acids (aa) protein of unknown function, has recently been linked to glaucoma. The protein is both intra- and extracellular and most known mutations map to its C-terminus, an olfactomedin-like domain. To investigate the properties of a TIGR/MYOC peptide lacking this important domain, we constructed a replication-deficient adenovirus with the first 344 aa and over-expressed the truncated protein in primary human trabecular meshwork cells and perfused human anterior segment cultures. The truncated mutant contains the entire N-terminus plus 98 aa of the olfactomedin-like domain. We found that the delivered truncated mutant accumulates inside the cell, reduces secretion of endogenous TIGR/MYOC and induces an increase in outflow facility at 48 h post-infection. Based on these findings, we hypothesize that TIGR/MYOC might have a dual role in trabecular meshwork function. This dual role might be that of an intracellular modulator of vesicular transport as well as that of a secreted protein involved in extracellular matrix conformation. Both functions could have a direct effect in maintaining aqueous humor outflow facility.

Adenoviridae↗
PubMed1994

[The effect of acute and persistent ocular hypertension on ultrastructure in rabbit tissues of anterior chamber angle].

Sclero-circular compression operation was performed on 20 unilateral rabbit eyes to make a model of intraocular hypertension which persisted for 3 days, 1, 3 and 4 weeks respectively in 4 groups of rabbits and another 4 rabbits were used as normal controls. The trabecular meshwork and aqueous plexus of the experimental rabbits' eyes were examined under transmission electron microscope on various post-operative days: 3 days, 1, 3 and 4 weeks. It was discovered that the corneo-scleral and uveal trabecular meshworks were obviously distended and their core collagenous and elastoid fibers were hyperplastic, the phenomenon being more marked with the prolongation of the duration of the intraocular hypertension. Many endothelial cells were detached from the trabecular cords and freely located in the intertrabecular spaces. Persistent hyperplasia of collagenous fibers and accumulation of extracellular plague materials occurred in the endothelial meshwork, leading to its compactness. The giant vacuoles in the endothelial cells lining the inner wall of the canal of aqueous plexus were gradually decreased in number and the cytoplasm of the cells became attenuated and some fenestrations appeared. The results show that the main site of the resistance of the aqueous outflow occurring during intraocular hypertension is at endothelial meshwork and the above experimental morphological changes are quite similar to those of glaucoma patients.

Animals↗
PubMed1994

Trabecular aspiration: a new surgical approach to improve trabecular facility in pseudoexfoliation glaucoma.

The main reason for elevation of intraocular pressure in pseudoexfoliation glaucoma is due to secondary plugging of the intertrabecular spaces by pigment and fibrillous material. The aim of the present study was to introduce a new surgical concept to clean trabecular meshwork in pseudoexfoliative glaucoma. 'Trabecular aspiration' was performed under the operating microscope prior to extracapsular cataract extraction in half of the chamber angle circumference using a specially designed irrigation-aspiration device. The hand-held instrument has three outlets, one for aspiration (400 microns wide and 45 degrees horizontally angulated to meet the slope of the meshwork) and two openings (650 microns) for irrigation to maintain a deep anterior chamber and to keep the iris away from suction. Trabecular debris and pigment was cleared with a suction force of 100-200 mmHg. The effect of trabecular aspiration was cross-checked by analysing the aspirate. Pigment granules, fibrillous protein and other forms of trabecular debris were identified in the aspirate using light- and scanning electron microscopy. The morphological analysis of the trabecular aspirate clearly indicates the efficacy of 'trabecular aspiration' for removing pre- and intratrabecular debris. The clinical relevance of this new procedure looks promising, a clinical trial, recently initiated, is warranted.

Cataract Extraction↗
PubMed1990

[Exfoliation syndrome: clinical study of the irido-corneal angle].

A prospective clinical study of the anterior chamber angle was performed in 263 eyes (152 patients) with pseudoexfoliation syndrome (PSX). No significant difference was found in width and depth of the anterior chamber in comparison with normal population. We found a significative correlation (p less than .0001) between the increase of intraocular pressure (IOP) and the degree of diffuse pigmentation in trabecular meshwork and the Sampaolesi line. When we confronted the two eyes of the same patient, we found that the IOP was greater in the eye with higher pigmentation; and the higher was the difference of pigmentation between the two eyes, the higher was the difference in IOP (p less than .0001). No relation was found between PSX material in angle and IOP. In 24.7% of eyes, we noticed large iris processes, without goniodysgenesis. The results show a significative correlation between the pigmentation of the trabecular meshwork and the IOP.

Aged↗
PubMed1994

Lectin binding in the anterior segment of the bovine eye.

Eleven different fluorescent lectin-conjugates were used to reveal the location of carbohydrate residues in frozen sections of the anterior segment of bovine eyes. The lectins were specific for the following five major carbohydrate groups: (1) glucose/mannose group (Concanavalin A (Con A)); (2) N-acetylglucosamine group (wheat germ agglutinin (WGA)); (3) galactose/N-acetylgalactosamine group (Dolichos biflorus agglutinin (DBA), Helix pomatia agglutinin (HPA), Helix aspersa agglutinin (HAA), Psophocarpus tetragonolobus agglutinin (PTA), Griffonia simplicifolia agglutinin-I-B4 (GSA-I-B4), Artocarpus integrifolia agglutinin (JAC), peanut agglutinin (PNA) and Ricinus communis agglutinin (RCA-I)); (4) L-fucose group (Ulex europaeus agglutinin (UEA-I)); (5) sialic acid group (wheat germ agglutinin (WGA)). All the studied lectins except UEA-I reacted widely with different structures and the results suggest that there are distinct patterns of expression of carbohydrate residues in the anterior segment of the bovine eye. UEA-I bound only to epithelial structures. Some of the lectins reacted very intensely with apical cell surfaces of conjunctival and corneal epithelia suggesting a different glycosylation at the glycocalyx of the epithelia. Also, the binding patterns of conjunctival and corneal epithelia differed with some of the lectins: PNA and RCA-I did not bind at all, and GSA-I-B4 bound only very weakly to the epithelium of the cornea, whereas they bound to the epithelium of the conjunctiva. In addition, HPA, HAA, PNA and WGA did not bind to the corneal basement membrane, but bound to the conjunctiva and vascular basement membranes. This suggests that corneal basement membrane is somehow different from other basement membranes. Lectins with the same carbohydrate specificity (DBA, HPA, HAA and PTA) reacted with the sections almost identically, but some differences were noticed: DBA did not bind to the basement membrane of the conjunctiva and the sclera and did bind to the basement membrane of the cornea, whereas other lectins with same carbohydrate specificities reacted vice versa. Also, the binding of PTA to the trabecular meshwork was negligible, whereas other lectins with the same carbohydrate specificities reacted with the trabecular meshwork. GSA-I-B4 reacted avidly with the endothelium of blood vessels and did not bind to the stroma, so that it made blood vessels very prominent and it might be used as an endothelial marker. This lectin also reacted avidly with the corneal endothelium. Therefore, GSA-I-B4 appears to be a specific marker in bovine tissues for both blood vessel and corneal endothelium cells.

Animals↗
PubMed1982

Short-term effects of Q-switched ruby laser on monkey anterior chamber angle.

Three Q-switched ruby laser pulses were applied to the trabecular meshwork of 10 monkey eyes. Pulse energies ranging from 20 to 110 mJ were studied. The spot size ranged from 100 to 200 micrometer (in air), and the pulse durations was 28 sec. Gonioscopic examinations showed a graded response from no appreciable change at 20 mJ per pulse to marked disruption of anterior chamber angle structures at 100 mJ or more per pulse. Perfusions done within an hour of treatment showed no consistent alteration of the outflow facility. Scanning electron microscopy demonstrated the graded anterior chamber angle response. No disruption of the angle structures was seen after the 20 mJ treatment, but discrete trabecular damage occurred after treatments with 25 mJ. After pulses equal to or greater than 45 mJ the anterior chamber angle structures were markedly altered. The power density causing extensive tissue disruption was equal to or greater than 150 X 10(8) watts/cm2. In each specimen with an identifiable trabecular lesion, tissue debris and endothelial edema were found on the adjacent inner surface of the cornea. Tearing of Descemet's membrane next to the trabecular meshwork occurred with the 100 mJ pulses.

Animals↗
PubMed1995

Dexamethasone-induced ocular hypertension in perfusion-cultured human eyes.

PURPOSE: Glucocorticoid administration can lead to the development of ocular hypertension and corticosteroid glaucoma in a subset of the population through a decrease in the aqueous humor outflow facility. The purpose of this study was to determine whether glucocorticoid treatment can directly affect the outflow facility of isolated, perfusion-cultured human eyes. METHODS: The anterior segments of human donor eyes from regional eye banks were placed in a constant flow, variable pressure perfusion culture system. Paired eyes were perfused in serum-free media with or without 10(-7) M dexamethasone for 12 days. Intraocular pressure was monitored daily. After incubation, the eyes were morphologically characterized by light microscopy, transmission and scanning electron microscopy, and scanning laser confocal microscopy. RESULTS: A significant increase in intraocular pressure developed in 13 of the 44 pairs of eyes perfused with dexamethasone with an average pressure rise of 17.5 +/- 3.8 mm Hg after 12 days of dexamethasone exposure. The contralateral control eyes, which did not receive dexamethasone, maintained a stable intraocular pressure during the same period. The outflow pathway of the untreated eyes appeared morphologically normal. In contrast, the dexamethasone-treated hypertensive eyes had thickened trabecular beams, decreased intertrabecular spaces, thickened juxtacanalicular tissue, activated trabecular meshwork cells, and increased amounts of amorphogranular extracellular material, especially in the juxtacanalicular tissue and beneath the endothelial lining of the canal of Schlemm. The dexamethasone-treated nonresponder eyes appeared to be morphologically similar to the untreated eyes, although several subtle dexamethasone-induced morphologic changes were evident. CONCLUSION: Dexamethasone treatment of isolated, perfusion-cultured human eyes led to the generation of ocular hypertension in approximately 30% of the dexamethasone-treated eyes. Steroid treatment resulted in morphologic changes in the trabecular meshwork similar to those reported for corticosteroid glaucoma and open angle glaucoma. This system may provide an acute model in which to study the pathogenic mechanisms involved in steroid glaucoma and primary open angle glaucoma.

Aged↗
PubMed1992

Outflow obstruction in pigmentary and primary open angle glaucoma.

To localize the site of outflow obstruction in glaucoma, we evaluated the trabecular meshwork tissues by morphometric methods. Thirty-three specimens from 27 patients with primary open angle glaucoma (n = 13), pigmentary glaucoma (n = 4), and pigment dispersion syndrome (n = 2), and from nonglaucomatous normal subjects (n = 8) were compared. In these specimens, the extent of aqueous channels and the area occupied by these channels where they terminate in cul-de-sacs were measured. In 32 nonglaucomatous normal specimens (six of the eight mentioned plus an additional 26), we discovered that 94% of the surface area of the cul-de-sacs is lined by trabecular cells. These measurements were used to calculate the resistance to aqueous outflow offered by cul-de-sacs. Three new concepts are advanced in this report: (1) the cul-de-sacs provide a major portion of the normal outflow resistance, (2) the cul-de-sac area is markedly reduced in pigmentary glaucoma and primary open angle glaucoma, accounting for a major portion of the increase in resistance in these conditions, and (3) macrophages are the major cell type responsible for trabecular meshwork clearance of pigment and debris. A common pathophysiologic sequence of events is proposed for the development of glaucoma in pigmentary glaucoma and primary open angle glaucoma.

Adult↗
PubMed2003

Protein expression, genomic structure, and polymorphisms of oculomedin.

PURPOSE: To elucidate protein expression, genomic structure, and genomic polymorphisms of a novel gene, 'oculomedin', that has been cloned as a mechanical stretch-response gene from human trabecular cells in culture. METHODS: Polyclonal antibody was prepared by immunizing rabbits with a chemically synthesized 15-mer peptide of oculomedin. Protein expression was revealed by Western blot analysis after polyacrylamide gel electrophoresis of extracts of mechanically stretched trabecular cells and control trabecular cells in culture as well as retinal tissue. Protein localization was studied immunohistochemically in the human eye section. Genomic structure was determined by searching the GenBank database. Genomic polymorphisms of the coding region in 163 glaucoma patients and 50 normal subjects were detected by PCR amplification and direct sequencing. RESULTS: Western blot analysis showed that oculomedin protein was expressed only in stretched trabecular cells, not in control trabecular cells in culture. Immunohistochemically, oculomedin protein was localized to the trabecular meshwork, Schlemm's canal endothelium, retinal photoreceptor cells, and corneal and conjunctival epithelium. The oculomedin gene (OCLM) consists of two exons which are located inside an intron of a different gene of unknown function (C1orf27) on chromosome 1q25, near and telomeric to myocilin (MYOC). Two types of heterozygous nucleotide substitutions resulting in amino acid changes were found in two of 75 patients with primary open-angle glaucoma, but not at all in patients with other types of glaucoma or in normal subjects. CONCLUSIONS: Oculomedin may play a role in the function of the trabecular meshwork and also in the development of primary open-angle glaucoma.

Animals↗
PubMed1997

[Application of proliferating cell nuclear antigen in the study of human trabecular cell proliferation].

OBJECTIVE: To evaluate the application of proliferating cell nuclear antigen (PCNA) in the study of human trabecular cell proliferation. METHODS: Immunohistochemical technique was used to observe the PCNA expressive level in the fourth passage of human trabecular meshwork, the effects of different concentrations of epinephrine, dexamethasone and epidermal growth factor on the level were also investigated, the results were compared with that of the normal cells cultured at the same time, and analyzed by a graphic device operation system in a computer. RESULTS: A stable proliferating curve was obtained according to the normal cell PCNA level, by which we could choose the best opportunity of drug application. Epinephrine and dexamethasone were found to significantly inhibit cell proliferation, while epidermal growth factor (EGF) could promote the proliferation. CONCLUSION: PCNA is considered to be a useful agent to observe the process of cell proliferation. The above mentioned methods are beneficial to the investigations of biochemical characteristics of trabecular meshwork cells and the pathogenic mechanisms of open-angle glaucoma.

Cell Division↗
PubMed2002

Expression of optineurin, a glaucoma-linked gene, is influenced by elevated intraocular pressure.

Optineurin (optic neuropathy inducing protein; OPTN) was recently linked to 16.7% of families with primary open-angle glaucoma. The function of OPTN in the eye is not known, but is present in the trabecular meshwork, which is responsible for maintenance of intraocular pressure (IOP). To gain insight into the role of OPTN in the development of glaucoma we studied its expression in response to factors known to be associated with the disease: elevated IOP, tumor necrosis factor-alpha (TNFalpha), and dexamethasone (DEX). We performed the treatments in human organ cultures under conditions mimicking physiological pressure. We find OPTN significantly upregulated after 2, 4, and 7 days of sustained elevated IOP. OPTN expression is also induced 2.3-fold by TNFalpha and 2.6-fold by prolonged DEX treatment. These results demonstrate that OPTN is part of the transcriptome responding to glaucomatous insults and support the protective role of this protein in the trabecular meshwork.

Aged↗
PubMed2004

In vitro and in vivo characterization of disulfide bond use in myocilin complex formation.

PURPOSE: Myocilin forms large complexes in aqueous humor. Part of this complex formation is due to myocilin-myocilin protein non-covalent interactions within the leucine zipper. However, additional covalent interactions also exist. We investigated the role of these covalent interactions in disulfide bond formation within myocilin. METHODS: Human aqueous humor was separated by denatured/non-reduced SDS-PAGE followed by Western blot analysis with myocilin specific antibodies. In part two of the study, site-directed mutagenesis was used to selectively mutate one, two, three, four, and all five cysteine residues in the mature myocilin protein expressed in an in vitro system. Products were immunoprecipitated with a hemagglutinin polyclonal antibody following in vitro transcription/translation and analyzed by SDS-PAGE. In part three of the study, glaucoma associated myocilin mutations Arg82Cys and Cys433Arg were created and complex formation analyzed in trabecular meshwork cells. RESULTS: Human aqueous humor showed myocilin in several distinct large complexes in non-reduced SDS-PAGE gels, indicating disulfide bonds occur. Similarly, in vitro expressed myocilin also produced large complexes. Mutation of all five cysteines (within the mature myocilin protein) eliminated this large complex formation. A combination of cysteine to alanine substitutions at amino acids 185, 245, and 433 had the most influence on myocilin complex formation under non-reducing conditions, however individual substitutions at each of the five cysteine amino acids had little influence on myocilin complexes. In trabecular meshwork cells, Arg82Cys was secreted but formed different sized complexes than wild type myocilin. Cys433Arg was not secreted and remained intracellular in a pattern that differed from wild type myocilin and Arg82Cys. CONCLUSIONS: Myocilin complexes present in human aqueous humor are in part due to disulfide bond formation between cysteine amino acids. Glaucoma associated mutations that affect the number of cysteine residues may alter covalent interactions.

Aqueous Humor↗
PubMed1998

[Human trabeculer cells in culture and cell identification].

PURPOSE: To establish the method of culturing human trabecular cells(HTCs) in vitro and to determine identifyng main points concerning cultured HTCs. METHODS: Trabecular meshwork, corneal and scleral tissue were collected from sixty-eight eyeballs of thirty-four human donors less than six years of age within 24 hours after death. The tissues were primarily cultured and the cells subcultured. Cultured cells were observed by light and electron microscopes. Fibronectin(FN), laminin(LN) and IV collagen (IV C) in extracellular matrix(ECM) of the cultured cells were immunohistocytochemically stained by S-P method. RESULTS: HTCs easily grew in the event of young donors and short-time drawing materials. Trabecular meshwork should be accurately separated. Techniques in primary culture, and time and proportion in subculture should be suitabled. Experimental studies should select the third to fifth passages. The growing characteristics, morphological features of cultured HTCs under light and electron microscopes differed from those of corneal and scleral cells adjoining them. Ultramicrostucture distinguished HTCs from corneal interstitial cells. ECM stained by S-P method distinguished HTCs from scleral fibroblast. CONCLUSIONS: These studies suggest that it is not difficult to culture HTCs in general culturing condition as long as the main points about culturing them are known-well. The identification of cultured HTCs must be combined with three aspects: the growing characteristics and morphological features of cultured cells under light and electron microscopes, and the immunohistochemical stained peculiarities of FN, LN and IV C in ECM of cultured cells. To establish the method of culturing HTCs in vitro and to determine identifyng main points concerning cultured HTCs become an essential condition and an important guarantee in order to investigate the pathogenesis and mechanism on primary open angle glaucoma(POAG) deeply.

Cells, Cultured↗
PubMed2001

Isopropyl unoprostone increases the activities of matrix metalloproteinases in cultured monkey ciliary muscle cells.

PURPOSE: The mechanism by which the prostaglandin F2alpha-related antiglaucoma compound isopropyl unoprostone (referred to as unoprostone) reduces intraocular pressure is largely unknown. Another prostaglandin F2alpha-related compound, latanoprost, influences the activities of matrix metalloproteinases in ciliary muscle. Unoprostone ophthalmic solution is metabolized to oxidized metabolites, mainly M1 and M2, in the eye. The aim of this study was to investigate whether intraocular metabolites of unoprostone, M1 and M2, change the metalloproteinase activity in cultured monkey ciliary muscle cells. MATERIALS AND METHODS: Monkey ciliary muscle cells and trabecular meshwork cells were grown separately to confluence in monolayer cell cultures. M1 (10 nM, 100 nM, or 1 microM), M2 (10 nM, 100 nM, or 1 microM), 100 nM prostaglandin F2alpha, or vehicle solutions were added to each culture medium for 48 hours. The media were then assayed to measure metalloproteinase activities quantitatively by means of substrate zymography. RESULTS: Compared with the vehicle controls, M1, M2, and prostaglandin F2alpha significantly increased the metalloproteinase-2 activity in cultured ciliary muscle cells in a dose-dependent manner, but did not affect the metalloproteinase-2 activity in cultured trabecular meshwork cells. All experimented prostaglandins slightly increased metalloproteinase-9 activity in ciliary muscle cells, although these changes were not significant. CONCLUSIONS: The current results show that unoprostone influences the metabolism of the extracellular matrix in the ciliary muscle and that remodeling of the extracellular matrix in the ciliary muscle may be a possible mechanism by which unoprostone ophthalmic solution reduces intraocular pressure.

Animals↗