Retinal proteomic changes following unilateral optic nerve transection and early experimental glaucoma in non-human primate eyes.

In this work we compared proteomic changes in non-human primate (NHP) retinas at the onset of early experimental glaucoma (EEG) and 3 weeks after optic nerve transection (ONT), as a means to identify regulators in the retina's response to EEG and ONT. Both eyes of 7 NHPs with either unilateral EEG (n = 4) or ONT (n = 3) were enucleated. Proteins were analyzed by a label-free quantitative mass spectrometry system and the abundance of identified retinal proteins was compared between the treated eye and its contralateral control for each NHP. Cellular processes associated with regulated proteins were identified using the MetaCore program. As a result, a total of 209 and 200 proteins were identified in EEG and ONT retinas, respectively. The EEG eyes exhibited two distinguishable levels of maximum IOP: the highest IOP <27 mmHg (n = 2) and >45 mmHg (n = 2), termed mild IOP EEG and high IOP EEG eyes, respectively. A limited overlap of proteins regulated in the same direction was seen between the high IOP EEG and either the mild IOP EEG eyes or ONT eyes. Most of the proteins that were up regulated in the high IOP EEG eyes were down regulated in the mild IOP EEG eyes; the latter showed an overall down regulation that was not seen in the other two conditions. An association with cytoskeleton regulation was recognized for up-regulated proteins in the high IOP EEG eyes. We conclude that mild IOP EEG, high IOP EEG and ONT retinas exhibited condition-specific proteomic changes with little overlap between conditions. Cytoarchitecture regulation appears to be a component of the early retinal response to chronic experimental IOP elevation.

Mesenchymal stem cells in rheumatoid synovium: enumeration and functional assessment in relation to synovial inflammation level.

OBJECTIVE: Achieving joint regeneration in rheumatoid arthritis (RA) represents a future challenge. Autologous synovial mesenchymal stem cells (MSCs) could be therapeutically exploited. However, the inflammatory milieu in the RA synovium could adversely affect endogenous MSC function. To test this hypothesis, the frequency and multipotency of RA synovial MSCs was evaluated in relation to existing synovial inflammation. METHODS: Synovial inflammation was measured using the arthroscopic visual analogue score (VAS) and further validated using immunohistochemistry and flow cytometry. Highly proliferative clonogenic in vivo MSCs were enumerated following fluorescence-activated cell sorting and expansion for 20 population doublings. MSC multipotency was quantified following standard in vitro culture expansion and trilineage differentiation assays. Real-time PCR, flow cytometry and ELISA were used to evaluate pro- and anti-chondrogenic molecules in standard polyclonal synovial MSCs. RESULTS: The arthroscopic VAS significantly correlated with synovial macrophage infiltration. In RA, synovial MSC chondrogenesis was inhibited in direct relation to VAS (r = -0.777, p<0.05) and reduced compared with control osteoarthritis (OA)-MSCs (p<0.05). In vivo, MSCs resided in the synovial fibroblastic/stromal fraction (CD45(-)CD31(-)) and were reduced in frequency in relation to VAS (r = -0.695, p<0.05). In RA-MSCs, CD44 levels correlated negatively with inflammation and positively with chondrogenesis (r = -0.830 and r = 0.865, respectively). Cytokine production and Sox9 expression was similar in RA-MSCs and OA-MSCs. CONCLUSIONS: There is a negative relationship between synovial MSC chondrogenic and clonogenic capacities and the magnitude of synovitis in RA. Effective suppression of joint inflammation is therefore necessary for the development of autologous MSC treatments aimed at cartilage regeneration in RA.

Abnormal T cell differentiation persists in patients with rheumatoid arthritis in clinical remission and predicts relapse.

OBJECTIVES: An abnormal CD4+ T cell subset related to inflammation exposure (inflammation-related cells, IRC) has been identified in rheumatoid arthritis (RA). Patients with inflammatory and non-inflammatory diseases were used to examine the relationship between inflammation and this T cell subset in vivo. METHODS: Blood was collected from healthy controls and patients with RA (active disease or in clinical remission), Crohn's disease and osteoarthritis. IRC and chemokine receptors were quantified by flow cytometry. Thymic activity and apoptotic factors were measured by real-time polymerase chain reaction. Circulating cytokines were measured by enzyme-linked immunosorbent assay. CXCR4 and SDF1 in synovial biopsies were measured using immunohistochemistry. RESULTS: IRC were identified in patients with RA (p<0.0001) and Crohn's disease (p = 0.005), but not in those with osteoarthritis. In RA in remission, IRC persisted (p<0.001). In remission, hyperproliferation of IRC was lost, chemokine receptor expression was significantly lowered (p<0.007), Bax expression dropped significantly (p<0.001) and was inversely correlated with IRC (rho = -0.755, p = 0.03). High IRC frequency in remission was associated with relapse within 18 months (OR = 6.4, p<0.001) and a regression model predicted 72% of relapse. CONCLUSIONS: These results suggest a model in which, despite the lack of systemic inflammation, IRC persist in remission, indicating that IRC are an acquired feature of RA. They have, however, lost their hyper-responsiveness, acquired a potential for survival, and no longer express chemokine receptors. IRC persistence in remission confirms their important role in chronic inflammation as circulating precursors of pathogenic cells. This was further demonstrated by much higher incidence of relapse in patients with high IRC frequency in remission.

A cellular solid model of the lamina cribrosa: mechanical dependence on morphology.

The biomechanics of the optic nerve head (ONH) may underlie many of the potential mechanisms that initiate the characteristic vision loss associated with primary open angle glaucoma. Therefore, it is important to characterize the physiological levels of stress and strain in the ONH and how they may change in relation to material properties, geometry, and microstructure of the tissue. An idealized, analytical microstructural model of the ONH load bearing tissues was developed based on an octagonal cellular solid that matched the porosity and pore area of morphological data from the lamina cribrosa (LC). A complex variable method for plane stress was applied to relate the geometrically dependent macroscale loads in the sclera to the microstructure of the LC, and the effect of different geometric parameters, including scleral canal eccentricity and laminar and scleral thickness, was examined. The transmission of macroscale load in the LC to the laminar microstructure resulted in stress amplifications between 2.8 and 24.5xIOP. The most important determinants of the LC strain were those properties pertaining to the sclera and included Young's modulus, thickness, and scleral canal eccentricity. Much larger strains were developed perpendicular to the major axis of an elliptical canal than in a circular canal. Average strain levels as high as 5% were obtained for an increase in IOP from 15 to 50 mm Hg.

Early rheumatoid arthritis is associated with a deficit in the CD4+CD25high regulatory T cell population in peripheral blood.

OBJECTIVE: Our aim was to test the hypothesis that there is a deficit in the CD4+CD25high regulatory T-cell population in early rheumatoid arthritis (RA), either in size or functional activity. METHODS: Peripheral blood mononuclear cells were examined from subjects with early active RA who had received no previous disease-modifying therapy (n = 43), from individuals with self-limiting reactive arthritis (n = 14), from subjects with stable, well-controlled RA (n = 82) and from healthy controls (n = 72). The frequencies of CD4+CD25high T-cells were quantified using flow cytometry, and function was assessed by the ability to suppress proliferation of CD4+CD25- T-cells. Paired blood and synovial fluid was analysed from a small number of RA and reactive arthritis patients. RESULTS: There was a smaller proportion of CD4+CD25high T-cells in the peripheral blood of early active RA patients (mean 4.25%) than in patients with reactive arthritis or in controls (mean 5.90 and 5.30%, respectively, P = 0.001 in each case). Frequencies in stable, well-controlled RA (mean 4.63%) were not significantly different from early active RA or controls. There were no differences in suppressor function between groups. Higher frequencies of CD4+CD25high T-cells were found in synovial fluid than blood in both RA and reactive arthritis. CONCLUSIONS: These data demonstrate a smaller CD4+CD25high regulatory T-cell population in peripheral blood of individuals with early active RA prior to disease-modifying treatment. This may be a contributory factor in the susceptibility to RA and suggests novel approaches to therapy.

Image analysis of optic nerve disease.

Existing methodologies for imaging the optic nerve head surface topography and measuring the retinal nerve fibre layer thickness include confocal scanning laser ophthalmoscopy (Heidelberg retinal tomograph), optical coherence tomography, and scanning laser polarimetry. For cross-sectional screening of patient populations, all three approaches have achieved sensitivities and specificities within the 60-80th percentile in various studies, with occasional specificities greater than 90% in select populations. Nevertheless, these methods are not likely to provide useful assistance for the experienced examiner at their present level of performance. For longitudinal change detection in individual patients, strategies for clinically specific change detection have been rigorously evaluated for confocal scanning laser tomography only. While these initial studies are encouraging, applying these algorithms in larger numbers of patients is now necessary. Future directions for these technologies are likely to include ultra-high resolution optical coherence tomography, the use of neural network/machine learning classifiers to improve clinical decision-making, and the ability to evaluate the susceptibility of individual optic nerve heads to potential damage from a given level of intraocular pressure or systemic blood pressure.

Anterior scleral canal geometry in pressurised (IOP 10) and non-pressurised (IOP 0) normal monkey eyes.

AIMS: To characterise lamina cribrosa and anterior scleral canal wall architecture in pressurised (IOP 10 mm Hg) and non-pressurised (IOP 0 mm Hg) normal monkey eyes. METHODS: Eight normal eyes from eight monkeys were enucleated before sacrifice and the optic nerve heads (ONH) trephined and immersion fixed in glutaraldehyde (IOP 0). Nine normal eyes from nine monkeys were perfusion fixed in situ with paraformaldehyde at IOP 10 mm Hg (IOP 10), and the ONHs trephined and stored in glutaraldehyde. Each ONH specimen was embedded in glycol methacrylate and cut into vertical or horizontal, 4 micro m thick, serial sections. Within digitised images of every sixth section, anterior laminar position and laminar thickness were measured at nine evenly spaced locations across the scleral canal opening. Additionally, scleral canal diameters at Bruch's membrane (SCD-B) and at the anterior laminar insertion (SCD-ALI) were measured within the 15 middle section images of each vertically sectioned ONH. RESULTS: Anterior laminar position was significantly more anterior (nearer Bruch's membrane) in the IOP 10 eyes, compared with the IOP 0 eyes (116 (+/-95% CI; 2) micro m v 184 (2) micro m, respectively). Also in the IOP 10 eyes, the lamina cribrosa was thinner (195 (2) micro m v 264 (2) micro m) and the scleral canal diameter was larger (SCD-B: 1751 (23) micro m v 1591 (19) micro m; SCD-ALI: 1961 (21) micro m v 1717 (17) micro m), compared with the IOP 0 eyes. CONCLUSION: The anterior scleral canal wall is expanded and the lamina cribrosa is thinned and more tautly stretched within pressurised (perfusion fixed at IOP 10) young monkey eyes, compared with non-pressurised (immersion fixed at IOP 0) young monkey eyes. The constricted scleral canal and the relaxed and thickened lamina in the non-pressurised eyes may represent phenomena that contribute to optic disc swelling in hypotonous eyes.

Fluvastatin induces apoptosis of vascular endothelial cells: blockade by glucocorticoids.

Statins block de novo synthesis of cholesterol by inhibiting the enzyme, HMG CoA reductase. The product of this reaction, mevalonic acid, is also a precursor of isoprenoids, molecules required for the activation of signaling G-proteins, such as Ras. Signal transduction pathways involving Ras are important for cell survival and this may be why statins induce apoptotic death of several cell types. Given that statins are used to treat vascular disease, surprisingly no studies have been conducted on vascular endothelial cells. Here we show that fluvastatin (FS), at concentrations from 1-2 microM, blocks growth and induces apoptosis of the endothelial cell line, EA.hy 926. Considerable redundancy is known to exist in cell signaling and in vivo toxicity of FS might be prevented by other signaling pathways, like those activated by adrenal or sex steroids. RT-PCR analysis revealed the expression of the androgen and glucocorticoid receptor in EA.hy 926 cells. Although the androgen, dihydrotestesterone (DHT) had no effect, the glucocorticoid, dexamethasone (Dex), blocked FS-induced apoptosis. Cell cycle analysis revealed that 24 h exposure to FS prevented cells from leaving G(1) and 24-48 h later a marked sub-G(1) peak was observed. Dex was able to reduce the sub-G(1) peak, but it failed to block accumulation of cells in G(1), indicating that it's effect was specific for blockade of apoptosis, and not specific to an effect on FS alone. This study strongly suggests that glucocorticoids have a role to play in preventing vascular injury and they may provide the reason why statins are not inherently toxic to vascular endothelial cells, in vivo.

Statin-induced apoptosis of vascular endothelial cells is blocked by dexamethasone.

Statins block de novo synthesis of cholesterol by inhibiting the enzyme, HMG CoA reductase. The product of this reaction, mevalonic acid, is also a precursor of isoprenoids, molecules required for the activation of signalling G-proteins, such as Ras. Signal transduction pathways involving Ras are important for cell survival and this may be why statins induce apoptotic death of several cell types. Given that statins are used to treat vascular disease, it is surprising that no studies have been conducted on vascular endothelial cells. For this reason, we have tested the effect of fluvastatin (FS) on the endothelial cell line EA.hy 926. Here we show that FS, at concentrations from 1 to 2 microM, blocks growth and induces apoptosis of the endothelial cell line, EA.hy 926. As considerable redundancy exists in cell signalling pathways for cell survival, toxicity of FS under more physiological conditions might be prevented by pathways that do not require Ras, such as those activated by adrenal or sex steroids. To test this hypothesis, first RT-PCR analysis was performed for nuclear receptor mRNA expression. This revealed the presence of mRNA for the androgen receptor (AR) and glucocorticoid receptor (GR). The effect of the AR agonist, dihydrotestosterone (DHT), and the GR agonist, dexamethasone (Dex), was then tested. Whilst DHT (100 nM) had no effect on FS-induced cell death, Dex (1 microM) blocked FS-induced apoptosis. Cell cycle analysis revealed that 24 h exposure to FS prevented cells from leaving G(1) and 24-48 h later a marked sub-G(1) peak was observed. Dex was able to reduce the sub-G(1) peak, but it failed to reduce accumulation of cells in G(1). Further studies revealed that, in addition to blocking FS-induced apoptosis, Dex was able to block apoptosis of EA.hy 926 cells induced by serum deprivation, tumour necrosis factor-alpha, oxidants, DNA damage and mitochondrial disruption. This study strongly suggests that glucocorticoids have a role to play in preventing vascular injury and they may provide a reason why statins are apparently not toxic to vascular endothelial cells in vivo.

Posterior scleral thickness in perfusion-fixed normal and early-glaucoma monkey eyes.

PURPOSE: To characterize posterior scleral thickness in the normal monkey eye and to assess the effects of acute (15- to 80-minute) and short-term chronic (3- to 7-week) intraocular pressure (IOP) elevations. METHODS: Both eyes of four normal monkeys (both eyes normal) and four monkeys with early glaucoma (one eye normal and one eye with induced chronic elevation of IOP) were cannulated. In each monkey, IOP was set to 10 mm Hg in the normal eye and 30 or 45 mm Hg in the contralateral eye (normal or early glaucoma) for 15 to 80 minutes. All eight monkeys were perfusion fixed, yielding eight low IOP-normal eyes, four high IOP-normal eyes, and four high IOP-early glaucoma eyes. Posterior scleral thickness was measured histomorphometrically at 15 measurement points within each eye, and the data were grouped by region: foveal, midposterior, posterior-equatorial, and equatorial. RESULTS: Overall, posterior scleral thickness was significantly different in the various regions and among the treatment groups (P < 0.0001). In the low IOP-normal eyes, the posterior sclera was thickest in the foveal region (307 microm) and thinner in the midposterior (199 microm), posterior-equatorial (133 microm), and equatorial (179 microm) regions. In the high IOP-normal and high IOP-early glaucoma eyes, the posterior sclera was thinner both overall and within specific regions, compared with the low IOP-normal eyes. CONCLUSIONS: The posterior sclera in the perfusion-fixed normal monkey eye thins progressively from the fovea to the equator and is thinnest just posterior to the equator. Acute and short-term chronic IOP elevations cause regional thinning within the posterior sclera of some monkey eyes, which significantly increases stresses in the scleral wall.

Slow release of the antimetabolite 5-fluorouracil (5-FU) from modified Baerveldt glaucoma drains to prolong drain function.

Drainage devices are routinely placed in the eyes of patients with glaucoma to reduce intraocular pressure (IOP) by providing controlled outflow of fluid (aqueous humor) via a filtering bleb. However, the natural wound healing response often interferes with fluid outflow by thickening the walls of the bleb over time, so that these devices rarely remain functional for more than 5 years. We investigated the use of controlled release of an antimetabolite, 5-fluorouracil (5-FU), within glaucoma drains to determine if the wound healing response could be reduced and the useful life span of the device increased. Collagen plugs containing 1.125 mg of 5-FU were placed in the silicone tubes of modified Baerveldt glaucoma drains. Eight drains with 5-FU and eight drains without 5-FU were implanted in one eye each of 16 New Zealand white rabbits: the contralateral eyes served as unoperated controls. Results were evaluated in terms of IOP, fibrous capsule thickness, macrophage density. and presence of type III collagen surrounding the drain plate, 3 and 6 months after implantation. In general, eyes implanted with antimetabolite-containing drains demonstrated significantly lower values for all evaluated parameters at 3 months and lower or equal values at 6 months, compared with the eyes not receiving 5-FU and the unoperated controls, indicating improved IOP-lowering function, reduced bleb wall thickness, and earlier achievement of a steady-state wound healing response. All eyes remained healthy throughout the 6-month duration of the study with no cytotoxicity complications in any of the eyes. Thus, biodegradable plugs placed within the silicone tubes of glaucoma drains can safely deliver 5-FU to filtering blebs over time, which could prolong the functional life of the bleb by decreasing the thickness of the anterior fibrous capsule and permitting sufficient fluid outflow to reduce IOP to physiological levels.

Optic disc surface compliance testing using confocal scanning laser tomography in the normal monkey eye.

PURPOSE: To determine the effect of acute, experimentally increased intraocular pressure on deformation of the surface of the optic nerve head (optic nerve head surface compliance testing) in normal monkey eyes using confocal scanning laser tomography. METHODS: A total of 156 compliance tests were performed on 48 normal eyes of 30 monkeys in three separate studies. Compliance testing involved obtaining confocal scanning laser tomographic images using a 10 degrees and/or 15 degrees and/or 20 degrees scan angle at various times after intraocular pressure was raised from 10 to 30 or 45 mm Hg. At each point, six images were analyzed to provide a value for a parameter, called mean position of the disc, which was used to express the amount of deformation the surface of the optic nerve head had undergone at that point. Statistical analysis (ANOVA) was performed to evaluate differences in the amounts of deformation in individual eyes at different intraocular pressures and at different compliance testing sessions (studies 1 and 2) and in the two eyes of individual monkeys under the same conditions (study 3). RESULTS: The majority of eyes showed posterior deformation of the surface of the optic nerve head ranging from 15 to 86 microm as early as 10 minutes after intraocular pressure was increased from 10 to 30 mm Hg. When pressure was increased from 30 to 45 mm Hg in a subset of these eyes, most showed additional deformation. Of the 12 eyes for which both 15 degrees and 20 degrees images were obtained at the same compliance test, 7 showed larger amounts of deformation in the 20 degrees images. Of the 18 monkeys tested in both eyes, 12 showed some differences and 4 showed substantial differences between the two eyes. CONCLUSIONS: In the normal monkey eye, the surface of the optic nerve head deforms rapidly (in as few as 10 minutes) in response to increased intraocular pressure. The amount of deformation varies between subjects and even within the two eyes of individual monkeys. Increasing the scan angle from 15 degrees to 20 degrees frequently increases the amount of deformation detected, suggesting that the peripapillary sclera and the optic nerve head may be involved in the deformation in some eyes.

Expanded polytetrafluoroethylene reinforcement material in glaucoma drain surgery.

PURPOSE: To determine whether a synthetic material, expanded polytetrafluoroethylene (E-PTFE), can be used successfully as a reinforcement material over the tubes of glaucoma drainage implants. METHODS: Patches of E-PTFE were sutured over the tubes of Baerveldt glaucoma drains implanted in the eyes of New Zealand white rabbits. Two material thicknesses were tested: 0.5 mm in four eyes and 0.25 mm in five eyes. Rabbit donor scleral patches were used in five eyes as the control. Total ocular health and intraocular pressure were monitored every 2 weeks after the procedure. Six months after implantation, the eyes were harvested and analyzed histologically. RESULTS: Two of the four eyes that received 0.5-mm thick E-PTFE patches showed some conjunctival melting over the anterior corners of the material close to the limbus. All five eyes that received 0.25-mm thick E-PTFE patches showed a healthy cellular wound healing response and no conjunctival melting. Cellular infiltration and collagen deposition in the E-PTFE materials showed integration of the patch material into the surrounding tissue. In the control eyes, marked thinning and resorption of the donor sclera immediately above the drainage tube was noted. CONCLUSION: Thin (0.25 mm) E-PTFE patches were well tolerated in all rabbit eyes tested. Thin E-PTFE should be investigated further as a functional alternative to donor sclera for reinforcement in glaucoma drain surgery.

The optic nerve head as a biomechanical structure: initial finite element modeling.

PURPOSE: To study the relationship between intraocular pressure (IOP) and the IOP-related stress (force/cross-sectional area) it generates within the load-bearing connective tissues of the optic nerve head. METHODS: Thirteen digital, three-dimensional geometries were created representing the posterior scleral shell of 13 idealized human eyes. Each three-dimensional geometry was then discretized into a finite element model consisting of 900 constituent finite elements. In five models, the scleral canal was circular (diameters of 0.50, 1.50, 1.75, 2.00, and 2.56 mm), with scleral wall thickness (0.8 mm) and inner radius (12.0 mm) held constant. In three models, the canal was elliptical (vertical-to-horizontal ratios of 2:1 [2.50 x 1.25 mm], 1.5:1 [2.1 x 1.4 mm], and 1.15:1 [1.92 x 1.67 mm]), with the same constant scleral wall thickness and inner radius. In five additional models, scleral canal size was held constant (1.92 x 1.67 mm), and either scleral wall thickness (three models, 0.5, 1.0, and 1.5 mm) or inner radius (two models, 13.0 and 14.0 mm) was varied. In all models, each finite element was assigned a single isotropic material property, either scleral (modulus of elasticity, 5500 kPa) or axonal (modulus of elasticity, 55 kPa). Maximum stresses within specific regions were calculated at an IOP of 15 mm Hg (2000 Pa). RESULTS: Larger scleral canal diameter, elongation of the canal, and thinning of the sclera increased IOP-related stress for a given level of IOP. For all models, maximum IOP-related stress ranged from 6 x IOP (posterior sclera) to 122 x IOP (laminar trabeculae). For each model, maximum IOP-related stress was highest within the laminar trabecular region and decreased progressively through the laminar insertion, peripapillary scleral, and posterior scleral regions. Varying the inner radius had little effect on the maximum IOP-related stress within the scleral canal. CONCLUSIONS: Initial finite element models show that IOP-related stress within the load-bearing connective tissues of the optic nerve head is substantial even at low levels of IOP. Although the data suggest that scleral canal size and shape and scleral thickness are principal determinants of the magnitude of IOP-related stress within the optic nerve head, models that incorporate physiologic scleral canal and laminar geometries, a more refined finite element model meshwork, and nonisotropic material properties will be required to confirm these results.

Functional inactivation of the oestrogen receptor by the antioestrogen, ZM 182780, sensitises tumour cells to reactive oxygen species.

Reactive oxygen species (ROS) play a fundamental role in both apoptotic and necrotic cell death. Their importance is highlighted by studies showing that they mediate cell death in response to radiotherapy and to some forms of chemotherapy. Here we provide the first evidence for a role of ROS in response to an antiendocrine agent currently undergoing clinical trials. Using the oestrogen receptor (ER) containing rat pituitary GH3 cell line, we show that cell death is induced by the pure steroidal antioestrogen, ZM 182780, and that this is blocked by the antioxidant, N-acetyl cysteine (NAC). By flow cytometry, we show that, prior to the onset of DNA breakdown measured by ELISA, ZM 182780 exposure has no significant effect on intracellular oxidant concentrations. In contrast, ZM 182780 exposure greatly increases sensitivity to oxidants generated by blocking cellular antioxidant pathways and from exogenous administration of hydrogen peroxide (H2O2). As both necrosis and apoptosis are controlled by mitochondrial function, further experiments conducted to determine mitochondrial membrane potential (Delta|gWm) have indicated that the ZM 182780-induced loss of ER function increases the ease with which oxidants collapse mitochondrial activity and, as a consequence, cell death.

Biocompatibility response to modified Baerveldt glaucoma drains.

Glaucoma implants are designed to increase fluid outflow from the eye in order to decrease intraocular pressure and prevent damage to the optic nerve. The implant consists of a silicone tube that is inserted into the anterior chamber at one end and is attached at the other end to a silicone plate that is sutured to the outside of the globe beneath the conjunctiva. The glaucoma "implant" becomes a "drain" over the first 3 to 6 postoperative weeks as the silicone plate is enclosed by a fibrous capsule that allows a space to form into which fluid can drain and from which fluid can be absorbed by the surrounding tissues. Ideally, the size and thickness of the capsule (the filtering bleb) that surrounds the plate is such that the amount of fluid that passes through the capsule is identical to the amount of fluid produced by the eye at an intraocular pressure of 8 to 14 mmHg. The most common long-term complication of these implants is failure of the filtering bleb 2 to 4 years after surgery due to the formation of a thick fibrous capsule around the device. Micromovement of the smooth drainage plate against the scleral surface may be integral to the mechanism of glaucoma implant failure by stimulating low-level activation of the wound healing response, increased collagen scar formation, and increased fibrous capsule thickness. To test this hypothesis, we modified seven Baerveldt implants by adding porous cellular ingrowth material to the posterior surface of the drainage plate. Seven modified and five unmodified implants were placed in adult rabbit eyes. After 6 months, we found that the fibrous capsule around the modified implants was significantly thinner than the capsule surrounding the unmodified implants (p < 0.05), particularly on the surface between the porous ingrowth material and the sclera (p < 0.05). Although type I collagen predominated in the fibrous capsules around both types of implants, the amount of type III collagen in the capsules around the modified implants was significantly less than the amount around the unmodified implants (p < 0.05). We believe that these data suggest a reduction in the wound healing response to the modified implants, with greater stability of capsule thickness. Long-term studies are needed to verify that the stability of the capsules around the modified implants persists over a period of years, in which case this type of modification may prove useful in prolonging the functional life of these devices in the surgical treatment of glaucoma.

Measurement of optic disc compliance by digitized image analysis in the normal monkey eye.

PURPOSE: To characterize the compliance of the normal monkey optic disc under conditions of induced short-term fluctuations in intraocular pressure (IOP). METHODS: In 10 monkeys, one eye was compliance tested on three separate days followed by a single test of the contralateral eye (40 compliance tests). In a testing session, the optic disc was imaged at 2 and 47 minutes (baseline time point) after IOP was lowered to 10 mmHg; then at 2, 17, 32, and 47 minutes after IOP was elevated to 45 mmHg; then at 2, 47, and, in some cases, 92 minutes after IOP was lowered back to 10 mmHg. Eight digitized images were analyzed at each time point, yielding two parameters to characterize the position of the disc: the Mean Position of the Disc (MPD) and the Change from MPDBaseline (the value of MPD at a given time point minus the value for MPD at the baseline time point of that testing session). Analysis of variance (ANOVA) testing was used to evaluate the overall effect of IOP on both parameters while taking into account the effects of variability due to different monkeys and repetitions of the test as well as differences between the two eyes of an individual monkey. With the addition of data from 11 compliance tests performed on eight additional monkeys, the overall results were calculated in terms of the mean Change from MPDBaseline at each time point for a total of 51 compliance testing sessions. RESULTS: The mean Change from MPDBaseline was -28 microns (95% confidence interval, -23 to -33 microns) 47 minutes after elevation of IOP. The disc surface returned to its baseline position 92 minutes after IOP was lowered back to 10 mmHg. Elevation of IOP within a compliance test had a significant effect on the position of the optic disc surface (P = 0.0002, ANOVA), as characterized by the parameter Change from MPDBaseline. Neither the difference in the amount of movement between the two eyes of an individual monkey nor the variability within the three repetitions of the test in a given eye was statistically significant. CONCLUSION: Small, reversible (elastic) posterior deformations of the optic disc surface follow acute elevations of IOP in the normal monkey eye. Detection of acute IOP-induced deformations of the optic disc surface may represent a means by which to mechanically test the deeper load-bearing tissues of the optic nerve head.

Early changes in optic disc compliance and surface position in experimental glaucoma.

PURPOSE: To detect changes in the compliance and baseline position (position at the baseline time point of a compliance test) of the monkey optic disc after the onset of chronic experimental glaucoma. METHODS: Sixty-six compliance tests were performed on 26 eyes of 13 monkeys. Longitudinal Study. In seven normal monkeys, compliance tests were performed three times in one eye (study eye) and once in the contralateral eye. In the study eye of five of these monkeys, chronic experimental glaucoma was then induced and compliance tests were performed at some or all of the following postglaucoma testing intervals: 1 to 2 weeks, 3 to 4 weeks, 5 to 8 weeks, 9 to 12 weeks, 13 to 18 weeks, and more than 18 weeks after the onset of elevated intraocular pressure (IOP). In the study eye of the remaining two monkeys, the optic nerve was transected, and compliance was tested at 5, 9, and 13 weeks after transection. An analysis of variance (ANOVA) was performed to detect an increase (hypercompliance) or decrease (rigidity) in the compliance of the glaucomatous eyes at each testing interval. A second ANOVA was performed to detect the onset of chronic posterior deformation of the baseline position of each disc. Cross-Sectional Study. In six additional monkeys with pre-existing experimental glaucoma, the glaucomatous study eye was compliance tested at one of the postglaucoma testing intervals used in the longitudinal study. The contralateral normal eye was compliance tested once. These data were then added to the data from the five longitudinally studied monkeys at the appropriate preglaucoma and postglaucoma testing intervals. A third ANOVA was done to compare the compliance of the expanded group of glaucomatous eyes at each postintervention testing interval with the compliance of the 13 normal contralateral eyes. RESULTS: Compliance. In the longitudinally (Pr > F = 0.0005) and cross-sectionally (Pr > F = 0.0001) studied glaucomatous eyes, optic disc compliance increased significantly by 1 to 2 weeks and then returned to a level statistically indistinguishable from normal within 13 to 18 weeks after the onset of glaucoma. In the transection eyes, the optic discs were significantly less compliant (more rigid) at 5 and 9 weeks after transection compared with the discs in either the normal or the glaucomatous eyes (Pr > F < 0.05). Baseline Optic Disc Position. Chronic posterior deformation of the disc was detected in one of three eyes tested 1 to 2 weeks and three of four eyes tested 3 to 4 weeks after the onset of glaucoma (Pr > F < 0.05). Chronic posterior deformation was not detected in the discs of either of the transection eyes at any of the post-transection testing intervals. CONCLUSION: Changes in optic disc compliance and surface position were detected by digitized image analysis within 2 to 4 weeks of the onset of experimental glaucoma in the monkey eye. These findings are unlikely to be due to axon loss alone, because they did not occur in optic nerve transection eyes (which constitute a model of axon loss in which intraocular pressures remain normal). The results suggest that IOP-related damage to the load-bearing connective tissues of the optic nerve head may occur early in the course of experimental glaucoma.