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Biomedical subjects

D H Geroski

Publications and source records attributed to D H Geroski.

At least 19 recordsLinked to original sources
PubMed2001

Transscleral drug delivery for posterior segment disease.

Exciting new treatments are being developed for retinal degenerations and posterior segment eye disease. The successful treatment of these visually devastating diseases will likely require delivering effective doses of pharmacologic agents to the posterior segment, possibly in conjunction with surgical or genetic interventions. Currently, the treatment of diseases affecting the posterior segment is limited by the difficulty in delivering effective doses of drugs to target tissues in the posterior vitreous, retina or choroid. This review summarizes recent laboratory and clinical studies that indicate that transscleral delivery of therapeutic solutes might be an effective means of achieving therapeutic concentrations of these agents in the posterior eye.

Animals↗
PubMed1999

The effect of intraocular pressure on human and rabbit scleral permeability.

PURPOSE: The purpose of this study was to evaluate the effects of intraocular pressure on the permeability of human and rabbit sclera to water, dexamethasone, and carboxyfluorescein. METHODS: Scleral sections excised from moist-chamber-stored human globes or eyes obtained from euthanatized New Zealand White rabbits were mounted in a perfusion chamber that can create a transscleral pressure that simulates an intraocular pressure. A small depot of drug (100 microl) was added to the episcleral surface while perfusing an irrigating solution slowly across the choroidal side. The perfusate was collected and scleral permeability calculated. Experiments were performed at 0, 15, 30, and 60 mm Hg for each compound in human and rabbit tissue. RESULTS: Analysis of variance showed a significant effect of intraocular pressure on both human and rabbit scleral permeability. Human scleral permeability was decreased by as much as a factor of two for water (P = 0.0004), dexamethasone (P<0.0001), and carboxyfluorescein (P = 0.0064) at elevated intraocular pressures. Rabbit scleral permeability was similarly affected by elevated intraocular pressure for water (P = 0.0039), dexamethasone (P = 0.0001), and carboxyfluorescein (P = 0.0016). CONCLUSIONS: This study shows that simulated intraocular pressure ranging from 15 to 60 mm Hg can decrease scleral permeability to small molecules by one half when compared with the sclera with no pressure applied.

Animals↗
PubMed1998

Human sclera: thickness and surface area.

PURPOSE: To assess the mean thickness and surface area of human sclera. METHODS: Fifty-five formalin-fixed eye bank eyes were hemisected from anterior to posterior. Cross-sectional slides were taken to include a millimeter scale ruler in each photograph. Slide photographs were projected and the scleral silhouette sketched. Mean scleral thickness measurements with standard deviation were obtained. Twenty-five human eye bank eyes were used to determine total scleral surface area by either a computerized tracing method (17 globes) or volumetric calculations (eight globes) using fluid displacement. RESULTS: Mean scleral thickness +/- SD was 0.53 +/- 0.14 mm at the corneoscleral limbus, significantly decreasing to 0.39 +/- 0.17 mm near the equator, and increasing to 0.9 to 1.0 mm near the optic nerve. The mean total scleral surface area by surface area computerized tracings was 16.3 +/- 1.8 cm2 and, by the volume displacement method, was 17.0 +/- 1.5 cm2. CONCLUSIONS: Scleral thickness and surface area measurements from cadaver eyes are important for ophthalmic surgeons and have implications for transscleral diffusion.

Body Surface Area↗
PubMed1998

Effects of laser in situ keratomileusis (LASIK) on the corneal endothelium.

PURPOSE: To assess the effects of laser in situ keratomileusis (LASIK) on the corneal endothelium. METHODS: In a prospective study, the corneal endothelium of 98 eyes of 65 consecutive patients (mean age, 41 years; range, 22 to 66 years) was photographed before, 2 weeks after, and 12 weeks after LASIK for the correction of 2.75 to 14.5 diopters of myopia. Theoretical ablation depths were 200 to 330 microm below the corneal surface. Cell density, coefficient of variation, and percent of hexagonal cells were determined using 150 to 200 cells from each image. Eighty-eight eyes (91%) of 59 patients had a history of contact lens wear. RESULTS: The mean+/-SD preoperative endothelial cell density was 2,549+/-365 cells per mm2, and the mean coefficient of variation was 0.35+/-0.06. There was no statistically significant change in the mean endothelial cell density or mean coefficient of variation of cell size at the 2-week (2,561+/-360 cells per mm2 and 0.35+/-0.06) or 12-week (2,541+/-364 cells per mm2 and 0.35+/-0.05) postoperative examinations. The percent of hexagonal cells was not significantly changed 2 weeks postoperatively; however, 12 weeks postoperatively (P=.0413, two-tailed t test), the percent of hexagonal cells was decreased by 1%. CONCLUSIONS: Corneal endothelial cell density and morphology were unchanged 2 and 12 weeks after LASIK for the correction of up to 14.5 diopters of myopia. In this LASIK study, the correction of up to 14.5 diopters of myopia appears to cause no clinically significant effect on corneal endothelial cell density or morphology.

Adult↗
PubMed1997

Corneal endothelial damage by air bubbles during phacoemulsification.

OBJECTIVE: To characterize the mechanism by which air bubbles damage the corneal endothelium during phacoemulsification. MATERIALS AND METHODS: A series of experiments was conducted to expose the corneal endothelium of New Zealand white rabbit and human eyes that were obtained from an eye bank to air under different conditions. Phacoemulsification at different power settings and irrigation with and without the introduction of air into the anterior chamber were performed. Corneal endothelial perfusion experiments were conducted with air bubbles that were introduced into the perfusion chamber for 2 seconds to 1 hour. Air was also injected into the anterior chambers of anesthetized rabbits for 2 minutes to 3 hours. Corneas were stained with nitrobenzo-xadiazole-phallacidin and examined with fluorescence microscopy. Selected corneas were also examined with scanning and transmission electron microscopy. RESULTS: Intracameral air bubbles during phacoemulsification, irrigation, and perfusion studies resulted in a severe injury to the corneal endothelium in as little as 20 seconds. Intracameral air bubbles in a living rabbit resulted in a slower injury that was morphologically different from the more rapid injury. CONCLUSIONS: Air bubbles in intraocular fluids with a high surface tension can cause a ring-shaped pattern of damage to the corneal endothelium. The mechanism that caused this pattern of damage appears to be a surface tension phenomenon.

Air↗
PubMed1997

Effect of intraocular irrigating solutions on the viability of cultured retinal vascular endothelial cells.

PURPOSE: To compare the abilities of balanced salt solution, BSS Plus and Hartmann's lactated Ringer's (HLR) solution to maintain the viability of retinal vascular endothelial cells (RVEC) in vitro. METHODS: Cultured retinal vascular endothelial cells were suspended in each irrigating solution for four hours. Viability was determined by trypan blue exclusion at 30 minute intervals. Regression analysis was used to determine the rate of viability loss. Additional studies were performed to determine the effectiveness of lactate in maintaining cell viability. RESULTS: Retinal vascular endothelial cells lost viability at a greater rate (p < 0.001) in BSS (8.7%/hr) compared with BSS Plus (3.3%/hr). Cells in Hartmann's lactated Ringer's lost viability at a significantly lower rate (4.4%/hr) than retinal vascular endothelial cells in lactate-free Hartmann's lactated Ringer's solution (8.4%/hr). Lactate was as effective as glucose in preserving RVEC viability. By comparison, the viability of corneal endothelial cells was not effectively maintained by lactate. For these cells, BSS Plus was clearly superior to Hartmann's lactated Ringer's solution in maintaining viability. CONCLUSIONS: BSS Plus and Hartmann's lactated Ringer's solution are both superior to balanced salt solution in maintaining retinal vascular endothelial cell viability. For retinal vascular endothelial cells, Hartmann's lactated Ringer's solution preserves cell viability as well as BSS Plus, since the retinal vascular endothelial cells, unlike corneal endothelial cells, can apparently utilize lactate as an energy source.

Animals↗
PubMed1996

The corneal epithelium after optisol-GS storage.

The objective of this study was to evaluate the epithelium of human corneas stored in Optisol-GS (Chiron Intraoptics, Irvine, CA) for extended periods (2-34 days). Human corneas stored in Optisol-GS (n = 64) were obtained from the Georgia Eye Bank. Corneal epithelial viability was assessed by using the Calcein-AM (Molecular Probes, Inc., Eugene, OR) ethidium homodimer stain, a fluorescent assay used to distinguish live from dead cells. Scanning and transmission electron microscopy was used to evaluate epithelial ultrastructure. The results showed that corneas stored up to 6 days in Optisol-GS had minimal damage of the epithelium. Calcein-AM ethidium homodimer staining showed 20-25% epithelial damage. Corneas stored 7-10 days had a further increase in epithelial damage (30-35%). Corneas stored for 11-15 days had marked increases in epithelial damage (40-50%), and corneas stored 16-34 days showed significant epithelial damage (60-70%). The data show that corneas stored in Optisol-GS are able to maintain the epithelium up to 6 days. A gradual decrease in epithelial viability and loss of epithelial cells occurs in corneas stored 6-10 days. Corneas stored for > 10 days have a marked loss of epithelial cells with extensive epithelial damage.

Cell Survival↗
PubMed1995

Viability of human corneal endothelium following Optisol-GS storage.

OBJECTIVES: To evaluate endothelial viability of human corneas stored in glass vials and in viewing chambers (Alcon) for extended periods, and to compare endothelial viability of Optisol-GS-stored corneas with corneas excised from moist chamber-stored globes. METHODS: Endothelial viability was assessed using two staining techniques. Endothelium from stored corneas was stained with trypan blue combined with alizarin red S or stained with calcein AM-ethidium homodimer. Both techniques were used to determine which method is a more sensitive indicator of cytotoxic change. RESULTS: Corneas stored 4 to 21 days in Optisol-GS had a rate (mean +/- SE) of endothelial cell damage of 0.57% +/- 0.30% per day in vials and 0.69% +/- 0.27% in chambers. After storage intervals from 4 to 21 days, the Optisol-GS endothelium had an average decrease in viability of 9.5% to 16%. The endothelium of moist chamber eyes had a 44% to 59% decrease in viability after 2 to 5 days. After 24 hours, corneal endothelium of moist chamber eyes had less than 15% decrease in viability. Optisol-GS corneas stored for 35 to 56 days had greater than 50% decrease in endothelial viability. After 67 days, 95% to 100% of endothelial viability was lost. CONCLUSIONS: Corneas stored in Optisol-GS through 21 days at 4 degrees C maintain a high percentage of viable endothelial cells. There was no significant difference of endothelial viability between corneas stored in glass vials or in viewing chambers (Alcon). A 50% loss of endothelial viability occurred in moist chamber-stored corneas after 2 days and by 35 days in corneas stored in Optisol-GS.

Aged↗
PubMed1995

Human scleral permeability. Effects of age, cryotherapy, transscleral diode laser, and surgical thinning.

PURPOSE: To determine the in vitro permeability of human sclera to compounds varying in molecular weight. To evaluate the effects of age, cryotherapy, transscleral diode laser, and surgical thinning on scleral permeability. METHODS: Scleral tissue from 97 human eye bank eyes was tested individually in a two-chamber Ussing apparatus with the following hydrophilic radiolabeled compounds on one side of the chamber: 5-fluorouracil, sucrose, dexamethasone, methotrexate, inulin, and three separate dextran polymers (MWt = 10,000, 40,000, and 70,000). Scleral hydration levels were obtained on 20 more scleral specimens. Additional groups of scleral specimens were treated with either a cryotherapy probe, a transscleral diode laser retinopexy probe, or partial thickness lamellar dissection, and specimens were mounted in the Ussing chambers for testing. Scleral tissue was digested to measure the amount of radioactivity present. Scleral sections were examined with electron microscopy. RESULTS: Scleral hydration was maintained during the perfusion. The mean scleral permeability (cm/second x 10(-6) +/- SD) was established for each of the above compounds. Age, cryotherapy, or diode laser treatment did not alter permeability or ultrastructure of the sclera. Surgical thinning significantly increased the scleral permeability to dexamethasone (P = 0.011) and methotrexate (P = 0.037). CONCLUSION: This study establishes baseline human scleral permeability to a series of hydrophilic compounds with various molecular weights. Age, cryotherapy, and diode laser treatment do not alter the permeability or ultrastructure of the sclera, whereas surgical thinning significantly increases permeability.

Adolescent↗
PubMed1994

Corneal endothelial permeability of human tissue after storage in Optisol.

The purpose of this study was to compare Optisol to moist chamber storage for maintaining human corneal endothelial barrier function. Human corneas preserved in Optisol were stored for up to 35 days at 4 C. Endothelial carboxyfluorescein permeability (P(ac)) was measured and endothelial ultrastructure was evaluated by electron microscopy. Endothelial P(ac) (x 10(-4) cm/min) of Optisol-stored corneas was 1.7, 2.0, and 3.1 at five, seven, and 14 days, respectively. The P(ac) increased to 6.5 at 35 days of storage. Endothelial P(ac) in moist chamber stored-eyes was 2.6 at two days, and increased to 13.5 14 days of storage. Multiple regressional analysis showed that storage time and donor age affected P(ac); but time from death to enucleation, time from enucleation to storage, or endothelial cell number did not. Electron microscopy showed that endothelial junctions were maintained through two weeks by Optisol. Large areas of cellular destruction were seen after five days of moist chamber storage. These results show that Optisol can preserve endothelial barrier function through 14 days; barrier function is lost by three days of moist chamber storage.

Adult↗
PubMed1994

Long-term observation of morphologic and functional features of cat corneal endothelium after wounding.

PURPOSE: (1) To test the hypothesis that corneas with enlarged endothelial cells (and thus less intercellular space) have decreased endothelial permeability to small polar solutes. (2) To measure corneal endothelial ouabain binding (Na+/K+ ATPase "pump site" density) and Descemet's membrane production after endothelial wounding. METHODS: Bilateral specular microscopy and anterior segment fluorophotometry were performed at 2-month intervals for 1 year in ten cats after mechanically damaging the corneal endothelium in one eye of each. The measurements were repeated at 2 years in four cats and at 3 years in two cats. Eighteen months after wounding, endothelial ouabain binding was measured in both eyes of six cats. Transmission electron micrographs of Descemet's membrane were analyzed in both eyes of six cats at 18 months, two cats at 2 years, and two cats at 3 years after wounding. RESULTS: From 6 to 12 months after wounding, the endothelial permeability to carboxyfluorescein was significantly decreased (P < 0.05), and the mean endothelial cell size was significantly increased (P < 0.001) in the damaged eyes. The enlarged endothelial cells persisted in the few cats observed 2 and 3 years after wounding. There was no significant difference in endothelial ouabain binding between the damaged and control corneas in six cats tested 18 months after wounding. On subsequent histologic examination, a layer of abnormal Descemet's membrane was present in all ten wounded eyes, with additional normal Descemet's membrane posterior to it, between the abnormal layer and the endothelial cells. CONCLUSIONS: The results are consistent with the hypothesis that corneal endothelial permeability to small polar solutes varies directly with the amount of intercellular space available for diffusion across the monolayer. The results also confirm clinical reports of decreased endothelial permeability in corneas with enlarged endothelial cells. In histopathologic specimens, a layer of abnormal Descemet's membrane can be a historical marker for a period of endothelial damage and corneal decompensation.

Animals↗
PubMed1993

Swelling in the isolated perfused cornea induced by 12(R)hydroxyeicosatetraenoic acid.

PURPOSE: To evaluate the effect of 12(R)hydroxyeicosatetraenoic acid (12(R)HETE) on corneal swelling when directly perfused to human and rabbit corneal endothelium. METHOD: Excised rabbit and human corneas were mounted in the in vitro specular microscope and the endothelium was perfused with 12(R)HETE at 10(-5), 10(-6), and 10(-7) mol/l. Both 12(R)HETE and 12(S)HETE were compared at equal molar (10(-6) mol/l) concentrations. The reversal of 12(R)HETE and ouabain corneal swelling was also compared. Endothelial permeability to carboxyfluorescein was measured after 12(R)HETE perfusion. High-performance liquid chromatographic analysis confirmed that 12(R)HETE remained in the perfusion media. RESULTS: 12(R)HETE caused a dose-dependent corneal swelling of 25 +/- 2, 24 +/- 1, and 14 +/- 0.5 microns/hr at 10(-5), 10(-6), and 10(-7) mol/l, respectively. Equal molar concentrations (10(-6) mol/l) of 12(S)HETE did not cause corneal swelling. Removal of the 12(R)HETE from the perfusion media resulted in reversal of corneal swelling whereas corneal swelling induced by ouabain did not reverse after ouabain removal. 12(R)HETE (10(-6) mol/l) perfused to the human corneal endothelium inhibited temperature reversal corneal thinning when compared to the paired corneal endothelium perfused with BSS Plus (Alcon Laboratories, Inc., Fort Worth, TX). Na/K adenosine triphosphatase activity was inhibited by 10(-6) mol/l ouabain by 35%, 10(-6) mol/l 12(R)HETE by 54%, and 10(-6) mol/l 12(S)HETE by 0.5%. Endothelial permeability to carboxyfluorescein was unaffected by 12(R)HETE. CONCLUSION: 12(R)HETE causes corneal swelling by inhibiting endothelial pump function. This inhibition of transport appears to be at least partly mediated by inhibition of endothelial Na/K adenosine triphosphatase.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1992

Corneal endothelial cytoskeletal changes in F-actin with aging, diabetes, and after cytochalasin exposure.

We investigated the changes in endothelial cytoskeletal F-actin that occur with aging, diabetes, and exposure to cytochalasin D. Rabbit corneas, human donor corneas (with or without polymegethism), and corneas of diabetic individuals were studied. Endothelial F-actin was stained using nitrobenzoxadiazole-phallacidin. Results of these experiments demonstrated that F-actin of the rabbit and human corneal endothelium was arranged in linear circumferential strands that formed a hexagonal array. After in vitro perfusion of cytochalasin D to the corneal endothelium, the F-actin became randomly distributed throughout the cytoplasm, the hexagonal shape of the endothelial cell was disrupted, and endothelial permeability to carboxyfluorescein increased. Changes in F-actin were also observed in the endothelium of the human corneas with polymegethism, and in donor tissue having had previous posterior chamber intraocular lens implantation. The corneas of diabetic individuals also showed marked irregular F-actin fibers crossing the endothelial cell cytoplasm. These abnormal patterns of F-actin may contribute in part to the polymegethism observed in the corneal endothelial cells and may be the result of constant stress in cell volume regulation, particularly in the corneas of diabetic individuals.

Actins↗
PubMed1992

Characterization of corneal endothelium cell cultured on microporous membrane filters.

In these experiments we characterize rabbit and bovine corneal endothelia cell cultured on microporous membrane filters (0.6cm2). Cell cultured bovine or rabbit corneal endothelial cells (subcultures 1-3) were seeded onto Millicell-HA filter inserts. Electrical resistance measured across the cultured monolayers increased steadily through 14 days of culture, reaching 34.2 +/- 0.8 ohm-cm2 (mean +/- SE) for rabbit cells and 33.1 +/- 1.1 ohm-cm2 for bovine cells. Alizarin red staining of the monolayers showed a polygonal morphology comparable to that observed in situ. Transmission electron microscopy showed well developed apical junctional complexes and flaps. Exposure of the monolayers to calcium-free medium resulted in the disruption of intercellular junctions, rounding-up of the cells and a decrease in electrical resistance (to near 0). Transmonolayer fluxes of inulin and dextran correlated well to the resistance measurements. Results of this study demonstrate that corneal endothelium, both bovine and rabbit, grown on filter inserts is comparable in morphology and ultrastructure to corneal endothelium in situ. The cells cultured in this system form functional apical junctional complexes that effect a barrier function comparable to that of the endothelium in situ.

Animals↗
PubMed1992

Comparison of corneal preservation media for corneal hydration and stromal proteoglycan loss.

Changes in the composition of stromal proteoglycans (PGs) have been previously demonstrated in corneal edema, wound healing, and disease. To examine if PGs are lost during corneal preservation, rabbit corneal PGs were radiolabeled in situ with 35S-sulfate and 3H-glucosamine, excised and stored in either modified McCarey-Kaufman medium (MMK), K-Sol (Coopervision-Cilco, Bellevue, WA, U.S.A.), corneal storage medium (CSM), or Dexsol (Chiron Ophthalmics, Irvine, CA, U.S.A.) for up to 14 days. The percentage of total radio-label lost was significantly greater from de-epithelialized corneas (p less than 0.05) and from corneas stored in CSM (p less than 0.05) or K-Sol (p less than 0.05). Corneas stored in CSM for 4 and 7 days were significantly more hydrated than corneas stored in MMK, K-Sol, or Dexsol. After 14 days of storage, all corneas were hydrated above control values with the Dexsol-stored cornea showing the least hydration. Results suggest that loss of stromal PGs during corneal storage is reduced with epithelial integrity and with preservation media containing dextran.

Animals↗
PubMed1991

The developing corneal endothelium: correlation of morphology, hydration and Na/K ATPase pump site density.

The physiology and anatomy of the cornea of the New Zealand white rabbit were studied from birth to young adulthood (3 months). The main objective of the study was to follow the ontogeny of the corneal endothelium and correlate its maturation with the establishment of stromal transparency. With maturity, central corneal thickness increases as do corneal diameter and surface area. Endothelial morphology undergoes marked changes including an increase in cell hexagonality and cell surface area, along with a decrease in cell density and coefficient of variation of cell area. Corneal hydration decreases from a high value at birth to the adult level by 20 days after birth, the time of the onset of stromal transparency. By transmission electron microscopy, corneas of newborn rabbits exhibit an endothelium of irregular cell height with some overlap at the bases of adjacent cells. Apical junctions are incomplete in the neonates. With time the endothelium thins and cells becomes more regular in height, overlap of adjacent cells diminishes, and apical junctions develop. Descemet's membrane is thin in newborns and thickens and becomes more homogenous in appearance with maturation. The abundance of Na/K ATPase pump sites per endothelial cell, as determined by 3H-ouabain binding, increases progressively with age even after the establishment of corneal transparency at 20 days. Scatchard and LIGAND analyses of 3H-ouabain binding data indicate that there is a progressive increase in Bmax with no change in the KD from 7 days to 3 months.

Age Factors↗
PubMed1991

Human corneal storage in modified McCarey-Kaufman and K-Sol media: effect on endothelial Na+/K+ ATPase pump site density and permeability.

Endothelial permeability (Pac) to carboxyfluorescein and Na+/K+ ATPase pump site density were determined in human corneas following storage for 4 or 7 days at 4 degrees C in either modified McCarey-Kaufman (mMK) or K-Sol media. Following 4 days of storage, Pac values for mMK- and K-Sol-preserved corneas were not significantly different from those of their prestorage mates. After 7 days of storage, however, corneas stored in K-Sol media showed a significant increase in Pac compared to their prestorage mates, whereas the mMK-stored corneas showed no change in Pac. Na+/K+ ATPase pump site density determined using [3H]ouabain was similar to a control group in the K-Sol-stored tissue but higher in the mMK-stored tissue following 7 days of storage. These studies suggest that mMK medium maintains endothelial barrier function and Na+/K+ ATPase pump site density at least as well as K-Sol medium through 7 days of corneal storage.

Adult↗