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Biomedical subjects

H F Edelhauser

Publications and source records attributed to H F Edelhauser.

At least 37 records · Page 2Linked to original sources

The effects of intraocular lidocaine on the corneal endothelium.

OBJECTIVE: The study aimed to evaluate the direct effect of intraocular lidocaine hydrochloride (HCl) 1% on corneal endothelial cell function, ultrastructure, and viability using an in vitro perfusion specular microscope system. DESIGN: Paired rabbit and human corneas were isolated and mounted in an in vitro specular microscope for endothelial perfusion evaluation. Corneas were perfused with a control solution (BSS Plus for humans, glutathione bicarbonate Ringer's [GBR] for rabbits) for a 1-hour stabilization period. After the stabilization period, one cornea of each matched pair was perfused with preservative-free lidocaine HCl 1% for 15 minutes followed by control solution for an additional 2 to 3 hours. The control cornea continued to receive either GBR or BSS Plus. Corneal thickness measurements were taken every 15 minutes throughout the perfusion period. Corneal swelling and deswelling rates were calculated by linear regression analysis. At the end of the experiment, corneas were fixed for scanning and transmission electron microscopy. In another group of corneas, the endothelial viability was assayed after direct perfusion with lidocaine HCl 1%. RESULTS: Lidocaine HCl 1% caused endothelial cell edema, which reversed on removal of lidocaine from perfusion media. Corneal swelling and deswelling rates did not differ significantly between the lidocaine and control groups. Electron microscopy showed the effects of transient endothelial cell edema with an otherwise normal mosaic pattern and ultrastructure for both treatment groups. Endothelial cell viability was maintained after the direct lidocaine exposure and a 2-hour washout. CONCLUSIONS: Lidocaine HCl 1% causes a transient endothelial cell edema to the in vitro perfused endothelium of human and rabbit corneas. Proper attention should be given to the type of lidocaine injected intraocularly (i.e., concentration, vehicle, preservatives, pH, osmolarity). Although lidocaine HCl 1% appears to be safe to both human and rabbit endothelium during short-term in vitro exposure, further in vivo and in vitro studies are needed to determine long-term effects of intraocular lidocaine on the corneal endothelium.

Aged↗

Comparison of rabbit and human corneas stored in Optisol-GS: changes in stromal sodium.

PURPOSE: To evaluate the hydration, and the levels of free, total and bound sodium in fresh rabbit corneal stromas and also those preserved for up to 21 days in Optisol-GS. The effect of epithelial removal on stromal sodium and hydration parameters was also evaluated. Trends in stromal hydration and sodium environment were compared to results we previously obtained using human eyes stored under identical conditions. METHODS: Stromal hydration was evaluated thermogravimetrically. A sodium-specific electrode and an atomic absorption spectrophotometer were used to determine the amounts of free and total stromal sodium, respectively. In one cornea of each pair, the epithelium was removed prior to placement in the storage media. After 3, 7, 14 or 21 days at 4 degrees C, corneas were removed from the Optisol-GS, at which time sodium and hydration measurements were obtained. RESULTS: With an intact epithelium, the hydration of the rabbit stromas was elevated significantly at each day of storage compared to fresh corneas. Free and total sodium levels of rabbit stromas did not differ statistically from fresh values, however the bound sodium values did increase during storage. In the absence of the epithelium, the stromal hydration and sodium content (free, total and bound) were significantly elevated and the increase was much greater than in corneas stored with an intact epithelium. These findings differ from those we measured previously using human tissue. CONCLUSIONS: Rabbit corneas responded differently from human corneas to storage in Optisol-GS. The hydration levels increased to a greater level in rabbit than human corneas under both storage conditions. The trends in amounts of both free and total sodium were similar between the species, although the absolute amounts differed. The largest discrepancy was observed in the amount of bound sodium, with the rabbit corneas experiencing large increases not documented in the human tissue. These results suggest that direct comparisons of stromal hydration and ionic environment between the species should be approached with caution.

Animals↗

Keratocyte repopulation in epikeratoplasty specimens.

PURPOSE: To study the histology and pattern of keratocyte repopulation of surgically removed human epikeratoplasty lenticules. METHODS: Removed epikeratoplasty lenticules and penetrating keratoplasty buttons that contained epikeratoplasty lenticules were evaluated for duration of epikeratoplasty, histologic and ultrastructural features, and average number of keratocytes per high-power microscopic field. The keratocyte density was compared with age-matched controls. RESULTS: Fifteen epikeratoplasty specimens from eight penetrating keratoplasties and seven removed lenticules were reviewed. The indications for keratoplasty were myopia, keratoconus, and aphakia. The lenticules were in place for 7-120 months, and the keratocyte count ranged from 14 to 40 per high-power field. Keratocyte density increased to 30-40 per high-power field, similar to age-matched controls, at approximately 48 months postoperatively, similar to the density of the controls. Keratocytes appeared to have migrated from the periphery to the center of the lenticules. CONCLUSIONS: Normal keratocyte density in epikeratoplasty lenticules is reached by approximately 48 months after surgery.

Adult↗

How might 12 (R) HETE cause the inhibition of Na,K-ATPase?

PURPOSE: 12 (R) hydroxy 5,8,10,14-eicosatetraenoic acid [12 (R) HETE] is a potent inhibitor of Na,K-ATPase. This study was an attempt to determine how the eicosanoid might inhibit the enzyme by using molecular modeling. METHODS: Models were generated using the program HyperChem 2.0 for Windows. Models of 12 (R) HETE, 12 (S) HETE (the "S" isomer of 12 (R) HETE), and 8 (R) hydroxy-hexadecatrienoic acid [8 (R) HHDTrE, a catabolic isomer of 12 (R) HETE] were formed and docked with phosphatidyl choline and the H3-H4 peptide of the alpha-subunit of Na,K-ATPase. In addition, models of 12 (R) HETE, and related compounds, were formed and complexed with calcium, and then docked with phosphatidyl choline. The energies of stabilization were calculated for each optimal docking. RESULTS: Optimal steric fitting and calculated energies of stabilization indicated that 12 (R) HETE and 8 (R) HHDTrE had the best fits when bound to the fatty acid portions of phosphatidyl choline. However, when Ca-HETE complexes were modeled, it was found that they formed even more stable complexes when bound to phosphatidyl choline. Calculated energies of 12 (S) HETE, whether complexed to calcium or not, were less favorable than the other HETE compounds. CONCLUSIONS: The results of the study indicate that plasma membrane lipids rather than Na,K-ATPase itself are more likely to be bound by 12 (R) HETE and its related compounds. Moreover, it was found that the calcium complexes of 12 (R) HETE and 8 (R) HHDTrE are even more likely to dock with plasma membrane lipids. This suggests that such complexes may be able to transport calcium into the cell and make it available for the inhibition of Na, K-ATPase at the enzyme's sodium binding site.

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

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↗

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↗

Pupillary response to tropicamide in patients with Alzheimer disease.

PURPOSE: To determine whether pupillary responses to dilute tropicamide could be used as a diagnostic test for Alzheimer disease (AD). The authors also investigated whether concurrent use of an oral acetylcholinesterase inhibitor (tacrine) alters the pupillary response to dilute tropicamide in patients with AD, and whether pupillary responses to dilute tropicamide differ in young versus older control subjects. METHODS: Pupillary diameter and area of both eyes were measured in light and darkness, at 10-minute intervals for 40 minutes after random instillation of 0.01% tropicamide to one eye. Four groups of subjects were studied: 9 patients with AD, 10 who were treated with tacrine, 11 older control subjects, and 10 young control subjects. RESULTS: Mean change in anisocoria was not significantly different among groups at any of the measurement time points. Mean percent change in diameter of the treated eyes showed a trend toward faster maximum dilatation in the AD groups, but change in pupillary measurements did not identify individuals with AD. CONCLUSION: Pupillary response to dilute tropicamide did not effectively distinguish individual patients with AD from young or older control subjects.

Adult↗

Long term changes in human corneal endothelium following toxic endothelial cell destruction: a specular microscopic and fluorophotometric study.

AIMS: To investigate the long term relation between corneal thickness, endothelial morphometric variables, and endothelial permeability in patients with endothelial cell counts under 900 cells/mm2 as a result of endothelial cell destruction after cataract surgery. METHODS: Eighteen patients developed the so called toxic endothelial cell destruction (TECD) syndrome following routine cataract surgery because of the intracameral injection of a toxic detergent residue. Ten patients with a mean (SEM) initial cell loss of 72% (2%) were followed for 4 years. Data were obtained at 6 months and 4 years postoperatively and compared between TECD eyes and contralateral control eyes. RESULTS: Mean (SEM) endothelial cell density of the TECD eyes increased from 642 (41) cells/mm2 to 849 (50) cells/mm2 at 4 years postoperatively (p = 0.005). There was no difference in coefficient of variation or percentage hexagonals between 6 months and 4 years postoperatively. Mean (SD) corneal thickness of the TECD eyes and control eyes was similar, 0.51 (0.02) mm and 0.49 (0.01) mm, respectively (p = 0.65). Mean (SD) endothelial permeability was also similar for TECD eyes and control eyes (4.3 (0.9) x 10(-4) cm/min and 4.4 (0.6) x 10(-4) cm/min, respectively (p = 0.57). There was no correlation between endothelial cell density, coefficient of variation, or percentage of hexagonal cells and endothelial permeability in the TECD eyes. In three patients a permanent corneal decompensation occurred. CONCLUSIONS: Four years after TECD corneal endothelial wound healing is stable and the barrier function has been restored.

Adult↗

Corneal diffusion and metabolism of 12(R)-hydroxyeicosatetraenoic acid (12(R)HETE).

PURPOSE: To quantify the corneal diffusion and metabolism of tritiated 12(R)-hydroxyeicosatetraenoic acid (12(R)HETE) in the in vitro-mounted rabbit cornea to determine if this compound or one of its metabolites can diffuse across the stroma to the corneal endothelium. METHODS: The studies were performed in a Lucite block perfusion chamber by placing tritiated 12(R)HETE on the tear side of the cornea under the following conditions: (A) cornea completely intact (endothelium and epithelium present); (B) cornea with epithelium removed; and (C) cornea with both epithelium and endothelium removed. Radioactivity of 12(R)HETE and metabolites were measured in the different corneal layers and in the corneal perfusates using scintillation spectroscopy. 12(R)-hydroxyeicosatetraenoic and its metabolites were then quantified in the tissue perfusates using high performance liquid chromatography (HPLC) analysis. RESULTS: 12(R)HETE is rapidly taken up and metabolized by the intact cornea to a number of more polar compounds including a metabolite which has spectral and retention time characteristics of 8(R)-hydroxyhexadecatrienoic acid (8(R)HHDTrE). Both 12(R)HETE and 8(R)HHDTrE can diffuse through the stroma to the endothelium. Corneas having the epithelium removed also allow diffusion of 12(R)HETE across the stroma; however, there is significantly less metabolism. When both the epithelium and endothelium are removed, 12(R)HETE is capable of diffusing across the stroma; however, there is little metabolism, which suggests that the majority of 12(R)HETE metabolism occurs in the epithelium and, to a lesser degree, in the endothelium and stroma. CONCLUSIONS: 12(R)HETE and its metabolites are capable of diffusing from the epithelium through the cornea where they may adversely affect the endothelium.

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

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↗

Effects of preservative-free artificial tear solutions on corneal epithelial structure and function.

OBJECTIVES: To test the efficacy of a bicarbonate-containing artificial physiologic tear solution (solution PT) in providing an environment in which the damaged corneal epithelium can recover its normal barrier function and to compare this solution with other available artificial tears. Also, to investigate the effects on the corneal mucin layer and epithelial ultrastructure. METHODS: The corneal epithelial permeability of anesthetized rabbits was increased by exposure to 0.1% benzalkonium chloride. The corneas were then exposed to solution PT, with or without bicarbonate, or one of four commercially available artificial tear solutions for 1.5 hours, followed by a 5-minute exposure to 5(6)-carboxyfluorescein. Frozen sections of the corneas were examined by fluorescence microscopy. The fluorescence intensity (FI) of the epithelium was measured by image analysis. Undamaged corneas exposed to tear solutions were examined by transmission electron microscopy after fixation of the mucin layer with cetylpyridinium chloride. RESULTS: The FI of corneas damaged by benzalkonium chloride was increased threefold above those of undamaged controls. Damaged corneas treated with either of two commercial isotonic tear solutions partially recovered their barrier function, but the FI did not reach control levels. Corneas treated with hypotonic solutions containing ethylenediaminetetraacetic acid (EDTA) did not recover. In contrast, the FI of corneas treated with solution PT returned to control levels. This effect was lost in the absence of bicarbonate. Solution PT and the two isotonic solutions maintained normal corneal ultrastructure and mucin layer. Lack of bicarbonate in solution PT resulted in focal damage to superficial epithelial cells, whereas the EDTA-containing solutions destroyed the first two cell layers and reduced the mucin thickness. CONCLUSIONS: Bicarbonate-containing solution PT is superior to the other tear solutions tested in promoting recovery of the damaged corneal epithelial barrier and maintaining normal ultrastructure. The presence of bicarbonate appears to be essential to this process.

Bicarbonates↗

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↗

Endothelial cell loss after 4 mm cataract surgery.

To assess whether a 4 mm scleral tunnel incision with a 1.5 mm internal corneal lip (three-step procedure) causes increased endothelial cell loss and damage to the cornea, we retrospectively evaluated the outcomes of 20 patients (40 eyes) who had a standard 4 mm scleral tunnel incision (two-step procedure) in one eye followed by a three-step incision in the second eye, with in situ phacoemulsification and insertion of a foldable silicone lens in each eye. Mean phacoemulsification time was 2.4 +/- 1.1 minutes for the two-step incisions and 3.4 +/- 1.4 minutes for the three-step incisions. Preoperative and postoperative endothelial cell counts were obtained to determine the effects of surgery on the corneal endothelium. Although the three-step procedure had a trend toward increased endothelial cell loss from the central corneal region compared with the two-step incision, the result was neither clinically nor statistically significant. The difference between the three-step and two-step incisions in postoperative endothelial cell counts from the superior corneal region was statistically significant. The difference in postoperative counts from the inferior region was not statistically significant. Although the three-step 4 mm incision does seem to affect the corneal endothelium, its clinical significance is unknown.

Aged↗

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↗

Ultrastructure in anterior and posterior stroma of perfused human and rabbit corneas. Relation to transparency.

PURPOSE: The authors sought to discover whether there are differences in the degree of spatial order in the fibrillar ultrastructure between anterior and posterior stroma. METHODS: Human corneas were obtained from eye bank eyes. Although they had been classified as normal, some swelling remained after 3 hours of deturgescence. Freshly excised, unswollen rabbit corneas also were used. Image analysis methods were applied to transmission electron micrographs of the anterior, middle, and posterior stroma of these corneas to determine the positions and radii of fibrils, the fraction of total area occupied by fibrils, and the fibril number density. Results were used to calculate the interference factor that appears in the direct summation of the fields for light scattering theory and to estimate the total scattering cross-section per fibril. The interference factor is a measure of the spatial order in the positions and sizes of the fibrils. RESULTS: Electron micrographs showed anterior-posterior variations in size and number density of fibrils. The interference factor at wavelengths of visible light was lower in posterior stroma than in anterior stroma for humans and rabbits. In some instances in humans, the anterior interference factor was characteristic of mildly swollen cornea. When averaged for the electron micrographs analyzed, the anterior stroma was predicted to scatter approximately twice as much light per unit depth as the posterior stroma in humans (at any given wavelength) and approximately three times as much in rabbits. CONCLUSIONS: Calculations of the interference factor showed that there were differences in the anterior-posterior spatial ordering of fibrils. In human corneas, the differences could have been caused by intrinsic in vivo differences between anterior and posterior stroma; however, possible anterior-posterior variations in swelling between the two regions in vitro also could have affected the results.

Aged↗

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↗