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Biomedical subjects

H Dwight Cavanagh

Publications and source records attributed to H Dwight Cavanagh.

At least 37 records · Page 2Linked to original sources

Quantitative assessment of ophthalmic viscosurgical device retention using in vivo confocal microscopy.

PURPOSE: To develop and apply a new laboratory method for in vivo quantitative assessment of the retention of ophthalmic viscosurgical devices (OVDs) following phacoemulsification. SETTING: Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, Texas, USA. METHODS: Studies of both eyes of New Zealand White rabbits were performed. Six OVDs were evaluated: Provisc and Healon (both sodium hyaluronate 1%), Healon5 (sodium hyaluronate 2.3%), Amvisc Plus (sodium hyaluronate 1.6%), Viscoat (sodium hyaluronate 3%-chondroitin sulfate 4%), and a new viscous-dispersive OVD, DisCoVisc (sodium hyaluronate 3%-chondroitin sulfate 4%). The OVD was injected to fill the anterior chamber and a phacoemulsification needle inserted with the tip positioned just anterior to the lens capsule. Simulated phacoemulsification was performed for 1 minute using flow rates of 20, 40, and 60 mL/min; a vacuum level of 300 mm Hg; and ultrasound power of 60% using a Legacy phacoemulsification unit. The needle was removed, and silicone oil (1000 centistokes) was injected into the anterior chamber. The distance between the corneal endothelium and the OVD-silicone oil interface was measured using in vivo confocal microscopy through-focusing (CMTF). RESULTS: Significant differences in residual thickness were found between the OVDs tested. Specifically, the residual thickness of both DisCoVisc (mean 324.5 microm +/- 163.7 [SD]) and Viscoat (251.4 +/- 100.9 microm) was significantly greater than that of Provisc (9.5 +/- 16.7 microm), Healon (3.8 +/- 11.3 microm), Healon5 (0.6 +/- 2.4 microm), and Amvisc Plus (65.6 +/- 134.0 microm) (P < .05, Dunn test). Ophthalmic viscosurgical device retention was greatest with DisCoVisc; however, there was no statistically significant difference between DisCoVisc and Viscoat in residual thickness. The flow rate did not have a significant effect on the residual thickness (Friedman 2-way analysis of variance by ranks). CONCLUSIONS: Residual OVD thickness following simulated phacoemulsification could be quantitatively measured using in vivo CMTF. The results are consistent with human surgical experience in that the dispersive OVD (Viscoat) was better retained than the cohesive compounds. DisCoVisc, a new viscous-dispersive OVD, also showed retention compared with Viscoat under the experimental conditions.

Animals↗

Central corneal thickness in patients with congenital aniridia.

PURPOSE: To compare the mean central corneal thickness (CCT) in patients with congenital aniridia to that of a group of age-matched control subjects. The findings of specular and confocal microscopy in a patient with aniridia are discussed. METHODS: The mean values of five consecutive pachymetry measurements of patients with aniridia and control subjects were used for analysis. Statistical analysis was performed with a Mann-Whitney rank sum test. Specular microscopy was performed on one patient with aniridia using a Konan Specular Microscope Noncon ROBO CA (Hyogo, Japan). Confocal microscopy through focusing was performed with the Tandem Scanning Confocal Microscope (Reston, VA). RESULTS: Mean CCT measured 691.8 +/- 75.4 mum for patients with aniridia (16 eyes of 10 patients) and 548.2 +/- 21.2 mum for control subjects (P < 0.001). Specular microscopy in a patient with aniridia showed normal endothelial cell counts and structure. Confocal microscopy through focusing of this patient showed normal-appearing keratocytes and a thick corneal stroma. CONCLUSIONS: Patients with congenital aniridia have significantly thicker corneas than do age-matched control subjects. This difference can have important implications for the treatment of those patients who develop secondary glaucoma. The increased CCT in patients with aniridia is not a result of endothelial dysfunction but appears to be the result of the production of a thickened but otherwise healthy cornea by the mutated PAX6 gene.

Adolescent↗

Pseudomonas aeruginosa keratitis in an atopic silicone hydrogel lens wearer with rosacea.

PURPOSE: This case report documents Pseudomonas aeruginosa corneal infection associated with daily wear of a silicone contact lens in a patient who also had bilateral, preexisting biomicroscopic findings of ocular rosacea, seborrheic blepharitis, keratoconjunctivitis sicca, and chronic punctate corneal epitheliopathy. CONCLUSIONS: Collectively, these problems produce increased risk of infection for wear of any contact lens; and underscore the importance of proper patient selection, education and post-fit monitoring even when hyper-oxygen transmitting silicone hydrogels are utilized.

Anti-Bacterial Agents↗

The current state of corneal reshaping.

PURPOSE: The application of contact lenses to alter the shape of the cornea and temporarily reduce or eliminate myopia is known as orthokeratology, corneal refractive therapy, or corneal reshaping. It was first introduced in the 1960s, but high oxygen permeable materials and more sophisticated designs allow patients to wear contact lenses only during sleep, while dramatically improving the predictability and rate of myopia reduction. Many studies have shown that most corneal reshaping patients achieve uncorrected visual acuity of 20/25 or better that lasts all day long in one to two weeks of nighttime wear. Treatment is primarily effective through central epithelial thinning and midperipheral epithelial and stromal thickening. Much remains to be learned about corneal reshaping contact lenses and their effects on the cornea. METHODS: The authors reviewed existing knowledge and determined what needs to be learned in order to provide patients with appropriate informed consent prior to corneal reshaping contact lens wear. RESULTS: While corneal reshaping contact lenses are effective at temporarily reducing or eliminating myopia, claims about the progress of myopia being controlled with corneal reshaping contact lenses should not be made until further studies are published in peer-reviewed literature. The incidence and prevalence of microbial keratitis related to corneal reshaping contact lens wear is not known. Any overnight wear of contact lenses increases the risk of infection, but it is not known whether the risks of microbial keratitis are greater for corneal reshaping overnight contact lens wearers than other form of overnight contact lens wear. It is also not known whether the risk of microbial keratitis is greater for children than adults, but we must determine if children are at greater risk than adults because many children are wearing corneal reshaping contact lenses. CONCLUSIONS: Finally, it is recommended that ongoing education be provided to practitioners and staff regarding safety, informed consent, and prevention of potential problems, with special emphasis on the critical need to properly and thoroughly disinfect lenses that will be worn overnight.

Child↗

Characterization of growth and differentiation in a telomerase-immortalized human corneal epithelial cell line.

PURPOSE: To develop and characterize a telomerase-immortalized human corneal epithelial cell line (hTCEpi) to serve as an in vitro model for studying the molecular mechanisms involved in regulating human corneal epithelial cell differentiation. METHODS: Primary cultures of human corneal epithelial cells were infected with a retroviral vector encoding human telomerase reverse transcriptase (hTERT). Infected hTCEpi cells were selected, cloned, and characterized to identify telomerase activity, proliferative capacity, karyotype, and differentiative potential in routine culture and under consecutive submerged and air-lifted conditions. Cells were evaluated to measure cell cycle kinetics (anti-Ki-67, anti-p16), stratification (phalloidin and anti-ZO-1), and differentiation (anti-K3, anti-BCL-2 and TUNEL labeling). RESULTS: hTCEpi cells exhibited telomerase activity, a normal karyotype and cell cycle kinetics at greater than 240 population doublings, and loss of p16 after passage 10. Air-lifting produced a well stratified epithelium (five to seven cell layers) with apical ZO-1-stained tight junctions. Submersed culture demonstrated increasing expression of stratification markers (K5/K14) with K3-corneal keratin marker expression in long-term, air-lifted culture. Anti-BCL-2 staining showed both nuclear and cytoplasmic localization with loss of nuclear BCL-2 expression in TUNEL-labeled surface epithelial cells. CONCLUSIONS: hTCEpi cells stratify, differentiate, and desquamate similar to normal human corneal epithelium. Further study of the hTCEpi cell line may be valuable in studying the molecular mechanisms regulating corneal epithelial cell differentiation and desquamation.

Blotting, Western↗

Internalization of Pseudomonas aeruginosa is mediated by lipid rafts in contact lens-wearing rabbit and cultured human corneal epithelial cells.

PURPOSE: The internalization of Pseudomonas aeruginosa (PA) in nasal and tracheal epithelium has recently been shown to involve the formation of cholesterol- and sphingolipid-rich plasma membrane domains (lipid rafts). The purpose of this study was to investigate the role of lipid rafts in PA internalization by corneal epithelium in vivo, in vitro, and after contact lens wear. METHODS: Lipid raft formation was evaluated in rabbit corneas with and without contact lens wear and a human corneal epithelial (hTCEpi) cell line before and after PA infection with cornea-pathogenic strains by staining with FITC-conjugated cholera toxin beta-subunit, known to bind the lipid raft component GM1. Bacterial internalization was assessed by gentamicin survival assay. The role of lipid rafts in PA internalization was evaluated by pretreatment of hTCEpi cells with cholesterol metabolism inhibitors. The interaction of PA with lipid rafts was confirmed by flow cytometry. RESULTS: Contact lens wear in rabbits induced lipid raft formation in occasional surface corneal epithelial cells. Subsequent PA exposure showed preferential binding to lipid raft-forming cells, leading to lipid raft aggregation and PA internalization. A similar sequence of lipid raft formation and PA internalization was also observed in hTCEpi for all PA strains. Internalization of all PA strains was blocked by three cholesterol metabolism inhibitors (P < 0.01). Flow cytometry showed an association of PA with rafts. CONCLUSIONS: These findings demonstrate that contact-lens-mediated PA internalization involves lipid raft formation. Also, hTCEpi cells may be used as an experimental model for studying further the molecular mechanism(s) of PA infection in the corneal epithelium.

Animals↗

Four-dimensional multiphoton confocal microscopy: the new frontier in cellular imaging.

This paper reviews new developments in microscopy that combine gene transfer technology, multiphoton confocal fluorescence microscopy, live cell imaging and digital imaging techniques that provide unique insights into the complex physiological processes involved in tissue function at the cellular and subcellular level. The evolution of this novel, new technology is discussed with particular attention to earlier achievements in noninvasive ocular surface imaging. The practical basis of confocal microscopy, multiphoton confocal fluorescence microscopy, and the vital fluorescent labeling of cells in living tissues are also discussed. Additionally, one application using retroviral gene transfer to express enhanced green fluorescent protein in living wound healing fibroblasts is presented as an example of how living biology can be studied in situ in four dimensions (x, y, z, time).

Journal Article↗

Evaluation of the corneal effects of topical ophthalmic fluoroquinolones using in vivo confocal microscopy.

PURPOSE: To compare the effects of several fluoroquinolone antibiotics on the corneal epithelium and stroma using in vivo confocal microscopy. METHODS: Five antibiotic solutions were evaluated: 1) 0.3% ofloxacin (Oflox) solution with 0.005% benzalkonium chloride (BAC); 2) 0.3% gatifloxacin (Gati) solution with 0.005% BAC; 3) 0.3% ciprofloxacin (Cipro) solution with 0.006% BAC; 4) 0.5% levofloxacin (Levo) with 0.005% BAC; and 5) 0.5% moxifloxacin (Moxi) solution with no BAC. Preservative-free artificial tears (Tears) were used as a control. New Zealand white rabbits were used for this study (six per solution group). Ten days prior to exposure to any solution, central corneal epithelial thickness and stromal thickness were measured using in vivo confocal microscopy through focusing. Images of the superficial epithelium were also acquired. Both eyes of each rabbit then received one drop of the assigned solution six times the first day and then four times per day for 6 days. On day 7, in vivo confocal microscopy was repeated. RESULTS: A significant decrease in epithelial thickness was induced by 7 days of exposure to Levo, Gati, Oflox, and Cipro (P < 0.05, two-way repeated-measures ANOVA, Tukey test). Tears and Moxi, which do not contain BAC, did not induce significant changes in epithelial thickness. No significant changes in stromal thickness were detected (P = 0.266), and no keratocyte activation was observed for any of the solutions evaluated. CONCLUSION: We have previously used confocal microscopy to establish a correlation between epithelial thinning (due to superficial cell loss) and slight ocular irritation. The results of this study suggest that Moxi induces less damage to the corneal epithelium than other antibiotic solutions, perhaps because it does not contain BAC.

Animals↗

Hyperopic laser-assisted in situ keratomileusis results with LADARVision, Visx Star S2, and Visx Star S3.

PURPOSE: To compare the LADARVision (LV), Visx Star S3 (S3), and Visx Star S2 (S2) excimer lasers in the treatment of spherical hyperopia and hyperopic astigmatism with laser-assisted in situ keratomileusis. SETTING: A parallel, consecutive, three-surgeon, cohort comparison was performed at Zale Lipshy University Hospital, Laser Center for Vision, The University of Texas Southwestern Medical Center at Dallas, Texas for consecutive cases between January 2001 and October 2001. METHODS: Treatment outcomes of 105 hyperopic eyes with manifest refraction spherical equivalents of up to +6.50 diopters and astigmatism up to 3.75 diopters were compared. Forty-one eyes were treated with LV; 25 eyes were treated with S3; and 39 eyes were treated with S2. Outcome measures included uncorrected visual acuity, best spectacle-corrected visual acuity, predictability, and stability of treatments at 1 week and 1, 3, and 6 months. RESULTS: All three lasers were comparable in terms of refractive stability and predictability. The LV group had consistently better uncorrected visual acuities than did the S2 and S3 groups during the follow-up period. At 6 months, the LV group had statistically significantly better visual results than the S3 group at the 20/25 and 20/20 levels, whereas the only significant difference between the LV and S2 groups was at the 20/25 level. Better visual outcomes were achieved with S2 than with S3, but the differences were not statistically significant. No eye lost more than one line of best spectacle-corrected visual acuity. CONCLUSIONS: Predictability and stability of hyperopic laser-assisted in situ keratomileusis treatments with all three lasers were comparable. Visual results with LV at 6 months were statistically significantly better than those with S3 and S2.

Astigmatism↗

Pseudomonas aeruginosa corneal binding after 24-hour orthokeratology lens wear.

PURPOSE: To examine the effect of short-term 24-hr orthokeratology lens (OKL) wear on Pseudomonas aeruginosa binding, epithelial surface cell morphology, epithelial sheet thickness, and stromal thickness in a rabbit model. METHODS: Seventeen New Zealand white rabbits were treated according to the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research. Partial membranectomy was performed on all rabbits 1 week before the experiments. Baseline values for epithelial and stromal thickness and epithelial surface cell size were determined by in vivo confocal microscopy in one randomly chosen eye (n = 6). One week later, rabbits were fitted in the same eye with a hyper oxygen-transmissible OKL. Twenty-four hours later, confocal microscopy was repeated. The second group of rabbits (n = 6) was fitted with an OKL in one randomly chosen eye for 24 hr. P. aeruginosa binding to the corneal epithelium was assessed for the control corneas and those exposed to the test lens. Scanning electron microscopy was performed on a third group of rabbits to assess epithelial surface damage (n = 5). RESULTS: There was a statistically significant difference (P<0.001) in P. aeruginosa binding between the control (1.11 +/- 0.74 x 10(5) colony-forming units per cornea) and the OKL-wearing eyes (2.74 +/- 0.69 x 10(5) colony-forming units per cornea). The central epithelium thinned by 6.5% after lens wear (48.2 +/- 1.9 microm to 45 +/- 1.7 microm, P=0.005); however, central stromal thickness increased by 7.3% (322 +/- 22 microm to 345 +/- 29 microm, P=0.006). Compared with the baseline value, central epithelial cell size increased significantly from 1,253 +/- 140 mm(2) to 1,627 +/- 393 mm(2) (29.4%, P=0.02). Scanning electron microscopy showed increased surface epithelial damage associated with OKL wear. CONCLUSIONS: This prospective, masked, pilot study showed that 24-hr hyper oxygen-transmissible OKL wear induced a statistically significant increase in P. aeruginosa binding to the epithelium of the rabbit cornea, accompanied by central epithelial thinning, stromal thickening, and surface cell damage assessed by scanning electron microscopy. Collectively, the data suggest that despite adequate lens oxygen transmissibility, the mechanical pressure inherent in the OKL design exerted on the corneal surface appears to be associated with increased adherence of P. aeruginosa to surface corneal epithelial cells, which may pose an increased risk for lens-related microbial keratitis, especially in overnight (i.e., closed-eye) wearing conditions. Future studies are needed to determine whether these results are similar in human wear and how P. aeruginosa binding during OKL wear compares with other lens-wearing modalities, such as daily or continuous soft lens wear.

Animals↗

Corneal intrastromal gatifloxacin crystal deposits after penetrating keratoplasty.

BACKGROUND: An 85-year-old man developed faint crystallike white precipitates in the mid peripheral stroma of his left cornea 3 weeks after undergoing penetrating keratoplasty. The patient had been initially treated with 1% prednisolone acetate ophthalmic suspension and 0.3% gatifloxacin eyedrops to his left eye from the first day postoperatively. Three weeks later, the precipitates were more numerous, larger, and diffuse in distribution. Gatifloxacin was discontinued and substituted with a neomycin-polymixin B-dexamethasone ophthalmic ointment. METHODS: A detailed history, physical examination, laboratory workup, and tandem scanning confocal microscopy were performed. RESULTS: Tandem scanning corneal confocal microscopy confirmed the presence of crystals in the cornea. CONCLUSIONS: Gatifloxacin, a fourth-generation fluoroquinolone, can cause intrastromal macroscopic crystalline deposits through a compromised corneal epithelium, similar to what has been described for ciprofloxacin, a second-generation fluoroquinolone.

Aged↗

In vivo fluorescent labeling of corneal wound healing fibroblasts.

Numerous studies have shown that fibroblasts play an important role in corneal wound healing, however, the dynamic cellular events underlying wound tissue organization and contraction remain unclear. The purpose of this study was to develop a system to enable live cell imaging of corneal wound healing fibroblasts in situ. To this end, concentrated preparations of an RD114 pseudotyped MLV-based vector expressing the enhanced green fluorescent protein (EGFP) were evaluated in vitro for gene transfer efficiency using cultured rabbit corneal keratocytes. Primary rabbit keratocytes were efficiently labeled in vitro (up to 50% EGFP(+)) at a low multiplicity of infection (MOI=10). To evaluate this gene transfer vector in vivo, rabbit corneal fibroblasts were transduced by direct application of vector supernatant to injured corneas following lamellar keratectomy. Fluorescent fibroblasts were then visualized in situ using epifluorescence microscopy and multiphoton confocal microscopy of excised fresh tissue at multiple time points from 14 days to four months following gene transfer. Fourteen days post-transduction, labeled fibroblasts expressing EGFP were readily detectable by fluorescence microscopy. Detectable fluorescence was noted up to eight weeks post-transduction. Labeled fibroblasts were detected in clusters located predominantly along the margin circumscribing the wound and to a lesser extent within the wound area. Cell growth in clusters was suggestive of the expansion of individual transduced clones. High-resolution imaging showed fluorescent fibroblasts to have a broad, flattened, dendritic morphology, distinct from the spindle shape of cultured fibroblasts. Utilizing multiphoton confocal microscopy, three-dimensional imaging of viable, labeled cells showed wound healing fibroblasts to be extensively interconnected and multi-layered within the corneal wound. These results demonstrate that rabbit corneal fibroblasts can be efficiently transduced in vitro and in vivo using RD114 pseudotyped MLV-based vectors and that these vectors direct long-term transgene expression without apparent toxicity, pathogenesis or perturbation of native fibroblast morphology. Our data further suggest that, in vivo, wound-healing fibroblasts have a defined life span within the wound.

Animals↗

Comparison of LADARVision and Visx Star S3 laser in situ keratomileusis outcomes in myopia and hyperopia.

PURPOSE: To compare visual outcomes with 2 commonly used excimer lasers, the Autonomous LADARVision (LV) (Alcon Laboratories Inc.) and the Visx Star S3 (S3), in the performance of myopic and hyperopic astigmatic laser in situ keratomileusis (LASIK). SETTING: Zale Lipshy University Hospital Laser Center for Vision, University of Texas Southwestern Medical Center, Dallas, Texas, USA. METHODS: In this retrospective single-center 3-surgeon cohort comparison, 100 consecutive LV-treated myopic eyes with a mean manifest refraction spherical equivalent (MRSE) of -3.22 diopters (D) (range 0 to -6.00 D) (mean astigmatism 0.95 D [range 0 to 4.00 D]) were compared with 100 consecutive S3-treated eyes with a similar MRSE (mean MRSE -3.19 D; mean astigmatism 0.66 D [range 0 to 2.75 D]). Forty-one consecutive LV-treated hyperopic eyes with a mean MRSE of +2.59 D (range +1.00 to +6.00 D) (mean astigmatism 0.89 D [range 0 to 3.50 D]) were compared with 25 consecutive S3-treated eyes with a similar MRSE (mean MRSE +2.70 D; mean astigmatism 0.87 D [range 0 to 2.75 D]). The following outcomes were compared: uncorrected visual acuity, refractive predictability and stability, gain or loss of best spectacle-corrected visual acuity, and need for enhancement. RESULTS: Statistically significant differences in outcomes were found between lasers in the treatment of hyperopia and hyperopic astigmatism. No outcome differences between lasers were found in the treatment of myopia and myopic astigmatism. CONCLUSION: The LV and S3 lasers yielded equivalent results in myopic LASIK between 0 and -6.00 D; however, the LADARVision yielded statistically significantly better results in hyperopic LASIK between +1.00 and +6.00 D.

Adult↗