Danielle M. Robertson

The TFOS International Workshop on Contact Lens Discomfort: Report of the Subcommittee on Epidemiology

This report characterizes the neurobiology of the ocular surface and highlights relevant mechanisms that may underpin contact lens-related discomfort. While there is limited evidence for the mechanisms involved in contact lens-related discomfort, neurobiological mechanisms in dry eye disease, the inflammatory pathway, the effect of hyperosmolarity on ocular surface nociceptors, and subsequent sensory processing of ocular pain and discomfort have been at least partly elucidated and are presented herein to provide insight in this new arena. The stimulus to the ocular surface from a contact lens is likely to be complex and multifactorial, including components of osmolarity, solution effects, desiccation, thermal effects, inflammation, friction, and mechanical stimulation. Sensory input will arise from stimulation of the lid margin, palpebral and bulbar conjunctiva, and the cornea.

Patient compliance during contact lens wear: perceptions, awareness, and behavior.

Objectives: Patient noncompliance with recommended hygienic practices in contact lens wear is often considered a significant risk factor for microbial keratitis and adverse contact lens-related events. Despite advancements in lens materials and care solutions, noncompliant behavior continues to hinder efforts to maximize contact lens safety. The objective of this pilot study was to assess the relationship between perceived and actual compliance with awareness of risk and behavior. Methods: One hundred sixty-two established contact lens wearers were sequentially evaluated after their routine contact lens examination at the Optometry Clinic at the University of Texas Southwestern Medical Center at Dallas, TX. Each patient was questioned by a single trained interviewer regarding his or her lens care practices and knowledge of risk factors associated with lens wear. Results: Eighty-six percent of patients believed they were compliant with lens wear and care practices; 14% identified themselves as noncompliant. Using a scoring model, 32% demonstrated good compliance, 44% exhibited average compliance, and 24% were noncompliant; age was a significant factor (P = 0.020). Only 34% of patients who perceived themselves as compliant exhibited a good level of compliance (P<0.001). Eighty percent of patients reported an awareness of risk factors, but awareness did not influence negative behavior. Replacing the lens case was the only behavior associated with a positive history for having experienced a prior contact lens-related complication (P = 0.002). Conclusions: Perceived compliance is not an indicator for appropriate patient behavior. A large proportion of patients remain noncompliant despite awareness of risk. Education alone is not a sufficient strategy to improve behavior; newer approaches aimed at improving compliance with lens care practices are urgently needed.

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 × 105 colony-forming units per cornea) and the OKL-wearing eyes (2.74 ± 0.69 × 105 colony-forming units per cornea). The central epithelium thinned by 6.5% after lens wear (48.2 ± 1.9 μm to 45 ± 1.7 μm, P=0.005); however, central stromal thickness increased by 7.3% (322 ± 22 μm to 345 ± 29 μm, P=0.006). Compared with the baseline value, central epithelial cell size increased significantly from 1,253 ± 140 mm2 to 1,627 ± 393 mm2 (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.

The Effects of Silicone Hydrogel Lens Wear on the Corneal Epithelium and Risk for Microbial Keratitis

Abstract: Previous studies using animal models and human clinical trials have demonstrated that the use of low-oxygen-transmissible contact lens materials produce corneal epithelial surface damage resulting in increased Pseudomonas aeruginosa (PA) adhesion and raft-mediated internalization into surface corneal epithelial cells. These findings led to the testable clinical predictions that (1) microbial keratitis (MK) risk is expected to be the greatest during the first 6 months of wear; (2) there is no difference between 6 and 30 night extended wear; and (3) that wear of hyperoxygen-transmissible lenses would reduce the reported incidence of infection. Subsequent epidemiologic studies have confirmed the first two predictions; however, increased oxygen transmissibility with silicone hydrogel (SiHy) lens wear has not altered the overall incidence of MK. In this review, more recent clinical and basic studies that investigate epithelial alterations and bacterial adhesion to corneal epithelial cells after the wear of SiHy lenses with and without concomitant exposure to chemically preserved multipurpose solutions (MPS) will be examined. The collective results of these studies demonstrate that even in the absence of lens-related hypoxia, MPS induce ocular surface changes during SiHy lens wear that are associated with a pathophysiologic increase in PA adherence and internalization in the corneal epithelium, and therefore, predict a greater risk for PA–MK. In addition, new data supporting an interactive role for inflammation in facilitating PA adherence and internalization in the corneal epithelium will also be discussed.

Non-compliance with contact lens wear and care practices: a comparative analysis

Purpose. To compare the effects of existing patient awareness of lens-related complications and underlying risk factors on actual patient behavior during contact lens wear and care practices in two different clinical study populations. Methods. Established contact lens wearers (n = 281) completed an anonymous written questionnaire on presenting to their habitual eye care practitioner in the Dallas-Fort Worth metroplex. Data were analyzed and compared against a second study population, which comprised established contact lens wearers (n = 152) who were sequentially evaluated after their routine contact lens examination at the University of Texas Southwestern Medical Center at Dallas, TX (UTSW). All patients were questioned regarding his or her lens care practices and knowledge of complications and risk factors associated with contact lens wear. Results. Fifty-eight percent of patients in the general community could identify by name a complication associated with lens wear compared with 91% within the medical center. The most frequent complications reported were related to comfort and handling (72%, Dallas-Fort Worth) and infection (47%, UTSW). The majority of patients could correctly identify risk factors associated with lens-related complications; awareness for topping-off solutions, tap water exposure, and hygiene varied between groups. Overall, 85% of patients perceived themselves as compliant with their lens wear and care practices. Using a standard scoring model to determine actual compliance, 2% of patients demonstrated good compliance; however, only 0.4% of patients were fully compliant with contact lens wear and care practices. Conclusions. The data reveal some study bias in complication and risk awareness between populations; however, despite this limitation, a significant proportion of patients exhibited actual non-compliant behavior despite acknowledged awareness of risk. Although most patients consider themselves to be complying with standard practitioner guidelines for lens wear and care practices, essentially all contact lens wearing patients exhibit behavioral non-compliance with resulting increased risk for significant complications.

Severing corneal nerves in one eye induces sympathetic loss of immune privilege and promotes rejection of future corneal allografts placed in either eye

Less than 10% of corneal allografts undergo rejection even though HLA matching is not performed. However, second corneal transplants experience a threefold increase in rejection, which is not due to prior sensitization to histocompatibility antigens shared by the first and second transplants since corneal grafts are selected at random without histocompatibility matching. Using a mouse model of penetrating keratoplasty, we found that 50% of the initial corneal transplants survived, yet 100% of the subsequent corneal allografts (unrelated to the first graft) placed in the opposite eye underwent rejection. The severing of corneal nerves that occurs during surgery induced substance P (SP) secretion in both eyes, which disabled T regulatory cells that are required for allograft survival. Administration of an SP antagonist restored immune privilege and promoted graft survival. Thus, corneal surgery produces a sympathetic response that permanently abolishes immune privilege of subsequent corneal allografts, even those placed in the opposite eye and expressing a completely different array of foreign histocompatibility antigens from the first corneal graft.

Castroviejo lecture 2009: 40 years in search of the perfect contact lens.

Purpose: To identify the pathophysiological changes produced by contact lens wear that predispose the cornea to infection and search for prospective modifiable risk factors that could reduce the incidence of this critical complication in millions of patients worldwide. Methods: Significant experimental and clinical publications are reviewed, and the results of ongoing studies are presented. Results: Pseudomonas aeruginosa (PA) is the most common pathogen causing lens-related infectious keratitis over 3 decades. Contact lens wear can increase the risk of infection by increasing surface cell PA binding, thereby promoting invasion between broken tight junctions and initiating direct intracellular invasion mediated by lens-induced membrane lipid rafts. Prevention of upregulation of specific surface-binding receptors for PA with concomitant increase in infection risk is a zero damage game where independent interactions among lens type, mode of wear, oxygen transmissibility, polymer, and toxic effects of associated care solutions ideally should collectively produce no increased ability for PA to attach and/or to invade, thus minimizing the risk for lens-associated infections. The specific hypothesis tested is, “no increased epithelial surface damage…no increased PA binding or invasion…no increased risk for infection.” Testing of this new paradigm has been performed in vitro and in animal and human clinical trials and correlated clinically with relative risk results from robust current epidemiological studies. Results to date clearly support the use of lens-related increases in PA binding (bench) as a noninvasive clinical predictor of risk for lens-related infection in subsequent large-scale population studies (bedside). Currently, results suggest that use of common commercial multipurpose lens care solutions with soft lenses may alone significantly increase infection risk by enhancing lens-related PA binding as compared with use of nonpreserved solutions (hydrogen peroxide). Clinical testing also shows that only peroxide solutions show significant disinfection capability against amoebic cysts. Further case-control studies to examine relative risk for infection by lens type and lens care solution are urgently needed. Conclusions: Millions of patients are dependent on contact lenses for vision worldwide; over 3 decades, lens use has increased, although risk for lens-related infection has remained stubbornly unchanged. Unfortunately, recent introduction of a new generation of hyper-oxygen transmissible lenses used with traditional multipurpose lens care solutions has not lowered overall risks for lens-related infections; however, similar lenses used with nonpreserved care solutions (peroxide) recently demonstrated no significant increases in PA binding in a 1-year clinical trial. Collectively, these findings along with the urgent need for amoebic cysticidal disinfection have led to a current recommendation to patients to use nonpreserved (hydrogen peroxide) care solutions in soft lens wear.

Elevated IGFBP3 levels in diabetic tears: a negative regulator of IGF-1 signaling in the corneal epithelium

To determine the ratio of IGFBP3:IGF-1 in normal and diabetic human tears, and in telomerase-immortalized human corneal epithelial cells (hTCEpi) cultured under elevated glucose conditions and to correlate these changes with total and phosphorylated levels of IGF-1R. Tear samples were collected noninvasively from diabetic subjects and non-diabetic controls; corneal sensitivity was assessed using a Cochet-Bonnet Aesthesiometer. Conditioned media were collected following culture of hTCEpi cells in normal (5 mM) and elevated (25 mM) glucose conditions; mannitol was used as an osmotic control. IGFBP3, IGF-1, and phosphorylated IGF-1R levels were assessed by ELISA. IGFBP3 and IGF-1R mRNA were assessed by real-time polymerase chain reaction (PCR). Total and phosphorylated IGF-1R expression in whole cell lysates was assessed by western blot. There was a 2.8-fold increase in IGFBP3 in diabetic tears compared to non-diabetic controls (P=0.006); IGF-1 levels were not significantly altered. No difference in corneal sensitivity was detected between groups. The concentration of IGFBP3 in tears was independent of IGF-1. Consistent with human tear measurements in vivo, IGFBP3 secretion was increased 2.2 fold (P<0.001) following culture of hTCEpi cells under elevated glucose conditions in vitro. Treatment with glucose and the mannitol control reduced IGFBP3 mRNA (P<0.001). Total IGF-1R levels were unchanged. The increase in the IGFBP3:IGF-1 ratio detected in diabetic tears compared to normal controls blocked phosphorylation of the IGF-1R by IGF-1 (P<0.001) when tested in vitro. Taken together, these in vivo and confirmatory in vitro findings suggest that the observed increase in IGFBP3 found in human tears may attenuate IGF-1R signaling in the diabetic cornea. A long-term increase in IGFBP3 may contribute to epithelial compromise and the pathogenesis of ocular surface complications reported in diabetes.

Disruption of Contact Lens-Associated Pseudomonas aeruginosa Biofilms Formed in the Presence of Neutrophils

PURPOSE To evaluate the capacity of neutrophils to enhance biofilm formation on contact lenses by an infectious Pseudomonas aeruginosa (PA) corneal isolate. Agents that target F-actin and DNA were tested as a therapeutic strategy for disrupting biofilms formed in the setting of neutrophils in vitro and for limiting the infectious bioburden in vivo. METHODS Biofilm formation by infectious PA strain 6294 was assessed in the presence of neutrophils on a static biofilm plate and on unworn etafilcon A soft contact lenses. A d-isomer of poly(aspartic acid) was used alone and with DNase to reduce biofilm formation on test contact lenses. The gentamicin survival assay was used to determine the effectiveness of the test compound in reducing subsequent intracellular bacterial load in the corneal epithelium in a contact lens infection model in the rabbit. RESULTS In a static reactor and on hydrogel lenses, PA biofilm density was enhanced 30-fold at 24 hours in the presence of neutrophils (P < 0.0001). The combination of DNase and anionic poly(aspartic acid) reduced the PA biofilms formed in the presence of activated neutrophils by 79.2% on hydrogel contact lenses (P < 0.001). An identical treatment resulted in a 41% reduction in internalized PA in the rabbit corneal epithelium after 24 hours (P = 0.03). CONCLUSIONS These results demonstrate that PA can exploit the presence of neutrophils to form biofilm on contact lenses within a short time. Incorporation of F-actin and DNA represent a mechanism for neutrophil-induced biofilm enhancement and are targets for available agents to disrupt pathogenic biofilms formed on contact lenses and as a treatment for established corneal infections.

Characterization of ΔNp63 isoforms in normal cornea and telomerase-immortalized human corneal epithelial cells

Previous reports have suggested that specific isoforms of the potential stem cell marker p63 may regulate corneal epithelial homeostatic renewal through control of cell proliferation. In this study, we characterized the presence of DeltaNp63 isoforms in telomerase-immortalized human corneal epithelial cells (hTCEpi) in comparison to normal human corneal epithelium to validate the hTCEpi cell line as a viable model for the study of p63 isoforms. We further examined roles for DeltaNp63 in proliferation and differentiation. For in vitro studies, hTCEpi cells were cultured in serum-free culture media and grown under 0.15 mM calcium or sequential 1.15 mM calcium/air-lifted culture. Fresh donor human corneal tissue was used to assess expression and localization in situ. mRNA and protein levels were assessed by real-time PCR, Immunofluorescence (IF) and Western blotting (WB). DeltaNp63 expression levels throughout the cell cycle were assessed by double-labeling with DeltaNp63 and Ki-67. In situ, DeltaNp63 localized to nuclei throughout the human corneal epithelium and was lost only in superficial cells. WB confirmed the presence of all three DeltaNp63 isoforms in the central corneal epithelium and in hTCEpi cells. DeltaNp63 mRNA levels decreased when grown on collagen substrate and under increased calcium/air-lifted culture. mRNA and protein levels increased as cells approached confluence, with a significant decrease in post-confluent culture. DeltaNp63 expression levels did not vary with the cell cycle, as assessed by Ki-67 labeling. Collectively, the presence of all three DeltaNp63 isoforms in hTCEpi cells and in intact cornea validates the use of this cell line for the study of individual isoforms in the corneal epithelium; and these data suggest that expression of DeltaNp63 isoforms are not altered as a function of the cell cycle or cell division in subconfluent hTCEpi cells cultured in serum-free media, but demonstrate reduced expression upon contact-inhibited growth down-regulation and differentiation. Significantly, the localization of DeltaNp63 in central corneal epithelial cells with a loss of expression in superficial cells suggests that DeltaNp63 may play a role in mediating desquamative events at the ocular surface.