Corneal confocal microscopy: a novel noninvasive test to diagnose and stratify the severity of human diabetic neuropathy.

OBJECTIVE: The accurate quantification of human diabetic neuropathy is important to define at-risk patients, anticipate deterioration, and assess new therapies. RESEARCH DESIGN AND METHODS: A total of 101 diabetic patients and 17 age-matched control subjects underwent neurological evaluation, neurophysiology tests, quantitative sensory testing, and evaluation of corneal sensation and corneal nerve morphology using corneal confocal microscopy (CCM). RESULTS: Corneal sensation decreased significantly (P = 0.0001) with increasing neuropathic severity and correlated with the neuropathy disability score (NDS) (r = 0.441, P < 0.0001). Corneal nerve fiber density (NFD) (P < 0.0001), nerve fiber length (NFL), (P < 0.0001), and nerve branch density (NBD) (P < 0.0001) decreased significantly with increasing neuropathic severity and correlated with NDS (NFD r = -0.475, P < 0.0001; NBD r = -0.511, P < 0.0001; and NFL r = -0.581, P < 0.0001). NBD and NFL demonstrated a significant and progressive reduction with worsening heat pain thresholds (P = 0.01). Receiver operating characteristic curve analysis for the diagnosis of neuropathy (NDS >3) defined an NFD of <27.8/mm(2) with a sensitivity of 0.82 (95% CI 0.68-0.92) and specificity of 0.52 (0.40-0.64) and for detecting patients at risk of foot ulceration (NDS >6) defined a NFD cutoff of <20.8/mm(2) with a sensitivity of 0.71 (0.42-0.92) and specificity of 0.64 (0.54-0.74). CONCLUSIONS: CCM is a noninvasive clinical technique that may be used to detect early nerve damage and stratify diabetic patients with increasing neuropathic severity.

Efficacy of 2 types of silicone hydrogel bandage contact lenses after photorefractive keratectomy.

PURPOSE: To compare the efficacy of 2 types of silicone hydrogel bandage contact lenses with high oxygen transmissibility after photorefractive keratectomy (PRK). SETTING: Institute of Vision and Optics, Faculty of Medicine, University of Crete, Heraklion, Crete, Greece. METHODS: In this prospective study, 1 eye of patients having bilateral PRK was randomly fitted with a bandage contact lens of lotrafilcon A (Night & Day) and the fellow eye, with a bandage contact lens of lotrafilcon B (O(2)Optix). The patients and the examiner were masked to which bandage contact lens type was in which eye. Patients were examined on the day of surgery and 1, 3, and 5 days postoperatively. Postoperative examinations included uncorrected distance visual acuity and slitlamp biomicroscopy to assess epithelial defect size. Subjective evaluation of pain and vision was recorded 1, 2, 3, and 4 days postoperatively. RESULTS: The study enrolled 44 patients (88 eyes). The mean epithelial defect size immediately after surgery was 47.0 mm(2) with both types of bandage contact lenses. There was no statistically significant difference in epithelial defect size between the 2 lenses at any postoperative visit. Three days postoperatively, reepithelialization was complete in 75.0% of eyes in the lotrafilcon A group and 72.7% of the eyes in the lotrafilcon B group. CONCLUSION: There were no differences in corneal reepithelialization or subjective measurements after PRK between the 2 types of silicone hydrogel bandage contact lenses.

Corneal confocal microscopy detects early nerve regeneration after pancreas transplantation in patients with type 1 diabetes.

OBJECTIVE: Corneal confocal microscopy (CCM) is a rapid, noninvasive, clinical examination technique that quantifies small nerve fiber pathology. We have used it to assess the neurological benefits of pancreas transplantation in type 1 diabetic patients. RESEARCH DESIGN AND METHODS: In 20 patients with type 1 diabetes undergoing simultaneous pancreas and kidney transplantation (SPK) and 15 control subjects, corneal sensitivity was evaluated using noncontact corneal esthesiometry, and small nerve fiber morphology was assessed using CCM. RESULTS: Corneal sensitivity (1.54 +/- 0.28 vs. 0.77 +/- 0.02, P < 0.0001), nerve fiber density (NFD) (13.8 +/- 2.1 vs. 42 +/- 3.2, P < 0.0001), nerve branch density (NBD) (4.04 +/- 1.5 vs. 26.7 +/- 2.5, P < 0.0001), and nerve fiber length (NFL) (2.23 +/- 0.2 vs. 9.69 +/- 0.7, P < 0.0001) were significantly reduced, and nerve fiber tortuosity (NFT) (15.7 +/- 1.02 vs. 19.56 +/- 1.34, P = 0.04) was increased in diabetic patients before pancreas transplantation. Six months after SPK, 15 patients underwent a second assessment and showed a significant improvement in NFD (18.04 +/- 10.48 vs. 9.25 +/- 1.87, P = 0.001) and NFL (3.60 +/- 0.33 vs. 1.84 +/- 0.33, P = 0.002) with no change in NBD (1.38 +/- 0.74 vs. 1.38 +/- 1.00, P = 1.0), NFT (15.58 +/- 1.20 vs. 16.30 +/- 1.19, P = 0.67), or corneal sensitivity (1.23 +/- 0.39 vs. 1.54 +/- 00.42, P = 0.59). CONCLUSIONS: Despite marked nerve fiber damage in type 1 diabetic patients undergoing pancreas transplantation, small fiber repair can be detected within 6 months of pancreas transplantation using CCM. CCM is a novel noninvasive clinical technique to assess the benefits of therapeutic intervention in human diabetic neuropathy.

Surrogate markers of small fiber damage in human diabetic neuropathy.

Surrogate markers of diabetic neuropathy are being actively sought to facilitate the diagnosis, measure the progression, and assess the benefits of therapeutic intervention in patients with diabetic neuropathy. We have quantified small nerve fiber pathological changes using the technique of intraepidermal nerve fiber (IENF) assessment and the novel in vivo technique of corneal confocal microscopy (CCM). Fifty-four diabetic patients stratified for neuropathy, using neurological evaluation, neurophysiology, and quantitative sensory testing, and 15 control subjects were studied. They underwent a punch skin biopsy to quantify IENFs and CCM to quantify corneal nerve fibers. IENF density (IENFD), branch density, and branch length showed a progressive reduction with increasing severity of neuropathy, which was significant in patients with mild, moderate, and severe neuropathy. CCM also showed a progressive reduction in corneal nerve fiber density (CNFD) and branch density, but the latter was significantly reduced even in diabetic patients without neuropathy. Both IENFD and CNFD correlated significantly with cold detection and heat as pain thresholds. Intraepidermal and corneal nerve fiber lengths were reduced in patients with painful compared with painless diabetic neuropathy. Both IENF and CCM assessment accurately quantify small nerve fiber damage in diabetic patients. However, CCM quantifies small fiber damage rapidly and noninvasively and detects earlier stages of nerve damage compared with IENF pathology. This may make it an ideal technique to accurately diagnose and assess progression of human diabetic neuropathy.

Assessment of stromal keratocytes and tear film inflammatory mediators during extended wear of contact lenses.

PURPOSE: To monitor quantitative changes in stromal keratocyte density and the level of tear film inflammatory mediators following extended contact lens wear. METHODS: Twenty-two subjects aged 32 +/- 11 years participated in this cross-sectional study. Eleven subjects had worn silicone hydrogel (Si-H) lenses on a 30-day continuous wear basis for 12 months. Eleven subjects had worn rigid gas permeable lenses on the same basis for 12 months. Eleven age-matched control subjects were also recruited. Ultrasound pachometry, confocal microscopy, and tear fluid sample collection were performed on all subjects. Tear samples were assayed for epidermal growth factor (EGF), hepatocyte growth factor (HGF) and interleukin (IL)-8. RESULTS: Corneal thickness was similar for all subject groups. Total keratocyte density was not different between the 3 groups; however, keratocyte density was lower for rigid lens wearers in the anterior to mid stroma and lower for Si-H lens wearers in the posterior stroma compared with control subjects. Rigid lens wearers exhibited an irregular keratocyte distribution across the corneal stroma. EGF concentration and rate of release was greater in the tears collected from the rigid lens wearers and Si-H lens wearers, and IL-8 concentration was higher in the samples collected from the rigid lens wearers compared with the samples collected from the control subjects. CONCLUSIONS: Mechanical stimulation of the corneal surface due to the physical presence of a contact lens and the consequent release of inflammatory mediators may account for a loss or redistribution of keratocytes.

On the etiology of keratocyte loss during contact lens wear.

PURPOSE: To employ confocal microscopy to investigate the etiology of keratocyte loss after short-term contact lens wear by monitoring quantitative changes in keratocyte density. METHODS: Twenty neophyte subjects aged 26 +/- 3 years participated in the study, which was conducted over the course of three experimental sessions. In the first session, one eye of each subject was fitted with a silicone hydrogel contact lens, and the other eye served as the control. Both corneas were exposed to an anoxic environment for 2 hours. Ultrasound pachometry and confocal microscopy were performed on both eyes at baseline, immediately after the experiment and 2 hours post experiment. This procedure was repeated after 72 hours, but in this case one eye of each subject was fitted with a hyper-Dk rigid contact lens, and the fellow eye served again as the control. In the third experimental session, each subject was asked to periodically rub one eye only. Tear samples collected from the rubbed and control eyes were assayed for epidermal growth factor (EGF), hepatocyte growth factor (HGF), and interleukin (IL)-8. RESULTS: The increase in corneal thickness was similar in the experimental and control eyes. Both anterior and posterior keratocyte densities decreased in the experimental eyes compared with the control eyes, in all sessions. EGF and IL-8 concentrations were increased in the rubbed eyes compared with the control eyes. CONCLUSIONS: It is hypothesized that the mechanical stimulation of the corneal surface, due to the physical presence of a contact lens, induces the release of inflammatory mediators that cause keratocyte dysgenesis or apoptosis.

Corneal nerve tortuosity in diabetic patients with neuropathy.

PURPOSE: Corneal confocal microscopy is a reiterative, rapid, noninvasive in vivo clinical examination technique capable of imaging corneal nerve fibers. Nerve fiber tortuosity may indicate a degenerative and attempted regenerative response of nerve fibers to diabetes. The purpose of this study was to define alterations in the tortuosity of corneal nerve fibers in relation to age, duration of diabetes, glycemic control, and neuropathic severity. METHODS: The cornea and collected images of the subbasal nerve plexus of 18 diabetic patients (stratified into mild, moderate, and severe neuropathic groups using conventional clinical measures of neuropathy) and 18 age-matched nondiabetic control subjects were scanned, and a novel mathematical paradigm was applied to quantify the extent of nerve tortuosity, which was termed the tortuosity coefficient (TC). RESULTS: TC was significantly different between the four clinical groups (F(3) = 12.2, P < 0.001). It was significantly greater in the severe neuropathic group than in control subjects (P < 0.003) and in the mild (P < 0.004) and moderate (P < 0.01) neuropathic groups. TC did not correlate significantly with the age (r = -0.003, P > 0.05), duration of diabetes (r = -0.219, P > 0.05), or hemoglobin A1c (HbA1c; r = 0.155, P > 0.05) of diabetic patients. CONCLUSIONS: Corneal confocal microscopy allows rapid, noninvasive in vivo evaluation of corneal nerve tortuosity. This morphologic abnormality relates to the severity of somatic neuropathy and may reflect an alteration in the degree of degeneration and regeneration in diabetes.