Panagiotis A. Kallinikos

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.

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/mm2 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/mm2 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.

Corneal confocal microscopy detects improvement in corneal nerve morphology with an improvement in risk factors for diabetic neuropathy

Diabet. Med. 28, 1261–1267 (2011)

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.

Corneal Sensitivity Is Reduced and Relates to the Severity of Neuropathy in Patients With Diabetes: Response to Dash

Dr. Dash (1) has correctly identified potential confounders when measuring corneal sensitivity. Accordingly, exclusion criteria in our study were as follows: previous ophthalmic surgery affecting the cornea (including refractive surgery), previous ocular trauma, current use of topical ophthalmic medications, contact lens wear, chronic dry eye, or other ocular disease that …