In Vivo Confocal Microscopy Evaluation in Patients with Keratoconus

Keratoconus is the most common primary corneal ectasia characterized by progressive focal thinning. Patients experience increased irregular astigmatism, decreased visual acuity and corneal sensitivity. Corneal collagen crosslinking (CXL), a minimally invasive procedure, is effective in halting disease progression. Historically, keratoconus research was confined to ex vivo settings. In vivo confocal microscopy (IVCM) has been used to examine the corneal microstructure clinically. In this review, we discuss keratoconus cellular changes evaluated by IVCM before and after CXL. Cellular changes before CXL include decreased keratocyte and nerve densities, disorganized subbasal nerves with thickening, increased nerve tortuosity and shortened nerve fibre length. Repopulation of keratocytes occurs up to 1 year post procedure. IVCM also correlates corneal nerve status to functional corneal sensitivity. Immediately after CXL, there is reduced nerve density and keratocyte absence due to mechanical removal of the epithelium and CXL effect. Nerve regeneration begins after 1 month, with nerve fibre densities recovering to pre-operative levels between 6 months to 1 year and remains stable up to 5 years. Nerves remain tortuous and nerve densities are reduced. Corneal sensitivity is reduced immediately postoperatively but recovers with nerve regeneration. Our article provides comprehensive review on the use of IVCM imaging in keratoconus patients.

Dissociation between Corneal and Cardiometabolic Changes in Response to a Time-Restricted Feeding of a High Fat Diet

Mice fed a high fat diet (HFD) ab libitum show corneal dysregulation, as evidenced by decreased sensitivity and impaired wound healing. Time-restricted (TR) feeding can effectively mitigate the cardiometabolic effects of an HFD. To determine if TR feeding attenuates HFD-induced corneal dysregulation, this study evaluated 6-week-old C57BL/6 mice fed an ad libitum normal diet (ND), an ad libitum HFD, or a time-restricted (TR) HFD for 10 days. Corneal sensitivity was measured using a Cochet-Bonnet aesthesiometer. A corneal epithelial abrasion wound was created, and wound closure was monitored for 30 h. Neutrophil and platelet recruitment were assessed by immunofluorescence microscopy. TR HFD fed mice gained less weight (p < 0.0001), had less visceral fat (p = 0.015), and had reduced numbers of adipose tissue macrophages and T cells (p < 0.05) compared to ad libitum HFD fed mice. Corneal sensitivity was reduced in ad libitum HFD and TR HFD fed mice compared to ad libitum ND fed mice (p < 0.0001). Following epithelial abrasion, corneal wound closure was delayed (~6 h), and neutrophil and platelet recruitment was dysregulated similarly in ad libitum and TR HFD fed mice. TR HFD feeding appears to mitigate adipose tissue inflammation and adiposity, while the cornea remains sensitive to the pathologic effects of HFD feeding.

The Corneal Changes in Diabetic Patients

Diabetes mellitus (DM) represents a systemic disorder which afects different organs. Ocular complications of the DM are the worldwide leading cause of blindness. The most common complications are diabetic retinopathy, diabetic cataract, neovascular glaucoma. Recently many investigations point out that DM can cause comlications at ocular surface as well. Condition such as decreased corneal sensitivity, dry eye or neurotrophic corneal ulceraction are the main clinical manifestations of the diabetic keratopathy (DK). Untreated, these conditions can lead to serious visual acuity decrease. Pathological processes, based on chronic inflammation, due to chronic hyperglycemia, are the main step in the process of DK development. Adequate treatment of the main disease - DM is an imperative in maintaining the healthy cornea without subjective sensations of diabetic patients.

Efficacy of rhNGF-loaded amniotic membrane transplantation for rabbit corneal epithelial and nerve regeneration

AIM: To evaluate the efficacy of recombinant human nerve growth factor-loaded amniotic membrane (rhNGF-AM) on corneal epithelial and nerve regeneration in rabbit model. METHODS: Freshly prepared human amniotic membrane (AM) were immersed into PBS buffer containing 100 or 500 μg/mL rhNGF for 15, 30, and 60min at 4℃. The in vitro release kinetics of rhNGF was measured with ELISA. For in vivo evaluation, the AM were immersed with 500 μg/mL rhNGF for 30min. Fifty-seven rabbits were selected to establish corneal epithelial defect model. In addition to the 19 rabbits in control group, 38 rabbits received AM transplantation with or without rhNGF after the removal of central epithelium. Corneal epithelial defect area, sub-epithelial nerve fiber density, corneal sensitivity, rhNGF contents in resident AM and corneas were measured after the surgery. RESULTS: rhNGF was sustained release from the AM within 14d in vitro, with the positive correlation with initial immersion concentration. The immersion of AM in 500 μg/mL rhNGF for 30min achieved the most stable release within 14d. After transplantation in rabbit cornea, a high concentration of rhNGF in resident rhNGF-AM and cornea was maintained within 8d. Corneal epithelial healing, nerve fiber regeneration and the recovery of corneal sensitivity were significantly accelerated after the rhNGF-AM transplantation when compared to simple AM transplantation (all P<0.05). CONCLUSION: Simple immersion of AM achieves the sustained release of rhNGF, and promotes corneal epithelial wound healing and nerve regeneration, as well as the recovery of corneal sensitivity in rabbit.

Epidemiology of Corneal Neovascularization and Its Impact on Visual Acuity and Sensitivity: A 14-Year Retrospective Study

Purpose: To quantify the severity and location of corneal neovascularization (cNV) and its impact on the visual acuity and corneal sensitivity in a cohort of the patients referred to a specialist cornea clinic and also to describe the etiology of cNV in the cohort.Methods: We retrospectively evaluated the charts of 13,493 subjects referred to the San Raffaele Cornea Unit between January 2004 and December 2018 to search for cNV diagnosis. The corneal neovascularization severity was measured in the quadrants (range: 1–4) and location was defined as superficial, deep, or both. Best spectacle corrected visual acuity (BSCVA) was measured in logMar. We used the multiple regression analysis to identify the independent predictors of logMAR, after adjusting for age, gender, keratoconus, herpes keratitis, penetrating keratoplasty, trauma, and cataract surgery.Results: Corneal neovascularization was diagnosed in 10.4% of the patients analyzed. The most prevalent etiology of cNV in our population was non-infectious corneal dystrophies/degenerations followed by herpes simplex virus infection. cNV affected OD, OS, or both eyes in 35.6, 40.2, and 24.2 of cases, respectively. Mean BSCVA (SD) was 0.59 (0.76), 0.74 (0.94), and 1.24 (1.08) in cNV one, two, and three or four of the quadrant groups. Superficial, deep, or mixed cNV occurred in 1,029, 348, and 205 eyes. Severe cNV (three or four of the quadrants) was a significant predictor of low visual acuity (p &lt; 0.001) and reduced corneal sensitivity (p &lt; 0.05). cNV location and its severity were associated (p &lt; 0.05). In addition, corneal anesthesia was associated with lower BSCVA (p &lt; 0.001).Conclusion: Severe and deep cNV are associated with the reduced visual acuity and corneal sensitivity. Our data strongly support the relevance of appropriate follow-up as cNV is a major risk factor for graft rejection.

Expert consensus on the identification, diagnosis, and treatment of neurotrophic keratopathy

Background Neurotrophic keratopathy (NK) is a relatively uncommon, underdiagnosed degenerative corneal disease that is caused by damage to the ophthalmic branch of the trigeminal nerve by conditions such as herpes simplex or zoster keratitis, intracranial space-occupying lesions, diabetes, or neurosurgical procedures. Over time, epithelial breakdown, corneal ulceration, corneal melting (thinning), perforation, and loss of vision may occur. The best opportunity to reverse ocular surface damage is in the earliest stage of NK. However, patients typically experience few symptoms and diagnosis is often delayed. Increased awareness of the causes of NK, consensus on when and how to screen for NK, and recommendations for how to treat NK are needed. Methods An 11-member expert panel used a validated methodology (a RAND/UCLA modified Delphi panel) to develop consensus on when to screen for and how best to diagnose and treat NK. Clinicians reviewed literature on the diagnosis and management of NK then rated a detailed set of 735 scenarios. In 646 scenarios, panelists rated whether a test of corneal sensitivity was warranted; in 20 scenarios, they considered the adequacy of specific tests and examinations to diagnose and stage NK; and in 69 scenarios, they rated the appropriateness of treatments for NK. Panelist ratings were used to develop clinical recommendations. Results There was agreement on 94% of scenarios. Based on this consensus, we present distinct circumstances when we strongly recommend or may consider a test for corneal sensitivity. We also present recommendations on the diagnostic tests to be performed in patients in whom NK is suspected and treatment options for NK. Conclusions These expert recommendations should be validated with clinical data. The recommendations represent the consensus of experts, are informed by published literature and experience, and may improve outcomes by helping improve diagnosis and treatment of patients with NK.

Correlation between Hyperalgesia and Upregulation of TNF-α and IL-1β in Aqueous Humor and Blood in Second Eye Phacoemulsification: Clinical and Experimental Investigation

The aim of this study was to explore the correlation between intraoperative hyperalgesia of the second eye and the dynamic changes of tumor necrosis factor (TNF)-α and interleukin (IL)-1β levels in aqueous humor (AH) of the second eye and whole blood after the first eye cataract surgery. A rabbit model of monocular phacoemulsification was established by administration of 0.3% levofloxacin. Whole blood and AH samples from non-surgical eyes in the experimental group (n =25) and second eye in the blank control group (n =15) were obtained and corneal sensitivity was examined after surgery (1, 3, 7, 14, and 21 days postoperatively). TNF-α and IL-1β levels in AH and TNF-α mRNA and IL-1β mRNA levels in whole blood were measured. In a clinical study, 30 patients who underwent bilateral phacoemulsification within 1 month were divided into six groups in accordance with the operation intervals (1, 3, 7, 10, 14, and 21days). TNF-α and IL-1β levels in AH were measured at the beginning of surgery and intraoperative pain was assessed immediately after surgery. Corneal sensitivity (F =244.910, P <0.05), TNF-α and IL-1β levels in AH (F =184.200, 82.900, P <0.05) of non-surgical eyes and in whole blood (F =272.800, 193.530, P <0.05) in the experimental group were significantly higher than the baseline levels after phacoemulsification. In the clinical study, NRS scores of second eye surgery were higher than those of the first eye(P =0.0025) and 19 (63.3%) patients reported more pain during the second eye surgery. TNF-α and IL-1β concentrations in AH of the second eye were significantly higher than those of the first eye (F =123.60, P <0.05; F =59.60, P <0.05). In conclusion, within 1 month after the first eye phacoemulsification, higher pain sensitivity (hyperalgesia) exists in the second eye, which may be related to dynamic changes in TNF-α, IL-1β levels in AH or whole blood.

CLINICAL PECULIARITIES OF BACTERIAL KERATITIS IN TYPE 1 DIABETIC PATIENTS

The purpose was to define the clinical peculiarities of bacterial keratitis in patients with type 1 diabetes mellitus (DM1) at visit 1. Methods. We retrospectively reviewed the results of 62 DM1 patients (62 eyes) with bacterial keratitis and 43 nondiabetic patients (43 eyes) with bacterial keratitis of the control group who were referred for visit 1 (before administering the treatment). Research methods were as follows: visual acuity, tonometry, slit-lamp biomicroscopy of anterior and posterior eye segments, bacteriological studies, fluorescein dye test, anterior eye OCT and non-contact corneal esthesiometry. Results. Compared to nondiabetic, DM1 patients with bacterial keratitis showed higher degree of inflammatory reaction in the anterior chamber of the eye at visit 1 (p<0.05) as well as 28.8% lower mean corneal sensitivity threshold (p<0.05). At visit 1, the degree of decreasing of corneal sensitivity in DM1 patients with bacterial keratitis was higher than in control group (p<0.05). Localization of bacterial keratitis, the degree of pericorneal injection, corneal ulcer defect size and depth, corneal infiltration as well as edema of the corneal tissue surrounding the ulcer did not depend on the presence of diabetes mellitus (р>0,05) at visit 1. Conclusions. There are clinical peculiarities of bacterial keratitis in patients with type 1 diabetes mellitus at visit 1.