[Chromatic Swept-Source Laser Scanning - Concept for a Cell-Resolving Confocal Laser Slit Lamp?] / Chromatic Swept-Source Laser Scanning ­ Konzept für eine zellauflösende konfokale Laserspaltlampe?

BACKGROUND: The in vivo characterisation of corneal epithelial tissue morphology is of considerable importance for diagnosis, disease prognosis, and the development of a treatment strategy for ocular surface diseases. In contrast to many alternative methods, in vivo corneal confocal microscopy (CCM) not only provides a macroscopic description of the corneal tissue but also allows its visualisation with cellular resolution. However, the translation of CCM from research to clinical practice is significantly limited by the complex and still largely manual operation of available CCM systems. In addition, for cross-sectional images, and analogously to conventional slit lamp microscopy, volume data must be acquired in time-consuming depth scans due to the frontal orientation of the image field in CCM, from which depth slices can subsequently be calculated. The pure acquisition time is already in the range of seconds, and additionally, motion artefacts have to be corrected in a sophisticated way. MATERIALS AND METHODS: This paper presents the concept and optics simulation of a new imaging technique based on a swept-source laser in combination with special chromatic optics. Here, the laser periodically changes its wavelength and is focused at different depths due to the wavelength-dependent aberration of the chromatic optics. RESULTS: The optics simulation results promise good optical resolution at a total imaging depth of 145 µm. CONCLUSION: The long-term goal is cell-resolving in vivo corneal confocal microscopy in real time with differently oriented sectioning directions.

Accurate In Vivo Bowman's Thickness Measurement Using Mirau Ultrahigh Axial Resolution Line Field Optical Coherence Tomography.

Purpose: The purpose of this study was to assess the accuracy, repeatability, and performance limits of in vivo Mirau ultrahigh axial resolution (UHR) line field spectral domain (LF-SD) optical coherence tomography (OCT) for the measurement of Bowman's and epithelial thickness, and to provide a reference range of these values for healthy corneas. Methods: Volunteers with no history and evidence of corneal disease were included in this study. An in vivo graph search image segmentation of the central cornea was obtained at the normal interface vector orientation. The Mirau-UHR-LF-SD-OCT system used has an axial resolution down to 2.4 µm in air (1.7 µm in tissue), with an A-scan speed of 204.8 kHz and a signal to noise ratio (sensitivity) of 69 (83) dB. Results: Nine volunteers were included, one of whom wore contact lenses. The repeatability of mean Bowman's and epithelial thicknesses were 0.3 and 1.0 µm, respectively. The measured 95% population range for healthy in vivo thickness was 13.7 to 19.6 µm for the Bowman's layer, and 41.9 to 61.8 µm for the epithelial layer. Conclusions: The measured thicknesses of Bowman's layer and the corneal epithelium using the Mirau-UHR-LF-SD-OCT were both accurate, with the range for healthy in vivo thicknesses matching prior confocal and OCT systems of varying axial resolutions, and repeatable, equaling the best value prior reported. Translational Relevance: T1. Development of a commercially viable clinical UHR OCT technology, enabling accurate measurement and interpretation of Bowman's and epithelial layer thickness in clinical practice.

Long-term Outcomes of Bowman's Membrane Relaxation for Enhancement of Femtosecond Intrastromal Lenticule Implantation Performed for the Management of High Hyperopia.

PURPOSE: To evaluate the feasibility and report long-term outcomes with Bowman's membrane relaxation (BMR) for enhancing the residual refractive error following femtosecond intrastromal lenticule implantation (FILI). METHODS: BMR was performed using a Hessburg-Barron trephine to create a circular incision into the Bowman's membrane and anterior corneal fibers up to the depth of approximately 120 to 130 µm. After enhancement, clinical outcomes were analyzed for a mean period of 36 months (range: 14 to 57 months). RESULTS: Four eyes of 3 patients (mean age: 29 years) underwent enhancement with BMR for a significant residual refractive error of +2.25 diopters (D) spherical equivalent following FILI for high hyperopia (mean spherical equivalent: +7.00 D). After BMR, the residual refraction reduced to +0.31 D, resulting in improvement in uncorrected distance visual acuity from 0.55 to 0.33 logMAR. The mean front keratometry values increased from 46.20 to 49.30 D, and the mean back keratometry values increased from -5.90 to -6.30 D following BMR, the latter returning to the baseline (pre-FILI) value of -6.30 D. An increase in Q-value with a corresponding increase in higher order aberrations was observed. Corneal biomechanics indicated reduction of stiffness and other parameters after enhancement. No intraoperative or postoperative complications were noted. CONCLUSIONS: BMR may be an effective technique for enhancement of residual hyperopia following tissue addition techniques such as FILI. BMR may reverse the posterior curvature changes, negating the steepening effect of the anterior cornea following tissue addition, potentially aiding in the enhancement. [J Refract Surg. 2022;38(2):134-141.].

Use of Bowman's membrane electrocautery in blind eyes with painful bullous keratopathy not amenable to corneal transplantation: a retrospective case series.

PURPOSE: This study evaluated the safety and efficacy of Bowman's membrane electrocautery in blind painful eyes with bullous keratopathy not amenable to corneal transplantation. METHODS: Eleven eyes of 11 subjects with painful bullous keratopathy and poor visual potential who underwent electrocautery of Bowman's membrane at a tertiary referral ophthalmology clinic were reviewed retrospectively. Subject demographics and preoperative and postoperative data were collected, including description of pain, slit lamp biomicroscopy, best corrected visual acuity, topical medication use, and complications. Efficacy of the procedure on pain reduction, bullae resolution, and topical medication use were assessed at post-operative visits. Safety was also evaluated based on any complications. RESULTS: Bowman's membrane electrocautery effectively resolved bullae in all eyes examined up to 6 months postoperatively; however, 2 eyes had recurrence by 1 year. Mean age at the time of surgery was 69.8 years and mean duration of follow-up was 15.4 months. Pain reduction was achieved in all eyes at 1 month, but 1 subject had pain recurrence by 6 months and another by 1 year. The median number of drops per day decreased from 6 preoperatively to 1.7 at 6 months. Two subjects who had underlying advanced ophthalmic disease had a mild reduction in vision. CONCLUSION: Bowman's membrane electrocautery is a safe and minimally invasive procedure for the management of painful bullous keratopathy in eyes with low vision potential and not amenable to corneal transplantation. Duration of effect appears to last at least 6 months and up to 3 years post-procedure.

Transcriptomic and Immunohistochemical Analysis of Progressive Keratoconus Reveal Altered WNT10A in Epithelium and Bowman's Layer.

Purpose: To identify global gene expression changes in the corneal epithelium of keratoconus (KC) patients compared to non-KC myopic controls. Methods: RNA-sequencing was performed on corneal epithelium samples of five progressive KC and five myopic control patients. Selected results were validated using TaqMan quantitative PCR (qPCR) on 31 additional independent samples, and protein level validation was conducted using western blot analysis on a subset. Immunohistochemistry was performed on tissue microarrays containing cores from over 100 KC and control cases. WNT10A transcript levels in corneal epithelium were correlated with tomographic indicators of KC disease severity in 15 eyes. Additionally, WNT10A was overexpressed in vitro in immortalized corneal epithelial cells. Results: WNT10A was found to be underexpressed in KC epithelium at the transcript (ratio KC/control = 0.59, P = 0.02 per RNA-sequencing study; ratio = 0.66, P = 0.03 per qPCR) and protein (ratio = 0.07, P = 0.06) levels. Immunohistochemical analysis also indicated WNT10A protein was decreased in Bowman's layer of KC patients. In contrast, WNT10A transcript level positively correlated with increased keratometry (Kmax ρ = 0.57, P = 0.02). Finally, WNT10A positively regulated COL1A1 expression in corneal epithelial cells. Conclusions: A specific Wnt ligand, WNT10A, is reduced at the mRNA and protein level in KC epithelium and Bowman's layer. This ligand positively regulates collagen type I expression in corneal epithelial cells. The results suggest that WNT10A expression in the corneal epithelium may play a role in progressive KC.

Bowman Layer Onlay Grafting: Proof-of-Concept of a New Technique to Flatten Corneal Curvature and Reduce Progression in Keratoconus.

PURPOSE: The aim of this study was to describe a new surgical technique for flattening the corneal curvature and to reduce progression in eyes with advanced progressive keratoconus (KC) by using Bowman layer (BL) onlay grafting and to report on the preliminary outcomes of this procedure. METHODS: In this prospective interventional case series, 5 patients with advanced progressive KC underwent BL onlay grafting. After removal of the epithelium, a BL graft was placed and "stretched" onto the stroma, and a bandage lens was placed to cover the BL graft. In 1 case, BL onlay grafting could be performed immediately after ultraviolet corneal crosslinking; all other eyes were ineligible for ultraviolet corneal crosslinking. Best spectacle- and/or best contact lens-corrected visual acuity, refraction, biomicroscopy, corneal tomography, anterior segment optical coherence tomography, and complications were recorded at 1 week and at 1, 3, 6, 9, and 12 to 15 months postoperatively. RESULTS: All 5 surgeries could be performed successfully. Average maximum keratometry went from 75 diopters (D) preoperatively to 70 D at 1 year postoperatively. All eyes showed a completely reepithelialized and a well-integrated graft. Best spectacle-corrected visual acuity improved at least 2 Snellen lines (or more) in 3 of 5 cases and best contact lens-corrected visual acuity remained stable, improving by 3 Snellen lines in case 1 at 15 months postoperatively. Satisfaction was high, and all eyes again had full contact lens tolerance. CONCLUSIONS: BL onlay grafting may be a feasible surgical technique, providing up to -5 D of corneal flattening in eyes with advanced KC.

Intrastromal Allogeneic Inclusions for the Management of Keratoconus: A Review of Current Literature.

ABSTRACT: A new therapeutic alternative has been developed in the past 6 years to treat severe keratoconus in young patients. Those patients had only corneal transplantation as an option, but now a variety of surgical alternatives in the form of allogeneic corneal inclusions have bloomed and are becoming more popular. Although Bowman layer transplantation is the most studied technique, recent studies have described different options with very promising preliminary results. Mostly all the techniques described improve corneal curvature, visual acuity, pachymetry, contact lens tolerance, and foremost, manage to avoid or postpone corneal transplantation. Very few complications have been described so far, which makes these techniques not only feasible but also safe. Herein, we focus on reviewing recently published studies describing these techniques and their first results.

In Vivo Confocal Microscopy Findings in Bowman Layer Transplantation.

PURPOSE: To report the in vivo confocal microscopy (IVCM) findings of a case who underwent femtosecond laser-assisted Bowman layer transplantation (BLT). METHODS: This is a case report. RESULTS: In a 25-year-old man with previous diagnosis of keratoconus in whom a BLT was performed using the femtosecond laser-assisted technique, IVCM (Confoscan 4; Fortune Technologies, Vigonza, Italy) was performed 3 months after surgery, obtaining a sequence of 4 digitalized images of the total thickness of the cornea, from the epithelium to the endothelium, and then backward. The images were analyzed with the Navis v. 3.5.0. system (NIDEK, Multi-Instrument Diagnostic System, Japan). In the anterior stroma, before arriving to the graft interphase, activated keratocytes could be observed with higher reflectivity. In this first image of the anterior interphase, a homogenous hyporreflective image could be observed. In the interior of the graft, no keratocytes were found and some hyperreflective particles were present. In addition to the anterior, the posterior interphase presented bright particles. CONCLUSIONS: This is the first report that documents a BLT using IVCM.

[Recurrent corneal erosion: modern view of the problem]. / Retsidiviruyushchaya eroziya rogovitsy: sovremennyi vzglyad na problemu.

Recurrent corneal erosion syndrome (RCES) is a common chronic disease associated with repeated episodes of epithelial disruption and is characterized by sudden onset of sharp pain in the eye, usually at night or upon waking, as well as redness, photophobia and lacrimation. The cause of the disease is considered to be impaired adhesion of the epithelial layer to the stroma, which is a consequence of abnormal regeneration of the basement membrane of corneal epithelium. A number of biological factors, including inflammatory mediators and proteinases in high concentrations, contribute to the destruction of the formed adhesion complexes. The basis of RCES diagnosis is characteristic medical history and biomicroscopy. An area of epithelium absence can be detected under direct illumination - more often in paracentral view - mainly in the lower third of the cornea, where an area of loose and irregular epithelium with microcysts or grayish inclusions can be seen. There may also be a card-like or «fingerprint¼ pattern. First of all, it is necessary to differentiate RCES from infectious lesions, which require a fundamentally different treatment regimen. It is believed that the first stage of treatment should include lubricants, drugs that promote epithelialization, inhibitors of matrix metalloproteinases and lipases. The «second line¼ of conservative treatment can include blood derivatives. If conservative therapy is ineffective, it is advised to use phototherapeutic keratectomy (PTK) or perform diamond burr polishing of Bowman's layer. The choice of a method for treating recurrent erosion should be based on the effectiveness of drug therapy, frequency of recurrence, duration of the disease, severity of clinical symptoms, availability of specific ophthalmic equipment and experience with it.

Identification of A Novel TGFBI Gene Mutation (p.Serine524Cystine) Associated with Late Onset Recurrent Epithelial Erosions and Bowman Layer Opacities.

BACKGROUND: Most transforming growth factor beta-induced (TGFBI) corneal dystrophies are associated with a characteristic phenotype, clinical course, and a conserved mutation in the TGFBI gene. However, we report a novel TGFBI missense mutation associated with a late-onset, variant Bowman layer dystrophy. METHODS: Participants underwent slit-lamp examination and multimodal imaging. Polymerase chain reaction amplification and Sanger sequencing were performed on saliva-derived genomic DNA to screen TGFBI exons 4 and 12 as well as COL17A1 exon 46. PolyPhen-2 and SIFT were used to predict the functional impact of any identified variants. RESULTS: A 56-year-old Thai woman reported a four-year history of decreased vision and intermittent eye irritation, suggestive of recurrent epithelial erosions, in both eyes. Slit-lamp exam revealed bilateral, irregular, limbal-sparing Bowman layer opacities, which were also noted on anterior segment optical coherence tomography. Phototherapeutic keratectomy was performed in the right eye, improving the best-corrected visual acuity from 20/50 to 20/30. Sequencing of the TGFBI gene revealed a novel heterozygous, missense mutation in exon 12 (c.1571 C > G; p.Ser524Cys), which was present in an affected son and absent in an unaffected son, and was predicted to be damaging by PolyPhen-2 and SIFT. The patient was diagnosed with a variant Bowman layer dystrophy given the late onset of an atypical phenotype and the identification of a novel TGFBI mutation. CONCLUSIONS: A novel TGFBI missense mutation is associated with a late-onset Bowman layer dystrophy. Given the atypical clinical appearance and course, molecular genetic analysis was utilized to establish a definitive diagnosis.

Severe corneal burn due to the accidental application of salicylic acid packed in a plastic dropper bottle / Quemadura corneal grave debida a la aplicación accidental de ácido salicílico envasado en un frasco gotero plástico

Eye burns due to the accidental application of pharmacological or nonpharmacological substances packaged in plastic dropper bottles have been described for more than three decades and continue to occur. These burns can cause potentially serious corneal injuries. We report the case of a patient who mistakenly applied salicylic acid to the right eye after confusing it with an eye lubricant, which caused him a severe corneal burn. Fortunately, after aggressive medical and surgical management (including oxygen therapy and amniotic membrane grafting), the visual results were good. We suggest conducting educational campaigns and taking legislative measures in our country to avoid packaging corrosive substances in this type of dropper bottle to reduce the risk of accidental burns.

Las quemaduras oculares por aplicación accidental de sustancias farmacológicas o no farmacológicas envasadas en frascos goteros plásticos, se han descrito desde hace más de tres décadas y siguen ocurriendo. Estas quemaduras pueden causar lesiones potencialmente graves de la córnea. Se presenta el caso de un paciente que se aplicó ácido salicílico en el ojo derecho al confundir el envase con el de un lubricante ocular, lo que le causó una grave quemadura corneal. Afortunadamente, tras un manejo médico y quirúrgico agresivo (incluida oxigenoterapia e injerto de membrana amniótica) los resultados visuales fueron buenos. Se sugiere hacer campañas educativas y adoptar normas en el país para evitar el envase de sustancias corrosivas en este tipo de frascos goteros, con el fin de disminuir el riesgo de quemaduras accidentales.

Gelatinous Drop-Like Corneal Dystrophy - 2 Clinical Cases. / Gelatinöse Hornhautdystrophie ­ 2 Verlaufsdarstellungen.

Gelatinous drop-like corneal dystrophy is a very rare autosomal recessive disease classified as an epithelial and subepithelial corneal dystrophy. Patients typically present under the age of 20 with drop-like corneal lesions showing high corneal fluorescein uptake. Their disease course is typically protracted and prone to frequent relapses. The condition is caused by a dysfunction of the epithelial barrier, leading to protein deposits most likely originating from tear fluid. Histology typically shows subepithelial amyloid deposits with corresponding defects of Bowman's layer and epithelial atrophy. Where topical lubricating and anti-inflammatory therapy proves insufficient, penetrating allogenic limbokeratoplasty can be considered in a curative approach. In this report, we present disease courses of 2 unrelated patients. Current findings on pathogenesis are discussed.

Bowman Layer Onlay Graft for Reducing Fluctuation in Visual Acuity After Previous Radial Keratotomy.

PURPOSE: To describe the clinical outcome of a first patient undergoing Bowman layer (BL) transplantation with an onlay graft to reduce fluctuation in visual acuity and refractive error after previous radial keratotomy (RK) surgery. METHODS: In 2018, a 66-year-old woman presented with complaints of long-standing diurnal fluctuation in best-spectacle corrected visual acuity (BSCVA) after RK in 1983. After the removal of host epithelium, a BL graft was positioned onto the host cornea. BSCVA, Scheimpflug-based corneal tomography, and anterior segment optical coherence tomography were evaluated up to 12 months postoperatively. RESULTS: The surgery and postoperative course were uneventful. After surgery, the subjective complaints of visual fluctuation were reduced from 10 to 3 on a scale from 1 to 10. BSCVA (20/40; 0.5) did not change from preoperative to postoperative. Corneal tomography showed an overall central corneal steepening of 5.9 diopters. Biomicroscopy, Scheimpflug imaging, and anterior segment optical coherence tomography showed a completely epithelialized and well-integrated graft, with some minor epithelial remnants located in the preexisting keratotomy incisions. CONCLUSIONS: BL onlay grafting may have the potential to manage patients with subjective complaints of diurnal fluctuation in visual acuity after previous RK.

Potential benefits of modified corneal tissue grafts for keratoconus: Bowman layer 'inlay' and 'onlay' transplantation, and allogenic tissue ring segments.

PURPOSE OF REVIEW: Keratoconus can be surgically challenging, especially in advanced cases. Classic corneal transplantation techniques, may often be associated with complications. New alternative procedures like isolated Bowman layer transplantation (as a corneal stromal inlay or as a corneal onlay) and corneal allogenic intrastromal ring segments (CAIRS) have recently shown promising results. The aim of this review is to describe the main new surgical developments for treating keratoconus. RECENT FINDINGS: Intrastromal Bowman layer transplantation has recently shown to be effective in halting keratoconus progression and maintaining visual acuity with contact lenses, at least up to 5-7 years postoperatively. Because intrastromal dissection can be challenging, we have recently developed a technique that allows using Bowman layer grafts as an onlay, that is positioned onto the patient's anatomical Bowman layer or anterior stroma, achieving comparable results as with Bowman layer inlay transplantation. CAIRS may also be an effective treatment, improving visual acuity, corneal surface parameters and keratoconus progression. SUMMARY: There are currently some new alternative treatments such as isolated Bowman layer inlay or onlay transplantation and CAIRS, both of which seem effective at treating keratoconus and which may offer a less invasive surgical approach.

Repeatability of OCT Anterior Surface and Bowman's Layer Curvature and Aberrations in Normal and Keratoconic Eyes.

PURPOSE: To study the repeatability of anterior surface and Bowman's layer curvature in normal and keratoconic eyes using optical coherence tomography (OCT). METHODS: In this study, 96 normal and 96 keratoconic eyes underwent corneal imaging using Pentacam (Oculus Optikgeräte, Wetzlar, Germany) and OCT (Triton, Topcon Corporation, Tokyo, Japan). The elevation data from segmented air-epithelium (A-E) and epithelium-Bowman's layer (E-B) interfaces in OCT scans were used to quantify curvature and aberrations. The wavefront aberrations were evaluated with the ray tracing method and 6th order Zernike polynomials. The intraclass correlation coefficient (ICC), within-subject standard deviation (Sw), and coefficient of variation (CoV) were used to assess repeatability. RESULTS: For curvatures, the Sw was less than 0.25 diopters (D) for the normal and keratoconic eyes. The Sw was highest for root mean square of lower order aberrations (0.14 µm) in keratoconic eyes. The CoV for curvatures was well below 0.5% for both groups. For some aberrations irrespective of groups, the CoV was greater because some individual aberrations (mean of three successive measurements) tended to be smaller in magnitude and even a small Sw resulted in a high CoV. For all variables, the ICC ranged between 0.80 and 0.99 for both the OCT and Pentacam measurements. Most variables were similar between the A-E and E-B interfaces (P > .05) for both groups. However, both differed significantly from all Pentacam variables (P < .05) in normal and keratoconic eyes. CONCLUSIONS: The repeatability of OCT curvatures and aberrations compared well with the Pentacam indices for normal and keratoconic eyes. [J Refract Surg. 2020;36(4):247-252.].

Accuracy of OCT Curvature and Aberrations of Bowman's Layer: A Prospective Comparison With Physical Removal of Epithelium.

PURPOSE: To compare optical coherence tomography (OCT) and Scheimpflug curvature and aberrations of the Bowman's layer before and after removal of the epithelium. METHODS: Bowman's layer was mapped with OCT (Optovue Inc., Irvine, CA) before and after removal of the epithelium in normal eyes undergoing photorefractive keratectomy (n = 14) and keratoconic eyes undergoing corneal cross-linking (n = 25). The anterior corneal surface before removal and the underlying Bowman's layer after removal of the epithelium were also mapped with Pentacam (Oculus Optikgeräte, Wetzlar, Germany), and the surface aberrations with ray tracing were computed. RESULTS: The agreement of OCT curvatures before and after removal of the epithelium was excellent (intraclass correlation coefficient [ICC] = 0.9). A similar trend was seen between OCT and Pentacam after removal of the epithelium. The agreement of surface wavefront aberrations of the Bowman's layer before and after removal of the epithelium was excellent (ICC = 0.9) between the devices for keratoconic eyes. However, this agreement was relatively inferior in normal eyes (ICC < 0.5). CONCLUSIONS: The virtual OCT curvature and aberrations of the Bowman's layer agreed well with its actual magnitudes on removal of the epithelium in the keratoconic eyes. In normal eyes, the agreement was inferior for aberrations but not for curvature. [J Refract Surg. 2020;36(3):193-198.].

In-vivo Three-dimensional Characteristics of Bowman's Layer and Endothelium/Descemet's Complex Using Corneal Microlayer Tomography in Healthy Subjects.

Purpose: To characterize the three-dimensional (3D) thickness profile and age-related changes of Bowman's layer (BL), and endothelium/Descemet's membrane (En/DM) complex among healthy individuals using Corneal Microlayer Tomography (CML-T), and to describe its repeatability and accuracy.Methods: Sixty-six eyes of 41 healthy volunteers; 27 eyes (< 40 years old), and 39 eyes (>40 years old) were imaged using HD-OCT. Automatic and manual segmentation of the corneal layers was performed, and 3D thickness maps were generated, using custom-built CML-T software. A regional analysis of mean thickness parameters between the 2 age groups was performed. A regression analysis was used to assess the correlation between age, and thickness maps. Intraclass Correlation Coefficients (ICC), Coefficients of Variation (COV), and Bland-Altman plots were used to assess the reliability of the repeated measurements in 198 locations.Results: CML-T successfully mapped the BL and En/DM in all included eyes. Thickness maps showed a significant increase in corneal thickness (CT), BL thickness (BMT), and En/DM complex thickness (DMT) toward the periphery with a mean difference 28 µm (p < .001), 1.1 µm (p < .001), and 1.4 µm (p < .001), respectively. There was a strong correlation between age and central DMT (r = 0.61; p < .001), while there was no correlation between age and both CT, and BMT. ICC values ranged from 0.9 (BMT) to 0.997 (DMT), and from 0.808 (BMT) to 0.979 (CT) for intraoperator repeatability of manual measurements, and the accuracy of auto matic measurements, respectively. COV values were lower than 7.5% in all cases.Conclusion: CML-T is a novel tool that can generate 3D-thickness maps of both BL and En/DM. CT, BMT, and DMT increase toward the periphery in healthy corneas. DMT increases with aging, while BMT does not. We also report excellent repeatability, accuracy and good agreement between automatic and manual measurements.

Multifunctional synthetic Bowman's membrane-stromal biomimetic for corneal reconstruction.

As the outermost layer of the eye, the cornea is vulnerable to physical and chemical trauma, which can result in loss of transparency and lead to corneal blindness. Given the global corneal donor shortage, there is an unmet need for biocompatible corneal substitutes that have high transparency, mechanical integrity and regenerative potentials. Herein we engineered a dual-layered collagen vitrigel containing biomimetic synthetic Bowman's membrane (sBM) and stromal layer (sSL). The sBM supported rapid epithelial cell migration, maturation and multilayer formation, and the sSL containing tissue-derived extracellular matrix (ECM) microparticles presented a biomimetic lamellar ultrastructure mimicking the native corneal stroma. The incorporation of tissue-derived microparticles in sSL layer significantly enhanced the mechanical properties and suturability of the implant without compromising the transparency after vitrification. In vivo performance of the vitrigel in a rabbit anterior lamellar keratoplasty model showed full re-epithelialization within 14 days and integration of the vitrigel with the host tissue stroma by day 30. The migrated epithelial cells formed functional multilayer with limbal stem cell marker p63 K14 expressed in the lower layer, epithelial marker K3 and K12 expressed through the layers and tight junction protein ZO-1 expressed by the multilayers. Corneal fibroblasts migrated into the implants to facilitate host/implant integration and corneal stromal regeneration. In summary, these results suggest that the multi-functional layers of this novel collagen vitrigel exhibited significantly improved biological performance as corneal substitute by harnessing a fast re-epithelialization and stromal regeneration potential.

Large Field of View Corneal Epithelium and Bowman's Layer Thickness Maps in Keratoconic and Healthy Eyes.

PURPOSE: To assess differences between epithelium thickness (ET) and Bowman's layer thickness (BLT) maps in keratoconic eyes and healthy eyes. DESIGN: Cross-sectional study. METHODS: Setting: institutional. STUDY POPULATION: 47 patients (1 eye) with keratoconus (KC) and 20 healthy subjects (1 eye). OBSERVATION PROCEDURE: epithelium and Bowman's layer measurements were performed by using custom-designed polarization-sensitive optical coherence tomography (PS-OCT) with a conical scanning optics design. En face corneal ET and BLT maps with a diameter of 11 mm were computed. Main outcome measurements were mean ET and BLT of 25 sectors; the thinnest (minET, minBLT) and thickest sectors (maxET, maxBLT) were assessed. Ratios between thinnest/thickest sectors (R1) and between mean ET and BLT of the inferior temporal quadrant/superior nasal quadrant (R2) were calculated (R1ET, R1BLT; R2ET, R2BLT). Receiver operator characteristic (ROC) curve analysis was used to assess the diagnostic power of statistically different parameters. RESULTS: In healthy eyes, smooth ET maps were observed. KC eyes showed a "doughnut pattern." The BLT maps of healthy eyes had a smooth appearance, but highly irregular "moth"-like damage pattern could be observed in keratoconic eyes. Highest area under the curve values were found for the thinnest sector of the BLT map, the R1ET, and the thinnest sector of the ET map. CONCLUSIONS: PS-OCT imaging enables the visualization of significant differences of the corneal epithelium and the Bowman's layer in en face maps covering almost the entire cornea. ET and BLT profiles could clearly show their diagnostic importance for the distinguishing of keratoconic eyes and healthy eyes.

[Morphological changes in the cornea of patients with recurrent corneal erosion after diamond burr polishing of Bowman's membrane]. / Morfologicheskie izmeneniia rogovitsy pri sindrome retsidiviruiushchei érozii posle shlifovki boumenovoi membrany almaznym borom.

PURPOSE: To evaluate morphological changes in the cornea of patients with recurrent corneal erosion (RCE) using laser confocal microscopy before and after abrasive polishing of Bowman's membrane with a diamond burr (APBMDB). MATERIAL AND METHODS: The study included 17 patients (20 eyes) with established RCE diagnosis; they underwent laser confocal microscopy on HRT III device with Rostock Cornea module (Heidelberg Engineering GmbH, Germany). Morphological analysis of epithelial cells, nerve fibers, stroma and corneal endothelium was conducted. After that, all patients underwent APBMDB. Patient examination was repeated after 1, 3, 6, and 12 months. RESULTS: Superficial epithelium was intact before treatment in almost all cases. Islets of hyper-reflective cells, drop-shaped arrangements and stripes characterizing abnormal basal membrane were found in basal epithelium layer. Activated keratocytes and anomalous extracellular matrix were observed in the anterior stroma. No changes could be found in the posterior stroma or endothelium. Subbasal nerve plexus had changes in the form of short and atypically formed corneal nerve fiber funiculi, reduced amount of long corneal nerve fiber funiculi, as well as their tendency for forming closed round shapes. Increase in the amount of Langerhans cells was observed. One month after the procedure, pathological changes in the epithelium and basal membrane were absent, reduction in the amount of Langerhans cells and activated keratocytes was observed. Three months after the polishing, continued reduction of the amount of Langerhans cells was noted. Regeneration of nerves partially restored after 6 months; after 12 months, confocal microscopy results were comparable to healthy volunteers. CONCLUSION: Confocal laser microscopy of the cornea of patients with RCE showed presence of changes in basal epitheliocytes, basal membrane, anterior stroma and corneal nerve fibers, as well as positive dynamics of these changes after abrasive polishing of Bowman's membrane with a diamond burr.