Mid Stroma Research Articles

Pre-Descemet Corneal Dystrophy – Changes in Corneal Morphology in Confocal Microscopy

PurposeTo identify and describe the in-vivo microstructural changes in the cornea of patients with pre-Descemet’s membrane corneal dystrophy.MethodsTen patients (20 eyes) were examined in the Laser Microsurgery Centre in Warsaw. The ophthalmic examination, including visual acuity and slit lamp biomicroscopy, was carried out. Because of suspicion of corneal dystrophy, in vivo confocal microscopy of the cornea was also performed.ResultsBiomicroscopy revealed bilateral, dust-like fine opacities in the deep parts of the corneal stroma. Confocal microscopy showed pleomorphic structures (enlarged keratocytes) containing dense, hyperreflective, granular inclusions in the posterior stroma, next to the Descemet’s membrane. In 3 patients (6 eyes) punctate particles were seen extracellularly also in the mid stroma. Superficial and basal epithelial layers, anterior stroma and endothelium cells appeared normal.Conclusions1. In vivo confocal microscopy findings of cornea with pre-Descemet’s membrane dystrophy are characteristic. 2. In vivo confocal microscopy is a useful tool in diagnosing rare corneal dystrophies in cases where classical examination methods give inconclusive results.

Influence of Opacity Depth on Big Bubble Formation During Deep Anterior Lamellar Keratoplasty in Corneal Stromal Scars.

To identify the key preoperative predictors of big bubble (BB) formation during deep anterior lamellar keratoplasty in patients with corneal stromal scars (CSS). This retrospective cohort study included consecutive patients with CSS after infective keratitis who underwent BB-deep anterior lamellar keratoplasty between January 2021 and July 2023 at a tertiary referral center. Topographic and tomographic data were collected to compare the rates and types of BB formations. Anterior segment optical coherence tomography (AS-OCT) was employed to assess the maximum depth of opacity by dividing the stroma into 3 zones of equal thickness: anterior (stage A), mid (stage B), and posterior stroma (stage C). Multivariate logistic regression analysis was performed to identify the potential preoperative predictors of bubble formation. Pneumatic dissection was achieved in 13 of 33 eyes (39.4%), with 11 BB type 1 eyes (33.3%) and 2 BB type 2 eyes (6.1%). According to AS-OCT grading, bubble formation was more frequent with CSS involving more superficial stromal layers (P <0.032). In the eyes with stage C, bubble formation failed 12 out of 14 times (85.7%, P <0.026). Spearman correlation showed that bubble formation was inversely associated with the AS-OCT grading (rho = -0.443, P = 0.001). After logistic regression analysis, AS-OCT grading was found to be the sole factor that predicted bubble formation (coeff. -1.58, confidence interval 95% -3.03 to -0.12, P = 0.034). Depth of opacity in CSS was the key determinant for predicting the success of pneumatic dissection, as advanced AS-OCT stages are strongly associated with BB failure.

Central versus peripheral thickness in the human cornea explained

PurposeThe human cornea is thicker in the periphery than the center and it has been suggested that this must be due to greater numbers of lamellae in the peripheral corneal stroma. The purpose of this study was to use high-resolution ultrastructural imaging to determine if the greater thickness of the peripheral cornea is due to the presence of more lamellae or if there is some other anatomical explanation. MethodsIn this study, full thickness corneas from three human donors were processed for light microscopy (LM) and transmission electron microscopy (TEM). Images were taken in three distinct stromal regions (anterior, middle, and posterior) from the central and peripheral cornea. Stromal thickness was evaluated by LM while TEM was used to evaluate numbers and thicknesses of lamellae, mean collagen fibril diameter, and mean collagen fibril density. ResultsMean stromal thickness was significantly thinner in the central (415 ± 34 µm) compared to the peripheral (536 ± 29 µm) cornea (P = 0.009). Numbers of lamellae were not significantly different between central (246 ± 14) and peripheral (251 ± 14) cornea. Average lamellar thickness was not different across all regions of the cornea, except for the peripheral posterior where the lamellae were approximately 50 % thicker (P < 0.05). Collagen fibril diameters were larger in the peripheral cornea by approximately 30 % when compared to the central cornea, in all regions (P < 0.01). ConclusionsThis study shows that it is an increase peripheral posterior lamellar thickness, rather than an increase in the number of lamellae, that accounts for the increase in corneal stromal thickness in the periphery of the human cornea. While collagen fibril diameters are greater throughout the peripheral stroma, the lamellae in the mid and anterior peripheral stroma are not thicker than centrally.

Chlorhexidine in the treatment of microsporidial stromal keratitis and the effect of host immunity: A case series and literature review.

Microsporidial stromal keratitis is an increasingly well-known vision-threatening disease. A large proportion of cases are initially misdiagnosed as herpes simplex keratitis and treated with topical steroids. In most of such cases, medical treatment failed, and corneal transplantation was required. This study reported the results of 0.02% topical chlorhexidine used to treat three cases of microsporidial stromal keratitis and reviewed the literature on the outcomes of microsporidial stromal keratitis treatment. In the first case, histopathology of a specimen from penetrating keratoplasty (PK) revealed severe chronic inflammation involving the entire stromal layer but no microorganism activity after the application of topical chlorhexidine for 10 months. The second case exhibited complete resolution of keratitis after topical chlorhexidine. The patient in the third case did not respond to medical treatment, and therapeutic PK was performed. Histopathological examination revealed numerous microsporidial spores that had colonized in the mid and deep stroma, where few inflammatory cells were observed. These findings explain the variable microsporidial susceptibility to chlorhexidine, suggesting the crucial role of host immunity. In cases of host immunity, topical chlorhexidine may represent a promising option for the treatment of microsporidial stromal keratitis.

Multidrug-resistant keratitis: challenging yet manageable

PurposeTo study the incidence, clinical features and outcomes of multidrug-resistant (MDR) bacterial keratitis.MethodsAll cases of MDR-bacterial keratitis presenting to our institute over a period of 2 years were retrospectively analysed....

Corneal biomechanical properties following corneal cross-linking: Does age have an effect?

PurposeTo explore the effect of age on corneal biomechanical properties following corneal cross-linking (CXL). MethodsA total of 12 pairs of human eye-banked corneas (24 corneas, from 14 females and 10 males) were used in the study. The mean donor age was 48.5 years (ranging from 26 to 71 years). Corneas were divided into three age groups: A (26–41 years), B (42–57 years) and C (58–71 years), with four pairs in each group. For each pair, the right corneas were cross-linked using accelerated CXL with UVA (10 mW/cm2) and riboflavin, while the left corneas served as controls and were not exposed to either UVA irradiation or riboflavin. The corneal elastic modulus of the anterior, mid and posterior corneal stroma was measured using nanoindentation. ResultsThe difference in the corneal elastic modulus following CXL was significant in the anterior (p = 0.00002) and mid stroma (p = 0.001); however, the difference was not significant in the posterior stroma (p = 0.27) when compared to control corneas. The corneal elastic modulus of the anterior stroma increased by 178.44% in Group A, 119.7% in Group B and 50.73% in Group C compared to control corneas. For the mid stroma, the elastic modulus increased by 47.35% in Group A, 25% in Group B and 24.56% in Group C. No differences were observed in the posterior stroma between age groups. ConclusionsCorneal elasticity showed a greater response to CXL in the younger group compared to older groups. CXL treatment showed effectiveness in enhancing stromal strength, and the effect was concentrated in the anterior and mid stroma with minimal impact on the posterior stroma in all age groups.

Investigation of corneal topographic and densitometric properties of Wilson's disease patients with or without a Kayser-Fleischer ring

BackgroundTo investigate the topographic measurements and densitometry of corneas in Wilson’s disease (WD) patients with or without a Kayser-Fleischer ring (KF-r) compared to healthy individuals.MethodsThis cross-sectional study included 20 WD patients without a KF-r (group I), 18 WD patients with a KF-r (group II), and 20 age-matched controls (group III). The Pentacam high resolution imaging system is used to determine corneal topographic measurements and densitometry.ResultsMean age for groups I, II and III was 25.40 ± 6.43 years (14–36 years), 25.38 ± 6.96 years (16–39 years), 23.60 ± 6.56 years (17–35 years), respectively (P = 0.623). There was no significant difference between the groups in terms of the anterior corneal densitometry values (P > 0.05), while the 6–10 mm and 10–12 mm mid stroma and the 2–6 mm, 6–10 mm, and 10–12 mm posterior corneal densitometry values in group II were significantly higher than those in groups I and III (for all values, P < 0.05). However, the 10–12 mm posterior corneal densitometry values in group I were also significantly higher than those in group III (P = 0.038). The central corneal thickness (CCT), thinnest corneal thickness (tCT), and corneal volume (CV) values in groups I and II were significantly lower than those in group III (for CCT values, P = 0.011 and P = 0.009; for tCT values, P = 0.010 and P = 0.005; for CV values, P = 0.043 and P = 0.029).ConclusionIn WD patients with a KF-r, corneal transparency decreased in the peripheral posterior and mid stromal corneal layers; for these patients, corneal transparency may be impaired not only in the peripheral cornea but also in the paracentral cornea.

Multimodal Imaging Features of Schnyder Corneal Dystrophy.

Objective To describe the multimodal imaging of Schnyder corneal dystrophy. Methods Seven eyes of seven patients (5 female and 2 male patients) aged 52 to 92 years were included in this prospective observational study. Diagnosis of SCD was confirmed by histology after keratoplasty. In vivo multimodal imaging consisted of spectral domain-optical coherence tomography with cross sections, en face scans, corneal pachymetry, and epithelial mapping, and in vivo confocal microscopy was recorded. Ex vivo full-field optical coherence tomography scans of two corneal buttons were analyzed. The seven corneal buttons obtained during penetrating or deep anterior lamellar keratoplasty were processed for light microscopy. Results Slit-lamp examination showed central stromal opacities, arcus lipoides, and midperipheral haze. Corneal crystals were found in 2 out of 7 eyes. SD-OCT cross sections and en face scans showed diffuse hyperreflectivity of the anterior, mid, and posterior stroma with a maximum in the anterior stroma, hyporeflective stromal striae, and epithelial hyperreflectivity. Central corneal thickness ranged from 507 to 635 μm. IVCM revealed hyperreflective deposits in the epithelium and throughout the stroma, thin subepithelial nerves, and needle-shaped and rectangular crystals. Keratocyte nuclei were rare or undetectable. FF-OCT scans confirmed the presence of small round and needle-shaped hyperreflective deposits in the epithelium and stroma. Histology revealed vacuolization of the basal epithelial cells and empty interlamellar stromal vacuoles. Conclusion High-resolution multimodal imaging demonstrates the characteristic features of SCD which involve both the corneal epithelium and stroma, and it provides diagnosis confirmation even in eyes with no visible corneal crystals at slit-lamp examination.

Multimodal imaging of pre-Descemet corneal dystrophy.

The aim of this study was to assess structural and histological changes associated with pre-Descemet corneal dystrophy with multimodal in vivo imaging. Retrospective case series including eight corneas from four unrelated male patients with pre-Descemet corneal dystrophy characterized by the presence of punctiform gray opacities located just anterior to the Descemet membrane at slit-lamp examination of both eyes. In vivo confocal microscopy images were obtained in the central, paracentral, and peripheral corneal zones from the superficial epithelial cell layer down to the corneal endothelium in both eyes. Spectral domain optical coherence tomography scans (central and limbal zones) and mapping of both corneas were acquired. Diffuse small extracellular stromal deposits, presence of enlarged hyperreflective keratocytes in the posterior stroma with either hyperreflective or hyporeflective intracellular dots, and presence of activated keratocytes in the very anterior stroma were observed in all corneas with in vivo confocal microscopy. Spectral domain optical coherence tomography scans showed a hyperreflective line anterior to Descemet's membrane running from limbus to limbus and associated with a second thinner hyperreflective line just beneath Bowman's layer. Fine hyperreflective particles were observed in the posterior, mid, and anterior stroma on optical coherence tomography scans. The clinical presentation and structural anomalies found in isolated sporadic pre-Descemet corneal dystrophy are in favor of a degenerative process affecting corneal keratocytes with no epithelial or endothelial involvement. The maximum damage is found just anterior to the Descemet membrane resulting in pre-Descemet membrane location of stromal opacities. Multimodal imaging of cornea reveals that the disorder affects the whole stroma and it permits better understanding of pre-Descemet corneal dystrophy pathophysiology together with ascertained diagnosis.

Predictive Factors for Successful Type 1 Big Bubble during Deep Anterior Lamellar Keratoplasty.

Objective Big bubble (BB)-deep anterior lamellar keratoplasty (DALK) has become the reference transplantation technique for corneal stromal disorders. Type 1 BB is the desired aspect but it is not constant. We aimed to determine the predictive factors of type 1 BB success. Methods Observational cohort study including 77 consecutive eyes of 77 patients undergoing DALK by one surgeon at a single reference center without any selection. Clinical and spectral domain optical coherence tomography data were collected pre- and postoperatively. Results Stromal scars were found in 91.8% of cases and were located in the anterior (90.9%), mid (67.5%), and posterior (36.4%) stroma. Type 1 BB (49.3% of cases) was significantly associated with the absence of scars in the posterior stroma, stage 1–3 keratoconus, and deep trephination. Among eyes with posterior scars, type 1 BB was associated with higher minimal corneal thickness, maximum-minimum corneal thickness < 220 μm, and diagnosis other than keratoconus. Eyes with type 1 BB featured significantly thinner residual stromal bed (22 ± 8 µm versus 61 ± 28 µm), thinner corneas at 12, 24, and 36 months, and better visual acuity at 12 months compared with eyes with no type 1 BB. Conversely, no significant differences between both groups were observed for graft survival, visual acuity at 24 and 36 months, and endothelial cell density at 12 and 36 months. Conclusion OCT assessment before DALK is useful for choosing trephination depth that should be as deep as possible and for looking for posterior scars. The BB technique may not be the most appropriate method in keratoconus with posterior scars. Follow-up data do not support the need for conversion to penetrating keratoplasty when type 1 BB cannot be obtained nor does it support the need for performing a penetrating keratoplasty as a first-choice procedure in eyes with posterior stromal scars.

Keratocyte Density Is Reduced and Related to Corneal Nerve Damage in Diabetic Neuropathy.

The purpose of this study was to assess the relationship between corneal keratocyte density (KD) and corneal nerve damage in patients with and without diabetic peripheral neuropathy. Eighty-six patients with type 1 and type 2 diabetes and 21 age-matched control subjects underwent assessment of the neuropathy disability score, quantitative sensory testing, electrophysiology, and corneal confocal microscopy and were divided into those without (DN-) (n = 22) and with (DN+) (n = 64) diabetic neuropathy. Corneal sub-basal nerve parameters and KD in the anterior, mid, and posterior stroma were quantified. Anterior, mid, and posterior stromal KD were significantly reduced in DN- (P = 0.02, P = 0.009, P = 0.01, respectively) and DN+ (all P < 0.0001) subjects compared to controls. Corneal nerve branch density (CNBD) (P < 0.0001, P < 0.0001) and corneal nerve fiber length (CNFL) (P = 0.03, P < 0.0001) were significantly reduced in DN- and DN+ subjects, respectively, and corneal nerve fiber density (CNFD) (P < 0.0001) was significantly reduced only in DN+ subjects compared to controls. Anterior, mid, and posterior stromal KD correlated significantly with CNFD (P = 0.008, P = 0.005, P = 0.01), CNBD (P = 0.01, P = 0.006, P = 0.001), and CNFL (P = 0.04, P = 0.008, P = 0.003), respectively. This study demonstrates a reduction in anterior, mid, and posterior KD, which is associated with corneal sub-basal plexus nerve damage in patients with diabetes.

Automated Detection and Classification of Corneal Haze Using Optical Coherence Tomography in Patients With Keratoconus After Cross-Linking.

To evaluate a proposed technology for offering objective grading and mapping of corneal haze as detected by corneal spectral domain optical coherence tomography after corneal cross-linking. This was a retrospective study to evaluate corneal optical coherence tomography images performed on 44 eyes of 44 patients who underwent corneal cross-linking between January 2014 and May 2015, at the American University of Beirut Medical Center. Overall average brightness of the cornea was markedly increased from 43.4% (±6.0) at baseline to 50.2% (±4.4) at 1 month, 47.9% (±4.4) at 3 months, and 46.4% (±5.7) at 6 months with P <0.001, <0.001, and 0.005, respectively. In the anterior stroma, the average brightness significantly increased at 1, 3, and 6 months with values of 54.8% (±3.9), 52.5% (±5.2), and 49.7% (±6.9) with P <0.001, <0.001, and 0.003, respectively. In the mid stroma, the change was clinically significant at 1 and 3 months, whereas in the posterior stroma, it was only significant at 1 month compared with baseline (P = 0.003). Overall, haze was mostly present at 1 month after surgery in all regions, especially in the anterior (32.1%; ±19.2) and mid stromal regions (9.1%; ±18.8), P <0.001 and 0.001, respectively. In contrast, haze in the posterior stromal region peaks at 3 and 6 months after surgery. Anterior stromal haze was the greatest in intensity and area and it was present for a longer time span than mid and posterior stromal haze. At 12 months, the anterior stroma had still more haze intensity than preoperatively. This image-based software can provide objective and valuable quantitative measurements of corneal haze, which may impact clinical decision-making after different corneal surgeries.

Assessment of stromal riboflavin concentration–depth profile in nanotechnology-based transepithelial corneal crosslinking

To determine the intrastromal concentration of riboflavin in nanotechnology-based transepithelial corneal crosslinking. Consiglio Nazionale delle Ricerche, Messina, Italy. Experimental study. Six human donor sclerocorneal tissues were used to evaluate penetration of nanotechnology-based riboflavin 0.1% solution in the stroma through the intact epithelium. Three additional tissues were deepithelialized and soaked with dextran 20.0%-enriched riboflavin 0.1% solution for 30minutes. After corneal soaking with riboflavin, all tissues were irradiated using a 10mW/cm2 device for 9minutes. Two-photon emission fluorescence (TPEF) axial scanning measurements were collected in all specimens before treatment and immediately after corneal soaking with riboflavin and ultraviolet-A (UVA) irradiation of the cornea. The absorbance spectra of each tissue were collected at the same time intervals. The TPEF signals and absorbance spectra were used to calculate the concentration-depth profile of riboflavin in the corneal stroma during treatments. The mean stromal riboflavin concentration was 0.008%±0.003% (SD) and 0.017%±0.001% after transepithelial soaking with the nanotechnology-based solution and standard soaking, respectively (P=.001). After UVA irradiation of the cornea, the mean consumption of riboflavin was 52%±13% and 67%±2% in the study group and control group, respectively (P<.01). The nanotechnology-based platform was effective in enriching the anterior stroma with riboflavin through the intact epithelium, although the riboflavin concentration-depth profile rapidly decreased across the mid and posterior stroma. The treatment-induced stiffening effect on the corneal stroma was not assessed in this study.

Corneal elasticity after oxygen enriched high intensity corneal cross linking assessed using atomic force microscopy

The purpose of this study was to assess anterior and mid corneal stromal elasticity after high intensity (HI) corneal cross linking (CXL), with and without oxygen (O2) enrichment, and compare these results to conventional CXL. Experiments were performed on 25 pairs of human cadaver eyes, divided into four different groups. Group 1 included corneas that did not receive treatment and served as controls; Group 2 included corneas that received conventional CXL treatment (Dresden Protocol: corneal epithelial debridement, 30 min of riboflavin pretreatment followed by 30 min of exposure to 3 mW/cm2 of ultraviolet light); Group 3 included corneas that received HI CXL treatment (corneal epithelial debridement, 30 min of riboflavin pretreatment followed by 3 min of exposure to 30mW/cm2 of ultraviolet light); and Group 4 included corneas that received the same treatment as Group 3, except that they were enriched with oxygen (4 L per minute pure O2 gas stream) during ultraviolet irradiation. In each group, corneas were subdivided to assess anterior stromal elasticity and mid stromal elasticity. Corneal stromal elasticity was quantified using Atomic Force Microscopy (AFM) through micro-indentation. Young's modulus for the anterior corneal stroma was 14.5 ± 6.0 kPa, 80.7 ± 44.6 kPa, 36.6 ± 10.5 kPa, and 30.6 ± 9.2 kPa, for groups 1, 2, 3 and 4 respectively. Young's modulus for the mid corneal stroma was 5.8 ± 2.0 kPa, 20.7 ± 4.3 kPa, 12.1 ± 4.9 kPa, and 11.7 ± 3.7 kPa, for groups 1, 2, 3 and 4, respectively. In the anterior stromal region, conventional CXL demonstrated a significantly different result from the control, whereas the two HI CXL protocols were not significantly different from the control. There were no statistical differences between the two HI CXL protocols, although only the HI CXL protocol with O2 enrichment was significantly different from the conventional CXL group. In the mid stromal region, once again only conventional CXL demonstrated a significantly different result from the control. There were no statistical differences between the two HI CXL protocols, and both HI CXL protocols were significantly different from the conventional CXL group. Oxygen enriched HI CXL seems to offer similar changes in corneal elasticity when compared to HI CXL without the presence O2. Conventional CXL increases corneal stiffness more than HI CXL both with and without O2 enrichment.

Variant lattice corneal dystrophy associated with compound heterozygous mutations in the TGFBI gene

Background/AimsTo report the clinical, histopathological and genetic features of a variant of lattice corneal dystrophy (LCD) associated with two pathogenic mutations in the transforming growth factor-B-induced (TGFBI) gene.MethodsClinical characterisation was...

Reasons to reconsider interpretation of densitometry for corneal transparency

To assess the effect of light scattering in the corneal epithelium on densitometric brightness of the stroma through mathematical modeling of the interaction between a light beam and the two-layer epithelium-stroma system. In order to study the scattering behavior of a plane-parallel non-coherent beam at the epithelium-stroma interface, a multi-age group was formed (87 patients, 174 eyes) that comprised two subgroups with equal number of assign participants: healthy patients with no systemic changes and mixed patients with undisturbed corneal transparency that, nevertheless, were under instillation therapy. In the first subgroup, the assessment of light scattering was done at random times, while in the second subgroup - within the first 200 seconds after the instillation in order to avoid a reaction of the ocular surface structures, including epithelium (rapid response of epithelial cells to instillations). Light scattering intensity was measured with Pentacam (USA). The densitometric curve and brightness maxima of the epithelium and neighboring stroma as well as the mid and upper stroma were recorded for each patient. A physico-mathematical model of the interaction between a light beam and the two-layer epithelium-stroma system was created. The study confirmed our theoretical CONCLUSION: on the effect size of light scattering intensity on the apparent densitometric brightness of the stroma. Direct densitometry of multilayered spherical objects, of which each layer possesses its own optical properties, has been proved inaccurate. Complex optical interactions in a multilayered model cause a false increase in brightness of more transparent layers that lie behind those with pronounced light scattering properties. The function that describes the false increase in scattering ability of the layer that is more transparent rises sharply at the border between the layers.

Characteristics of Pre-Descemet Membrane Corneal Dystrophy by Three Different Imaging Modalities—In Vivo Confocal Microscopy, Anterior Segment Optical Coherence Tomography, and Scheimpflug Corneal Densitometry Analysis

To evaluate the characteristics of pre-Descemet membrane corneal dystrophy by 3 different imaging modalities: in vivo confocal microscopy (IVCM), anterior segment optical coherence tomography (ASOCT), and Scheimpflug corneal densitometry analysis. A 32-year-old male patient with pre-Descemet membrane corneal dystrophy was subjected to imaging by IVCM, ASOCT, and Scheimpflug tomography. Slit-lamp biomicroscopy showed the presence of tiny pleomorphic opacities in the posterior stroma, immediately anterior to Descemet membrane bilaterally. On IVCM, pleomorphic, hyperreflective punctate particles were seen both intracellularly and extracellularly in the anterior and mid stroma with increased reflectivity of some keratocytes that, however, were of normal size. These changes increased in severity from the anterior to mid stroma. The posterior stroma had grossly enlarged hyperreflective keratocytes with prominent processes. The hyperreflective particles were also seen scattered on the endothelium. ASOCT revealed a well-delineated homogenous band of increased reflectivity of approximately 70 μm width in the posterior stroma of both eyes with a normal-appearing anterior and mid stroma. Corneal densitometry measured by Scheimpflug optical analysis revealed a higher amount of backscattered light from the posterior stroma with a posterior to anterior ratio of 0.8. In pre-Descemet membrane corneal dystrophy, although the structural changes seem to be limited to the posterior stroma as seen clinically and on ASOCT, IVCM demonstrates that the pathology is more extensive involving the entire corneal stroma and endothelium.

Confocal Microscopy Analysis of Corneal Changes After Photorefractive Keratectomy Plus Cross-Linking for Keratoconus: 4-Year Follow-up

To analyze corneal confocal microscopy changes after combined photorefractive keratectomy (PRK) plus the cross linking (CXL) procedure. Prospective interventional case series. At the Department of Basic Medical Sciences, Neuroscience, and Sense Organs of the University of Bari, Bari, Italy, 17 eyes of 17 patients with progressive keratoconus underwent confocal microscopy examination before and after 1, 3, 6, 12, 18, and 48 months following PRK plus the CXL procedure. The main outcome measures were mean superficial epithelial cell density; mean basal epithelial cell density; mean anterior, mid and posterior keratocyte density; qualitative analysis of stromal backscatter; sub-basal and stromal nerve density parameters; and mean endothelial cell density. During the 4-year follow-up, the mean superficial epithelial cell density, mean basal epithelial cell density and mean endothelial cell density remained unchanged (P > 0.05). The anterior mid-stromal keratocyte density showed a significant decrease (P < 0.05) as compared with preoperative values, and the posterior stromal keratocyte density showed a significant increase at 1 and 3 months of follow-up. Sub-basal and stromal nerve density parameters were significantly decreased until postoperative month 6 (P < 0.05 at 1, 3, and 6 months) and then tended to increase up to preoperative values by the 18th postoperative month. Corneal changes after the PRK plus CXL procedures seem to be pronounced and long lasting as far as keratocyte density of the anterior and mid stroma is concerned. Sub-basal nerve densities tend to reach preoperative values 6 months after surgery.

Laboratory evaluation of selective in situ refractive cornea collagen shrinkage with continuous wave infrared laser combined with transepithelial collagen cross-linking: a novel refractive procedure

BackgroundThis research comprised a laboratory evaluation of a novel refractive surgery technique involving sequential corneal subsurface shrinkage-driven reshaping using a continuous wave mid-infrared laser application followed by stiffening via rapid transepithelial higher fluence collagen cross-linking for shape persistence/longevity on cadaver corneas.Materials and methodsTen cadaver corneas were used in this study. During use, all were affixed in an artificial chamber. Thermal delivery entailed a continuous wave laser at 2013 nm wavelength, approximately 650 mW power under scanner control (about 3 mm/sec linear draw speed), with a planoconcave sapphire applanation lens cooled to 8°C. Group 1 (n = 5, myopic treatment) eyes were exposed to three concentric annuli with diameters of 3 mm, 4 mm, and 5 mm. Group 2 (n = 5, hyperopic treatment) eyes were exposed to three concentric annuli with diameters of 6 mm, 7 mm, and 8 mm. The clinical change in shape of the cornea was visualized immediately under a slit-lamp. A transepithelial higher fluence corneal collagen cross-linking step followed each thermal treatment, comprising 0.1% riboflavin drops with 0.02% benzalkonium chloride and 0.2% carboxymethlycellulose in deuterated water (D2O) applied on the epithelium corneal surface for 10 minutes. Next, each cornea was exposed to 10 mW/cm2 of ultraviolet A (365 nm) light for 10 minutes. The corneas were evaluated before and after thermal remodeling and cornea collagen cross-linking for corneal topography, corneal optical coherence tomography, cornea Scheimpflug tomography, and clinical photographs.ResultsThe histopathology effect was noted clinically as concentric white rings underneath the epithelium and Bowman’s membrane, with the lesion depth extending down to 400 μm in the mid stroma of the cornea. This was confirmed by corneal anterior segment optical coherence tomography. The topographic change noted was a 4–8 diopter hyperopic shift in group 1 and a 2–6 diopter myopic shift in group 2.ConclusionThis novel refractive surgery technique appears to generate a significant refractive change (+/−) in the cornea, without affecting the epithelium or Bowman’s membrane and without any visible epithelial defect. The transepithelial collagen cross-linking used as a second step in the same procedure aims to stabilize this effect in the long term. Further clinical studies are planned to validate these initial clinical results.

Assessment of Corneal Thickness and Keratocyte Density in a Rabbit Model of Laser In Situ Keratomileusis Using Scanning Laser Confocal Microscopy

To determine the repeatability of corneal thickness and keratocyte density using in vivo confocal scanning laser microscopy in a rabbit model of laser in situ keratomileusis. Prospective, experimental animal study. En face tomographic images of corneal tissue were captured from 5 New Zealand white rabbits. Central corneal thickness was compared with conventional ultrasonic pachymetry. Keratocyte density was measured as a function of stromal depth at baseline and 6 weeks after a 130-μm lamellar incision in the following regions: first countable stromal image (30 to 39 μm), anterior stroma (40 to 75 μm), incision zone (76 to 150 μm), mid stroma (151 to 250 μm), and deep stroma (251 to 400 μm). The mean residual difference between ultrasonic and confocal corneal thickness measurements was 2.1 μm (95% confidence interval [CI], -7.0 to 11.2 μm; P = .61). Before the lamellar incision, keratocyte density was highest in the first countable frame of the anterior stroma, 53 800 cells/mm(3) (95% CI, 35 000 to 72 000 cells/mm(3)) and was least in deep stroma, 27 100 cells/mm(3) (95% CI, 22 400 to 32 000 cells/mm(3)). Six weeks after stromal lamellar incision, keratocyte density was unchanged in the first countable frame of the anterior stroma, 43 700 cells/mm(3) (95% CI, 31 800 to 55 500 cells/mm(3); P = .29). There were no changes in cell density in deeper stromal regions. There was excellent agreement between ultrasonic and confocal microscopy measurements of corneal thickness. In vivo repeatability of keratocyte density estimation using scanning laser confocal microscopy is comparable with the results of previous reports using tandem-scanning confocal microscopy. Keratocyte density was more varied, but not significantly different, in the anterior-most corneal stroma 6 weeks after a lamellar incision.