Evaluation of perioperative pachymetry of thin corneas during corneal crosslinking using hypo-osmolar riboflavin and hydroxypropyl methylcellulose

ABSTRACT Purpose: This study aimed to analyze variations in intraoperative corneal thickness during corneal cross-linking in patients with keratoconus and to investigate its possible correlation with presurgical maximal keratometry (Kmax) and pachymetry. Methods: This was a prospective case series. We used a method similar to the Dresden protocol, with the application of hydroxypropyl methylcellulose 0.1% hypo-osmolar riboflavin in corneas between 330 and 400 µm after epithelium removal. Corneal thickness was measured using portable calipers before and immediately after epithelium removal, and 30 and 60 min after the procedure. Results: The 30 patients in this study were followed up for one year. A statistically significant difference was observed in pachymetry values during the intraoperative period (p<0.0001) and an increase of 3.05 µm (95%C1: 0.56-5.54) for each diopter was seen after epithelium removal (p0.019). We found an average Kmax difference of —2.12 D between men and women (p0.013). One year after treatment, there was a statistically significant reduction in pachymetry (p<0.0001) and Kmax (p0.0170) values. Conclusions: A significant increase in pachymetry measurements was seen during the procedure, and most patients showed a regression in Kmax and pachymetry values one year after surgery.

Inflammatory and genomic interactions within keratoconus susceptible patients: a nationwide registered case-control study.

PURPOSE: This study aimed to investigate the association between variants in the interleukin (IL)-1 gene cluster and susceptibility to keratoconus (KC) in an Iranian population. METHODS: In the case group, there were 188 KC patients diagnosed by clinical findings and corneal tomography. The control group included all 205 healthy controls with no personal or family history of eye-related, metabolic, or immune system-related disease. Using the standard salting out extraction procedure, genomic DNA was isolated from peripheral blood leukocytes. The genotypes were determined by applying agarose gel electrophoresis for the IL-1RN 86 bp VNTR and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rs16944 and rs1143634. RESULTS: The results showed a significant association between the IL-1ß rs1143634 (rs1143634 T allele, P = 0.008) and IL-1RN 86 bp VNTR polymorphisms (LL and LS genotype, P = 0.048 and 0.012 respectively) and susceptibility to KC in the Iranian population. The genotype distributions of rs1143634 (P = 0.004) and rs2234663 (P = 0.042) significantly differed between case and control groups, with certain genotypes demonstrating a protective effect against KC. Logistic regression analysis revealed a protective effect of the IL-1RN L allele [odds ratio (OR) = 0.367, 95% confidence interval (CI): 0.240-0.562; P = 0.000] and certain haplotypes (OR = 0.628, 95% CI: 0.447-0.884; P = 0.007) against KC. However, no significant association was found for the IL-1ß rs16944 polymorphism. CONCLUSION: This study provides evidence for an association between variants in the IL-1 gene cluster and susceptibility to KC in an Iranian population. Further research on larger and more diverse populations is warranted to validate these findings and explore the underlying mechanisms involved.

The development of a machine learning model to train junior ophthalmologists in diagnosing the pre-clinical keratoconus.

Purpose: This study aims to evaluate the diagnostic performance of a machine learning model (ML model) to train junior ophthalmologists in detecting preclinical keratoconus (PKC). Methods: A total of 1,334 corneal topography images (The Pentacam HR system) from 413 keratoconus eyes, 32 PKC eyes and 222 normal eyes were collected. Five junior ophthalmologists were trained and annotated the images with or without the suggestions proposed by the ML model. The diagnostic performance of PKC was evaluated among three groups: junior ophthalmologist group (control group), ML model group and ML model-training junior ophthalmologist group (test group). Results: The accuracy of the ML model between the eyes of patients with KC and NEs in all three clinics (99% accuracy, area under the receiver operating characteristic (ROC) curve AUC of 1.00, 99% sensitivity, 99% specificity) was higher than that for Belin-Ambrósio enhanced ectasia display total deviation (BAD-D) (86% accuracy, AUC of 0.97, 97% sensitivity, 69% specificity). The accuracy of the ML model between eyes with PKC and NEs in all three clinics (98% accuracy, AUC of 0.96, 98% sensitivity, 98% specificity) was higher than that of BAD-D (69% accuracy, AUC of 0.73, 67% sensitivity, 69% specificity). The diagnostic accuracy of PKC was 47.5% (95%CI, 0.5-71.6%), 100% (95%CI, 100-100%) and 94.4% (95%CI, 14.7-94.7%) in the control group, ML model group and test group. With the assistance of the proposed ML model, the diagnostic accuracy of junior ophthalmologists improved with statistical significance (p < 0.05). According to the questionnaire of all the junior ophthalmologists, the average score was 4 (total 5) regarding to the comprehensiveness that the AI model has been in their keratoconus diagnosis learning; the average score was 4.4 (total 5) regarding to the convenience that the AI model has been in their keratoconus diagnosis learning. Conclusion: The proposed ML model provided a novel approach for the detection of PKC with high diagnostic accuracy and assisted to improve the performance of junior ophthalmologists, resulting especially in reducing the risk of missed diagnoses.

Air Pollution and the Prevalence of Keratoconus: Is There a Connection?

PURPOSE: Keratoconus is a progressive, asymmetrical corneal ectasia with multifactorial origin. Three identified risk factors for keratoconus include exposure to ultraviolet (UV) rays, eye rubbing, and atopy. Other factors like pollution would play a role in the physiopathology of keratoconus. In this study we investigate the effects of particles matter (PM) of 2.5 and 10 µm, but also nitrogen dioxide (NO2) and the correlation with the prevalence in the scientific literature. METHOD: A literature review was performed using four databases (PubMed, Research gate, Google scholar and International Journal of Keratoconus and Ectatic Corneal Diseases) according to strict selection criteria. Levels of fine particles and nitrogen dioxide were extracted from available World Health Organization (WHO) databases and correlated with prevalences from epidemiological studies. RESULTS: The mean pollution rate in the selected studies was 26.88 ± 25.26 µg/m3 for PM2.5 , 58.23 ± 60.98 µg/m3 for PM10 and 24.79 ± 12.58 µg/m3 for NO2. Pearson correlation tests revealed a significant positive correlation between prevalence of keratoconus with particles rate of PM2.5 (R = 0.58; p < 0.001), PM10 (R = 0.67; p < 0.001) and NO2 (R = 0.64; p = 0.00016). CONCLUSION: Fine particles appear to be a risk factor for keratoconus. These pollutants may act indirectly by exacerbating known risk factors such as atopy and eye rubbing. Atmospheric pollution may also have a direct effect on the cornea, by disturbing the structure of the epithelium and increasing cell apoptosis.

Comparison of a Scheimpflug imaging with other screening indices in diagnosing keratoconus and keratoconus suspect.

Keratoconus (KC) is an irreversible blinding eye disease; therefore, early screening of KC suspects (KCS) is crucial for protecting patients' quality of life. Scheimpflug imaging is a commonly used screening device in clinical practice. We aimed to evaluate the diagnostic ability of a Scheimpflug imaging device (Scansys) for KC and KCS and compared it with other Scheimpflug-based devices (Pentacam and Corvis ST). This prospective case-control study included 107 normal eyes, 72 KCS, and 57 KC. Scansys screening index Keratoconus probability (KCP) showed excellent performance in diagnosing KC at a cutoff value of 16.4 (area under the receiver operating characteristic [AUROC] = 1.000), with 100% sensitivity and 98.11% specificity. KCP had a better KCS diagnostic ability at a cutoff value of 8.9 (AUROC = 0.813) than Corvis biomechanical index (CBI, AUROC = 0.764), reaching 67.61% sensitivity and 85.85% specificity. Pentacam screening index Belin/Ambrósio enhanced ectasia display deviation (BAD-D) showed the best performance with 92.96% sensitivity and 89.62% specificity at a cutoff value of 1.525 (AUROC = 0.970) in diagnosing KCS. Scansys provides accurate KCP parameters in diagnosing KC; however, the efficiency of diagnosing KCS should be further optimized.

Is there a relationship between the keratoconus and the peripapillary choroidal vasculature?

PURPOSE: To evaluate the peripapillary/parapapillary choroidal vascular parameters in the keratoconus (KC) and to determine the relationship between topography parameters and the peripapillary/parapapillary choroidal vascular parameters. METHOD: Ninety eyes of ninety patients with different stages of KC and 29 eyes of twenty-nine patients without KC were enrolled in the study. Patients with KC were divided into three groups according to the Amsler-Krumeich classification scale. The choroidal vasculature was assessed by choroidal vascular parameters [such as parapapillary choroidal microvascular density (pCMVd) and peripapillary choroidal vascularity index (pCVI)]. These parameters were also evaluated for correlation with other parameters. RESULT: The retinal nerve fibre layer thickness (RNFLT) of the superior-temporal area and the pCVI were decreased in group 3 compared to the control group (superiror-temporal RNFLT: 122.27 ± 21.43 vs 139.90 ± 21.7, p = 0.01 and pCVI: 67.04 ± 4.14 vs 69.99 ± 4.38, p = 0.04). The superior-temporal RNFLT was decreased in group 3 compared to group 2 (122.27 ± 21.43 vs 141.83 ± 25.58, p = 0.006). There was a negative correlation between pCVI and average simulated keratometry (mean sim K), but this association was weak (r = - 0.29 p = 0.001). CONCLUSION: This study demonstrated that there may be changes in pCVI in patients with grade 3 KC and that there may be an association between pCVI and mean sim K. As KC grade increases, pCVI may decrease. Furthermore, pCVI may have a negative correlation with mean sim K.

Alport Syndrome With a Rare Collagen Type IV Alpha-4 (COL4A4) Gene Mutation: A Case Report.

Alport syndrome (AS) is a rare, progressive hereditary kidney disease characterized by sensorineural hearing loss and visual abnormalities. It is caused by a mutation in the collagen type IV alpha-4 (COL4A4) gene, which produces type IV collagen, and often manifests in individuals with hematuria, proteinuria, edema, and hypertension. Here, we present a case of AS in a 15-year-old boy with a COL4A4 gene mutation, with renal and extrarenal findings. The patient presented with subnephrotic proteinuria and microscopic hematuria, autoimmune hypothyroidism, and keratoconus. Light microscopy examination of renal biopsy revealed three globally sclerosed renal cortical parenchyma areas with periglomerular fibrosis, and electron microscopy showed variable thickness of glomerular basement membrane with festooned appearance, as well as splitting of lamina densa giving basket weave and criss-cross pattern.

Mendelian randomization reveals that abnormal lipid metabolism mediates the causal relationship between body mass index and keratoconus.

Previous studies suggest that a high body mass index (BMI) may be a risk factor for keratoconus (KC), but the causal relationship remains unclear. This study used Mendelian randomization (MR) to investigate this connection and explore the mediating role of circulating serum metabolites and inflammatory factors in this association. Two-sample MR analysis was conducted to assess the relationship between BMI and KC. The study employed a two-step MR approach to evaluate the mediating roles of 91 inflammatory markers and 249 serum metabolites in the BMI-KC relationship. Inverse variance weighting (IVW) was the primary method, and multiple sensitivity analyses were performed to ensure robustness. IVW analysis revealed a positive causal relationship between BMI and KC (OR IVW = 1.811, 95% CI 1.005-3.262, P = 0.048). Although IL-12ß and IL-4 were causally associated with KC, they did not mediate the BMI-KC relationship. Five serum metabolites were identified as potential mediators, with HDL cholesterol and triglyceride ratios showing significance. This study clarified the causal relationship between high BMI and KC, suggesting that high BMI may induce KC through lipid metabolism abnormalities. These findings underscore the importance of managing BMI for KC prevention.

Experimental Models for Keratoconus: Insights and challenges.

Keratoconus, a progressive corneal disorder characterized by the thinning and conical protrusion of the cornea because of collagen degradation, poses significant challenges to patients and clinicians. Most successful animal models in keratoconus are based on genetic mutations and knock-outs in mice and rats that hinder normal corneal stromal architecture, thickness, or strength. While mice and rat models are suitable to study the molecular mechanism and physiological changes to the cornea, they are not suitable for experimental research; especially for surgical interventions like: Deep anterior lamellar keratoplasty (DALK), Stromal lenticule addition keratoplasty etc. using advanced therapies. This review article comprehensively examines recent advancements in experimental models for keratoconus, focusing on their potential for translational research and the challenges ahead. It explores the historical context of experimental models, focusing on animal-based models, mainly rabbits in particular. These advancements enable researchers to mimic the biomechanical and biochemical alterations observed in keratoconic corneas. While these models offer valuable insights into disease mechanisms and treatment development, several challenges remain in transforming experimental findings into clinical applications.

Transcriptomic analysis of Keratoconus in Han Chinese patients: Insights into differential gene expression and ethnic-specific patterns.

Keratoconus (KC) is a progressive corneal ectatic disorder with a high prevalence among Asians. This study aimed to explore the differential gene expression patterns in Han Chinese patients with KC, focusing on mRNAs and long noncoding RNAs (lncRNAs), to provide insights into the pathogenesis of the disease. Corneal tissues from KC patients and healthy controls were collected, and RNA sequencing was performed to profile mRNA and lncRNA expression. A total of 1973 differentially expressed mRNAs (DEGs) and 386 differentially expressed lncRNAs (DELs) were identified in KC-affected corneas. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed significant enrichment in pathways related to ECM modulation, PI3K-Akt pathway and calcium signaling pathway. Furthermore, protein-protein interaction (PPI) network highlighted hub genes involved in ECM remodeling and inflammatory responses. Co-expression analysis of lncRNAs and mRNAs further prioritized 13 DELs linked to these hub genes. RT-qPCR validation confirmed the differential expression of select candidates. A meta-analysis integrating seven datasets from diverse ethnic backgrounds was performed and it suggested ethnic-specific differences in gene expression patterns. This study sheds new light on the molecular mechanisms underlying KC in the Han Chinese population, pinpointing potential therapeutic targets. It also emphasizes the critical role of ethnic-specific gene expression patterns in KC research, highlighting a need for tailored approaches in disease management and treatment.

Pediatric Keratoconus.

Keratoconus is a common pediatric corneal disease, leading to vision impairment and amblyopia. Compared to its adult counterpart, pediatric keratoconus has an advanced presentation, rapid progression, higher incidence of complications such as corneal hydrops, and potential impact on the child's quality of life. It typically manifests during puberty and can evolve rapidly to more severe stages if left untreated. This rapid progression underscores the importance of early diagnosis through regular screening in pediatric populations and vigilant monitoring of pediatric keratoconus suspects. Concomitant ocular allergies, ocular anomalies, systemic diseases (eg. syndromes), and poor compliance with contact lenses might impede prompt intervention and frequently postpone rehabilitation. Corneal collagen crosslinking is a crucial intervention in the management of pediatric keratoconus because it strengthens the corneal microstructure and halts the disease's progression. When conservative measures fail, keratoplasty remains a viable option with generally favorable outcomes, though with unique challenges in post-operative care, including concerns related to sutures, long-term graft survival and need for repeated examinations under anesthesia. A multidisciplinary approach involving ophthalmologists, optometrists, pediatricians, and other healthcare professionals, focusing on early diagnosis and timely intervention, is essential for the comprehensive management of pediatric keratoconus and to mitigate its impact on children's lives.

Patient evaluated economic impact of keratoconus in New Zealand.

CLINICAL RELEVANCE: Few studies have investigated the economic burden of keratoconus to patients and society including a patient's lifetime expenditure. Analysing disease cost is important in advocating funding and planning for appropriate management and treatment. BACKGROUND: Keratoconus can be an expensive disease, including costs associated with visual rehabilitation and surgery. This study aimed to assess direct and indirect expenditures incurred by keratoconus subjects, to estimate the lifetime per capita cost and the total costs of keratoconus per year in Auckland, New Zealand. METHODS: A cross-sectional study where participants with keratoconus were recruited from public and private clinics to complete an anonymous questionnaire. A keratoconus health expenditure questionnaire was used to assess direct and indirect expenditures for participants. Estimated lifetime per capita costs and total direct and indirect costs associated with keratoconus were calculated. RESULTS: Responses from 110 participants were recorded. Most participants (55.5%) were between the ages of 21 and 30, 59.1% were male, and Pacific People were over-represented (14.5%) compared to the national population. The prevalence of visual rehabilitation was low with 74.5% of participants never wearing contact lenses and 32.7% of subjects wearing spectacles purchased over 24 months prior. Few participants (26.5%) had subsidised care, including private health insurance with optical cover or the contact lens subsidy. Total direct and indirect costs are estimated to be NZD 30.9 million per year with an estimated lifetime per capita out-of-pocket cost of NZD 79,254. CONCLUSION: Subjects with keratoconus experience significant direct and indirect costs with limited subsidisation from the government and private health insurance. The significant out-of-pocket costs required for managing keratoconus are likely a significant barrier to accessing appropriate visual rehabilitation in New Zealand.

Crosslinked All-Femtosecond Laser-Cut Corneal Allogenic Intracorneal Ring Segments (AFXL CAIRSs): Pilot Ex Vivo Study and Report of First Two Cases Performed in Italy.

Objectives: This pilot ex vivo study and first clinical experience in Italy evaluate the impact of using pre-implantation crosslinking on all-femtosecond laser-cut corneal allogenic intracorneal ring segments (AFXL CAIRSs). Methods: Six human donor eye-bank corneas were used for this preclinical ex vivo human study. Three donor (D) corneas were used for AFXL CAIRSs. First, they were prepared with an IntraLase™ femtosecond laser (Johnson & Johnson, New Brunswick, NJ, USA). The allogenic tissue rings were crosslinked before implantation with Riboflavin-UV-A accelerated crosslinking protocol (ACXL) with a 0.1% HPMC Riboflavin isotonic solution (Vibex Rapid, Glaukos-Avedro, Burlington, MA, USA) and a new KXL UV-A emitter (Glaukos-Avedro, USA). Three corneas were used as recipients (Rs) of the AFXL CAIRSs. After completing the ex vivo phase, IRB approval and signing a specific informed consent, the first two Italian patients were treated. A single ACXL CAIRS was implanted in a 51-year-old male with 53.53 D K steep, 363 µm minimum corneal thickness (MCT) and a double ACXL CAIRS was implanted in a 46-year-old male patient with 58.30 D K steep, 443 µm MCT. The longest follow-up was at three months. Results: Crosslinking of the segments enhanced tissue stiffness and grip, facilitating manipulation and CAIRS insertion into the recipient tunnels, and the yellowish color of the crosslinked segments improved visibility. The segment's thickness and volume remained unaltered during the follow-up. Both patients improved UDVA and BSCVA. K steep and High-Order Aberrations (HOAs) were reduced and MCT increased. Conclusions: Pre-implantation ACXL facilitated CAIRS insertion preserving dimensions and volume during the follow-up, rendering this important step a promising candidate in method standardization. Functional data and MCT improved significantly without adverse events.

One-Year Results of a Posterior Chamber Toric Phakic Intraocular Lens Implantation in Patients with Keratoconus.

Purpose: To determine clinical and refractive results after the implantation of EyeCryl Phakic Toric intraocular lens in patients with stable keratoconus. Methods: The study included all patients diagnosed with keratoconus who underwent implantation of an EyeCryl Phakic Toric intraocular lens (Biotech Healthcare Holding; Ahmedabad, India) in at least one eye and had a follow-up of at least 12 months. Visual and refractive data were collected for all patients, along with corneal tomography measurements using Pentacam, and vault measurement using optical coherence tomography. This retrospective study was conducted at a high-volume private refractive surgery center in Medellín, Colombia. Results: A total of 83 eyes from 47 patients were included in the study. The majority (71.1%) were female, with a mean age of 31.2 ± 5.1 years. After 12 months of follow-up post-surgery, the spherical equivalent improved significantly from -8.19 ± 4.04 D to -0.06 ± 0.48 D (p < 0.001). Furthermore, 77% of eyes had a post-surgical spherical equivalent within ±0.50 D, while 92% had residual astigmatism ≤0.50 D. Twelve months after surgery, mean manifest astigmatism was -0.28 ± 0.27 D. Uncorrected visual acuity also showed improvement, from 1.11 ± 0.35 LogMAR to 0.14 ± 0.11 LogMAR. Moreover, 52.4% of eyes demonstrated an improvement of at least one line in best-corrected visual acuity. Notably, no intraoperative or postoperative complications were observed in the study population. Conclusion: The implantation of EyeCryl Phakic Toric intraocular lenses represents a highly effective and safe option for correcting refractive errors in patients with a history of keratoconus. Refractive accuracy is excellent, and a significant proportion of patients experienced an improvement in their best-corrected vision by at least one line.

Associations between endothelial cell characteristics and corneal topography findings in different stages of keratoconus.

PURPOSE: This study aimed to compare and correlate specular microscope indices and corneal topography indices in different stages of keratoconus. METHODS: Two hundred forty-six eyes of 123 participants were enrolled in the study. Corneal topography was performed using Sirius (CSO, Italy), with a rotating Scheimpflug camera and a Placido disc topographer. Corneal endothelial cell indices were assessed using a specular microscope (Nidek CEM-530, Japan). Eyes were graded as keratoconus stages 0-4 according to the Amsler-Krumeich classification. Corneal topography and endothelial cell indices were compared among the groups, and the correlations between them were analyzed. RESULTS: The mean age of the patients was 23.26 ± 6.75 years (range, 14-47 years). Forty-eight cases were male (39%) and 75 were female (61%). There were no statistically significant age (p = 0.578) or sex ratio (p = 0.529) differences between the groups. Twenty-nine eyes were included in the control group (11.78%), while 41 (16.67%) had stage 1 keratoconus, 88 (35.77%) had stage 2, and 88 (35.77%) had stage 3. Measurement was not possible in stage 4 keratoconus. No statistically significant difference was determined in specular microscopy values according to the stage of keratoconus, except for the number of analyzed cells (NUM) (p > 0.05). The lowest NUM values were observed in stages 1, 2, and 3, with values of 184.34 ± 67.62 cells/mm2, 155.07 ± 59.48 cells/mm2, and 127.06 ± 64.39 cells/mm2, respectively (p = 0.001). In the keratoconus group, weak statistically significant negative correlations were observed between NUM and SimK1, SimK2, KVf, BCVf, KVb, and BCVb, while a weak positive correlation was noted between NUM and central corneal thickness (p < 0.05). CONCLUSIONS: NUM seems to decrease, while endothelial cell density exhibits no significant changes, with the progression of keratoconus. It appears that as keratoconus index values increase, NUM may decrease in different stages of keratoconus.

Risk Factors for Glaucoma and Ocular Hypertension and Post-Transplant Complications in Keratoconus: A Multivariable Analysis.

Background: The purpose of this study was to investigate the risk factors for glaucoma in patients with keratoconus following keratoplasty and to identify potential factors that could affect post-transplant complications. Methods: A retrospective analysis was conducted on the medical records of 192 keratoconus patients who underwent keratoplasty. Data on treatment history, perioperative information, tissue bank data, postoperative regimens, complications, and infections were collected and analyzed. Statistical analysis was performed to identify risk factors associated with glaucoma and transplant complications. Results: There was a 41.6% incidence of glaucoma (high IOP) (p < 0.001), with the median time to glaucoma development being 314 ± 67 days post-transplant. A significant number of patients required surgical intervention, accounting for 48.05%. Our analysis revealed a 14% graft failure rate. On univariable Cox proportional hazard analysis, the following factors demonstrated statistically significant associations with the risk of glaucoma after transplantation: donor endothelial cell density, the use of a single continuous suture type, and the surgeon (performing the surgery). For many variables, the only factor that remained significant was the surgeon. Among the factors analyzed for risks of failure post-transplantation, significant associations were observed for the donor age, the time from harvest to transplant, and the surgeon. In the multivariable analysis, donor age emerged as a significant predictor of post-transplant complications. Conclusions: Risk factors such as donor endothelial cell density, suture type, surgeon, host and donor size, and host gender were found to increase the risk of developing glaucoma post-keratoplasty. Surgeon type was identified as a significant risk factor, while donor age was predictive of post-transplant complications.

Challenging corneal diseases and microRNA expression: Focus on rare diseases and new therapeutic frontiers.

MicroRNAs (miRNAs) function as posttranscriptional regulators of gene expression by targeting specific messenger RNA (mRNA). This interaction modulates mRNA stability or translational efficiency, ultimately impacting the level of protein production. Emerging evidence suggests that miRNAs act as critical regulators in corneal diseases. These molecules finetune key processes like cell proliferation, differentiation, inflammation, and wound healing. We reviewed the literature to understand the role that miRNAs may play in the development of challenging and poorly understood corneal diseases. We focused on vernal keratoconjunctivitis, neurotrophic keratitis, keratoconus, Fuchs endothelial corneal dystrophy, and limbal stem cell deficiency. Furthermore, we explored currently studied agonists or antagonists of miRNAs that share similar pathways with ocular diseases and could be employed in ophthalmology in the future. The distinct miRNA expression profiles observed in different ocular surface pathologies, combined with the remarkable stability and relatively easy access of miRNA sampling in biofluids, present possibilities for the development of noninvasive and highly accurate diagnostic tools. Furthermore, comprehending miRNA's pathophysiological role could open new frontiers to a more comprehensive understanding of the pathophysiology underlying ocular surface diseases, thereby paving the way for the creation of novel therapeutic strategies.

Corneal higher-order aberrations in different types of irregular cornea.

OBJETIVE: To assess differences in corneal higher-order aberrations (i.e., HOAs) between six different types of irregular corneas (i.e, keratoconus (KC), keratoglobus (KG), pellucid marginal degeneration (PMD), laser-assisted in situ keratomileusis (LASIK), penetrating keratoplasty (PK), and intrastromal corneal ring segments (ICRS)). METHODS: Corneal aberration data from the 3rd and 4th order together with secondary vertical and horizontal coma, and secondary spherical aberration were retrospectively retrieved from a corneal tomographer and compared between irregular cornea types and for 3-, 5-, and 7-mm pupil diameters. RESULTS: Significant differences were observed in vertical coma, oblique trefoil, secondary oblique astigmatism, primary spherical, and secondary spherical between irregular cornea types across all three pupil sizes (all p≤0.025). Vertical coma consistently exhibited the greatest magnitude of HOA across all irregular cornea types and pupil diameters. For the larger pupil diameters (5 and 7 mm), the 3rd order aberrations (mainly coma-like), the 4th order aberrations (mainly spherical-like), and total HOA RMS were significantly lower in the LASIK group compared to all other irregular cornea types (all p≤0.003). Additionally, at pupil sizes of 5 and 7 mm, the 3rd, 4th, coma-like, spherical-like, and total HOA RMS were significantly lower in KC compared to the ICRS and PK groups (all p≤0.01). Furthermore, the 4th, spherical-like, and total HOA RMS were significantly lower in KC compared to KG at pupil diameters of 3 and 7 mm, 5 and 7 mm, and 7 mm, respectively (all p≤0.03). CONCLUSIONS: Vertical coma was the most elevated HOA across all irregular cornea types and pupil diameters. Corneal aberrations were lower in LASIK and KC, and higher in KG and PK.

Static magnetic field effects on the secondary structure and elasticity of collagen molecules; a possible biophysical approach to treat keratoconus.

Type I collagen is among the major extracellular proteins that play a significant role in the maintenance of the cornea's structural integrity and is essential in cell adhesion, differentiation, growth, and integrity. Here, we investigated the effect of 300 mT Static Magnetic Field (300 mT SMF) on the structure and molecular properties of acid-solubilized collagens (ASC) isolated from the rat tail tendon. The SMF effects at molecular and atomic levels were investigated by various biophysical approaches like Circular Dichroism Spectropolarimetery (CD), Fourier Transform Infrared Spectroscopy (FTIR), Zetasizer light Scattering, and Rheological assay. Exposure of isolated type I collagen to 300 mT SMF retained its triple helix. The elasticity of collagen molecules and the keratoconus (KCN) cornea treated with SMF decreased significantly after 5 min and slightly after 10, 15, and 20 min of treatments. The exposure to 300 mT SMF shifted the Amid I bond random coil to antiparallel wave number from 1647 to 1631 cm-1. The pH of the 300 mT SMF treated collagen solution increased by about 25 %. The treatment of the KCN corneas with 300 mT SMF decreased their elasticity significantly. The promising results of the effects of 300 mT SMF on the collagen molecules and KCN cornea propose a novel biophysical approach capable of manipulating the collagen's elasticity, surface charges, electrostatic interactions, cross binding, network formation and fine structure. Therefore, SMF treatment may be considered as a novel non-invasive, direct, non-chemical and fast therapeutic and manipulative means to treat KCN cornea where the deviated physico-chemical status of collagen molecules cause deformation.

Multi-dataset identification of innovative feature genes and molecular mechanisms in keratoconus.

This study aimed to identify feature genes and explore the molecular mechanisms of keratoconus (KC). We downloaded data files from NCBI GEO public database. The Limma package was used for differential expression analysis of gene profiles. Lasso regression was used to identify the feature genes. The CIBERSORT algorithm was used to infer the proportion of immune-infiltrating cells and analyse the correlation between gene expression levels and immune cells. Related transcription factors and miRNAs of key genes were predicted using the Cistrome DB and Mircode databases. Analysis of expression differences in disease genes was based on the GeneCards database. The CMap was used to analyse targeted therapeutic drugs. IHC was performed to verify the expression levels of ATOH7 and MYRF in corneas. Exactly 593 upregulated and 473 downregulated genes were identified. Lasso regression analysis identified ATOH7, DBNDD1, RNF217-AS1, ARL11, MYRF and SNORA74B as feature genes for KC. All key genes were correlated with immune infiltration and the levels of activated memory CD4+ T cells and plasma cells were significantly increased. miRNA, IRF and STAT families were correlated to feature genes. The expression levels of key genes were significantly correlated to KC-related genes. Entinostat, ochratoxin-a, diphencyprone and GSK-3-inhibitor-II were predicted as potential KC medications. The expression of MYRF was significantly higher in the KC samples, contrary to the expression of ATOH7. KC is related to both immune infiltration and genetic factors. MYRF and ATOH7 were newly identified and verified feature genes of KC.