Corneal stromal structure replicating humanized hydrogel patch for sutureless repair of deep anterior-corneal defect.

A critical shortage of donor corneas exists worldwide. Hydrogel patches with a biological architecture and functions that simulate those of native corneas have garnered considerable attention. This study introduces a stromal structure replicating corneal patch (SRCP) composed of a decellularized cornea-templated nanotubular skeleton, recombinant human collagen, and methacrylated gelatin, exhibiting a similar ultrastructure and transmittance (above 80 %) to natural cornea. The SRCP is superior to the conventional recombinant human collagen patch in terms of biomechanical properties and resistance to enzymatic degradation. Additionally, SRCP promotes corneal epithelial and stromal cell migration while preventing the trans-differentiation of stromal cells into myofibroblasts. When applied to an ocular surface (37 °C), SRCP releases methacrylated gelatin, which robustly binds SRCP to the corneal stroma after activation by 405 nm light. Compared to gelatin-based photocurable hydrogel, the SRCP better supports the restoration of normal corneal curvature and withstands deformation under an elevated intraocular pressure (100 mmHg). In an in vivo deep anterior-corneal defect model, SRCP facilitated epithelial healing and vision recovery within 2 weeks, maintained graft structural stability, and inhibited stromal scarring at 4 weeks post-operation. The ideal performance of the SRCP makes it a promising humanized corneal equivalent for sutureless clinical applications.

Controlled Release of Molecules to Enhance Cell Survival and Regeneration.

Controlled release or controlled drug delivery comprises the set of techniques and approaches to improve bioavailability through improved safety and/or efficacy using a carrier material for the molecule of interest. The predictability and tunability of these carriers make them ideal for protection, localization, and sustained presentation of a wide range of therapeutics, including growth factors implicated in cell survival and regeneration. Here we provide a method for encapsulating epidermal growth factor in a degradable polymer matrix for delivery to the cornea. Additional notes are included to demonstrate the wide-ranging capabilities of such methods for other materials, therapeutic agents, and sites of action within the eye.

Application and progress of corneal confocal microscopy in the evaluation of diabetes-related peripheral neuropathy.

Diabetic peripheral neuropathy (DPN) is one of the most common chronic complications of diabetes. As a new detection method for DPN, corneal confocal microscopy (CCM) is characterised by rapid, non-invasive, sensitive, and quantitative characteristics, as well as good repeatability. By detecting changes in the corneal nerves, DPN can be diagnosed early, and the severity of neuropathy evaluated. It is currently an ideal DPN evaluation method and has good clinical application prospects. This paper reviews the application and progress of CCM in the evaluation of DPN and summarises the evaluation methods of CCM, corneal nerve, and DPN to provide new ideas for the clinical diagnosis and treatment of DPN.

Clinical characteristics and viral colonization of corneal donors with coronavirus disease 2019: a comprehensive analysis before and after corneal transplantation.

PURPOSE: To evaluate the clinical characteristics and viral Colonization of corneas donated by volunteers with coronavirus disease 2019 (COVID-19) before and after corneal transplantation. METHODS: We retrospectively compared the characteristics and clinical outcomes of patients who received corneas from donors with and without a history of COVID-19 after corneal transplantation. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate the expression of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA on ocular surfaces in corneal preservation solutions as well as the recipients' tears. Immunofluorescence was also performed to evaluate the expression of viral spike proteins in the corneas. Intraocular pressure (IOP) measurement, optical coherence tomography (OCT), and slit-lamp inspection at each follow-up examination were performed to assess the surgical efficacy. RESULTS: The RT-PCR results of eye surface swabs before corneal extraction, the corneal preservation solutions before transplantation as well as the recipients' tears were negative, thereby indicating the suitability for transplantation. No significant differences in IOP measurements, OCT findings, or in the incidence of post transplantation complications were observed between donors with and without COVID-19. CONCLUSIONS: Corneal transplantation using corneas from COVID-19 infected donors does not alter clinical outcomes when compared to controls receiving corneas from non-infected donors.

Revealing regional variations in scleral shear modulus in a rabbit eye model using multi-directional ultrasound optical coherence elastography.

The mechanical properties of the sclera play a critical role in supporting the ocular structure and maintaining its shape. However, non-invasive measurements to quantify scleral biomechanics remain challenging. Recently introduced multi-directional optical coherence elastography (OCE) combined with an air-coupled ultrasound transducer for excitation of elastic surface waves was used to estimate phase speed and shear modulus in ex vivo rabbit globes (n = 7). The scleral phase speed (12.1 ± 3.2 m/s) was directional-dependent and higher than for corneal tissue (5.9 ± 1.4 m/s). In the tested locations, the sclera proved to be more anisotropic than the cornea by a factor of 11 in the maximum of modified planar anisotropy coefficient. The scleral shear moduli, estimated using a modified Rayleigh-Lamb wave model, showed significantly higher values in the circumferential direction (65.4 ± 31.9 kPa) than in meridional (22.5 ± 7.2 kPa); and in the anterior zone (27.3 ± 9.3 kPa) than in the posterior zone (17.8 ± 7.4 kPa). The multi-directional scanning approach allowed both quantification and radial mapping of estimated parameters within a single measurement. The results indicate that multi-directional OCE provides a valuable non-invasive assessment of scleral tissue properties that may be useful in the development of improved ocular models, the evaluation of potential myopia treatment strategies, and disease characterization and monitoring.

TRPV1 corneal neuralgia mutation: Enhanced pH response, bradykinin sensitization, and capsaicin desensitization.

The factors that contribute to pain after nerve injury remain incompletely understood. Laser-assisted in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK) are common surgical techniques to correct refractive errors. After LASIK or PRK, a subset of patients suffers intense and persistent pain, of unknown origin, described by patients as feeling like shards of glass in their eye. Here, we evaluated a TRPV1 variant, p.V527M, found in a 49-y-old woman who developed corneal pain after LASIK and subsequent PRK enhancement, reporting an Ocular Surface Disease Index score of 100. Using patch-clamp and Ca2+ imaging, we found that the V527M mutation enhances the response to acidic pH. Increasing proton concentration induced a stronger leftward shift in the activation curve of V527M compared to WT, resulting in channel activity of the mutant in acidic pH at more physiological membrane potentials. Finally, comparing the responses to consecutive applications of different agonists, we found in V527M channels a reduced capsaicin-induced desensitization and increased sensitization by the arachidonic acid metabolite 12-hydroxyeicosatetraenoic acid (12-HETE). We hypothesize that the increased response in V527M channels to protons and enhanced sensitization by 12-HETE, two inflammatory mediators released in the cornea after tissue damage, may contribute to the pathogenesis of corneal neuralgia after refractive surgery.

Application of eye organoids in the study of eye diseases.

The eyes are one of the most important sensory organs in the human body. Currently, diseases such as limbal stem cell deficiency, cataract, retinitis pigmentosa and dry eye seriously threaten the quality of people's lives, and the treatment of advanced blinding eye disease and dry eye is ineffective and costly. Thus, new treatment modalities are urgently needed to improve patients' symptoms and suffering. In recent years, stem cell-derived three-dimensional structural organoids have been shown to mimic specific structures and functions similar to those of organs in the human body. Currently, 3D culture systems are used to construct organoids for different ocular growth and development models and ocular disease models to explore their physiological and pathological mechanisms. Eye organoids can also be used as a platform for drug screening. This paper reviews the latest research progress in regard to eye organoids (the cornea, lens, retina, lacrimal gland, and conjunctiva).

Keratin 8/18a.1 Expression Influences Embryonic Neural Crest Cell Dynamics and Contributes to Postnatal Corneal Regeneration in Zebrafish.

A complete understanding of neural crest cell mechanodynamics during ocular development will provide insight into postnatal neural crest cell contributions to ophthalmic abnormalities in adult tissues and inform regenerative strategies toward injury repair. Herein, single-cell RNA sequencing in zebrafish during early eye development revealed keratin intermediate filament genes krt8 and krt18a.1 as additional factors expressed during anterior segment development. In situ hybridization and immunofluorescence microscopy confirmed krt8 and krt18a.1 expression in the early neural plate border and migrating cranial neural crest cells. Morpholino oligonucleotide (MO)-mediated knockdown of K8 and K18a.1 markedly disrupted the migration of neural crest cell subpopulations and decreased neural crest cell marker gene expression in the craniofacial region and eye at 48 h postfertilization (hpf), resulting in severe phenotypic defects reminiscent of neurocristopathies. Interestingly, the expression of K18a.1, but not K8, is regulated by retinoic acid (RA) during early-stage development. Further, both keratin proteins were detected during postnatal corneal regeneration in adult zebrafish. Altogether, we demonstrated that both K8 and K18a.1 contribute to the early development and postnatal repair of neural crest cell-derived ocular tissues.

Triple-crosslinked double-network alginate/dextran/dendrimer hydrogel with tunable mechanical and adhesive properties: A potential candidate for sutureless keratoplasty.

An ideal adhesive hydrogel must possess high adhesion to the native tissue, biocompatibility, eligible biodegradability, and good mechanical compliance with the substrate tissues. We constructed an interpenetrating double-network hydrogel containing polysaccharides (alginate and dextran) and nanosized spherical dendrimer by both physical and chemical crosslinking, thus endowing the hydrogel with a broad range of mechanical properties, adhesive properties, and biological functions. The double-network hydrogel has moderate pore sizes and swelling properties. The chelation of calcium ions significantly enhances the tensile and compressive properties. The incorporation of dendrimer improves both the mechanical and adhesive properties. This multicomponent interpenetrating network hydrogel has excellent biocompatibility, tunable mechanical and adhesive properties, and satisfied multi-functions to meet the complex requirements of wound healing and tissue engineering. The hydrogel exhibits promising corneal adhesion capabilities in vitro, potentially supplanting the need for sutures in corneal stromal surgery and mitigating the risks associated with donor corneal damage and graft rejection during corneal transplantation. This novel polysaccharide and dendrimer hydrogel also shows good results in sutureless keratoplasty, with high efficiency and reliability. Based on the clinical requirements for tissue bonding and wound closure, the hydrogel provides insight into solving the mechanical properties and adhesive strength of tissue adhesives.

Comparison of long-term changes in the effective optical zone following SMILE and FS-LASIK for moderate and high myopia.

BACKGROUND: Visual quality after corneal refractive surgery is linked to the postoperative effective optical zone (EOZ). This study aims to compare long-term changes in the EOZ following small incision lenticule extraction (SMILE) and femtosecond laser-assisted in-situ keratomileusis (FS-LASIK) for moderate and high myopia. METHODS: This study included 42 patients (72 eyes) who underwent either SMILE (36 eyes) or FS-LASIK (36 eyes). A custom software program based on the tangential curvature difference map of the Pentacam HR (Oculus Optikgeräte GmbH) was used to define the EOZ at 3 and 7 years postoperatively. The EOZ, its chronological changes compared to the programmed optical zone (POZ), and the corneal wavefront aberrations following SMILE and FS-LASIK were analyzed. Correlations between the EOZ changes and relevant parameters were evaluated. RESULTS: Three years postoperatively, EOZ following SMILE and FS-LASIK were 5.13 ± 0.27 mm and 4.70 ± 0.24 mm (P < 0.001), respectively. Seven years postoperatively, EOZ following SMILE and FS-LASIK decreased to 5.03 ± 0.28 mm and 4.63 ± 0.23 mm (P < 0.001), respectively. At postoperative 7 years, the percentages of EOZ/POZ were negatively correlated with Q-value changes (ß = -5.120, P = 0.009) following SMILE and positively correlated with the cylinder correction (ß = 1.184, P = 0.004) following FS-LASIK. The induced spherical aberrations in the SMILE group were less than those in the FS-LASIK group (P < 0.05) and were negatively correlated with the EOZ/POZ (ß = -16.653, P < 0.001). CONCLUSIONS: The EOZ following SMILE was larger than that following FS-LASIK in the long postoperative term for moderate and high myopia. Furthermore, a continual reduction in the EOZ was noted after both surgical modalities.

Repeatability of a Combined Adaptive Optics Visual Simulator and Hartman-Shack Aberrometer in Pseudophakic Eyes With and Without Previous Corneal Refractive Surgery.

PURPOSE: To evaluate the intrasession repeatability of wavefront aberrations obtained by a combined adaptive optics visual simulator and Hartman-Shack aberrometer in pseudophakic eyes with and without previous corneal refractive surgery. METHODS: Three consecutive measurements were performed in one eye of each individual. Total ocular aberrations were recorded up to the 5th Zernike order for a 4.5-mm pupil. Repeatability was assessed by calculating the within-subject standard deviation (Sw), the repeatability limit (R), and the intraclass correlation coefficient (ICC). Vector analysis was performed to assess astigmatism variability between scans. RESULTS: The study enrolled 32 normal individuals and 24 individuals with a history of refractive surgery. In normal and eyes that had previous refractive surgery, respectively, the Sw values were 0.155 and 0.176 diopters (D) for sphere and 0.184 and 0.265 D for cylinder. The Sw values for all 3rd order terms ranged from 0.037 to 0.047 µm in normal eyes and 0.044 to 0.063 µm in eyes that had previous refractive surgery. The Sw for primary spherical aberration was 0.020 µm in normal eyes and 0.026 µm in eyes that had previous refractive surgery. ICC values for measurements of astigmatism yielded larger variability (ICC = 0.751 and 0.879). However, both groups demonstrated excellent repeatability (ICC > 0.9) for root mean square higher order aberrations (RMS-HOA) and total RMS values. CONCLUSIONS: In pseudophakic eyes, the adaptive optics Hartmann-Shack device demonstrated acceptable repeatability for measurement of sphere and 3rd and 4th order HOAs with higher variability for astigmatism measurements, especially in eyes with a prior history of corneal refractive surgery. [J Refract Surg. 2024;40(9):e645-e653.].

Effect of Posterior Keratometry and Corneal Radius Ratio on the Accuracy of Intraocular Lens Formulas After Myopic LASIK/PRK.

PURPOSE: To investigate the impact of back-to-front corneal radius ratio (B/F ratio) and posterior keratometry (PK) on the accuracy of intraocular lens power calculation formulas in eyes after myopic laser in situ keratomileusis (LASIK)/photorefractive keratectomy (PRK) surgery. METHODS: A retrospective, consecutive case series study included 101 patients (132 eyes) with cataract after myopic LASIK/PRK. Mean prediction error (PE), mean absolute PE (MAE), median absolute error (MedAE), and the percentage of eyes within ±0.25, ±0.50, and ±1.00 diopters (D) of PE were determined. RESULTS: The Barrett True K-TK formula exhibited the lowest MAE (0.59 D) and MedAE (0.48 D) and the highest percentage of eyes within ±0.50 D of PE (54.55%) in total. In eyes with a B/F ratio of 0.70 or less and PK of -5.70 D or greater, the Potvin-Hill formula displayed the lowest MAE (0.46 to 0.67 D). CONCLUSIONS: The Barrett True-TK exhibited the highest prediction accuracy in eyes after myopic LASIK/PRK overall. However, for eyes with a low B/F ratio and flat PK, the Potvin-Hill performed best. [J Refract Surg. 2024;40(9):e635-e644.].

Misclassification of Eyes With Progressive Keratoconus Using the KISA% Index.

PURPOSE: To determine the misclassification rate of the keratoconus percentage (KISA%) index efficacy in eyes with progressive keratoconus. METHODS: This was a retrospective case-control study of consecutive patients with confirmed progressive keratoconus and a contemporaneous normal control group with 1.00 diopters or greater regular astigmatism. Scheimpflug imaging (Pentacam HR) was obtained for all patients. KISA% index and inferior-superior (IS) values were obtained from the Pentacam topometric/keratoconus staging map. Receiver operating characteristic curves were generated to determine the area under the receiver operating characteristic curve (AUROC), sensitivity, and specificity values. RESULTS: There were 160 eyes from 160 patients evaluated, including 80 eyes from 80 patients with progressive keratoconus and 80 eyes from 80 control patients. There were 20 eyes (25%) with progressive keratoconus misclassified by the KISA% index, with 16 eyes (20%) of the progressive keratoconus cohort classified as normal (ie, KISA% < 60). There were 4 eyes (5%) with progressive keratoconus that would classify as having "normal topography" using the published criteria for very asymmetric ectasia with normal topography of KISA% less than 60 and IS value less than 1.45. All controls had a KISA% index value of less than 15. The optimal cut-off value to distinguish cohorts was 15.31 (AUROC = 0.972, 93.75% sensitivity). KISA% index values of 60 and 100 achieved low sensitivity (80% and 73.75%, respectively). CONCLUSIONS: The KISA% index misclassified a significant proportion of eyes with progressive keratoconus as normal. Although highly specific for clinical keratoconus, the KISA% index lacks sensitivity, does not effectively discriminate between normal and abnormal topography, and thus should not be used in large data analysis or artificial intelligence-based modeling. [J Refract Surg. 2024;40(9):e614-e624.].

[Comparative assessment of the state of the cornea after YAG-laser interventions on anterior segment structures]. / Sravnitel'naya otsenka sostoyaniya rogovitsy posle YAG-lazernykh vmeshatel'stv na strukturakh perednego segmenta glaza.

YAG-laser interventions are associated with the risk of complications, including in the cornea. PURPOSE: This study evaluates the condition of the cornea after laser discission (LD) of secondary cataracts (SC) and laser iridectomy (LI) using corneal confocal microscopy (CCM). MATERIAL AND METHODS: Group 1 included patients with the diagnosis "Pseudophakia, secondary cataract", they underwent LD of SC. Patients of group 1 were divided into 2 subgroups depending on the initial state of the cornea: group 1A included patients with unaltered corneas; group 1B - with changes in the corneas. Group 2 included patients diagnosed with angle-closure glaucoma (ACG) or suspected ACG, they underwent LI. CCM was performed on the Heidelberg HRT-III system. Laser treatment was performed using the Nd:YAG-laser LPULSA SYL-9000, λ=1.064 µm. RESULTS: Immediately after treatment, subgroup 1A exhibited singular hyperreflective deposits and negligible endothelial cell loss (ECL). After 1 month, CMM findings revealed no changes in this subgroup. In subgroup 1B, a post-LD reduction in endothelial cell density led to increased polymegathism, decreased pleomorphism, heightened endothelial cell nucleus reflectivity, and moderate hyperreflective deposits after 1 month. In the second group, significant hyperreflective deposits of various sizes, increased nucleus reflectivity, and notable endothelial cell density reduction were observed immediately and 1 month after LI. CONCLUSION: The results of this study show that the possibility of developing corneal complications after photo destructive laser interventions is to a certain extent related to the initial state of the cornea. The risk of developing corneal damage increases with decreasing distance between the cornea and the irradiated structure. An increase in the level of laser radiation energy and its total values also contributes to damage to the cornea, which is possible with dense secondary cataracts and thick irises.

Vermamoeba vermiformis as the etiological agent in a patient with suspected non-Acanthamoeba keratitis.

Vermamoeba vermiformis (V. vermiformis) is one of the most common free-living amoeba (FLA) and is frequently found in environments such as natural freshwater areas, surface waters, soil, and biofilms. V. vermiformis has been reported as a pathogen with pathogenic potential for humans and animals. The aim is to report a case of non-Acanthamoeba keratitis in which V. vermiformis was the etiological agent, identified by culture and molecular techniques. Our case was a 48-year-old male patient with a history of trauma to his eye 10 days ago. The patient complained of eye redness and purulent discharge. A slit-lamp examination of the eye revealed a central corneal ulcer with peripheral infiltration extending into the deep stroma. The corneal scraping sample taken from the patient was cultured on a non-nutritious agar plate (NNA). Amoebae were evaluated according to morphological evaluation criteria. It was investigated by PCR method and confirmed by DNA sequence analysis. Although no bacterial or fungal growth was detected in the routine microbiological evaluation of the corneal scraping sample that was cultured, amoeba growth was detected positively in the NNA culture. Meanwhile, Acanthamoeba was detected negative by real-time PCR. However, V. vermiformis was detected positive with the specific PCR assay. It was confirmed by DNA sequence analysis to be considered an etiological pathogenic agent. Thus, topical administration of chlorhexidine gluconate %0.02 (8 × 1) was initiated. Clinical regression was observed 72 h after chlorhexidine initiation, and complete resolution of keratitis with residual scarring was noticed in 5 weeks. In conclusion, corneal infections due to free-living amoebae can occur, especially in poor hygiene. Although Acanthamoeba is the most common keratitis due to amoeba, V. vermiformis is also assumed to associate keratitis in humans. Clinicians should also be aware of other amoebic agents, such as V. vermiformis, in keratitis patients.

Optical microsensing reveals spatiotemporal oxygen dynamics in cornea wounds that affect healing via reactive oxygen species.

Oxygen (O2) metabolism plays a critical role in cornea wound healing, regeneration, and homeostasis; however, the underlying spatiotemporal mechanisms are poorly understood. Here we used an optical sensor to profile O2 flux in intact and wounded corneas of mouse eyes. Intact corneas have unique centrifugal O2 influx profiles, smallest flux at the cornea center, and highest at the limbus. Following cornea injury, the O2 influx profile presents three distinct consecutive phases: a "decreasing" phase from 0 to 6 h, a "recovering" phase from 12 to 48 h, and a 'peak' phase from 48 to 72 h, congruent to previously described healing phases. Immediately after wounding, the O2 influx drops at wound center and wound edge but does not change significantly at the wound side or limbus. Inhibition of reactive oxygen species (ROS) in the decreasing phase significantly reduces O2 influx, decreases epithelial migration and consequently delays healing. The dynamics of O2 influx show a positive correlation with cell proliferation at the wound side, with significantly increased proliferation at the peak phase of O2 influx. This study elucidates the spatiotemporal O2 dynamics in both intact and wounded rodent cornea and shows the crucial role of O2 dynamics in regulating cell migration and proliferation through ROS metabolism, ultimately contributing to wound healing. These results demonstrate the usefulness of the micro-optrode in the characterization of spatiotemporal O2 dynamics. Injury-induced changes in O2 metabolism and ROS production modulate O2 dynamics at wound and control cell migration and proliferation, both essential for proper wound healing.