Purpose: While the first murine endothelial keratoplasty (EK) model established surgical feasibility, manual lamellar dissection and endothelial scraping introduce significant procedural variability. This study aims to develop a standardized murine EK model implementing a controlled stromal peeling technique and quadrant-based host bed preparation to minimize inconsistency. Methods: We introduced 2 critical methodological refinements. First, donor corneas were prepared using a controlled stromal peeling approach to ensure uniform graft thickness. Second, traditional circular scraping of the recipient endothelium was replaced with quadrant-based mechanical removal to standardize the denuded area. Postoperative outcomes were quantified using AS-OCT and a novel time-resolved opacity scoring system designed to distinguish early technical failure from immunologic rejection. Results: Controlled stromal peeling produced posterior lamellar grafts with significantly higher thickness consistency compared with traditional needle dissection. The quadrant-based removal resulted in highly reproducible postoperative corneal edema, reflecting standardized disruption of host endothelial function. Furthermore, the time-resolved grading system allowed more granular, objective differentiation between early graft failure and delayed rejection compared with traditional binary survival assessments. Conclusions: These refinements overcome the inherent variability of the initial murine EK model. By standardizing both the surgical procedure and the interpretive framework, this optimized model provides a robust, reproducible platform for investigating cellular mechanisms of graft outcomes in experimental keratoplasty.
Purpose: To describe and evaluate a pilot technique using a sterile bandage contact lens (CL) as a spacer to overcome the minimum-depth limitations of applanation-based femtosecond laser (FSL) platforms when harvesting thin donor Bowman layer (BL) grafts, with preliminary clinical application in BL transplantation for advanced keratoconus (KC). Methods: This experimental and interventional case series included 9 in vitro donor corneas and 4 in vivo eyes with progressive KC. A ∼70-μm plano CL was applied to the bare donor stroma. The FSL dissection depth was programmed by adding the intended graft thickness to the CL thickness (110 μm for a ∼40-μm BL graft). Graft thickness and morphology were evaluated via optical coherence tomography (OCT). For the clinical cases, grafts were implanted into FSL-created stromal pockets; in vivo outcomes were limited to anterior segment OCT assessment of graft integration, graft thickness, and central corneal thickness over 12 months of follow-up. Results: The combined success rate for graft preparation was 92.3% (12/13). In vitro BL grafts yielded a median central thickness of 70 μm. The in vivo BL grafts achieved a median central thickness of 46 μm, with a median deviation of 1.5 μm from the individualized targets. Over the 12-month follow-up, anterior segment OCT showed all grafts to remain well integrated within the stromal pocket, with no displacement or interface complications. Conclusions: In this pilot study, the CL-assisted FSL protocol proved a reproducible method for harvesting BL grafts. By bypassing the minimum depth constraints of widely used applanation-based FSL platforms, this technique may minimize tissue loss and expand surgical access to BL transplantation for managing advanced KC, pending confirmation in larger series.
Purpose: Herpetic stromal keratitis (SK) caused by herpes simplex virus (HSV) or varicella zoster virus (VZV) can cause sight-threatening corneal scarring. Given high recurrence rates of herpetic SK with steroid reduction and limited consensus on treatment duration, we examine outcomes of an approach we term the supervised reduction protocol (SRP). Methods: Among 162 patients with ocular HSV or VZV evaluated between September 2016 and June 2025, 53 had SK managed with SRP. Under the SRP, patients underwent frequent slit-lamp evaluation allowing for early detection and management of stromal inflammation. Topical steroids and systemic antivirals were only reduced under direct physician supervision to ensure quiescence before further reduction. Results: Thirty-three patients had HSV SK and 20 had VZV SK. The median time from diagnosis to quiescence was 72 days [interquartile range (IQR) 35.5–145)] for HSV SK and 51 days (IQR 24.5–77) for VZV SK. The median time from quiescence to recurrence of clinical SK was 63 days (IQR 44–250) for HSV SK and 175 days (IQR 124.3–358) for VZV SK. Most recurrences occurred after steroid discontinuation; none occurred on QID dosing. Recurrence rates were 33% (HSV) and 50% (VZV), and most were asymptomatic at the time of recurrence (73% HSV, 50% VZV). Visual acuity remained stable from initial diagnosis to quiescence. Conclusions: Close follow-up under the SRP allows for detection and treatment of early, often asymptomatic recurrences before they progress to symptomatic or vision-threatening disease. This was a single-site retrospective study of one physician's outcomes; thus, these findings do not establish that SRP is superior to others management approaches. Prospective, multisite comparative effectiveness studies are needed to validate the role of SRP for herpetic SK.
Purpose: To describe multifocal eyelid margin and canthal calcinosis cutis in a child with chronic recalcitrant vernal keratoconjunctivitis (VKC), and to review the reported ophthalmic cases of periocular calcinosis cutis. Methods: The clinicopathological data of a 14-year-old immunocompetent boy with a 9-year history of chronic VKC and inconsistent treatment adherence were reviewed retrospectively. Eyelid-margin papules and a gelatinous canthal mass were excised and examined by light microscopy with hematoxylin and eosin and with calcium-specific staining. A PubMed/MEDLINE review of periocular calcinosis cutis and subepidermal calcified nodules was additionally conducted, and individual patient data were synthesized. Results: Histopathology showed keratinized stratified squamous epithelium with epithelial atrophy; the stroma contained calcific lobules, focal metaplastic ossification, and hyaline-fibrinoid degeneration, with calcium confirmed on Von Kossa and Alizarin red staining. Immunosuppression and dysmetabolic states were excluded (HIV, HBsAg, HCV negative; serum calcium and phosphate normal). After tacrolimus withdrawal, the right canthal mass enlarged over 6 months and produced mechanical ptosis, prompting definitive excision with cryotherapy. Best-corrected visual acuity improved from 20/200 to 20/80 and from 20/50 to 20/30, with resolution of ptosis, remission of VKC, and no recurrence. Across the 12 reported cases and the present one, all were painless upper lid nodules in healthy young patients, cured by excision; none occurred in VKC. Conclusions: Eyelid dystrophic calcification, or calcinosis cutis, is diagnosed histopathologically and cured by excision and can arise in chronic poorly controlled VKC. Sustained Th2-driven inflammation with repeated low-grade injury to the lid margin and tarsus is the most plausible mechanism; calcinosis cutis warrants consideration when a child with long-standing VKC develops firm lid margin papules or a canthal mass.
