The fundamental problem of corneal transplantation is a severe shortage donor tissues worldwide, resulting in approximately 1.5 million new cases blindness annually. To explore an alternative to corneas, we conducted clinical study which implanted acellular porcine stromas (APCSs) replaced the pathologic anterior corneas 47 patients who had experienced fungal infections. Subsequently, demonstrated safety and efficacy APCSs human keratoplasty for minimum follow-up period 6 months, during time...

In human corneal epithelium, self-renewal and fate decision of stem cells are highly regulated in a niche microenvironment called palisades Vogt the limbus. Herein, we discovered that digestion with dispase, which cleaves off basement membrane, did not remove entire basal epithelial progenitor cells. contrast, collagenase isolated on cluster consisting only but also their closely associated mesenchymal because better preservation some membrane matrix. Collagenase more cells, were p63α+ small...

The 2019 novel coronavirus disease (COVID-19) caused by severe acute respiratory syndrome 2 (SARS-CoV-2) has spread globally, while the routes of transmission this virus are still controversial. We enrolled 33 patients, without any ocular manifestation, with their surface swabs collected for detection. RNA was detected strong positive in samples both eyes from two patients. Therefore, SARS-CoV-2 may exist normal COVID-19 suggesting that might be through conjunctival contact.

We investigated whether human limbal niche cells generate mesenchymal stem cells.Limbal were isolated from the stroma by collagenase alone or following dispase removal of epithelium (D/C), and cultured on plastic in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS), coated three-dimensional Matrigel embryonic cell leukemia inhibitory factor basic fibroblast growth factor. Expression markers, colony-forming units-fibroblast, tri-lineage differentiation, ability...

Abstract Corneal epithelial stem cells (SCs) are an ideal model for investigating how adult lineage-committed SCs regulated by anatomically defined and accessible niche, that is, limbal palisades of Vogt, located between the cornea conjunctiva. We have used collagenase digestion to isolate entire subjacent mesenchymal cells, we demonstrated their close association is crucial promoting clonal growth, implying latter serves as niche (NCs). After was disrupted trypsin/EDTA, single NCs could...

Limbal stromal niche cells heterogeneously express embryonic stem cell (SC) markers. This study was conducted to isolate and expand them prove that their phenotype is critical for supporting SCs.Human limbus isolated by dispase or collagenase. Single were seeded on coated, 2D, 3D Matrigel serially passaged in modified SC medium (MESCM), supplemented hormonal epithelial (SHEM), Dulbecco's Eagle's plus 10% fetal bovine serum (DF) before they Matrigel. Sphere growth achieved mixing expanded...

Data on long-term outcomes of users inhibitors the mammalian target rapamycin (mTORI) are lacking in kidney transplantation. In an analysis 139 370 US transplant recipients between 1999 through 2010, we compared clinical among mTORIs versus calcineurin (CNI) their primary immunosuppresive regimen. During first 2 years posttransplantation, use without CNIs (N = 3237) was associated with greater risks allograft failure and death a CNI-based regimen 125 623); hazard ratio (HR) composite outcome...

Abstract In this article, human limbal niche cells (LNC) or bone marrow derived mesenchymal stem (BMMSC) were used to prevent cell deficiency (LSCD) in an alkali burn rabbit model and their results compared. The epithelial defect area, corneal neovascularization, the print cytometry quantified grade severity of LSCD. Three months after burn, a partial LSCD was observed control group (no treatment) indicated by chronic defects, positive fluorescein staining, neovascularization goblet...

The perivascular localization of stem cell (SC) niches suggests the presence a vascular niche. We aimed to determine angiogenesis potential limbal niche cells (NCs).Human NCs were isolated and serially passaged on plastic or coated Matrigel in embryonic SC medium containing BFGF leukemia inhibitory factor before being reseeded 3D Matrigel. Expression markers was assessed by RT-qPCR immunofluorescence staining. Their measured differentiation into endothelial supporting tube network formed...

Abstract The meibomian gland (MG) is a sebaceous that secretes through holocrine process. Because such secretion requires the destruction of MG acinar epithelial cells, they need constant renewal and differentiation. processes promote these regenerative events in human are unknown, nor it known how to distinguish progenitor differentiated cells. We discovered Lrig1 DNase2 serve as biomarkers for respectively. expressed basal cells periphery, location where originate glands. central acinus....

Purpose: Meibomian glands are essential in maintaining the integrity and health of ocular surface. gland dysfunction (MGD), mainly induced by ductal occlusion, is considered as major cause dry eye disease. In this study, a novel vitro model was established for investigating role inflammation process MGD. Methods: Mouse tarsal plates were removed from eyelids after dissection explants cultured during various time ranging 24 to 120 hours. epithelial cells further enzymatically digested...

Purpose: To explore the role of substrate stiffness and mechanism beneath corneal endothelial cells' (CECs') stemness maintenance differentiation. Methods: CECs were divided into central zone (8 mm trephined boundary) peripheral edge with attached limbal). Two zones analyzed by hematoxylin-eosin staining scanning electron microscopy for anatomic structure. The elastic modulus Descemet's membrane (DM) was atomic force microscopy. Compressed type I collagen gels different constructed as an in...

To develop a method for enriching keratinocyte progenitor cells (KPCs) and establish limbal niche (LN)-mediated transdifferentiation protocol of KPCs into corneal epithelial cells. Limbal (LNCs) were isolated from tissues through enzymatic digestion characterized. Conditioned medium LNCs cultures was collected. enriched by rapid adhesion Matrigel subsequently cultured in either an LNCs-conditioned supplemented with KSFM (LN-KS) or SHEM (LN-SH) 14 days. Corneal-specific marker expression...

As a consequence of the natural aging process, lacrimal glands may become dysfunctional. The present study aimed to investigate potential role melatonin (MLT) in alleviation age-related gland dysfunction and elucidate underlying mechanism. In this study, 2-month-old, 18-month-old, MLT intraperitoneally injected 18-month-old mice were obtained for immunofluorescence, immunohistochemistry experiments, Western blotting detect inflammatory factors AQP5 expression, electron microscopy...

AIM: To investigate the pathological features of ocular surface squamous neoplasia (OSSN) and evaluate synergistic therapeutic effects interferon-α2b (IFNα2b) 5-fluorouracil (5-FU) on cellular proliferation, migration, apoptosis, cell cycle human oral carcinoma line Cal27. METHODS: Tissue specimens from OSSN were processed with hematoxylin-eosin (HE) immunofluorescence (IF) staining to characterize changes. We analyzed expression levels four pivotal proteins involved in 5-FU metabolism:...

AIM: To investigate the demographics, risk factors, clinical presentation, treatments and outcomes of Acanthamoeba keratitis (AK) in a tertiary referral hospital central China. METHODS: In this noncomparative, retrospective study, total 21 AK patients were enrolled between April 2020 March 2023. All underwent anterior segment photography, vivo confocal microscopy (IVCM), best-corrected visual acuity (BCVA) optical coherence tomography (AS-OCT). RESULTS: The most common potential predisposing...

Purpose: Meibomian glands play a vital role in maintaining ocular surface stability. This study aimed to investigate whether Hedgehog signaling is involved the regulation of meibomian gland epithelial cells. Methods: Rat cells (RMGECs) were isolated from ducts and ductules, then cultivated passage two on Matrigel coated wells medium (MGECM). Cells switched MGECM differentiation (DM) or DM added 10 µg/mL azithromycin (DM + AZM) when reached 50% 60% confluence. The effects Smoothened (Smo)...

Tissue-engineered cornea endothelial sheets (TECES), created using a biocompatible thin and transparent carrier with corneal cells, could alleviate the shortage of donor corneas provide abundant functional cells. In our previous clinical trials, effectiveness safety acellular porcine stroma (APCS) applied in lamellar keratoplasty have been confirmed. this study, we optimized method to cut APCS into multiple 20 μm ultrathin lamellae by cryostat microtome investigated feasibility TECES seeding...

Meibomian glands (MGs) are vital for ocular surface health. However, the roles of inflammation in progression meibomian gland dysfunction (MGD) largely unknown. In this study, factor interleukin-1β (IL-1β) via p38 mitogen-activated protein kinases (MAPK) signaling pathway on rat epithelial cells (RMGECs) were explored. Eyelids from adult mice at 2 months and years age stained with specific antibodies against IL-1β to identify levels. RMGECs exposed and/or SB203580, a inhibitor MAPK pathway,...

To investigate the characteristics of in vitro culture and vivo confocal microscopy (IVCM) patients with fungal keratitis (FK) presented a tertiary referral hospital central China.In this noncomparative retrospective study, diagnosis FK between October 2021 November 2022 were reviewed. An IVCM (corneal scraping specimens) performed, analyzed.During 2022, 85 diagnosed FK. From 63 culture-positive cases, 8 species fungus identified. The proportions isolated Fusarium Aspergillus equally...