ABSTRACT
We assessed the maintenance and distribution of epithelial stem/progenitor cells after corneal reconstruction using tissue-engineered oral mucosal cell sheets in a rat model. Oral mucosal biopsy specimens were excised from green fluorescent protein (GFP) rats and enzymatically treated with Dispase II. These cells were cultured on inserts with mitomycin C-treated NIH/3T3 cells, and the resulting cell sheets were harvested. These tissue-engineered cell sheets from GFP rats were transplanted onto the eyes of a nude rat limbal stem cell deficiency model. Eight weeks after surgery, ocular surfaces were completely covered by the epithelium with GFP-positive cells. Transplanted corneas expressed p63 in the basal layers and K14 in all epithelial layers. Epithelial cells harvested from the central and peripheral areas of reconstructed corneas were isolated for a colony-forming assay, which showed that the colony-forming efficiency of the peripheral epithelial cells was significantly higher than that of the central epithelial cells 8 weeks after corneal reconstruction. Thus, in this rat model, the peripheral cornea could maintain more stem/progenitor cells than the central cornea after corneal reconstruction using oral mucosal epithelial cell sheets.
Subject(s)
Epithelial Cells/transplantation , Epithelium, Corneal/physiology , Mouth Mucosa/cytology , Regeneration/physiology , Stem Cells/cytology , Animals , Epithelial Cells/cytology , Green Fluorescent Proteins/metabolism , Mice , NIH 3T3 Cells , Rats, NudeABSTRACT
To report clinical and histopathologic findings in a case of a failed AlphaCor artificial cornea explanted due to corneal stromal melting. We describe the case of a 77-year-old man who received multiple penetrating keratoplasties (PKPs) and subsequent placement of an AlphaCor artificial cornea. Examination showed total corneal infiltration as well as an AlphaCor that was partially dehisced from the host cornea. After explantation, the implant and adjacent host tissue underwent hematoxylin and eosin staining and high-resolution scanning electron microscopy (HR-SEM). Histopathologic analysis of the specimens revealed infiltration of the skirt pores by reactive corneal fibroblasts. Although the AlphaCor implant is an established method of treating multiple failed PKPs, in this case, HR-SEM imaging strongly suggests that the strength of the interface between the implant and corneal tissue is highly dependent on collagen deposition between the pores found in the implant skirt. Collagen deposition then increases the mechanical strength of the cornea-skirt interface.
Subject(s)
Corneal Diseases/surgery , Corneal Stroma/pathology , Keratoplasty, Penetrating/adverse effects , Prostheses and Implants/adverse effects , Prosthesis Failure/etiology , Aged , Corneal Diseases/pathology , Corneal Stroma/surgery , Corneal Stroma/ultrastructure , Humans , Male , Microscopy, Electron, Scanning , Prostheses and Implants/ultrastructureSubject(s)
Cicatrix/surgery , Corneal Stroma/surgery , Corneal Transplantation/methods , Cryopreservation , Keratomileusis, Laser In Situ , Postoperative Complications , Cicatrix/microbiology , Corneal Stroma/pathology , Corneal Topography , Corneal Ulcer/drug therapy , Corneal Ulcer/microbiology , Corneal Ulcer/surgery , Eye Infections, Bacterial/drug therapy , Eye Infections, Bacterial/microbiology , Eye Infections, Bacterial/surgery , Humans , Male , Middle Aged , Pseudomonas Infections/drug therapy , Pseudomonas Infections/microbiology , Pseudomonas Infections/surgery , Surgical Flaps , Visual Acuity/physiologyABSTRACT
The aim of this study was to compare angiogenesis-induction capabilities of cultured corneal epithelial cells (CCE) and cultured oral mucosal epithelial cells (COE) in vitro, and identify candidate factors that induce corneal neovascularization after transplantation of COE sheets. Rabbit corneal and oral mucosal epithelial cells were co-cultured with mitomycin C-treated NIH/3T3 cells on culture plates and inserts. After CCE and COE were multilayered, culture medium was replaced by basal medium and incubated. Angiogenic potential was examined by invasion, migration and tube formation assays with human umbilical vein endothelial cells (HUVECs). Protein secretion of fibroblast growth factor 2 (FGF2), vascular endothelial growth factor (VEGF), angiopoietin-1 and transforming growth factor beta1 was assessed in conditioned medium by ELISA. Gene expression of FGF2 and VEGF was also quantified by real-time RT-PCR and neutralizing antibodies against FGF2 and VEGF were employed for blocking assays. COE induced significantly greater invasion, migration and tube formation of HUVECs, when compared to CCE. CCE secreted a significantly lower amount of FGF2 than COE, while amounts of VEGF were approximately equal in both culture media. Similarly, significantly higher expression of FGF2 mRNA was observed with COE, while no significant difference in VEGF mRNA expression was observed between COE and CCE. Only anti-FGF2 neutralizing antibody significantly suppressed HUVEC invasion and migration induced by COE, without suppression in CCE. In conclusion, angiogenic potential of COE is greater than that of CCE and FGF2 is a candidate involved in the induction of corneal neovascularization after COE sheet transplantation.
Subject(s)
Epithelium, Corneal/cytology , Mouth Mucosa/cytology , Neovascularization, Physiologic/physiology , Angiogenesis Inducing Agents/metabolism , Animals , Cell Movement/physiology , Cells, Cultured , Coculture Techniques , Corneal Neovascularization/etiology , Corneal Neovascularization/metabolism , Culture Media, Conditioned , Endothelium, Vascular/cytology , Epithelium, Corneal/metabolism , Fibroblast Growth Factor 2/genetics , Fibroblast Growth Factor 2/immunology , Fibroblast Growth Factor 2/metabolism , Gene Expression , Humans , Male , Mouth Mucosa/metabolism , Mouth Mucosa/transplantation , RNA, Messenger/genetics , Rabbits , Reverse Transcriptase Polymerase Chain Reaction/methods , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/immunology , Vascular Endothelial Growth Factor A/metabolismABSTRACT
BACKGROUND: Vogt-Koyanagi-Harada disease (VKH), an inflammatory ocular disorder characterized by bilateral granulomatous panuveitis and a variety of extraocular manifestations, has been reported to be associated with various immune disorders but has not been linked to malignant lymphoma (ML). CASE: We present here a case of VKH associated with a recurrence of ML. OBSERVATIONS: A 69-year-old man who initially had ML presented with a history of sudden bilateral visual acuity loss. Funduscopy showed papilloedema and serous retinal detachment in both eyes, and a diagnosis of VKH was reached soon thereafter. Chest X-ray and an abdominal computed tomography scan indicated the metastatic focus of the ML. A recurrence was suspected because the ML-associated soluble interleukin-2 receptor (sIL-2R) in the serum was highly elevated. Treatment successfully resolved both the ML and the VKH. The inflammatory activities of VKH and ML were found to correlate with the serum levels of sIL-2R. CONCLUSIONS: This case suggests an association between sIL-2R levels and disease activity in VKH and ML, and provides additional evidence that VKH can be induced by immune disorders caused by high sIL-2R levels in ML.
