fibrin and Corneal-Injuries

The shortage of corneal donors has prompted the development of tissue-engineered corneal grafts as an alternative solution. Currently, amniotic membranes with good biocompatibility are widely used as scaffolds for loading stem cells in the treatment of corneal injury. However, this approach has its limitations. In this study, BMSCs were induced to differentiate into corneal epithelial cells

Topics: Animals; Bone Marrow Cells; Burns; Corneal Injuries; Epithelial Cells; Fibrin; Fibrosis; Inflammation; Mesenchymal Stem Cells; Rats

Formation of new vessels in granulation tissue during wound healing has been assumed to occur solely through sprouting angiogenesis. In contrast, we show here that neovascularization can be accomplished by nonangiogenic expansion of preexisting vessels. Using neovascularization models based on the chick chorioallantoic membrane and the healing mouse cornea, we found that tissue tension generated by activated fibroblasts or myofibroblasts during wound contraction mediated and directed translocation of the vasculature. These mechanical forces pulled vessels from the preexisting vascular bed as vascular loops with functional circulation that expanded as an integral part of the growing granulation tissue through vessel enlargement and elongation. Blockade of vascular endothelial growth factor receptor-2 confirmed that biomechanical forces were sufficient to mediate the initial vascular growth independently of endothelial sprouting or proliferation. The neovascular network was further remodeled by splitting, sprouting and regression of individual vessels. This model explains the rapid appearance of large functional vessels in granulation tissue during wound healing.

Topics: Animals; Biomechanical Phenomena; Chickens; Collagen; Cornea; Corneal Injuries; Fibrin; Mice; Neovascularization, Pathologic; Neovascularization, Physiologic; Rats; Time Factors; Wound Healing

In this study, temporal and spatial distribution of three TGF-beta isoforms and their downstream signaling pathways including pSmad2 and p38MAPK were examined during fibrotic wound repair. In normal chick corneas, TGF-beta1, -2, and -3 were weakly detected in Bowman's layer (BL). In healing corneas, TGF-beta1 was primarily deposited in the fibrin clot and the unwounded BL. TGF-beta2 was highly expressed in healing epithelial and endothelial cells, and numerous active fibroblasts/myofibroblasts. TGF-beta3 was mainly detected in the unwound region of basal epithelial cells. alpha-Smooth muscle actin (alpha-SMA) was initially appeared in the posterior region of repairing stroma at day 3, and was detected in the entire healing stroma by day 7. Notably, alpha-SMA was absent in the central region of healing stroma by day 14, and its staining pattern was similar to those of TGF-beta2 and p38MAPK. By contrast, pSmad2 was mainly detected in the fibroblasts. In normal cornea, laminin was mainly detected in both epithelial basement membrane (BM) and Descemet's membrane (DM). By contrast to reconstitution of the BM in the wound region, the DM was not repaired although endothelial layer was regenerated, indicating that high levels of TGF-beta2 were released into the posterior region of healing stroma on day 14. High levels of alpha-SMA staining, shown in cultured repair stromal cells from healing corneas on day 14 and in TGF-beta2 treated normal stromal cells, were significantly reduced by p38MAPK inhibition. Collectively, this study suggests that TGF-beta2-mediated myofibroblast transformation is mediated, at least partly, by the p38MAPK pathway in vivo.

Topics: Ablation Techniques; Actins; Aging; Animals; Basement Membrane; Bowman Membrane; Cells, Cultured; Chickens; Cornea; Corneal Injuries; Descemet Membrane; Fibrin; Fibrosis; Intracellular Signaling Peptides and Proteins; Laminin; p38 Mitogen-Activated Protein Kinases; Protein Isoforms; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Smad2 Protein; Stromal Cells; Time Factors; Transforming Growth Factor beta; Wound Healing

The reepithelialization of the corneal surface is an important process for restoring the imaging properties of this tissue. The purpose of the present study was to characterize and validate a new human in vitro three-dimensional corneal wound healing model by studying the expression of basement membrane components and integrin subunits that play important roles during epithelial cell migration and to verify whether the presence of exogenous factors could accelerate the reepithelialization.. Tissue-engineered human cornea was wounded with a 6-mm biopsy punch, and the reepithelialization from the surrounding margins was studied. Biopsy samples of the reepithelialized surface were harvested 3 days after wounding and were processed for histologic, electron microscopic, and immunofluorescence analyses. The effects of fibrin and epithelial growth factor (EGF) on wound reepithelialization were also studied.. Results demonstrated that this in vitro model allowed the migration of human corneal epithelial cells on a natural extracellular matrix. During reepithelialization, epithelial cell migration followed a consistent wavelike pattern similar to that reported for human corneal wound healing in vivo. This model showed a histologic appearance similar to that of native tissue as well as expression and modulation of basement membrane components and the integrin subunits known to be main actors during the wound healing process. It also allowed quantification of the reepithelialization rate, which was significantly accelerated in the presence of fibrin or EGF. The results indicated that alpha v beta6 integrin expression was increased in the migrating epithelial cells compared with the surrounding corneal tissue.. The similarity observed with the in vivo wound healing process supports the use of this tissue-engineered model for investigating the basic mechanisms involved in corneal reepithelialization. Moreover, this model may also be used as a tool to screen agents that affect reepithelialization or to evaluate the effect of growth factors before animal testing.

Topics: Basement Membrane; Cells, Cultured; Corneal Injuries; Epidermal Growth Factor; Epithelium, Corneal; Fibrin; Fibroblasts; Fluorescent Antibody Technique, Indirect; Humans; Integrins; Models, Biological; Tissue Engineering; Wound Healing

Tissue plasminogen activator (tPA) has been used to treat severe postcataract and vitrectomy fibrinous membranes, but intraocular bleeding has occurred with doses of 25 micrograms or higher. We report three patients, one with nonclearing total hyphema and uncontrollable intraocular pressure and two with severe fibrinous membrane formation, who had treatment with low-dose (4 micrograms to 6 micrograms) intraocular tPA. Although the fibrinous membranes or hyphema resolved in all three patients, they recurred and bleeding that required additional treatment occurred in one patient. Intraocular low-dose tPA may minimize the risk of corneal and retinal toxicity and may be considered an alternative treatment in intractable cases. However, secondary intraocular hemorrhage can occur, and the timing between the initial vascular injury, treatment with tPA, and subsequent bleeding may reduce the risk of further hemorrhaging.

Topics: Adult; Aged; Anterior Chamber; Cataract Extraction; Child; Cornea; Corneal Injuries; Eye Diseases; Eye Injuries, Penetrating; Fibrin; Humans; Hyphema; Injections; Keratoplasty, Penetrating; Male; Postoperative Complications; Recombinant Proteins; Tissue Plasminogen Activator

Fibronectin, a glycoprotein present in plasma and extracellular matrix, is believed to be involved in cell-cell and cell-matrix interactions. Using immunofluorescence techniques, we studied the time course of appearance and distribution of fibronectin in healing rabbit corneal epithelial wounds and compared fibronectin to other selected proteins in the cornea. In the normal cornea, fibronectin was detected only in Descemet's membrane and not in the epithelial basement membrane. Shortly after wounding, fibronectin deposited on the denuded corneal surface and was a continuous prominent layer by 8 hours. The epithelium had begun to migrate over the deposited fibronectin by 22 hours and by 52 hours had completely covered the denuded surface. Fibrinogen/fibrin was also detected on the initial bare wound surface. Once the wound was reepithelialized, the subepithelial fibronectin and fibrin layer then progressively disappeared, so that by 2 weeks only a small amount was detected. Fibronectin also appeared in the deep stroma of corneas after wounding in elongated patches, a pattern suggestive of keratocyte association. Fibronectin had an inverse relationship to bullous pemphigoid antigen which was used as a marker for the lamina lucida of the epithelial basement membrane. The bullous pemphigoid antigen, which was found in the normal corneal epithelial basement membrane, was removed with the epithelium and reappeared during the wound healing when fibronectin was diminishing. IgG and albumin did not localize on the wound surface, and the diffuse staining seen in the stroma did not change during healing. These findings are compatible with the hypothesis that fibronectin and fibrin play a role in epithelial migration and temporary adhesion to the surface during corneal wound healing, at a time when the normal anchoring mechanism is lost.

Topics: Albumins; Animals; Basement Membrane; Cornea; Corneal Injuries; Fibrin; Fibronectins; Fluorescent Antibody Technique; Immunoglobulin G; Rabbits; Surface Properties; Tears; Wound Healing

Topics: Animals; Cornea; Corneal Injuries; Descemet Membrane; Endoplasmic Reticulum; Epithelium; Fibrin; Fibroblasts; Microscopy, Electron; Rabbits; Ribosomes; Time Factors; Wound Healing

Topics: Animals; Corneal Injuries; Epithelial Cells; Fibrin; Inflammation; Microscopy; Microscopy, Electron; Rats; Time Factors; Wound Healing

Topics: Cornea; Corneal Injuries; Fibrin; Humans; Wounds and Injuries

Topics: Adhesives; Cornea; Corneal Injuries; Fibrin; Humans; In Vitro Techniques; Wound Healing