重视角膜基质细胞的作用

位于角膜基质胶原纤维板层之间的角膜基质细胞处于静息状态,可以分泌胶原以及硫酸角质素,对角膜透明性的维持发挥着重要作用。当角膜受到损伤时,角膜基质细胞可发生凋亡,向成纤维细胞以及肌成纤维细胞等修复细胞表型转化,进而促进细胞再生,诱导角膜纤维瘢痕形成。此外,大量角膜基质细胞还具有干细胞特性。目前已知角膜基质细胞是多种机体功能的积极参与者,应重视角膜基质细胞的作用。     

Formation of stromal collagen fibrils and proteoglycans in the developing zebrafish cornea

Purpose: Collagen fibrils and proteoglycans are the main components of the corneal extracellular matrix and corneal transparency depends crucially on their proper arrangement. In the present study, we investigated the formation of collagen fibrils and proteoglycans in the developing cornea of the zebrafish, a model organism used to study vertebrate embryonic development and genetic disease. Methods: We employed thin‐section electron microscopy to investigate the ultrastructure of the zebrafish cornea at different developmental stages. Results: The layering of the zebrafish cornea into an epithelium, a Bowman’s layer, stroma and endothelium was observed starting at 72 hr post‐fertilization. At this stage, the stroma contained orthogonally arranged collagen fibrils and small proteoglycans. The density of proteoglycans increased gradually throughout subsequent development of the cornea. In the stroma of 2‐week‐old larvae, the collagen fibrils were organized into thin lamellae and were separated by very large, randomly distributed proteoglycans. At 4 weeks, a regular arrangement of proteoglycans in relation to the collagen fibrils was observed for the first time and the lamellae were also thickened. Conclusion: The present study, for the first time, provides ultrastructural details of collagen fibril and proteoglycan development in the zebrafish cornea. Furthermore, it directly correlates the collagen fibril and proteoglycan composition of the zebrafish cornea with that of the human cornea. The similarities between the two species suggest that the zebrafish could serve as a model for investigating the genetics of human corneal development and diseases.     

Heterogeneity, polydispersity, and physiologic role of corneal proteoglycans

Gel chromatography, affinity chromatography, ultracentrifugation, enzymic fragmentation, and analysis of amino acids, hexosamines and neutral sugars were used to characterize a heterogeneous fraction of proteoglycans from bovine corneal stroma. The results indicate that the fraction largely consists of a mixture of the 2 main types of corneal proteoglycans described earlier, namely keratan sulfate proteoglycans and chondroitin sulfate-rich proteoglycans with covalently bound oligosaccharides. Models for the structure of proteoglycans are suggested, an it is concluded that the molecular size of corneal proteoglycans makes them appropriate as 'spacers' between the collagen fibrils, a property important for corneal transparency. Cornea is softer than cartilage because corneal proteoglycans are less underhydrated than cartilage proteoglycans.     

Corneal stromal wound healing: Major regulators and therapeutic targets

Corneal stromal wound healing is a complex event that occurs to restore the transparency of an injured cornea. It involves immediate apoptosis of keratocytes followed by their activation, proliferation, migration, and trans-differentiation to myofibroblasts. Myofibroblasts contract to close the wound and secrete extracellular matrix and proteinases to remodel it. Released proteinases may degenerate the basement membrane allowing an influx of cytokines from overlying epithelium. Immune cells infiltrate the wound to clear cellular debris and prevent infections. Gradually basement membrane regenerates, myofibroblasts and immune cells disappear, abnormal matrix is resorbed, and transparency of the cornea is restored. Often this cascade deregulates and corneal opacity results. Factors that prevent corneal opacity after an injury have always intrigued the researchers. They hold clinical relevance as they can guide the outcomes of corneal surgeries. Studies in the past have shed light on the role of various factors in stromal healing. TGFβ (transforming growth factor-beta) signaling is the central player guiding stromal responses. Other major regulators include myofibroblasts, basement membrane, collagen fibrils, small leucine-rich proteoglycans, biophysical cues, proteins derived from extracellular matrix, and membrane channels. The knowledge about their roles helped to develop novel therapies to prevent corneal opacity. This article reviews the role of major regulators that determine the outcome of stromal healing. It also discusses emerging therapies that modulate the role of these regulators to prevent stromal opacity.     

Distribution of types I, II, III, IV and V collagen in normal and keratoconus corneas

By using type-specific antibodies to types I, II, III, IV and V collagens, distribution of distinct types of collagen in normal human cornea as well as keratoconus cornea were examined by indirect immunofluorescence microscopy. In normal human cornea, immunohistochemical evidence supported the previous biochemical finding that type I collagen was the major type of collagen in human corneal stroma. No reaction was observed to anti-type II collagen antibody in the whole cornea. Anti-type III collagen antibody reacted with the corneal stroma in a similar fashion as that of anti-type I collagen antibody. Type IV collagen was observed in the basement membrane of the corneal epithelium and in Descemet's membrane. Anti-type V collagen antibody also reacted with the corneal stroma diffusely. Bowman's membrane was strongly stained only with he anti-type V collagen antibody. For further details of the distribution of type I, type III and V collagens in human corneal stroma, immunoelectron microscopic study was undertaken. The positive reaction products of anti-type I and anti-type III collagen antibodies were located on the collagen fibrils, while that of anti-type V collagen antibody was either on or close to collagen fibrils. In keratoconus cornea, no difference was observed in terms of the distribution of type I, III and V collagens, while the disruptive and excrescent distribution of type IV collagen was noted in the basement membrane of the corneal epithelium.     

Clinical and morphological features including expression of betaig-h3 and keratan sulphate proteoglycans in Maroteaux-Lamy syndrome type B and in normal cornea

AIM: To carry out a detailed morphological study of the cornea of a 16 year old female with a Maroteaux-Lamy syndrome (MLS). METHODS: Following a penetrating keratoplasty in July 1999, ultrastructural changes in the cornea were examined using electron microscopy. Proteoglycans were visualised using cuprolinic blue dye; and betaig-h3 and keratan sulphate were detected by immunoelectron microscopy. RESULTS: The epithelial cells were degenerate and contained apoptotic nuclei. Proteoglycans were present in epithelial cells, intercellular spaces, and in swollen desmosomes. An abnormally large quantity of proteoglycans was present throughout the stroma. Keratocytes throughout the stroma had no cell organelles, were vacuolated, and contained a large quantity of abnormal proteoglycans. Labelling for betaig-h3 was intense around electron lucent spaces in stroma. No labelling was seen in keratocytes or endothelial cells. In normal cornea, keratan sulphate labelling was regular throughout the stroma. In MLS VI type B cornea, keratan sulphate labelling was weak in the anterior stroma but very intense in the posterior stroma and in keratocyte lysosomes and vacuoles. CONCLUSION: A deficiency of aryl sulfatase B results in the deposition of keratan sulphate proteoglycan and other proteoglycans in lysosomes, causing the death of keratocytes and an abnormal build-up of proteoglycans in the stroma. This might be responsible for the lateral aggregation of collagen fibrils and impaired fibrillogenesis in MLS VI. Degenerate swollen keratocytes, together with gross changes in epithelial, stromal, and endothelial cells, would be expected to increase light scattering significantly in these corneas.     

Surface ultrastructure of collagen fibrils and their association with proteoglycans in human cornea and sclera by atomic force microscopy and energy-filtering transmission electron microscopy

PURPOSE: We aimed to investigate the possible association of proteoglycans with D-periodic collagen fibrils in the human cornea and sclera, using energy-filtering transmission electron microscopy (EF-TEM) and atomic force microscopy (AFM). METHODS: Human cornea and sclera were digested with keratanase to eliminate keratan sulfate proteoglycans (KSPGs). For EF-TEM observation, surface proteoglycans were detected by cupromeronic blue (CB) staining. For AFM observation, cornea and sclera were treated with sodium hydroxide before and after keratanase digestion, and the surface topology of collagen fibrils was analyzed. RESULTS: With CB staining, numerous CB-positive short filaments of surface proteoglycans (proteoglycan filaments) were observed in the interfibrillar spaces of cornea and sclera associated with collagen fibrils. AFM imaging showed that the depth and periodicity of D-periodic collagen fibrils in keratanase-treated corneal collagens were deeper and more regular than in untreated ones. Moreover, the depth and periodicity of keratanase-untreated corneal collagens were shallow and irregular in comparison with keratanase-untreated scleral collagens. On the other hand, there was no difference in depth or regularity between keratanase-treated and -untreated scleral collagen fibrils. Using AFM imaging, additional thin grooves sub-bands were detected on the surface of keratanase-treated corneal collagen fibrils. The grooves were not detected in keratanase-untreated collagen fibrils nor in scleral collagen fibrils with or without keratanase digestion. Comparing densitometry waves, the grooves of D-periodic corneal collagen sub-bands corresponded to a and c bands. CONCLUSION: Using AFM and EF-TEM to study corneal and scleral collagen fibrils and their association with proteoglycans, we conclude that KSPG is found in ample amounts in the human cornea in comparison with sclera. Moreover, we topologically detected KSPG attached to a and c bands of collagen fibrils.     

Corneal cells: chatty in development,homeostasis, wound healing,and disease

PURPOSE: To provide an overview of cell-cell interactions in the cornea that have a critical role in corneal development, homeostasis, wound healing, and disease. DESIGN: Review. METHODS: Review of the literature. RESULTS; Cell-cell interactions make critical contributions to development, homeostasis, and wound healing in the cornea. Many of these interactions are mediated by cytokines, growth factors, and chemokines. The best characterized are stromal-epithelial interactions between epithelial cells and stromal cells such as keratocytes, keratoblasts, and myofibroblasts. However, interactions also occur between corneal nerves and epithelial cells and between corneal cells (epithelial cells and stromal cells) and corneal immune cells. Although investigations are limited, it is likely that there are interactions between corneal endothelial cells and keratocytes in the posterior stroma. CONCLUSIONS: Cellular communications in the cornea are critical during development, homeostasis, and wound healing. Disorders of cellular communication likely contribute to many corneal diseases.     

Loss of alpha3(IV) collagen expression associated with corneal keratocyte activation

PURPOSE: To determine whether changes in the expression of type IV alpha1, alpha2, or alpha3 collagen isoforms are stringently associated with corneal stromal cell activation. METHODS: Keratocytes isolated from rabbit corneal stroma by collagenase digestion were plated in serum-free or insulin-, bFGF/heparin sulfate (HS)-, TGF-beta1-, or fetal bovine serum (FBS)-supplemented DMEM/F12 medium. Expression of type IV collagen isoforms and keratan sulfate proteoglycans (KSPGs) was evaluated by immunocytochemical analysis, Western blot analysis, or both. Concentrations of mRNAs were estimated by quantitative RT-PCR using SYBR Green RT-PCR reagents. RESULTS: Immunohistochemical analysis indicated that type IV alpha1, alpha2, and alpha3 collagens were expressed in normal rabbit corneal stroma and in keratocytes cultured in serum-free and insulin-supplemented media. However, alpha3(IV) collagen was not detectable in the regenerating stroma after photorefractive keratectomy (PRK) in rabbit or in corneal stromal cells cultured in media supplemented with FBS, bFGF/HS, or TGF-beta1. alpha3(IV) collagen mRNA levels were also diminished in the stromal cells cultured in these growth factor-supplemented media. KSPGs (lumican and keratocan) were expressed and secreted in serum-free medium. Although the expression of KSPGs was promoted by insulin, the expression and intracellular levels of lumican and keratocan mRNAs were downregulated by TGF-beta1 and FBS. bFGF/HS promoted the downregulation of intracellular keratocan but not lumican mRNA levels. CONCLUSIONS: The loss in the expression of alpha3(IV) collagen is a stringent phenotypic change associated with activation of keratocytes in vivo and in vitro. This phenotypic change in activated corneal stromal cells is induced by bFGF/HS and by TGF-beta1, and it accompanies the downregulation of keratocan expression.     

Thy-1 distinguishes human corneal fibroblasts and myofibroblasts from keratocytes

After corneal injury, keratocytes become activated and transform into repair phenotypes-corneal fibroblasts or myofibroblasts, however, these important cells are difficult to identify histologically, compromising studies of stromal wound healing. Recent studies indicate that expression of the cell surface protein, Thy-1, is induced in fibroblast populations associated with wound healing and fibrosis in other tissues. We investigated whether keratocyte transformation to either repair-associated phenotype induced Thy-1 expression. Human corneal keratocytes were isolated by collagenase digestion. The cells were either processed immediately (i.e. freshly isolated keratocytes) or were cultured in the presence of 10% fetal bovine serum or transforming growth factor-beta to induce transformation to the corneal fibroblast and myofibroblast phenotypes, respectively. Thy-1 mRNA and protein expression by freshly isolated keratocytes and corneal fibroblasts were assessed by RT-PCR and Western blotting. mRNA also was extracted from the whole intact stroma and assessed by RT-PCR. Thy-1 was localised immunocytochemically in cultured human corneal fibroblasts, myofibroblasts, and in keratocytes in normal human corneal tissue sections. Thy-1 mRNA and protein were detectable in cultured human corneal fibroblasts, but not freshly isolated keratocytes. Whole uninjured stroma showed no detectable Thy-1 mRNA expression. Cultured human corneal fibroblasts and myofibroblasts both labelled for Thy-1, but keratocytes in the stroma of normal human cornea did not. We conclude that Thy-1 expression is induced by transformation of keratocytes to corneal fibroblasts and myofibroblasts, suggesting a potential functional role for Thy-1 in stromal wound healing and providing a surface marker to distinguish the normal keratocyte from its repair phenotypes.     

Corneal alkali burn in the rabbit. Waterbalance, healing and transparency.

Healing following a standardized central corneal alkali wound was studied morphologically in New Zealand white rabbits up to one year after the initial wound. Clinical examination, light and transmission electron microscopy was performed. The study was focused on how permanent scar tissue formed. Following the penetrating alkali injury, all cells (epithelium, keratocytes and endothelium) in the wound area disappeared. The fibroblasts/keratocytes repopulated an extensively swollen central corneal stroma. Cells and extracellular matrix filled stromal lacunae in an irregular fashion and upon deswelling the lacunae remained as irregularities in the stroma, reducing the transparency. In the periphery of the wound repopulation occurred in a less swollen stroma and normal cytoarchitecture and transparency resumed. It appears that the degree of swelling decides the degree of scar tissue formation in the corneal stroma following alkali wound healing.     

Distribution and isoform characterization of type XII collagen in bovine cornea

Streptavidin peroxidase immunohistochemistry and immunoelectron microscopy were used to determine the localization of type XII collagen in sections from bovine corneas. Type XII collagen extracted from bovine cornea and skin was assayed by Western blotting. Immunohistochemical experiments showed that type XII collagen was restricted to the corneal stroma; it was not present in corneal epithelium, epithelial basement membrane, Descemet's membrane or endothelium. Type XII collagen was distributed throughout the corneal stroma, and it was prominently localized at the superficial stroma. Immunoelectron-microscopic examination demonstrated that type XII collagen was regularly found along the surface of banded collagen fibrils with a periodic distribution. By Western blot analysis, we observed that extracts from bovine cornea contained both the long and short isoforms of type XII collagen, whereas extracts from bovine skin contained only the short isoform. The homogeneous distribution and/or presence of the long isoform of type XII collagen may be related to the characteristically regular arrangement of collagen fibrils and thereby the transparency of corneal tissue.     

糖胺聚糖与角膜透明度关系的研究

对12例正常角膜、12例斑块状角膜营养不良和1例全身性糖胺聚糖贮积症角膜进行形态学和胶性铁超微组化研究。结果表明正常角膜上皮下基底膜和前弹力层有硫酸软骨素,实质层胶原纤维间有硫酸软骨素和硫酸角质素,内皮细胞表面有透明质酸,各种细胞膜上有硫酸肝素存在。斑块状角膜营养不良实质层胶原纤维间硫酸软骨素明显增多,硫酸角质素缺乏,角膜实质细胞和内皮细胞胞质内合成了大量的细纤维状物质和异常的糖胺聚糖。全身性糖胺聚糖贮积症为硫酸肝素在细胞内的蓄积,病变细胞膜上硫酸肝素缺乏。作者认为角膜组织中一定量的糖胺聚糖维持了角膜透明,其分布部位、性质和数量变化均可导致角膜混浊。     

Morphological characteristics and intercellular connections of corneal keratocytes

PURPOSE: To investigate the morphological characteristics of keratocytes and the interconnection of keratocytes with adjacent keratocytes using the flat preparation method and scanning electron microscopy with a frontal section of the human corneal stroma. METHODS: The thin, corneal collagen lamellae were carefully dissected from the cornea (n=7), which had been stained by the flat preparation method. The remaining tissue was fixed in 3% glutaraldehyde and observed by transmission electron microscopy following the frontal section. RESULTS: The flat preparation revealed the corneal fibroblasts between the lamellae of the collagen fibers and showed that the ramifying cellular processes of the keratocytes were in contact with the cytoplasmic processes or cell bodies of neighboring fibroblasts. Two types of discrete subpopulations of keratocytes were identified: a smaller, cellular type of keratocyte with spindle-shaped nucleus with heterochromatin, and a larger, cellular type with a large indented nucleus with relatively scanty cytoplasm. Collagen fibers ran parallel to each other toward the fenestration of the cytoplasmic wall of the keratocyte. CONCLUSIONS: These flat preparation method results showed that the keratocytes within the corneal stroma are interconnected with the adjacent keratocytes, which indicates the presence of a functional communicating network through the keratocyte circuits within the stroma. A smaller, cellular type of keratocyte with spindle-shaped nucleus was morphologically differentiated from a larger, cellular type with a large, indented nucleus by flat preparation and transmission electron microscopy.     

Spatial and temporal alterations in the collagen fibrillar array during the onset of transparency in the avian cornea

In the latter stages of development, the embryonic avian cornea undergoes significant changes in structure, composition and transparency. The rearrangement of stromal collagen fibrils at this time is important because it is believed to play a key role in the acquisition of corneal transparency. Here, we investigate spatial alterations in the internal fine structure of the avian cornea during development. Chicken corneas at developmental days 14, 16 and 18 were examined by transmission electron microscopy and quantitative image analysis. For anterior and posterior regions we determined fibril number densities, two-dimensional distribution functions, and, where appropriate, radial distribution functions. Stromal collagen fibrils became more closely spaced over the developmental range studied here. Changes in fibril number density indicated that fibrils became compacted first in the anterior stroma, and later (i.e. after day 16) in the posterior stroma. By day 18 collagen fibril number densities were essentially the same in superficial and deep tissue regions. At day 14, two-dimensional distribution functions of collagen fibrils in the posterior stroma pointed to a fibrillar array that was unlike that in the anterior stroma because there was no clear radial symmetry. Rather, in the deep stroma at day 14 there was evidence of different nearest neighbour spacings in two orthogonal directions. By day 18, fibril distributions in the anterior and posterior stroma were spatially homogeneous and radially symmetric, with radial distribution functions typical of those ordinarily found in mature cornea. Corneal transparency requires the stromal matrix to have some degree of regularity in the arrangement of its uniformly thin collagen fibrils. The chicken cornea becomes progressively transparent between days 14 and 18 of development as the stroma dehydrates and thins. We show that over this time period collagen fibrils in the anterior stroma become configured in advance of fibrils in deeper stromal regions, leading to questions over the potential roles of sulphated proteoglycans in different regions of the corneal stroma during morphogenesis.     

Vitronectin or fibronectin is required for corneal fibroblast-seeded collagen gel contraction

PURPOSE: The wound healing process in the corneal stroma involves the activation of corneal keratocytes and the expression of associated phenotypes (fibroblasts and myofibroblasts). One of these phenotypes, the myofibroblasts, synthesizes alpha-smooth muscle actin in order to affect wound closure by contracting the surrounding matrix. Excessive contraction results in the formation of unresolvable scars that are undesirable in the corneal stroma. The authors tested the effect of vitronectin and fibronectin on the contraction process associated with corneal wound healing. METHODS: Collagen gels were prepared and were exposed to different treatments of fetal calf serum (FCS). The FCS used was either depleted of fibronectin and vitronectin or contained a known concentration of fibronectin, vitronectin, or both at 50 microg/ml. Contraction was measured using image analysis and cross sections of contracted gels were examined for alpha-smooth muscle actin expression using laser confocal microscopy. RESULTS: Fibroblasts seeded in collagen gels paralleled the morphologic characteristics and cell distribution of keratocytes in unwounded cornea. Matrix contraction was dependent on the presence of fibronectin and/or vitronectin where myofibroblasts were present. The cell-mediated contraction process was maximal at 0.5 x 10(5) fibroblasts/ml. CONCLUSIONS: These studies showed that vitronectin or fibronectin is required for the myofibroblast-associated contraction to occur in this in vitro model of stromal wound healing. This model system shows a distinct potential for further studies relating to the corneal wound healing process.     

The cornea of the sand lance, Limnichthyes fasciatus (Creeiidae)

The sand lance, Limnichthyes fasciatus, is a small fish, 15-30 mm in length, found on the Great Barrier Reef. The eyes, which are not covered by a spectacle, are externally placed, move independently, and measure about 1 mm in diameter. The cornea, unusual even in the teleost world, is about 0.14 mm thick, which represents one-seventh of the length of the eye. The layers of the cornea include an epithelium with a complex pattern of surface microplicae, a grossly enlarged basal cell layer, and a thick basement membrane. Structures (iridophores), which may represent the vestigial remnants of a secondary spectacle, are present in the peripheral corneal epithelium. The stroma, which has no Bowman's layer, is composed of lamellae of collagen fibrils but contains no keratocytes. Posterior to the stroma is a thick (0.1 mm) cellular layer that may represent the autochthonous layer seen in some teleost species. An iridescent layer consisting of approximately 70 parallel cytoplasmic plates oriented at right angles to the visual axis lies anterior to a thick Descemet's membrane. There is a single layer of endothelial cells on the posterior surface.     

Stimulation of collagen synthesis by insulin and proteoglycan accumulation by ascorbate in bovine keratocytes in vitro

PURPOSE: Ascorbate is required for the hydroxylation of collagen that is present in the corneal stroma. The keratan sulfate proteoglycans (KSPGs) lumican and keratocan are also present, and they interact with collagen and modulate its assembly into fibrils. In this study, ascorbate was added to a defined medium containing insulin, and its effects on the synthesis of collagen and KSPGs by keratocytes were determined. METHODS: Collagenase-isolated keratocytes were cultured with or without insulin with or without ascorbate. Collagen and glycosaminoglycan synthesis was determined by collagenase digestion of incorporated 3H-glycine and by chondroitinase ABC or endo-beta-galactosidase digestion of incorporated 35SO4. KSPGs were detected by Western blot. Collagen stability was determined by pepsin digestion. Ethyl-3,4-dihydroxybenzoate (EDB) was used to inhibit collagen hydroxylation. RESULTS: Insulin stimulated the synthesis of collagen but did not affect the accumulation of lumican and keratocan. Insulin plus ascorbate, however, stimulated the synthesis of collagen and increased the accumulation of these proteoglycans. The accumulation of PGDS, a KSPG that does not interact with collagen, was not affected by ascorbate. Only the collagen synthesized in the presence of ascorbate was pepsin resistant. EDB overrode the effects of ascorbate on pepsin resistance and proteoglycan accumulation. CONCLUSIONS: The results of this study indicate that the accumulation of lumican and keratocan depends in part on the level of collagen synthesis and its hydroxylation. The interaction of lumican and keratocan with the stably folded triple helix provided by hydroxylation may also serve to stabilize these proteoglycans.     

Immunolocalization of type VI collagen in developing and healing rabbit cornea

We have localized type VI collagen in normal developing and corneal scar tissue. Indirect immunofluorescence showed that type VI collagen was distributed throughout the normal stroma and most of the scar. No fluorescence was detected along the posterior margin of the scar and in a retrocorneal membrane continuous with the scar. Since the corneal endothelium in rabbits contributes to the formation of scar tissue and retrocorneal membrane, our observations suggest that the endothelium does not synthesize type VI collagen. Indirect immunoelectron microscopy showed that type VI collagen was located abundantly between collagen fibrils as fine filamentous structures containing beads with a periodicity of 100 nm, consistent with published observations of other tissues. Because these filaments are more prominent when stained with ruthenium red, and predigestion of tissue with Chondroitinase ABC enhances binding of monoclonal antibody to type VI collagen, proteoglycans probably are associated with this collagen in the cornea. Ultrastructural observations supported by previous biochemical analyses show that the proportion of type VI collagen to fibrillar collagen is smaller in scar tissue compared with fetal cornea. The abundance of type VI collagen and its distribution and association with proteoglycans in rabbit corneal tissues suggest that this macromolecule plays a role in the tensile strength and transparency of the stroma.     

Expression and function of fibroblast growth factor-inducible 14 in human corneal myofibroblasts

The interaction of fibroblast growth factor-inducible 14 (Fn14) and, its ligand tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is known to be important in wound healing of tissues. However, to our knowledge, expression and function of Fn14 in corneal myofibroblasts, which have a crucial role in wound healing of corneal stroma, has not been investigated. In this study, we investigated the expression and function of Fn14 in corneal myofibroblasts. Expression of Fn14 protein was assessed by flow cytometry. Corneal myofibroblasts showed strong expression of Fn14 protein, while keratocytes did not. TGF-β₁ promoted the differentiation of keratocytes into corneal myofibroblasts, and induced Fn14 expression. These data reveal that keratocytes phenotype determines the level of Fn14 expression. ELISA was used to detect chemokines and matrix metalloproteinases in the supernatant of corneal myofibroblasts cultured with or without stimulation by TWEAK and/or TGF-β₁. TWEAK increased the production of IL-8, MCP-1, and RANTES by corneal myofibroblasts via Fn14. TGF-β₁ augmented the TWEAK-induced production of these chemokines. TWEAK also increased the production of MMP-1 and -3 by corneal myofibroblasts via Fn14, while TGF-β₁ inhibited this effect of TWEAK on MMP production. TWEAK-induced phosphorylation of NF-κB and MAP kinase in corneal myofibroblasts. Furthermore, TWEAK partially inhibited the differentiation of keratocytes into corneal myofibroblasts promoted by TGF-β₁. These data suggest that the Fn14/TWEAK system may have several roles in wound healing by corneal myofibroblasts. In the future, modulation of the TWEAK/Fn14 system may become a novel approach for control corneal wound healing.