Effect of varied beam diameter of picosecond laser on Foturan glass volume microprocessing.

Foturan glass is a photosensitive transparent material which has attracted much interest for microfluidic applications due to possibility of volume processing by ultrafast lasers. In this work, we have investigated the effect of picosecond laser on volume processing in Foturan glass when varying the beam diameter incident on a lens. To this end, specific laser focusing configurations have been designed using raytracing models and an analysis protocol has been developed in the lens focusing region in order to describe the focal point displacement occurring at the variation of the incident laser beam diameter. The numerically simulated results were explained in association with Rayleigh length and found to be in good agreement with the experimental data obtained at well-defined conditions. Specifically, it was found that the hollow microstructures developed by thermal treatment and chemical etching after laser irradiation were significantly displaced along the propagation direction when the incident beam diameter varied in the range of 1-3.5 times. This approach aims to bring an essential contribution to the field of ultrashort pulse lasers micro- and nanoprocessing in transparent materials proving that the laser beam focus position and its size can be precisely controlled with high precision by automated optics for the variation of incident laser beam diameter in predefined conditions. This approach has the potential for laser multi-beam processing at various volume depths using the same optics setup and may even be applicable to two-photon excitation microscopy. On the other hand, the processing protocol in Foturan glass may allow understanding transparent material modification by tailoring laser beam characteristics.

Substance P promotes transforming growth factor-ß-induced collagen synthesis in human corneal fibroblasts.

Corneal fibroblasts maintain homeostasis of the corneal stroma by mediating the synthesis and degradation of extracellular collagen, and these actions are promoted by transforming growth factor-ß (TGF-ß) and interleukin-1ß (IL-1ß), respectively. The cornea is densely innervated with sensory nerve fibers that are not only responsible for sensation but also required for physiological processes such as tear secretion and wound healing. Loss or dysfunction of corneal nerves thus impairs corneal epithelial wound healing and can lead to neurotrophic keratopathy. The sensory neurotransmitter substance P (SP) promotes corneal epithelial wound healing by enhancing the stimulatory effects of growth factors and fibronectin. We have now investigated the role of SP in collagen metabolism mediated by human corneal fibroblasts in culture. Although SP alone had no effect on collagen synthesis or degradation by these cells, it promoted the stimulatory effect of TGF-ß on collagen type I synthesis without affecting that of IL-1ß on the expression of matrix metalloproteinase-1. This effect of SP on TGF-ß-induced collagen synthesis was accompanied by activation of p38 mitogen-activated protein kinase (MAPK) signaling and was attenuated by pharmacological inhibition of p38 or of the neurokinin-1 receptor. Our results thus implicate SP as a modulator of TGF-ß-induced collagen type I synthesis by human corneal fibroblasts, and they suggest that loss of this function may contribute to the development of neurotrophic keratopathy.NEW & NOTEWORTHY This study investigates the role of substance P (SP) in collagen metabolism mediated by human corneal fibroblasts in culture. We found that, although SP alone had no effect on collagen synthesis or degradation by corneal fibroblasts, it promoted the stimulatory effect of transforming growth factor-ß on collagen type I synthesis without affecting that of interleukin-1ß on the expression of matrix metalloproteinase-1.

Urokinase-type plasminogen activator promotes corneal epithelial migration and nerve regeneration.

Urokinase-type plasminogen activator (uPA) is a serine protease that plays a central role in the pericellular fibrinolytic system, mediates the degradation of extracellular matrix proteins and activation of growth factors, and contributes to the regulation of various cellular processes including cell migration and adhesion, chemotaxis, and angiogenesis. The corneal epithelium responds rapidly to injury by initiating a wound healing process that involves cell migration, cell proliferation, and tissue remodeling. It is innervated by sensory nerve endings that play an important role in the maintenance of corneal epithelial homeostasis and in the wound healing response. We here investigated the role of uPA in corneal nerve regeneration and epithelial resurfacing after corneal injury with the use of uPA-deficient mice. Both the structure of the corneal epithelium and the pattern of corneal innervation in uPA-/- mice appeared indistinguishable from those in uPA+/+ mice. Whereas the cornea was completely resurfaced by 36-48 h after epithelial scraping in uPA+/+ mice, however, such resurfacing required at least 72 h in uPA-/- mice. Restoration of epithelial stratification was also impaired in the mutant mice. Fibrin zymography revealed that the expression of uPA increased after corneal epithelial scraping and returned to basal levels in association with completion of re-epithelialization in wild-type animals. Staining of corneal whole-mount preparations for ßIII-tubulin also revealed that the regeneration of corneal nerves after injury was markedly delayed in uPA-/- mice compared with uPA+/+ mice. Our results thus demonstrate an important role for uPA in both corneal nerve regeneration and epithelial migration after epithelial debridement, and they may provide a basis for the development of new treatments for neurotrophic keratopathy.

Towards understanding the mechanism of 3D printing using protein: Femtosecond laser direct writing of microstructures made from homopeptides.

Femtosecond laser direct write (fs-LDW) is a promising technology for three-dimensional (3D) printing due to its high resolution, flexibility, and versatility. A protein solution can be used as a precursor to fabricate 3D proteinaceous microstructures that retain the protein's native function. The large diversity of protein molecules with different native functions allows diverse applications of this technology. However, our limited understanding of the mechanism of the printing process restricts the design and generation of 3D microstructures for biomedical applications. Therefore, we used eight commercially available homopeptides as precursors for fs-LDW of 3D structures. Our experimental results show that tyrosine, histidine, glutamic acid, and lysine contribute more to the fabrication process than do proline, threonine, phenylalanine, and alanine. In particular, we show that tyrosine is highly beneficial in the fabrication process. The beneficial effect of the charged amino acids glutamic acid and lysine suggests that the printing mechanism involves ions in addition to the previously proposed radical mechanism. Our results further suggest that the uneven electron density over larger amino acid molecules is key in aiding fs-LDW. The findings presented here will help generate more desired 3D proteinaceous microstructures by modifying protein precursors with beneficial amino acids. STATEMENT OF SIGNIFICANCE: Femtosecond laser direct write (fs-LDW) offers a three-dimensional (3D) printing capability for creating well-defined micro-and nanostructures. Applying this technology to proteins enables the manufacture of complex biomimetic 3D micro-and nanoarchitectures with retention of their original protein functions. To our knowledge, homopeptides themselves have never been used as precursor for fs-LDW so far. Our study gains several new insights into the 3D printing mechanism of pure protein for the first time. We believe that the experimental evidence presented greatly benefits the community of 3D printing of protein in particular and the biomaterial science community in general. With the gained insight, we aspire to expand the possibilities of biomaterial and biomedical applications of this technique.

Urokinase-type plasminogen activator negatively regulates α-smooth muscle actin expression via Endo180 and the uPA receptor in corneal fibroblasts.

Corneal fibroblasts are embedded within an extracellular matrix composed largely of collagen type 1, proteoglycans, and other proteins in the corneal stroma, and their morphology and function are subject to continuous regulation by collagen. During wound healing and in various pathological conditions, corneal fibroblasts differentiate into myofibroblasts characterized by the expression of α-smooth muscle actin (α-SMA). Endo180, also known as urokinase-type plasminogen activator (uPA) receptor-associated protein (uPARAP), is a collagen receptor. Here we investigated whether targeting of Endo180 and the uPA receptor (uPAR) by uPA might play a role in the regulation of α-SMA expression by culturing corneal fibroblasts derived from uPA-deficient (uPA-/-) or wild-type (uPA+/+) mice in a collagen gel or on plastic. The expression of α-SMA was upregulated, the amounts of full-length Endo180 and uPAR were increased, and the levels of both transforming growth factor-ß (TGF-ß) expression and Smad3 phosphorylation were higher in uPA-/- corneal fibroblasts compared with uPA+/+ cells under the collagen gel culture condition. Antibodies to Endo180 inhibited these effects of uPA deficiency on α-SMA and TGF-ß expression, whereas a TGF-ß signaling inhibitor blocked the effects on Smad3 phosphorylation and α-SMA expression. Our results suggest that uPA deficiency might promote the interaction between collagen and Endo180 and thereby increase α-SMA expression in a manner dependent on TGF-ß signaling. Expression of α-SMA is thus negatively regulated by uPA through targeting of Endo180 and uPAR.

Plasmonic Superstructure Arrays Fabricated by Laser Near-Field Reduction for Wide-Range SERS Analysis of Fluorescent Materials.

Surface-enhanced Raman scattering (SERS) enables trace-detection for biosensing and environmental monitoring. Optimized enhancement of SERS can be achieved when the energy of the localized surface plasmon resonance (LSPR) is close to the energy of the Raman excitation wavelength. The LSPR can be tuned using a plasmonic superstructure array with controlled periods. In this paper, we develop a new technique based on laser near-field reduction to fabricate a superstructure array, which provides distinct features in the formation of periodic structures with hollow nanoclusters and flexible control of the LSPR in fewer steps than current techniques. Fabrication involves irradiation of a continuous wave laser or femtosecond laser onto a monolayer of self-assembled silica microspheres to grow silver nanoparticles along the silica microsphere surfaces by laser near-field reduction. The LSPR of superstructure array can be flexibly tuned to match the Raman excitation wavelengths from the visible to the infrared regions using different diameters of silica microspheres. The unique nanostructure formed can contribute to an increase in the sensitivity of SERS sensing. The fabricated superstructure array thus offers superior characteristics for the quantitative analysis of fluorescent perfluorooctanoic acid with a wide detection range from 11 ppb to 400 ppm.

Promotion of conjunctival fibroblast-mediated collagen gel contraction by mast cells through up-regulation of matrix metalloproteinase release and activation.

Mast cells and conjunctival fibroblasts contribute to conjunctival wound healing and allergic ocular inflammation. The number of mast cells in the conjunctiva is increased in individuals with cicatricial fibrosis-causing ocular surface diseases and after glaucoma filtering surgery, suggesting that these cells may contribute to the scarring observed after such surgery. We studied the potential mechanism of fibroblast-mast cell interaction in the healing of conjunctival wounds using a three-dimensional collagen gel culture system. We found that mast cells derived from the bone marrow of mice embedded in a collagen gel did not induce gel contraction. However, an increase in mast cells was associated with increased collagen gel contraction mediated by mouse conjunctival fibroblasts. The extent of collagen degradation was not affected by the co-culture of mast cells and conjunctival fibroblasts. Gelatin zymography disclosed that mast cells increased the amounts of both the pro form of matrix metalloproteinase (MMP)-9 and the active form of MMP-2 in supernatants of conjunctival fibroblast cultures. Furthermore, the potentiating effect of mast cells on contraction of the collagen gel through conjunctival fibroblasts was attenuated by the addition of a synthetic MMP inhibitor. Thus, current results suggest that mast cells accelerate the conjunctival fibroblast-dependent contraction of collagen gel by increasing the release as well as activation of MMPs. Therefore, the interaction between mast cells and conjunctival fibroblasts may contribute to conjunctival scar formation after glaucoma filtering surgery.

Identification of Interphotoreceptor retinoid-binding protein in the Schisis cavity fluid of a patient with congenital X-linked Retinoschisis.

BACKGROUND: This case report describes the surgical outcome in a patient with congenital X-linked retinoschisis (CXLRS) and the results of proteomic analysis of surgically extracted samples from both vitreous and intraschisis cavities by mass spectrometry. CASE PRESENTATION: A 3-month-old boy presented with extensive retinoschisis involving macula and retinal periphery in both eyes. Genetic analysis confirmed retinoschisin 1 mutation (c.554C > T), and an electroretinogram showed significant reduction of b-wave and decreased cone and rod responses, which led to a diagnosis of CXLRS. By performing pars plana vitrectomy, including inner wall retinectomy, clear visual axes with stable retinal conditions and functional vision in both eyes were obtained during the 4 years of follow-up. Proteomic analysis of surgically retrieved fluid from the intraschisis cavity revealed a higher expression of interphotoreceptor retinoid-binding protein (IRBP) than that from the vitreous humor. However, both samples showed equal levels of albumin, transferrin, and pigment epithelium-derived factor. CONCLUSIONS: Cellular adhesive imperfection in CXLRS may cause IRBP diffusion from the interphotoreceptor matrix, resulting in the strong expression of IRBP in the intraschisis cavity. An impaired retinoid cycle caused by an absence of IRBP in the retina may potentially underlie the pathology of CXLRS.

Real-time capture of single particles in controlled flow by a rapidly generated foci array with adjustable intensity and pattern.

We propose a new, to the best of our knowledge, technique to capture single particles in real-time in a microfluidic system with controlled flow using micro-pillar traps fabricated by one-step. The micro pillars are fabricated in parallel by femtosecond multi-foci laser beams, which are generated by multiplexing gratings. As the generation process does not need integration loops, the pattern and the intensity distribution of the foci array can be controlled in real-time by changing the parameters of gratings. The real-time control of the foci array enables rapidly fabricating microtraps in the microchannel with adjustment of the pillar spaces and patterns according to the sizes and shapes of target particles. This technology provides an important step towards using platforms based on single-particle analysis, and it paves the way for the development of innovative microfluidic devices for single-cell analysis.

Pivotal Role of Corneal Fibroblasts in Progression to Corneal Ulcer in Bacterial Keratitis.

The shape and transparency of the cornea are essential for clear vision. However, its location at the ocular surface renders the cornea vulnerable to pathogenic microorganisms in the external environment. Pseudomonas aeruginosa and Staphylococcus aureus are two such microorganisms and are responsible for most cases of bacterial keratitis. The development of antimicrobial agents has allowed the successful treatment of bacterial keratitis if the infection is diagnosed promptly. However, no effective medical treatment is available after progression to corneal ulcer, which is characterized by excessive degradation of collagen in the corneal stroma and can lead to corneal perforation and corneal blindness. This collagen degradation is mediated by both infecting bacteria and corneal fibroblasts themselves, with a urokinase-type plasminogen activator (uPA)-plasmin-matrix metalloproteinase (MMP) cascade playing a central role in collagen destruction by the host cells. Bacterial factors stimulate the production by corneal fibroblasts of both uPA and pro-MMPs, released uPA mediates the conversion of plasminogen in the extracellular environment to plasmin, and plasmin mediates the conversion of secreted pro-MMPs to the active form of these enzymes, which then degrade stromal collagen. Bacterial factors also stimulate expression by corneal fibroblasts of the chemokine interleukin-8 and the adhesion molecule ICAM-1, both of which contribute to recruitment and activation of polymorphonuclear neutrophils, and these cells then further stimulate corneal fibroblasts via the secretion of interleukin-1. At this stage of the disease, bacteria are no longer necessary for collagen degradation. In this review, we discuss the pivotal role of corneal fibroblasts in corneal ulcer associated with infection by P. aeruginosa or S. aureus as well as the development of potential new modes of treatment for this condition.

Magnetically driven rotary microfilter fabricated by two-photon polymerization for multimode filtering of particles.

In this Letter, a magnetically driven rotary microfilter that enables switching the modes of filtering and passing is fabricated in microfluidic devices via two-photon polymerization using a femtosecond laser for selective filtering of particles. The high-quality integration of a microfilter is ensured by accurately formulating the magnetic photoresist and optimizing the processing parameters. By changing the direction of the external magnetic field, the fabricated microfilter can be remotely manipulated to rotate by desired angles, thereby achieving the "filtering" or "passing" mode on demand. Taking advantage of this property, the magnetically rotary microfilter realizes multi-mode filtering functions such as capturing 8 µm particles/passing the 2.5 µm particles and passing both particles. More importantly, the responsive characteristic increases the reusability of the microchip. The lab-on-chip devices integrated with remotely rotary microfilters by the femtosecond laser two-photon polymerization with the functional photoresist will offer extensive applications in chemical and biological studies.

FACTORS ASSOCIATED WITH REACTIVATION AFTER INTRAVITREAL BEVACIZUMAB OR RANIBIZUMAB THERAPY IN INFANTS WITH RETINOPATHY OF PREMATURITY.

PURPOSE: To investigate the efficacy and risk factors of intravitreal antivascular endothelial growth factor injection (anti-VEGF therapy) for retinopathy of prematurity (ROP). METHODS: We retrospectively reviewed 80 consecutive eyes of 43 patients with Type 1 ROP or worse who received anti-VEGF therapy during January 2012-February 2018. Patients were divided into those who were injected with 0.25 mg of bevacizumab (IVB group, 37 eyes) and 0.25 mg of ranibizumab (IVR group, 43 eyes). Serum VEGF concentrations of 18 patients were measured before and after IVR. RESULTS: Antivascular endothelial growth factor injection therapy reduced ROP activity in all eyes; however, 14 eyes (17.5%) exhibited reactivation. The reactivation rates of the IVB and IVR groups were 13.5% and 20.9%, respectively (P = 0.556). Multivariate logistic regression analysis showed that postmenstrual age ≤35 weeks at anti-VEGF therapy (P = 0.014) and aggressive posterior ROP (P = 0.044) was significantly associated with reactivation. Serum VEGF was significantly suppressed at Days 1 (P < 0.001) and 7 (P = 0.012) after IVR and returned to the preinjection level by Day 14 (P = 0.210). CONCLUSION: Both IVR and IVB seemed effective in reducing ROP activity. Reactivation after anti-VEGF therapy may be associated with younger postmenstrual age at anti-VEGF therapy and aggressive posterior ROP.

Microfabrication of cellulose nanofiber-reinforced hydrogel by multiphoton polymerization.

The mechanical strength of hydrogel microstructures is crucial for obtaining the desired flexibility, robustness, and biocompatibility for various applications such as cell scaffolds and soft microrobots. In this study, we demonstrate the fabrication of microstructures composed of cellulose nanofibers (CNFs) and poly(ethylene glycol) diacrylate (PEGDA) hydrogels by multiphoton polymerization. The stress of the fabricated microstructure during tensile testing increased with an increase in the CNF concentration, indicating that the mechanical strength of the microstructure was enhanced by using CNFs as fillers. Moreover, the swelling ratio of the microstructure increased with increasing CNF concentration in the PEGDA hydrogel. Our results show the potential of the technique for the microfabrication of advanced cell scaffolds and soft microrobots with the desired mechanical strength.

The fibrinolytic system in the cornea: A key regulator of corneal wound healing and biological defense.

The cornea is a relatively unique tissue in the body in that it possesses specific features such as a lack of blood vessels that contribute to its transparency. The cornea is supplied with soluble blood components such as albumin, globulin, and fibrinogen as well as with nutrients, oxygen, and bioactive substances by diffusion from aqueous humor and limbal vessels as well as a result of its exposure to tear fluid. The healthy cornea is largely devoid of cellular components of blood such as polymorphonuclear leukocytes, monocytes-macrophages, and platelets. The location of the cornea at the ocular surface renders it susceptible to external insults, and its avascular nature necessitates the operation of healing and defense mechanisms in a manner independent of a direct blood supply. The fibrinolytic system, which was first recognized for its role in the degradation of fibrin clots in the vasculature, has also been found to contribute to various biological processes outside of blood vessels. Fibrinolytic factors thus play an important role in biological defense of the cornea. In this review, we address the function of the fibrinolytic system in corneal defense including wound healing and the inflammatory response.

Attomolar Sensing Based on Liquid Interface-Assisted Surface-Enhanced Raman Scattering in Microfluidic Chip by Femtosecond Laser Processing.

Surface-enhanced Raman scattering (SERS) is a multidisciplinary trace analysis technique based on plasmonic effects. The development of SERS microfluidic chips has been exploited extensively in recent times impacting on applications in diverse fields. However, despite much progress, the excitation of label-free molecules is extremely challenging when analyte concentrations are lower than 1 nM because of the blinking SERS effect. In this paper, a novel analytical strategy which can achieve detection limits at an attomolar level is proposed. This performance improvement is due to the use of a glass microfluidic chip that features an analyte air-solution interface which forms on the SERS substrate in the microfluidic channel, whereby the analyte molecules aggregate locally at the interface during the measurement, hence the term liquid interface-assisted SERS (LI-SERS). The microfluidic chips are fabricated using hybrid femtosecond (fs) laser processing consisting of fs laser-assisted chemical etching, selective metallization, and metal surface nanostructuring. The novel LI-SERS technique can achieve an analytical enhancement factor of 1.5 × 1014, providing a detection limit below 10-17 M (<10 aM). The mechanism for the extraordinary enhancement afforded by LI-SERS is attributed to Marangoni convection induced by the photothermal effect.

Multiscale Hierarchical Micro/Nanostructures Created by Femtosecond Laser Ablation in Liquids for Polarization-Dependent Broadband Antireflection.

In this work, we present the possibility of producing multiscale hierarchical micro/nanostructures by the femtosecond laser ablation of transition metals (i.e., Ta and W) in water and investigate their polarization-dependent reflectance. The hierarchical micro/nanostructures are composed of microscale-grooved, mountain-like and pit-rich structures decorated with hybrid laser-induced periodic surface structures (LIPSSs). The hybrid LIPSSs consist of low/high and ultrahigh spatial frequency LIPSSs (LSFLs/HSFLs and UHSFLs). LSFLs/HSFLs of 400-600 nm in a period are typically oriented perpendicular to the direction of the laser polarization, while UHSFLs (widths: 10-20 nm and periods: 30-50 nm) are oriented perpendicular to the curvatures of LSFLs/HSFLs. On the microstructures with height gradients, the orientations of LSFLs/HSFLs are misaligned by 18°. On the ablated W metasurface, two kinds of UHSFLs are observed. UHSFLs become parallel nanowires in the deep troughs of LSFLs/HSFLs but result in being very chaotic in shallow LSFLs, turning into polygonal nanonetworks. In contrast, chaotic USFLs are not found on the ablated Ta metasurfaces. With the help of Fourier transform infrared spectroscopy, it is found that microgrooves show an obvious polarization-dependent reflectance at wavelengths of 15 and 17.5 µm associated with the direction of the groove, and the integration of microstructures with LSFs/HSFLs/UHSFLs is thus beneficial for enhancing the light absorbance and light trapping in the near-to-mid-infrared (NIR-MIR) range.

Corneal Perforation during Combination Chemotherapy including Cetuximab in a Patient with a History of Herpetic Keratitis.

PURPOSE: To report a case of corneal perforation, in a patient with a history of herpetic keratitis, during combination chemotherapy including cetuximab. CASE: We report the case of a 71-year-old man who was diagnosed with a hypopharyngeal carcinoma and received radiation therapy combined with cetuximab, the epidermal growth factor receptor (EGFR) inhibitor monoclonal antibody. He was referred to us because of ocular hyperemia and corneal perforation in his left eye. In spite of conservative therapy, his corneal perforation was exacerbated, with iris incarceration into the wound site and exposure to the surface of the cornea. He therefore discontinued treatment with the combination chemotherapy and underwent lamellar keratoplasty using a preserved donor cornea. After treatment with cetuximab resumed, there was no recurrence of the corneal perforation. CONCLUSION: We have presented the first case of cetuximab-related corneal perforation in a patient who had a history of recurrent herpetic keratitis. EGFR inhibitors, such as cetuximab, can induce corneal perforation in cases with a history of herpetic stromal keratitis.

Rapid fabrication of high-resolution multi-scale microfluidic devices based on the scanning of patterned femtosecond laser.

Femtosecond-laser-induced two-photon polymerization has distinct advantages in micro-nanofabrication due to its intrinsic three-dimensional processing capability and high precision with sub-100 nanometer fabrication resolution. However, the high resolution causes a drawback in fabricating large-scale structures due to unacceptably long processing times. To solve this problem, we applied the patterned focus as the basic element for scanning processing. Theoretically, the relationship between patterned-focus scanning parameters and the uniformity of scanned light field was analyzed and optimized. Experimentally, we quantitatively investigated the relationship between the microstructure surface quality and the parameters of patterned-focus scanning. Based on above, we put forward a hybrid method that combines the femtosecond laser patterned exposure with direct-writing fabrication to rapidly fabricate large-scale microfluidic devices for various practical applications.

Hybrid femtosecond laser fabrication of a size-tunable microtrap chip with a high-trapping retention rate.

In this Letter, we propose a new (to the best of our knowledge), promising concept of a hybrid femtosecond (fs) laser processing method composed of single-point scanning and holographic light modulation fabrication for manufacturing a tunable-size microtrap chip. The hybrid method not only ensures key microfluidic device precision but also greatly improves the fabrication speed. By using a new asymmetry-bracket-shaped microtrap design with a mechanical strain stretching method, real-time size-tunable trapping is obtained, and a 100% particle trapping retention is realized, ignoring the flow fluctuation. Finally, the microtrap array is successfully applied to trap single yeast cells and hold them for $\sim{10}\;{\rm h}$∼10h without escaping.

Requirement for the collagen receptor Endo180 in collagen gel contraction mediated by corneal fibroblasts.

The interaction of keratocytes with extracellular matrix components plays an important role in the maintenance of corneal transparency and shape as well as in the healing of corneal wounds. In particular, the interaction of these cells with collagen and cell-mediated collagen contraction contribute to wound closure. Endo180 is a receptor for collagen that mediates its cellular internalization. We have now examined the role of Endo180 in collagen contraction mediated by corneal fibroblasts (activated keratocytes). Antibodies to Endo180 inhibited the contractile activity of mouse corneal fibroblasts embedded in a three-dimensional collagen gel and cultured in the presence of serum, with this effect being both concentration and time dependent and essentially complete at an antibody concentration of 0.2 µg/ml. Whereas corneal fibroblasts cultured in a collagen gel manifested a flattened morphology with prominent stress fibers under control conditions, they showed a spindlelike shape with few stress fibers in the presence of antibodies to Endo180. Antibodies to Endo180 had no effect on the expression of α-smooth muscle actin or the extent of collagen degradation in collagen gel cultures of corneal fibroblasts. Immunohistofluorescence analysis did not detect the expression of Endo180 in the unwounded mouse cornea. However, Endo180 expression was detected in keratocytes migrating into the wound area at 3 days after a corneal incisional injury. Together, our results suggest that Endo180 is required for the contraction of collagen matrix mediated by corneal fibroblasts and that its expression in these cells may contribute to the healing of corneal stromal wounds.