Effect of topical beta blockers on corneal epithelial wound healing in the rabbit.

Topical beta adrenergic receptor blocking agents (beta blockers) are among the most frequently prescribed ophthalmic drugs. It has been suggested that some of these agents have a toxic effect on the corneal epithelium. In the present study, four beta blockers in common therapeutic concentrations, as well as their vehicles, were applied to rabbit corneas that had undergone mechanical removal of epithelium from a 6 mm diameter corneal wound. The tested drugs (0.25% timolol, 0.25% levobunolol, 0.25% betaxolol, and 0.3% metipranolol) were found significantly to accelerate wound closure, compared with saline treatment in controls. Eyes treated with two of the vehicles (betaxolol vehicle and metipranolol vehicle) also demonstrated more rapid healing than controls, but the magnitude of the effect was not as great as that seen with the drugs. Only the beta blockers were responsible for wound closure before 60 hours, whereas the saline treated controls and vehicle treated eyes required longer times for wound closure. In this model of wound healing, beta blockers appear to have no deleterious effect on corneal epithelial wound healing.     

Influences of methylcellulose on corneal epithelial wound healing.

We performed a study to evaluate the influence of methylcellulose (MC) on the corneal epithelial wound healing. A precise epithelial wound 2.0 mm in diameter and 70 microm in depth was created by an excimer laser on the cornea of a pig's eyeball. After treatment, the eyeballs were cultured in the incubator and perfused with TC-199 medium into the vitreous cavity. Topical MC with eight different combinations of concentrations and viscosities for experimental groups and BSS for the control group were applied on the epithelial defects three times. Wounds were evaluated 24 hours later with fluorescein stain and recorded with a scoring system. All of the experimental groups except one showed a more statistically significant superior wound-healing rate than the control group. MC possesses the effect to accelerate corneal epithelial wound healing.     

Effects of all-trans retinoic acid nanoparticles on corneal epithelial wound healing

Background

We have developed inorganically-coated all-trans retinoic acid (atRA) nanoparticles, nano-sized egg-like particles of atRA (NANOEGG®-atRA). The purpose of this study was to determine the effects of NANOEGG®-atRA on corneal wound healing in vivo and in vitro.

Methods

A rabbit corneal epithelial wound healing model was exposed to different concentrations of NANOEGG®-atRA. Wound healing was serially quantified as the ratio of fluorescein-stained area at the selected times to that at baseline. After wound closure, the barrier function of the cornea was determined using low concentrations of tropicamide. At the completion of the experiments, the corneal epithelium was histologically examined. For the in vitro studies, linear scratch wounds were made on cultured SV40-immortalized human corneal epithelial cells (HCE-T). Then, the cells were exposed to different concentrations of NANOEGG®-atRA, and wound healing was determined by the degree of closure of the scratch wound. In addition, the effects of NANOEGG®-atRA on the proliferation of HCE-T cells were determined by WST-8 assays.

Results

Exposure to NANOEGG®-atRA decreased the injured area 24 hrs after the ablation. The maximum effect of NANOEGG®-atRA was observed at a concentration of 33 mM. Histologically, no abnormal or differentiated corneal epithelial cells were observed in the histological sections treated with NANOEGG®-atRA. The tropicamide-induced pupillary dilation was significantly slowed in the eyes treated with NANOEGG®-atRA. NANOEGG®-atRA at concentrations of 3.3 and 33 nM induced earlier wound closure in vitro, but did not induce proliferation of HCE-T cells.

Conclusion

NANOEGG®-atRA promotes wound healing and should be considered for the treatment of wounds of the corneal epithelium.

     

Concentration-dependent effects of lidocaine on corneal epithelial wound healing.

Local anesthetic toxicity is a recognized clinical problem that has limited the use of topical corneal anesthetics for pain relief after corneal abrasion. Studies have shown clinically administered concentrations (0.5-2%) of local anesthetics impair corneal reepithelialization. Unfortunately, instillation of local anesthetic drops into an eye does not provide a measurable, steady-state concentration of drug. Thus, it has not been possible to evaluate whether there is an analgesic concentration of local anesthetic that does not impair corneal wound healing. Using the new in vitro rabbit cornea wound healing model, the effect of steady-state lidocaine concentrations on epithelial wound healing was examined. At lidocaine concentrations below 100 micrograms/ml, wound healing was not impaired. Higher concentrations (250-1000 micrograms/ml) resulted in dose-dependent impairment of epithelial wound healing. Combined with electrophysiologic evidence that corneal nerve injury discharge can be abolished by lidocaine concentrations less than 100 micrograms/ml, this research suggests that topical lidocaine in low concentration may be a safe topical corneal analgesic.     

Effects of capsaicin on corneal wound healing.

This study examined whether the depletion of neuropeptides from sensory nerve terminals induced by capsaicin modifies the healing rate of experimental corneal wounds in adult rabbits. Capsaicin (33 or 3.3 mM solutions) was administered topically and/or by a single retrobulbar injection to one eye while the fellow eye, treated with the vehicle, served as a control. After 1-3 weeks of treatment, an epithelial wound was made in the center of the cornea of both eyes with n-heptanol. Migration rates of epithelial cells surrounding the wound and estimated wound closure times were calculated by measuring the reduction in wound size. Combined treatment with 33 mM retrobulbar and 3.3 mM topical capsaicin for 3 weeks induced a significant delay in epithelial migration rates and in wound closure times (P less than 0.05). Topical or retrobulbar capsaicin alone for 3 weeks and combined treatment lasting only 1 week were not sufficient to modify wound healing times. The substance P antagonist, spantide (3 mM), applied topically for 1-3 weeks before or immediately after corneal wounding was also ineffective in changing wound closure rates. These findings suggest that the delayed wound healing observed after prolonged treatment with capsaicin could be due to a sustained depletion of neuropeptides from corneal sensory endings, supporting the hypothesis that trophic effects of sensory nerves on corneal epithelium are, at least in part, mediated by neuropeptides contained in peripheral nerve terminals.     

Alteration of epithelial paracellular permeability during corneal epithelial wound healing.

We studied the paracellular permeability to mannitol of corneas with epithelium of corneal, limbal, or conjunctival origin. Corneas with epithelial defects reepithelialized by corneal or limbal epithelium were nonvascularized; the corneal permeability was initially increased and returned to normal 3 days later. When epithelial defects extended beyond the limbus, they were healed by conjunctival epithelium. If corneas remained avascular or minimally vascularized, the conjunctiva-derived epithelium underwent a transdifferentiation process into a cornealike morphology in which the corneal permeability was initially increased upon complete reepithelialization, and gradually decreased to a level similar to that of normal cornea, 4 weeks after healing. However, when corneas became vascularized, the conjunctiva-derived epithelium retained its original phenotype, and corneal permeability remained increased throughout the 8-month period of study. The deranged barrier functions noted in the above vascularized cornea were demonstrated further by horseradish peroxidase tracer, which was found in the intercellular spaces of conjunctiva-derived epithelium of vascularized corneas but not in the avascular corneas with epithelia of corneal or limbal origin, or transdifferentiated conjunctival epithelium. To study further the effect of subsequent ocular surface trauma, conjunctival biopsy was performed on transdifferentiated avascular corneas 3 months after initial wounding. The biopsy resulted in extensive vascularization in three of eight previously nonvascularized corneas. Two weeks later, the corneal permeability was increased to a level similar to that of conjunctiva. These results indicate that corneal epithelial paracellular permeability correlates well with the status of the epithelial phenotype.     

小鼠角膜上皮创伤再上皮化过程基本特征的动力学研究

目的 研究小鼠角膜上皮机械性创伤再上皮化过程炎性细胞迁移、血小板聚集的动力学特征.方法 计算小鼠角膜2 mm上皮缺损创面改变;免疫荧光染色观察角膜基底细胞分裂和中性粒细胞、血小板、γδT细胞及巨噬细胞迁移特征;电子显微镜观察角膜上皮细胞形态学、基质PMNs和角膜缘血小板的变化.结果 创伤后24 h基本再上皮化,96 h基底细胞恢复正常;伤后18 h和30 h分裂细胞达峰值;PMNs迁移峰为12 h和30 h,血小板聚集峰于12 h;γδT细胞的两峰为6 h和24 h,MΦ两峰滞后为24 h和42 h.结论 角膜上皮创伤修复过程分3期.基底细胞分裂增生和PMNs、MΦ以及γδT细胞迁移呈双相式;早期PMNs迁移与血小板聚集同步.     

Clinical patterns of corneal epithelial wound healing

We studied the reepithelialization of corneal abrasions in 21 patients. All abrasions, irrespective of the nature of injury, followed a consistent pattern during reepithelialization. Three to six convex leading fronts of migrating epithelial sheets developed along the circumference of the defect and progressed toward the center. Neighboring fronts met along their sides, resulting in the formation of various geometric shapes. In the final stage of the healing process a contact line shaped like a "Y" or two "Y's" placed with their long axes end to end was seen where the advancing fronts of migrating epithelial sheets met. The rate of healing of the abrasions was determined by measuring the area of the abrasions at daily intervals from serial photographs. The area of the epithelial defects decreased exponentially with time, indicating a constant rate of epithelial cell migration.     

Effect of topical anti-inflammatory drugs on corneal and limbal wound healing

PURPOSE: To examine the effect of topical treatment with either steroidal or nonsteroidal anti-inflammatory drugs (NSAIDs) on the wound healing of corneal and limbal incisions using histologic criteria. SETTING: Loyola University Medical Center, Maywood, Illinois, USA. METHODS: Eighteen eyes of 9 cats were used for the study. All right eyes received a temporal limbal incision and the left eyes, a clear corneal incision. All eyes were treated with ofloxacin 0.3% postoperatively for 3 days. The animals were equally divided among 3 groups. Group 1 received topical steroid drops (prednisolone acetate 1%) for 7 days maximum; Group 2 received topical NSAID drops (ketorolac tromethamine 0.5%) for 7 days maximum; Group 3 received no additional treatment. One animal from each group was killed and the eye enucleated 3, 7, or 28 days postoperatively. The eyes were processed for staining with hematoxylin-eosin and smooth muscle actin, and corneal sections were evaluated in a blind fashion by an ophthalmic pathologist. RESULTS: Three and 7 days after surgery (1) the limbal incision exhibited more prominent wound healing than the similarly treated clear corneal incision, and (2) the steroid-treated corneas had less wound healing than untreated or NSAID-treated corneas. At 28 days, the wounds in all eyes were almost completely healed. CONCLUSIONS: Limbal incisions heal faster than clear corneal incisions. Steroids, but not NSAIDs, inhibit wound healing. Cataract surgery using limbal incisions and postoperative topical NSAIDs may result in faster wound healing and provide a reduced risk of related postoperative complications.     

血小板源性生长因子与角膜损伤修复

血小板源性生长因子(PDGF)在机体伤口愈合过程中具有重要的调节作用。由于角膜的特殊结构和功能,其伤口愈合的调节显得至关重要。本文简述PDGF的结构、受体、生物学活性及其在角膜伤口愈合中的作用。     

P-选择素对角膜上皮创伤修复及其中性粒细胞迁移的影响

目的观察P-选择素对角膜上皮创伤修复及其中性粒细胞迁移的影响。方法选用P-选择素基因敲除小鼠与野生型小鼠进行对比,观察角膜上皮创伤修复过程,同时用免疫组织化学方法观察中性粒细胞在角膜的迁移,并用酶联免疫吸附测定方法检测角膜中细胞因子和趋化因子的变化。结果与野生型小鼠相比,P-选择素基因敲除小鼠的角膜创伤修复延迟,向角膜迁移的中性粒细胞数量减少;中性粒细胞的迁移与趋化因子和细胞因子有密切的关系。结论P-选择素表达缺陷可导致角膜上皮创伤修复的延迟。这种延迟可能与创伤后中性粒细胞向创伤区迁移数量减少有关。     

Lack of effect of pilocarpine on corneal epithelial wound healing

The effect of a commercially available pilocarpine preparation on corneal epithelial healing rates was evaluated in rabbit eyes. Epithelial defects were created with filter paper discs soaked in n-heptanol. After creation of the defects, the corneas were stained with fluorescein and photographed using a Wratten blue filter. Follow-up examinations, drop administration, and photography were carried out every six hours for three days. Defect size was determined by computerized planimetry. The first phase of this study compared the effects of non-preserved saline and a commercially available methylcellulose preparation. The second phase compared a commercially available 2% pilocarpine preparation with 2% pilocarpine solution in non-preserved saline. We found that 2% pilocarpine had no significant effect on corneal epithelial healing rates.     

角质细胞生长因子促进角膜上皮损伤修复的研究

目的寻找促进角膜上皮损伤修复，治疗持续性角膜上皮缺损的有效方法。方法用３Ｈ－胸腺嘧啶核苷（３Ｈ－ＴｄＲ）掺入及液体闪烁技术，观察不同浓度的外源性角质细胞生长因子（ｋｅｒａｔｉｎｏｃｙｔｅｇｒｏｗｔｈｆａｃｔｏｒ，ＫＧＦ）对体外培养的人角膜上皮细胞生长的影响，由此推算出有效滴眼液浓度并应用于兔眼。用计算机图形分析系统计算角膜上皮愈合速率；用光镜和电镜评估愈合的质量。结果０．１～１００ｎｇ／ｍｌＫＧＦ有明显促进体外培养的人角膜上皮细胞生长的作用（增长率为２７．６６％～７６．７３％），且呈剂量依赖性（ｒ＝０．９２３３，Ｐ＜０．００１）。１μｇ／ｍｌＫＧＦ滴兔眼，加速了角膜上皮损伤修复（愈合速率，ＫＧＦ组为１．７７±０．２３ｍｍ２／ｈ，与对照组１．４９±０．２４ｍｍ２／ｈ比较，Ｐ＜０．０５）。结论外源性ＫＧＦ对体外培养的人角膜上皮细胞有明显的促生长作用，其滴眼液有加速兔眼角膜上皮创伤修复的作用。     

Cell-matrix and cell-cell interactions during corneal epithelial wound healing

The corneal epithelium serves as a barrier and contributes to the maintenance of corneal transparency and rigidity. In most instances, corneal epithelial defects caused by simple injury are resurfaced promptly. However, in individuals with certain clinical conditions, such as herpes simplex virus infection, neurotrophic keratopathy or diabetic keratopathy, corneal epithelial defects persist and do not respond to conventional treatment regimens because of delayed epithelial wound healing. After the corneal epithelium is removed by injury, the remaining epithelial cells migrate over the denuded surface of the cornea in a manner that is dependent both on the interaction of the cells with the underlying substrate and on cell-cell adhesion. In this review, we describe the specific roles of cell-matrix and cell-cell interactions during the course of corneal epithelial wound healing. The clinical implications of the basic research findings are also discussed.