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Here, animal models based on mouse and rabbit are developed for mechanical and chemical injury of corneal epithelium to screen new therapeutics and the underlying mechanism.
Corneal injury to the ocular surface, including chemical burn and trauma, may cause severe scarring, symblepharon, corneal limbal stem cells deficiency, and result in a large, persistent corneal epithelial defect. Epithelial defect with the following corneal opacity and peripheral neovascularization result in irreversible visual impairment and hinder future management, especially keratoplasty. Since the animal model can be used as an effective drug development platform, models of corneal injury to the mouse and alkali burn to rabbit corneal epithelium are developed here. New Zealand white rabbit is used in the alkali burn model. Different concentrations of sodium hydroxide can be applied onto the central circular area of the cornea for 30 s under intramuscular and topical anesthesia. After copious isotonic normal saline irrigation, residual loose corneal epithelium was removed with corneal burr deep down to the Bowman's layer within this circular area. Wound healing was documented by fluorescein staining under Cobalt blue light. C57BL/6 mice were used in the traumatic model of murine corneal epithelium. The murine central cornea was marked using a skin punch, 2 mm in diameter, and then debrided by a corneal rust ring remover with a 0.5 mm burr under a stereomicroscope. These models can be prospectively used to validate the therapeutic effect of eye drops or mixed agents such as stem cells, which potentially facilitate corneal epithelial regeneration. By observing corneal opacity, peripheral neovascularization, and conjunctival congestion with stereomicroscope and imaging software, therapeutic effects in these animal models can be monitored.
The human cornea consists of five major layers and plays a pivotal role in ocular refraction to maintain visual acuity and structural integrity for protecting intraocular tissues1. The outermost part of the cornea is the corneal epithelium, composed of five to six layers of cells that sequentially differentiate from the basal cells and move upward to shed from the ocular surface1. Compared to the cornea in humans and New Zealand rabbits, mouse cornea has a similar corneal structure, but thinner periphery than the central part due to a reduced thickness in the epithelium and the stroma2. Because of....
All of the experimental procedures in animal studies were approved by the Research Ethics Committee at the Chang Gung Memorial Hospital and adhered to the ARVO statement for use of animals in ophthalmic and vision research.
1. Ex vivo wound healing model of the mouse corneal epithelium
Ex vivo wound healing model of the mouse corneal epithelium:
After in vivo debridement of mouse corneal epithelium with hand-held corneal rust ring remover, a mildly depressed central corneal area with positive fluorescein stain can be found in the central 2 mm area (Figure 3A-B). After harvesting the mouse eyeball, it was easily fixed onto a wax-coated 48-well culture plate without significant rotati.......
Mouse and rabbit models of corneal injury provide a useful ex vivo and in vivo platform for monitoring wound healing, testing new therapeutics, and studying underlying mechanisms of wound healing and treatment pathways. Different animal models can be used for a short-term or long-term experiment, depending on the purpose of the research. For instance, after creating an epithelial defect on mouse cornea in vivo, a confined epithelial defect could be used to monitor liquid therapeutic agents in a.......
The study was funded by the Atomic Energy Council of Taiwan (Grant No. A-IE-01-03-02-02), Ministry of Science and Technology (Grant No. NMRPG3E6202-3), and Chang Gung Medical Research Project (Grant No. CMRPG3H1281).
....Name | Company | Catalog Number | Comments |
6/0 Ethicon vicryl suture | Ethicon | 6/0VICRYL | tarsorrhaphy |
Barraquer lid speculum | katena | K1-5355 | 15 mm |
Barraquer needle holder | Katena | K6-3310 | without lock |
Barron Vacuum Punch 8.0 mm | katena | K20-2108 | for cutting filter paper |
C57BL/6 mice | National Laboratory Animal Center | RMRC11005 | mouse strain |
Castroviejo forceps 0.12 mm | katena | K5-2500 | |
Corneal rust ring remover with 0.5 mm burr | Algerbrush IITM; Alger Equipment Co., Inc. Lago Vista, TX | CHI-675 | for debridement of the corneal epithelium |
Filter paper | Toyo Roshi Kaisha,Ltd. | 1.11 | |
Fluorescein sodum ophthalmic strips U.S.P | OPTITECH | OPTFL100 | staining for corneal epithelial defect |
Ketamine hydrochloride | Sigma-Aldrich | 61763-23-3 | intraperitoneal or intramuscular anesthetics |
New Zealand White Rabbits | Livestock Research Institute, Council of Agriculture,Executive Yuan | Rabbit models | |
Normal saline | TAIWAN BIOTECH CO., LTD. | 100-120-1101 | |
Proparacaine | Alcon | ALC2UD09 | topical anesthetics |
Skin biopsy punch 2mm | STIEFEL | 22650 | |
Sodium chloride (NaOH) | Sigma-Aldrich | 1310-73-2 | a chemical agent for alkali burn |
Stereomicroscope | Carl Zeiss Meditec, Dublin, CA | SV11 | microscope for surgery |
Westcott Tenotomy Scissors Medium | katena | K4-3004 | |
Xylazine hydrochloride 23.32 mg/10 mL | Elanco animal health Korea Co., LTD. | 047-956 | intraperitoneal or intramuscular anesthetics |
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