Moxifloxacin

25G PPV was performed using the CONSTELLATION Vision System (Alcon Laboratories, Inc.). In all eyes, a central vitrectomy was performed. The posterior vitreous was separated from the retina by active aspiration with the vitrectomy probe, and any visible vitreous strands that were adherent to the retina were removed. Intravitreous triamcinolone (40 mg/mL, Triesence^®, Alcon, Forth Worth, Texas, USA) was systematically used in all cases as a marker to facilitate visualization and removal of the adherent posterior cortical vitreous. triamcinolone was fully washed out before performance of ILM-peeling in all cases. ILM-peeling was systematically performed using vision blue G (0,125 mg Brilliant Blue G, Fluoron, Ulm, Germany) to stain and then remove the ILM. Postoperatively, topical antibiotic (Vigamox, moxifloxacin 0.5%, Alcon, USA) and antiinflammatory therapy (Pred Forte, prednisolone acetonide 1%, Allergan, Ireland) were administered 4 times daily over 1 month.

For injury experiments, adult 60-day-old male mice were anesthetized with intraperitoneal injection of ketamine (100 mg/kg; Mylan Pharmaceuticals, Rockford, IL, USA) and xylazine (10 mg/kg; Mylan Pharmaceuticals), and subcutaneous analgesia was administered as described above. Once the mouse was under general anesthesia, 0.1 ml of 0.5% proparacaine hydrochloride (Alcon Labs, Dallas, TX, USA) and 1% atropine (Alcon Labs) were placed on the ocular surface. By visualization under a microscope, a full-thickness corneal incision, 1 mm in length, was made centrally in the cornea with a 15-degree blade. The stromal matrix was labeled with 5[(4,6-dichlorotriazin-2yl)-amino]fluorescein (DTAF, Sigma-Aldrich) in six I-KeramTmG corneas to demarcate the areas of tissue repair (Sun et al., 2020a (link)). Immediately after the procedure, drops of moxifloxacin (Alcon Labs) and artificial tear ointment (Alcon Labs) were applied to the ocular surface. Mice were housed and treated in accordance with the National Institutes of Health's Guide for the Care and Use of Laboratory Animals. Eyes were evaluated at 4, 9 and 14 weeks after injury.

The experiment used 60 human eyeballs that did not meet the safety serological criterion for corneal transplantation. Only 1 eye of each donor was used. To be included in the study, the eyes could not have any of the following conditions: gross epithelial defects, gross alterations of transparency, congenital disorders, ectasias, corneal guttata, keratitis, and more than 72 h of preservation in the wet chamber. The study protocol was approved by the ethical board of our institution.
A set of numbers, ranging from 1 to 60, was randomly divided into three subsets of 20. Each eye received a number corresponding to its order of retrieval, which automatically allocated it to one of those subsets. The eyeballs were fully immersed for 10 min in 15 mL of one of the following commercial solutions: 0.5% moxifloxacin (Vigamox^©, Alcon, Brazil), 0.3% ciprofloxacin (Ciloxan^©, Alcon, Brazil), and 0.3% ofloxacin (Oflox^©, Allergan, Brazil). The globes were then washed with 100 mL of saline to remove all traces of antibiotics. Next, whole corneas and samples of aqueous humor (collected with 30G tuberculin syringes at volumes of at least 100 µL) were collected and stored in non-transparent Eppendorf plastic vials. After identification, they were immediately stored in liquid nitrogen at –70°C until analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS).

The following FLQs were used in this study: ciprofloxacin (Hikma Pharmaceutical: Amman, Jordan), gatifloxacin (Allergen Pharmaceuticals: Dublin, Ireland), levofloxacin (Sanofi-Aventis Deutschland GmbH: Frankfurt, Germany), moxifloxacin (Alcon Laboratories: Fort Worth, TX, USA), norfloxacin (Amman Pharmaceutical industries: Amman, Jordan), and ofloxacin (Allergen Pharmaceuticals, Ireland).

Prior to surgical procedures, the rabbits were anesthetized by an intramuscular injection of ketamine hydrochloride (30 mg/kg) and xylazine hydrochloride (5 mg/kg). Pupils were dilated using 2.5% phenylephrine/0.5% tropicamide eye drops. Povidone iodine 5% was applied to the eyelids. In addition, topical anesthesia was administered using several drops of lidocaine 0.4%. Two 23ga trocars, inserted in the superotemporal and superior-nasal quadrants (1.0 mm posterior to the corneoscleral limbus were used to insert a lighted infusion cannula and a vitrector (Millennium, Bausch & Lomb, Bridgewater, NJ). Under constant infusion of a balanced saline solution, as much vitreous was removed as possible, while avoiding damage to the lens. After fluid-air exchange, the vitreous cavity was filled with silicone oil. Intraocular pressure was not measured.
Subconjunctival injection of gentamicin and betamethasone depot (Celestone Chronodose, MSD) was administered. Moxifloxacin (Vigamox, Alcon, Fort Worth, TX, USA) drops and tobramycin eye ointment (Alcon, USA) were applied locally.