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Effects of Topical Human Amniotic Fluid and Human Serum in a Mouse Model of Keratoconjunctivitis Sicca

Quinto, Guilherme G. MD; Camacho, Walter MD; Castro-Combs, Juan MD; Li, Li MD; Martins, Suy Anne R. MD, PhD; Wittmann, Priscila MD; Campos, Mauro MD, PhD; Behrens, Ashley MD

doi: 10.1097/ICO.0b013e31823f0a64
Basic Investigation

Purpose To compare the effects of topical human amniotic fluid (HAF), topical human serum (HS), and topical artificial tears in a mouse model of dry eye.

Methods Thirty C57BL/6 mice were divided into 3 treatment groups: HAF, HS, and preservative-free artificial tears. Dry eye was induced by an injection of botulinum toxin B (BTX-B) into the lacrimal gland. Tear production and ocular surface fluorescein staining were evaluated in each mouse at 6 time points during a 4-week period. Goblet cell density was assessed in stained histological sections. Apoptotic keratocytes were evaluated by terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling test assay.

Results A significant decrease in tear production was observed 3 days after BTX-B injection in all groups. At week 1, the HAF and HS groups had improved tear production compared with the control group (P < 0.001 and P = 0.003, respectively). HAF had a significantly improved fluorescein staining score compared with the HS (P = 0.043) and control (P = 0.007) groups at week 2. Goblet cell density was significantly decreased in the control group compared with the HAF and HS groups (P < 0.001). No difference in the amount of terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling-positive keratocytes was observed among the groups.

Conclusion HAF was superior to HS and artificial tears for improving corneal staining within 2 weeks of therapy in this induced mouse model of keratoconjunctivitis sicca. Clinical studies are needed to ascertain the benefits of these therapies in patients with ocular surface disorders associated with dry eye.

*Wilmer Ophthalmological Institute, Department of Ophthalmology, Johns Hopkins University, Baltimore, MD

Vision Institute, Department of Ophthalmology, Federal University of São Paulo, São Paulo, Brazil.

Reprints: Ashley Behrens, The Wilmer Ophthalmological Institute, Department of Ophthalmology, Johns Hopkins University, 255 Woods Bldg, 600 North Wolfe St, Baltimore, MD 21287-0005 (e-mail:

Funding/Support: This study was supported by Research to Prevent Blindness Inc., New York, NY.

The authors state that they have no proprietary interest in the products named in this article.

Received July 2, 2011

Accepted October 14, 2011

Copyright © 2012 Wolters Kluwer Health, Inc. All rights reserved.