Solvent/Detergent Virally Inactivated Serum Eye Drops Restore Healthy Ocular Epithelium in a Rabbit Model of Dry-Eye Syndrome.

Ching-Li Tseng,Shun-Hung Hsiao,Thierry Burnouf,Ting-Yi Renn,Zhi-Yu Chen,Andrew W Taylor

doi:10.1371/journal.pone.0153573

Abstract

Application of autologous serum eye drops (SEDs) is a recognized means to treat severe dry-eye syndrome (DES). Due to the inconvenience and difficulty of preparing SEDs from some patients, producing SEDs from allogeneic blood donations is gaining popularity. A major safety concern associated with allogeneic blood is virus transmission. We therefore herein evaluated the possibility of applying a solvent/detergent (S/D) treatment to inactivate viruses and studied the impacts of such treatment of SEDs to resolve DES in a rabbit model. Sera prepared from the blood of five rabbits were pooled and divided into two sub-pools. One was untreated (SEDs), while the other was virally-inactivated with 1% Tri-n-butyl phosphate/1% Triton X-45 at 31°C for 1 h (S/D-SEDs). DES was induced in rabbits using 0.1% benzalkonium chloride (BAC). Rabbits were divided into five groups of two rabbits each. One group was untreated (control), three were treated twice daily for 3 weeks using PBS, SEDs, or S/D-SEDs, and the last received an additional 0.1% BAC (as the negative control). The DES condition was determined by measuring aqueous tear secretion (Schirmer’s test), corneal fluorescein staining, a corneal histologic examination, TUNEL stain apoptosis, and corneal inflammatory marker (tumor necrosis factor-α, interleukin (IL)-1β, IL-8, and IL-6) expressions. We first confirmed that SEDs and S/D-SEDs had similar protein profiles and transforming growth factor (TGF)-β contents. Animal experiments showed that tear secretion did not significantly differ between the SED and S/D-SED groups but was significantly higher than in the PBS group. Eye fluorescein staining revealed dramatic improvements in epithelial defects in groups treated with SEDs or S/D-SEDs, and hematoxylin/eosin staining revealed microscopic epithelial layers similar to those of the untreated controls. Inflammatory markers and TUNEL studies showed that healthy epithelium had been restored in groups treated with SEDs or S/D-SEDs. In conclusion, this preclinical study supports the possibility of using S/D virally inactivated SEDs to treat DES and restore a normal epithelium.

Highlights

Dry-eye syndrome (DES) is a multifactorial disease of the ocular surface characterized by rapid evaporation and insufficient production of tears
SDS-PAGE revealed no substantial differences in protein compositions regardless of the molecular mass, with an expected prominent band corresponding to albumin (Fig 2)
DES is associated with inflammation of the ocular surface [31,36,37]

Summary

Introduction

Dry-eye syndrome (DES) is a multifactorial disease of the ocular surface characterized by rapid evaporation and insufficient production of tears. Its severity is worsened by pathological conditions such as Sjögren's syndrome, which induces disruption of the ocular surface epithelium [3], or chronic graft-versus-host disease (GVHD) [4]. Serum eye drops (SEDs) made from human blood have recently emerged as an alternative and apparently more-efficient method than conventional treatment to improve the ocular surface health, tear film stability, and subjective comfort in refractory DES [5,6,7,8,9,10,11]. A major serum component, may contribute to ameliorating ocular surface damage [13]

Methods

Results

Conclusion