PURPOSE: To determine the abundance of extracellular DNA (eDNA) in tear fluid of patients with dry eye disease (DED) and to report clinical outcomes after DNase I eyedrops use to reduce excessive tear fluid eDNA. METHODS: Tear fluid was collected from healthy control subjects and patients with DED. The eDNA abundance was determined with the PicoGreen dye assay. The DED symptoms and clinical signs were recorded and correlated with eDNA abundance. Two patients with DED having excessive eDNA in tear fluid were treated with DNase I eyedrops. RESULTS: The PicoGreen dye assay measures tear fluid eDNA abundance after a 2-minute incubation time. With longer incubations, admixed cells also contribute to eDNA measurements. The mean (SE) eDNA abundance in healthy control subjects' tear fluid was 1.4 (0.2) μg/mL. The mean (SE) eDNA abundance in tear fluid of patients with nonautoimmune DED, autoimmune DED, and graft versus host disease was significantly higher: the values were 2.9 (0.6), 5.2 (1.2), and 9.1 (2.3) μg/mL, respectively (P < 0.05). In most of these patients, the PicoGreen dye kinetic assay of tear fluid showed an increase in fluorescence signal due to the presence of viable cells in tear fluid. Tear fluid eDNA had the best correlation with corneal Rose Bengal staining (r = 0.55). Treatment of patients having DED with DNase I eyedrops reduced eDNA abundance, abrogated signal increase, and improved comfort. CONCLUSIONS: Excessive eDNA is present in tear fluid of patients with dry eyes. A novel therapeutic approach for managing DED may be to measure eDNA abundance in tear fluid with the PicoGreen dye assay and reduce excessive amounts with DNase I eyedrops.
PURPOSE: To determine the abundance of extracellular DNA (eDNA) in tear fluid of patients with dry eye disease (DED) and to report clinical outcomes after DNase I eyedrops use to reduce excessive tear fluid eDNA. METHODS: Tear fluid was collected from healthy control subjects and patients with DED. The eDNA abundance was determined with the PicoGreen dye assay. The DED symptoms and clinical signs were recorded and correlated with eDNA abundance. Two patients with DED having excessive eDNA in tear fluid were treated with DNase I eyedrops. RESULTS: The PicoGreen dye assay measures tear fluid eDNA abundance after a 2-minute incubation time. With longer incubations, admixed cells also contribute to eDNA measurements. The mean (SE) eDNA abundance in healthy control subjects' tear fluid was 1.4 (0.2) μg/mL. The mean (SE) eDNA abundance in tear fluid of patients with nonautoimmune DED, autoimmune DED, and graft versus host disease was significantly higher: the values were 2.9 (0.6), 5.2 (1.2), and 9.1 (2.3) μg/mL, respectively (P < 0.05). In most of these patients, the PicoGreen dye kinetic assay of tear fluid showed an increase in fluorescence signal due to the presence of viable cells in tear fluid. Tear fluid eDNA had the best correlation with corneal Rose Bengal staining (r = 0.55). Treatment of patients having DED with DNase I eyedrops reduced eDNA abundance, abrogated signal increase, and improved comfort. CONCLUSIONS: Excessive eDNA is present in tear fluid of patients with dry eyes. A novel therapeutic approach for managing DED may be to measure eDNA abundance in tear fluid with the PicoGreen dye assay and reduce excessive amounts with DNase I eyedrops.
Authors: Patrick M Ladage; James V Jester; W Matthew Petroll; Jan P G Bergmanson; H Dwight Cavanagh Journal: Invest Ophthalmol Vis Sci Date: 2003-03 Impact factor: 4.799
Authors: Kavitha R Sivaraman; Renu V Jivrajka; Ketki Soin; Charles S Bouchard; Asadolah Movahedan; Ellen Shorter; Sandeep Jain; Deborah S Jacobs; Ali R Djalilian Journal: Ocul Surf Date: 2016-05-12 Impact factor: 5.033
Authors: Yoshihiro Inamoto; Nuria Valdés-Sanz; Yoko Ogawa; Monica Alves; Luigi Berchicci; John Galvin; Hildegard Greinix; Gregory A Hale; Biljana Horn; Debra Kelly; Hien Liu; Scott Rowley; Helene Schoemans; Ami Shah; Maria Teresa Lupo Stanghellini; Vaibhav Agrawal; Ibrahim Ahmed; Asim Ali; Neel Bhatt; Michael Byrne; Saurabh Chhabra; Zachariah DeFilipp; Kristina Fahnehjelm; Nosha Farhadfar; Erich Horn; Catherine Lee; Sunita Nathan; Olaf Penack; Pinki Prasad; Seth Rotz; Alicia Rovó; Jean Yared; Steven Pavletic; Grzegorz W Basak; Minoo Battiwalla; Rafael Duarte; Bipin N Savani; Mary E D Flowers; Bronwen E Shaw; Igor Petriček Journal: Biol Blood Marrow Transplant Date: 2018-11-24 Impact factor: 5.742
Authors: Daniel Wolff; Vedran Radojcic; Robert Lafyatis; Resat Cinar; Rachel K Rosenstein; Edward W Cowen; Guang-Shing Cheng; Ajay Sheshadri; Anne Bergeron; Kirsten M Williams; Jamie L Todd; Takanori Teshima; Geoffrey D E Cuvelier; Ernst Holler; Shannon R McCurdy; Robert R Jenq; Alan M Hanash; David Jacobsohn; Bianca D Santomasso; Sandeep Jain; Yoko Ogawa; Philipp Steven; Zhonghui Katie Luo; Tina Dietrich-Ntoukas; Daniel Saban; Ervina Bilic; Olaf Penack; Linda M Griffith; Meredith Cowden; Paul J Martin; Hildegard T Greinix; Stefanie Sarantopoulos; Gerard Socie; Bruce R Blazar; Joseph Pidala; Carrie L Kitko; Daniel R Couriel; Corey Cutler; Kirk R Schultz; Steven Z Pavletic; Stephanie J Lee; Sophie Paczesny Journal: Transplant Cell Ther Date: 2021-06-10