Wahaj Saleem1, Angela H Benton2, Mary E Marquart2, Shuli Wang1, Waqas Saleem1, Randy Vigil1, Bo Huang3, Anjal C Sharma1. 1. Experimental Therapeutics and Diagnostics Group, Lynntech, Inc., 2501 Earl Rudder Fwy S., College Station, TX 77845, USA. 2. Department of Microbiology and Immunology, University of Mississippi Medical Center, Jackson, MS 39216, USA. 3. Department of Ophthalmology, University of Mississippi Medical Center, Jackson, MS 39216, USA.
Abstract
PURPOSE: To evaluate the effect of application of 3% air in helium cold atmospheric plasma jet, using an inexpensive device termed iCAP, in corneal scratch wound closure in vitro and the treatment of Pseudomonas aeruginosa (P. aeruginosa) keratitis in vivo. METHODS: Thermal imaging to measure temperature of surfaces to which iCAP was applied and UV energy density delivered by iCAP were measured. Scratch wounds inflicted on in vitro cultures of a human corneal epithelial cell line were treated with iCAP and wound widths at various times post-application were measured. Rabbit eyes infected with P. aeruginosa were treated with iCAP and slit lamp biomicroscope examination conducted to determine corneal health outcomes 25h post infection. Corneal homogenates were plated on agar and viable bacterial colonies enumerated to determine the effect of iCAP on bacterial load in vivo in P. aeruginosa keratitis. RESULTS: iCAP was shown to operate in the non-thermal regime and also shown to deliver much lower UV energy density than that necessary to cause harmful effects on ocular tissue. iCAP treatment significantly improved the rate of scratch wound gap closure in vitro in a human corneal epithelial cell line compared to controls. In vivo, iCAP treatment of P. aeruginosa keratitis infection in the rabbit eyes (N = 20) significantly reduced the incidence of corneal ulcer (P = 0.003) and corneal edema (P = 0.011) and significantly improved total cornea health (P = 0.034) compared to untreated (N = 10). Finally, in vivo iCAP treatment of P. aeruginosa keratitis infection in the rabbit eyes (N = 19) significantly reduced bacterial loads (P = 0.012) compared to untreated (N = 9). CONCLUSION: Our results strongly suggest that iCAP treatment was effective in improving corneal epithelial defect closure in vitro, reducing ulcer formation and decreasing inflammation in P. aeruginosa infected corneas in vivo and decreasing bacterial loads in P. aeruginosa infected corneas in vivo which led to improved overall cornea health outcomes in vivo. Further studies to investigate iCAP's safety and efficacy against other infectious microbes responsible for causing ulcerative keratitis, with and without co-treatment with antimicrobial therapies are warranted.
PURPOSE: To evaluate the effect of application of 3% air in helium cold atmospheric plasma jet, using an inexpensive device termed iCAP, in corneal scratch wound closure in vitro and the treatment of Pseudomonas aeruginosa (P. aeruginosa) keratitis in vivo. METHODS: Thermal imaging to measure temperature of surfaces to which iCAP was applied and UV energy density delivered by iCAP were measured. Scratch wounds inflicted on in vitro cultures of a human corneal epithelial cell line were treated with iCAP and wound widths at various times post-application were measured. Rabbit eyes infected with P. aeruginosa were treated with iCAP and slit lamp biomicroscope examination conducted to determine corneal health outcomes 25h post infection. Corneal homogenates were plated on agar and viable bacterial colonies enumerated to determine the effect of iCAP on bacterial load in vivo in P. aeruginosa keratitis. RESULTS: iCAP was shown to operate in the non-thermal regime and also shown to deliver much lower UV energy density than that necessary to cause harmful effects on ocular tissue. iCAP treatment significantly improved the rate of scratch wound gap closure in vitro in a human corneal epithelial cell line compared to controls. In vivo, iCAP treatment of P. aeruginosa keratitis infection in the rabbit eyes (N = 20) significantly reduced the incidence of corneal ulcer (P = 0.003) and corneal edema (P = 0.011) and significantly improved total cornea health (P = 0.034) compared to untreated (N = 10). Finally, in vivo iCAP treatment of P. aeruginosa keratitis infection in the rabbit eyes (N = 19) significantly reduced bacterial loads (P = 0.012) compared to untreated (N = 9). CONCLUSION: Our results strongly suggest that iCAP treatment was effective in improving corneal epithelial defect closure in vitro, reducing ulcer formation and decreasing inflammation in P. aeruginosa infected corneas in vivo and decreasing bacterial loads in P. aeruginosa infected corneas in vivo which led to improved overall cornea health outcomes in vivo. Further studies to investigate iCAP's safety and efficacy against other infectious microbes responsible for causing ulcerative keratitis, with and without co-treatment with antimicrobial therapies are warranted.
Authors: Mary E Marquart; Kathryn S Monds; Clare C McCormick; Sherrina N Dixon; Melissa E Sanders; Julian M Reed; Larry S McDaniel; Armando R Caballero; Richard J O'Callaghan Journal: Invest Ophthalmol Vis Sci Date: 2007-06 Impact factor: 4.799
Authors: Christopher R Henry; Harry W Flynn; Darlene Miller; Richard K Forster; Eduardo C Alfonso Journal: Ophthalmology Date: 2012-08-01 Impact factor: 12.079
Authors: R J O'Callaghan; L S Engel; J A Hobden; M C Callegan; L C Green; J M Hill Journal: Invest Ophthalmol Vis Sci Date: 1996-03 Impact factor: 4.925