Efficacy of three different methods for side port incision wound sealing

PURPOSE: To evaluate wound leakage and bacteria-sized particle influx in differently corneal sealed side port incisions. METHODS: Four 1.5mm tunnel squared incisions were created in each of four cadaveric human eyes. In each cornea, one incision was left unsealed, whereas the other three incisions were sealed using a 10-0 nylon suture, cyanoacrylate glue, or stromal hydration, respectively. A Seidel and an India ink test were performed on each eye. During each Seidel test, flourescein was applied, the IOP increased from 15 to 80mmHg, and the IOP at which each incision started to leak recorded. During each India ink test, ink was placed on the eye and rinsed out with balanced salt solution (BSS). Ink penetration was then measured by planimetry at physiologic conditions and after an IOP plunge from 80mmHg to 0mmHg. RESULTS: Regardless of IOP variations, no leakage or ink inflow was observed through the glued incisions. In contrast, leakage did occur in the other three sealing methods, albeit at significantly different IOP levels in each one (p=0.013). Ink inflow occurred in these sealing methods at physiologic IOP and, to a significantly greater extent, after the IOP challenge (p<0.05). At both of these IOP conditions, the differences in ink influx among these three incision-sealing methods were deemed statistically insignificant. CONCLUSION: This study showed that glue was more effective at preventing wound leakage and bacteria-sized particle influx than other commonly used methods especially hydrosealing.

OBJETIVO: Avaliar o sinal de Seidel positivo e a penetração de tinta da Índia em incisões corneanas acessórias seladas por diferentes métodos. MÉTODOS: Quatro incisões de 1,5 x 1,5mm foram criadas em cada um dos quatro olhos provindos do banco de olhos. Em cada córnea, uma incisão foi mantida sem manipulação, enquanto que as outras três incisões foram seladas respectivamente com Nylon 10-0, cola de cianoacrilato e hidratação estromal. Foram realizados dois testes: Sinal de Seidel positivo e penetração da tinta da Índia. No primeiro, foi aplicado fluoresceína gotas e a pressão intraocular (PIO) elevada de 15 para 80mmHg. No segundo, a tinta da Índia foi aplicada sobre o olho estudado em duas situações: sob PIO fisiológica e sob variação súbita da pressão, de 80 para 0mmHg. RESULTADOS: Na incisão selada com cola, apesar da variação da PIO, não houve vazamento e nem penetração de partículas de tinta. Por outro lado, o sinal de Seidel foi positivo nas outras três incisões em diferentes níveis de PIO (p=0,013). A penetração da tinta da Índia ocorreu nestas três incisões sob pressão fisiológica e com maior extensão após a variação da PIO (p<0,05). Esta diferença, no entanto não foi considerada estatisticamente significante quando comparadas as incisões entre si. CONCLUSÃO: No presente estudo, a cola foi mais eficaz em prevenir Seidel e entrada de partículas do que outro método comumente usado especialmente, hidratação estromal.

Assessment of fungal viability after long-wave ultraviolet light irradiation combined with riboflavin administration.

BACKGROUND: Corneal collagen cross-linking (CXL), a technique that combines riboflavin administration with long-wave ultraviolet light irradiation, was primarily developed to increase the biomechanical strength of collagen fibrils of the cornea to avoid the progression of keratoconus. Recently, this method has been proposed to treat selected cases of infectious keratitis. METHODS: To test the protocol used for progressive keratoconus in infectious keratitis, Candida albicans, and Fusarium solani, strains were exposed to irradiation using a wavelength of 365 nm at a power density of 3 mW/cm(2) for 30 min in the presence of riboflavin photosensitizer. All experiments were performed in triplicate. Qualitative and quantitative measurements of fungal viability used plate cultures and an automated trypan blue dye exclusion method respectively. Fungal cell diameter was also assessed in all groups. Statistical analyses were performed using the triplicate values of each experimental condition. RESULTS: Experimental findings of photodynamic therapy applied to the cell inactivation of both yeasts and filamentous fungi were compared with control groups. Qualitative results were corroborated with quantitative findings which showed no statistical significance between challenged samples (experimental groups) and the control group (p-value = 1). In comparison with a control group of live cells, statistical significance was observed when riboflavin solution alone had an effect on the morphologic size of filamentous fungi, while ultraviolet light irradiation alone showed a slight decrease in the cell structure of C. albicans. CONCLUSIONS: The impact of long-wave ultraviolet combined with riboflavin photosensitizer showed no antifungal effect on C. albicans and F. solani. The significant decrease in cell morphology of both filamentous fungi and yeasts submitted to photosensitizing riboflavin and exposure to ultraviolet light, respectively, may be promising in the development and standardization of alternatives for fungal cell inactivation, because of their minimal cytotoxic effects on the corneal surface. The methodological improvement in the preparation and application of individual chemical compounds, such as riboflavin, or physical systems, such as a long-wave light source, as antifungal agents may also assist in establishing promising therapeutic procedures for keratomycosis.

Assessing efficacy of combined riboflavin and UV-A light (365 nm) treatment of Acanthamoeba trophozoites.

PURPOSE: To assess the Acanthamoeba trophozoite viability in vitro and treatment of Acanthamoeba keratitis in a hamster model using ultraviolet light A (UV-A) and riboflavin (B2). METHODS: A sample of Acanthamoeba sp. cultured was transferred to a 96-well plate and exposed to B2 and the UV-A light (365 nm wavelength) at a power density of 3 mW/cm(2), 8 mm spot diameter, for 30 minutes. The exposure was done in triplicate. Control groups were prepared in triplicate as well: blank control, UV-A only, riboflavin only, and dead control. Cell viability assessment was done using the trypan blue dye exclusion method. Acanthamoeba keratitis was induced in Chinese hamsters; who were randomly assigned to one of the animal groups: UV-A + B2, propamidine isethionate (Brolene; Sanofi-Aventis, Ellerslie, Auckland, Australia), UV-A + B2 + propamidine isethionate (Brolene), only UV-A, only B2, and blank. Throughout the 14 days after treatment the animals were examined clinically. Histology and clinical scores of all groups were compared. RESULTS: The in vitro study showed no difference between the treatment group UV-A + B2 and the control groups. In the hamster keratitis model a significant improvement of clinical score was observed for the groups propamidine isethionate (Brolene) and UV-A + B2 + propamidine isethionate (Brolene) (P = 0.0067). Also a significant worsening of clinical score was observed in the other groups: UV-A + B2 group (P = 0.0084), only UV-A (P = 0.0078), B2 only (P = 0.0084), and blank (P = 0.0082). No difference was observed between propamidine isethionate (Brolene) and UV-A + B2 + propamidine isethionate (Brolene). CONCLUSIONS: The adjunctive use of UV-A and B2 therapy did not demonstrate antitrophozoite activity; in vivo UV-A and B2 did not demonstrate efficacy in this model.