Ophthalmic adverse effects of nasal decongestants on an experimental rat model / Efeitos oftálmicos adversos de descongestionantes nasais em modelo experimental com ratos

ABSTRACT Purpose: To investigate the potential effects of chronic exposure to a nasal decongestant and its excipients on ocular tissues using an experimental rat model. Methods: Sixty adult male Wistar rats were randomized into six groups. The first two groups were control (serum physiologic) and Otrivine® groups. The remaining four groups received the Otrivine excipients xylometazoline, benzalkonium chloride, sorbitol, and ethylene diamine tetra acetic acid. Medications were applied into both nostrils twice a day for 8 weeks. Before the rats were sacrificed, epithelial staining, the Schirmer test, and intraocular pressure measurements were performed under ketamine/xylasine anesthesia (50 and 5 mg/kg, respectively). Results: Epithelial defects and dry eye were common findings in all study groups. Cataracts developed in two cases clinically. Histopathological evaluation revealed many different pathological alterations in all parts of the ocular tissues such as corneal edema, polypoid proliferation and hyalinization of the vessel wall, cystic formation of the lens, retinal nerve fiber layer degeneration, and corpora amylacea formation of the lacrimal gland. Conclusions: Prolonged usage of the nasal decongestant xylometazoline and its excipients may cause ophthalmic problems such as dry eyes, corneal edema, cataracts, retinal nerve fiber layer, and vascular damage in rats. Although these results were obtained from experimental animals, ophthalmologists should keep in mind the potential ophthalmic adverse effects of this medicine and/or its excipients and exercise caution with drugs containing xylometazoline, ethylene diamine tetra acetic acid, benzalkonium chloride and sorbitol for patients with underlying ocular problems.

RESUMO Objetivo: Investigar os possíveis efeitos da exposição crônica de descongestionante nasal e seus excipientes em tecidos oculares, utilizando um modelo experimental com ratos. Métodos: Sessenta ratos Wistar adultos machos foram divididos aleatoriamente em seis grupos. Os primeiros dois grupos foram controle (soro fisiológico) e Otrivina®. Os quatro grupos restantes receberam os excipientes de Otrivina, tais como Xilometazolina, Benzalcônio, Sorbitol e Ácido Etilenodiamino Tetracético (EDTA). Os medicamentos foram aplicados em ambas as narinas dos ratos, duas vezes ao dia, durante 8 semanas. Antes que os ratos fossem sacrificados, a coloração epitelial, o teste de Schirmer e a medida da pressão intraocular foram realizados sob anestesia com Ketamina/Xilasina (50 e 5 mg/kg, respectivamente). Resultados: Defeitos epiteliais e olho seco foram achados comuns nos grupos de estudo. A catarata desenvolveu-se clinicamente em dois casos. A avaliação histopatológica revelou a existência de alterações em todas as partes dos tecidos oculares, tais como edema de córnea, proliferação polipoide e hialinização da parede vascular, formação cística da lente, degeneração da camada de fibra nervosa da retina (RNFL) e formação de corpos amiláceos da glândula lacrimal. Conclusões: O uso prolongado do descongestionante nasal Xilometazolina e seus excipientes pode causar vários problemas oftalmológicos, como olho seco, edema de córnea, catarata, RNFL e dano vascular em ratos. Embora esses resultados tenham sido obtidos a partir de animais experimentais, os oftalmologistas devem ter em mente os potenciais efeitos oftalmológicos adversos desse medicamento e/ou de seus excipientes.

Efeito midriático da fenilefrina a 10% na comparação entre a autoinstilação de gota em olhos abertos e a vaporização em olhos fechados / Phenylephrine 10% mydriatic effect: comparison between self instillation of eye drop in open eyes and spray in closed eyes

RESUMO Objetivo: Comparar a eficácia fenilefrina a 10% aplicada pelo próprio paciente por vaporização em olho fechado em relação à instilação de gota em olho aberto em indivíduos que irão realizar exame de fundoscopia e avaliar o nível de dificuldade e a adequação técnica entre os métodos de administração. Métodos: Ensaio clínico controlado, randomizado e pareado realizado em 2014 envolvendo 100 olhos de 50 pacientes na Policlínica Ronaldo Gazolla - RJ, sem doenças oculares ou sistêmicas que comprometiam a dilatação pupilar. Os pacientes foram submetidos à instilação de 1 gota de fenilefrina a 10% e aplicação de vaporizador do mesmo midriático no olho contralateral. O olho em que se instilou o colírio permaneceu aberto, enquanto o olho vaporizado ficou fechado durante as aplicações da medicação. O diâmetro pupilar foi medido antes da aplicação, 10, 20 e 30 minutos após. O processo de instilação ou vaporização foi observado quanto a sua adequação técnica por um dos autores. Após o processo foi perguntado ao paciente questões pré-formuladas sobre a praticidade de ambos os métodos. Resultados: A diferença de midríase média entre os grupos de olhos avaliados em um determinado tempo foi no máximo 0,3 mm , o que não foi clinicamente ou estatisticamente significativo (ANOVA: F = 1,97 e p = 0,163609) . Porém, ao longo do tempo, a diferença entre o diâmetro da pupila no tempo inicial e no tempo de 30 minutos foi 1,15 mm para os olhos vaporizados e 1,58 mm para os olhos instilados com gotas (ANOVA: F = 129,22 e p ≤ 0,0001). Percentual de 60% dos pacientes tocaram a ponta do frasco de colírio nos olhos, enquanto que 12% tocaram o orifício na ponta do vaporizador com os dedos (p < 0,000001). Setenta de dois por cento (72%) consideraram a instilação de gotas fácil ou muito fácil enquanto 62% consideraram a vaporização em olho fechado fácil ou muito fácil (p = 0,238). Conclusão: A instilação de gotas em olhos abertos e a vaporização de olhos fechados da fenilefrina a 10% apresentou eficácia clínica semelhante. A vaporização foi mais segura e apresentou nível de dificuldade um pouco maior do que a instilação, apesar dos pacientes serem experientes para instilar gotas e inexperientes para vaporizar a medicação em olho fechado.

ABSTRACT Objective: To compare the effectiveness of phenylephrine 10% applied by a spray onto the eye closed over drop instillation onto an open eye on patients who will perform ophthalmoscopy and assess the level of difficulty and technical adequacy of the administration methods. Methods: The study was a clinical trial, controlled, randomized and paired, performed in 2014, involving 100 eyes of 50 patients in the Polyclinic Ronaldo Gazolla - RJ, with no ocular or systemic diseases that compromised the pupillary dilation. Patients underwent 10% phenylephrine eye drop instillation onto one open eye and spray application onto the other eye, which was closed. Pupillary diameter was measured before application and 10, 20, 30 minutes after. The process of instillation or vaporization was observed for its technical correctness by one of the authors. A questionnaire was asked to the patient about the difficulty of both methods after topical administration. Results: The average mydriasis difference between the eye groups assessed at a given time was at most 0.3 mm, which was not clinically or statistically significant (ANOVA: F = 1.97 and p = 0.163609). However, over time, the difference between the average pupil diameter before application and after 30 minutes was 1.15 mm to vaporized eyes and to 1.58 mm in eyes instilled with drops (ANOVA: F = 129, 22 and p ≤ 0.0001). Sixty per cent of patients touched the tip of the eye drop bottle onto the eye, while 12% touched the tip of the vaporizer with their fingers (p <0.000001). Seventy two percent (72%) considered the drops instillation easy or very easy, while 62% considered vaporization in a closed eye easy or very easy (p = 0.238). Conclusion: The instillation of drops phenylephrine 10% in open eyes and the vaporization onto closed eyes showed similar clinical efficacy. Vaporization was safer and a little more difficult than instillation, despite the patients being experienced for instilling drops and inexperienced to vaporize the medication in a closed eye.

Percepção de autoinstilação de gotas oculares em idosos com ou sem dispositivo de apoio facial / Self instillation perception of eye drops in elderly patients with and without facial support device

RESUMO Objetivo: Determinar o grau de dificuldade para instilação tópica ocular em idosos, com ou sem o auxílio de dispositivo de apoio facial, por meio de questionário. Observar qual método foi tecnicamente melhor para aplicação tópica ocular de gotas. Métodos: O estudo foi um ensaio clínico, controlado, randomizado e pareado, realizado em 50 pacientes idosos de setembro de 2015 a junho de 2016 na Policlínica Ronaldo Gazolla, Lapa-Rio de Janeiro. Um frasco de colírio Optive® foi acoplado ao dispositivo de apoio facial denominado Eyedrop®. Cada participante instilou o colírio com ou sem o auxílio do dispositivo em cada um dos olhos, sendo que a seleção ocular foi feita aleatoriamente. Foi perguntado ao paciente questões pré-formuladas sobre a dificuldade de ambos os métodos e a técnica de administração tópica ocular foi avaliada. Resultados: A instilação de gotas foi considerada difícil ou muito difícil por 10% dos idosos com o auxílio do dispositivo e por 36% sem o auxílio (p = 0,0047). Houve toque da ponta do colírio com os tecidos oculares em 64% dos pacientes que não usaram o Eyedrop® e em 4% dos que o utilizaram (p=0,000001). A maior dificuldade descrita na instilação tradicional foi acertar o olho com a gota (32%) e com o dispositivo de apoio foi entender seu uso(4%). Conclusão: A maioria dos idosos instila colírios erroneamente, tocando a ponta do frasco em tecidos oculares, o que favorece sua contaminação. O dispositivo de apoio facial tornou mais segura e fácil a instilação.

ABSTRACT Objective: To determine the degree of difficulty for topical ocular instillation in the elderly, through a questionnaire, with or without the aid of facial support device. Observe which method was technically better to topical ocular application of drops. Methods: The study was a clinical trial, controlled, randomized and paired, which was conducted in 50 elderly patients from September 2015 to June 2016 at the Polyclinic Ronaldo Gazolla, Lapa, Rio de Janeiro. A Optive® eyedrop bottle was attached to a facial support device called Eyedrop®. Each participant instilled an eye drop with or without the device help in each of both eyes, wherein the eye selection was made randomly. He was asked to answer pre-formulated questions about the difficulty of both methods and the topical ocular administration technique was evaluated. Results: Eye drop instillation was difficult or very difficult for 10% of the elderly with the device aid and for 36% without it (p = 0.0047). There were bottle tip touch onto the ocular tissues in 64% of patients who did not use Eyedrop® and 4% who used it (p = 0.000001). The greatest difficulty described in traditional instillation was to head properly the eye drop (32%) and when the support device was used, it was to understand how to use it (4%). Conclusion: Most elderly instills eye drops mistakenly, touching the tip of the bottle onto ocular tissues, which favors contamination. The facial support device increased security and facility in instillation.

Induction of docosahexaenoic acid to expression of heme oxygenase-1 in retinal pigment epithelial cells / 中华实验眼科杂志

Background Oxydative stress is an important pathogenesis of age-related macular degeneration.Resent evidences indicate that docosahexaenoic acid (DHA) plays an important role during the development of retinal photoreceptor cells and protect the cells against oxydative stress by inducing the expression of heme oxygenase-1 (HO-1).However,whether DHA can induce the expression of HO-1 in human retinal pigment epithelium (RPE) cells is unelucidated.Objective This study was to investigate the effect of DHA on the expression of HO-1 in RPE cells and its molecular mechanism.Methods Human RPE cell line ARPE-19 was cultured in vitro and treated with 30,50,100 and 120 μmol/L DHA for 4 to 24 hours,respectively,and the cells were cultured without DHA as the control group.The cytotoxicity of DHA was detected by lactate dehydrogenase(LDH),and the expression of HO-1 mRNA and protein were detected by real-time PCR and Western blot assay,respectively.The enzymatic activity of HO-1 was detected by colorimetry.The reactive oxygen species (ROS) proportion in the cells was detected using fluorescence probe H2 DCFDA,and immunofluorescence technology was adopted to detect the nuclear translocation of nuclear facotor-E2-related factor 2 (Nrf2).The expression of Nrt2 protein in the cells was detected by Western blot after intervention of ROS inhibitor N-acetylcysteine (NAC) and transfection of Nrf2 small interfering RNA (siRNA).Results The LDH leakage rate was significantly different after 0,3,50,100 and 120 μmol/L DHA treated the cells for 24 hours (F=8.14,P＜0.05),and the LDH leakage rate in the 120 μmol/L DHA group was significantly higher than that of 0,30,50 and 100 μmol/L DHA group (all at P＜0.05).The relative expression levels of HO-1 mRNA and HO-1 protein or HO-1 enzymatic activity in the cells were significantly different among different concentrations of DHA group in 8 hours after treatment (F=16.24,P＜0.05;F=11.34,P＜0.05;F=11.81,P＜0.05),and the expressions of these factors were considerably higher in the 30,50 and 100 μ mol/L DHA group than those in the 0 μmol/L DHA group (all at P＜0.05).The ROS relative fluorescence intensity and nuclear Nrf2 positive cells proportion were statistically significant among different concentrations of DHA groups (F =11.08,P ＜ 0.05;F=16.42,P＜0.05),and the ROS relative fluorescence intensity and nuclear Nrf2 positive cells proportion were evidently higher in the 30,50 and 100 μmol/L DHA group than those in the 0 μmol/L DHA group (all at P＜0.05).The relative expression levels of HO-1 protein and the proportion of nuclear Nrf2 positive cells were significantly lower in the NAC pretreated 100 μmol/L DHA group than those in the 100 μmol/L DHA group.In addition,the HO-1 relative expression level and the positive cells proportion of nuclear Nrf2 were significantly lower in the of Nrf2 siRNA transfection group than those in the blank siRNA transfection group (both at P＜0.05).Conclusions DHA with concentration below 100 μ mol/L can protect RPE cells from oxidative stress by inducting the expression of HO-1 in the cells via ROS/Nrf2 pathway.

Percepção da aplicação tópica ocular de drogas: comparação entre instilação de gotas em olhos abertos e vaporização em olhos fechados / Perception of topical ocular drug delivery: comparison between eyedrop instillation in open eyes and vaporisation in closed eyes

Objetivo: Avaliar por questionário qual o nível de facilidade ou dificuldade para aplicação tópica de medicações oculares: vaporização em olho fechado ou instilação de gotas em olho aberto e constatar por meio da observação de pacientes pelos autores qual o método que foi utilizado com maior adequação técnica para aplicação de drogas tópicas oculares. Métodos: A pesquisa foi um ensaio clínico pareado e randomizado, realizada nos meses de agosto e setembro de 2012 no ambulatório de Oftalmologia da Policlínica Ronaldo Gazolla (Campus Arcos da Lapa, Faculdade de Medicina da Universidade Estácio de Sá, RJ) em 50 pacientes conveniados de planos de saúde ou do SUS. Foi utilizado um frasco de colírio e um de vaporizador com solução Optive®. Cada participante aplicou em um dos olhos a solução por vaporização ou instilação de gotas através de um processo randomizado. Foi perguntado aos pacientes questões pré-formuladas sobre a praticidade de ambos os métodos e observada à técnica de aplicação. Resultados: 32% acharam difícil ou muito difícil a vaporização em olho fechado e 34% a instilação de colírio (p=0,9562). A dificuldade mais comum para ambos os métodos foi "acertar o olho" e ocorreu em 53% dos pacientes que tiveram dificuldades para vaporização e por 65%dos que apresentaram dificuldade para aplicação de colírio. 38% dos pacientes necessitaram de mais de uma instilação para aplicação do colírio, enquanto 30% dos pacientes precisaram de mais de uma aplicação para que a droga vaporizada tivesse contato com o olho (p=0,5224). Em 74% dos pacientes houve toque da ponta do colírio com os cílios, já com ...

Objective: Evaluate how difficult it is to apply ocular topical medications based on patient observation and answers to a questionnaire. Eye drops in open eyes were compared to vaporization in closed eyes. Methods: The study was a randomized clinical trial paired and held in the months of august and september of 2012 in the ophthalmological department of Polyclinic Ronaldo Gazolla (Arcos da Lapa Campus, Faculty of Medicine, University Estáciode Sá, RJ) in 50 patients. The Optive® ophthalmic solution was applied topically via an eyedrop bottle or a vaporizer through a randomized process. Patients were asked pre-formulated questions about the practicality of both methods and the technique of topical ocular drug delivery was observed. Results: 32% informed that it was difficult or very difficult to vaporize and 34% to use eye drops (p=0,9562). The major problem described by patients was to direct the eye drop to the eye surface. This difficulty was considered by 53% for vaporization and by 65% for topical eye drop use. 38% of the patients needed more than one eye drop application to have eye drop contact, while 30% of the patients needed more than one application of vaporization in order to get drug eye contact (p=0,5224). In 74% of patients there were an eyedropper tip contact with cilia, however there was one eye finger contact when the medicine was vaporized (p=0,5433). Conclusion: The ease perceived by patients to instil eye drops in open eyes was equivalent compared to the vaporization in closed eyes; the method of spraying was performed more appropriately due to the high frequency of eyedrop tip touches on the ocular surface. .

Percepção da administração tópica ocular de drogas: vaporização x gotas / Topical drug administration perception: spray x eyedrop

OBJETIVO: Avaliar por questionário qual o nível de dificuldade para aplicação tópica de medicações oculares: vaporização ou gotas através da observação do paciente e qual método foi tecnicamente melhor utilizado para aplicação de drogas tópicas oculares. MÉTODOS: A pesquisa foi realizada no decorrer de 2008 e 2009 no ambulatório de oftalmologia da Policlínica Ronaldo Gazolla. Foi utilizado um frasco de colírio e um de vaporizador com solução de carboximetilcelulose sódica a 0,5 por cento. Cada participante aplicou em um dos olhos a solução por vaporização ou instilação de gotas através de um processo randomizado. Foi perguntado ao paciente questões pré-formuladas sobre a praticidade de ambos os métodos. RESULTADOS: Considerando o grau de dificuldade de administração tópica ocular: 36 por cento acharam difícil ou muito difícil a vaporização e 14 por cento a instilação de colírio. As dificuldades descritas pelos pacientes foram relatadas por 64 por cento dos pacientes para vaporização e por 34 por cento para aplicação de colírio (p= 0,0027). Já 42 por cento dos pacientes necessitaram de mais de uma instilação para aplicação do colírio, enquanto 36 por cento dos pacientes precisaram de mais de uma aplicação para que a droga vaporizada tivesse contato com o olho (p= 0,49). Em 56 por cento dos pacientes houve toque da ponta do colírio com os cílios, já com o vaporizador não houve toque (p=0,0001). CONCLUSÃO: A vaporização foi o método mais seguro para evitar a contaminação do frasco. A maior facilidade percebida pelos pacientes ao instilar o colírio em relação a vaporização foi devido a terem apoiado a ponta do frasco nos tecidos oculares.

OBJECTIVE:Evaluate how difficult it is to apply ocular topical medications based on patient observation and answers to a questionnaire. Eye drops were compared to vaporization. METHODS: The study was performed in 2008 and 2009 in Policlinica Ronaldo Gazolla ophthalmological ambulatory. An eyedropper and a vaporizer with carboximetilcelulose 0,5 percent were used. Each individual tested applied randomly on one of their eyes an eye drop or vaporization. The patients had to answer questions about the practice concerning both forms of topical eye drug application. RESULTS: 36 percent informed that it was difficult or very difficult to vaporize and 14 percent to use eye drops. Problems described by patients were considered by 64 percent for vaporization and by 34 percent for topical eye drop use (p= 0,0027). 42 percent of the patients needed more than one eye drop application to have eye drop contact , while 36 percent of the patients needed more than one application of vaporization in order to get drug eye contact (p= 0,49). In 56 percent of patients there were an eyedropper tip contact with cilia, however there was not contact when the medicine was vaporized (p=0,0001). CONCLUSION: Vaporization was the safest method to avoid topical ocular drug contamination by manipulation; the greater facility noticed for patients when they administered eye drops was achieved using eye tissues as an eyedropper support.

Effect of triamcinolone acetonide on the activation and barrier function of human retinal pigment epithelium / 中华眼底病杂志

Objective To observe the effect of triamcinolone acetonide(TA) on activation and barrier function of human retinal pigment epithelium (RPE). Methods ARPE-19 cells were cultured in 96-well tissue culture plate. Four weeks later, TA with different concentration (0.02 and 0.05 mg/ml)was added to the cells and culture for 3 or 7 days. The activation of ARPE-19 cells was assessed by methyl thiazolyl tetrazolium (MTT). ARPE-19 cells were cultured on polyester microporous filters for 4 weeks, and the transepithelial resistance (TER) was recorded. TA (0.02 and 0.05 mg/ml) was added to the culture fluid respectively, and after cultured for 1 week TER was measured again. The RPE permeability was detected by enzyme-linked immunosorbent assay (ELISA) with horse radish peroxidase as the tracer. [WTHZ]Results In the culture fluid with 0.02 mg/ml TA cultured for 3 or 7 days, the average survival rate of ARPE-19 cells was 93.70% and 90.63% respectively, without statistic difference compared with the control (P=0.147, 0.091). While in the 0.05 mg/ml TA group after cultured for the same duration, the activation of ARPE-19 cells decreased significantly compared with the control (with the average survival rate of 87.75% and 88.98%; P=0.025, 0.043). One week after cultured with TA, TER decreased significantly while permeability improved obviously in the 2 TA groups compared to the control (P

Effects of cytokines on the expression of syndecan-1 in cultured human retinal pigment epithelial cells / 中华眼底病杂志

Objective To investigate the effects of cytokines on the expression of syndecan-1 in cultured human retinal pigment epithelial (RPE) cells and the signal transduction pathway. Methods Reverse transcription polymerase chain reaction and immunofluorescence staining were used to detect the expression of syndecan-1 mRNA and protein in normal RPE cells. The expression of syndecan-1 in RPE cells stimulated by different cytokines was detected and quantitatively analyzed by image process of immunofluorescence. The stimulation included 7 and 35 ng/ml tumor necrosis factor (TNF)-? for 24 hours, 1 and 6 ?g/ml lipopolysaccharide (LPS) for 11 hours, 7 ng/ml TNF-? for 0 to 24 hours (once per 2 hours, and 13 times in total), and 30% supernatant of monocyte/macrophage strain (THP-1 cells) for 3, 14 and 43 hours. The effect of 30% supernatant of THP-1 cells was assayed after pretreated by PD098059 [the specific inhibitor of extracellular signal regulated kinase(ERK) 1/2] for 2 hours. After exposed to 30% supernatant of THP-1 cells for 3 hours and treated by 0.25% trypsin for 5 minutes, RPE cells attaching was evaluated by methyl thiazolyl tetrazolium assay. Results In normal human RPE cells, expressions of syndecan-1 mRNA and protein were detected, and strong syndecan-1 positive yellowish green fluorescence was found in the cell membrane and cytoplasm while light green fluorescence was in the nucleus. As the concentration and stimulated time of TNF-? or LPS increased, the fluorescence intensity decreased(P

Effect of hypericin on the activity of protein kinase C in cultured human retinal pigment epithelial cells in vitro / 中华眼底病杂志

Objective To investigate the effect of hypericin on the activity of protein kinase C (PKC) in cultured human retinal pigment epithelium (RPE) cells in vitro. Methods RPE cells were cultured in standard medium with 10% serum concentrations containing 0.5 to 5.0 ?mol/L hypericin with or without preincubation of phorbol 12 myristate 13 acetate (PMA). The activities of cytosolic PKC (c PKC) and membranous PKC (m PKC) were assayed by PKC kit. Results The original activities of c PKC and m PKC of RPE cells were (35.34?4.10) pmol?min 1 ?mg 1 and (62.52?8.80) pmol?min 1 ?mg 1 . The activity of c PKC in RPE cells with PMA preincubation decreased rapidly in 5 minutes, with a subsequent slow decrease after 20 minutes and a decrease to 18% of the activity of c PKC in RPE cells without PMA preinubation after 60 minutes. While the activity of m PKC in RPE cells with PMA preincubation increased gradually after 5 minutes and reduced after reached the peak at 40 minutes, and then returned to baseline after 60 minutes, eventually decreased below 30% of the control group. When RPE cells were cultured with PMA for 48 hours, the activities of c PKC and m PKC were hardly detectable, while RPE cells were cultured with both PMA and hypericin, hypericin could counteract most of down regulation by PMA. Conclusion Hypericin may inhibit the translocation of PKC in RPE cells,change the activity of PKC, promote the apoptosis of RPE cells likely,and then prevent proliferative vitreoretinopathy.