ABSTRACT Purpose: This study investigates the protective effect of cilostazol on the development and evolution of diabetic retinopathy in rats. Methods: Sixty male rats were divided into four groups: untreated nondiabetic rats, untreated diabetic rats, cilostazol-treated nondiabetic rats, and cilostazol-treated diabetic rats. The thickness of the internal limiting membrane to the outer limiting membrane, inner plexiform layer, inner nuclear layer, and outer nuclear layer were measured. The number of cell nuclei per 50-μm length in retinal sections was counted to quantify the degree of retinal cell loss. Results: The number of nuclei in the ganglion cell layer was significantly higher in untreated nondiabetic rats (p<0.05). The mean number of nuclei in the cilostazol-treated nondiabetic rats was significantly higher than that in the cilostazol-treated diabetic rats (p<0.05). The cilostazol-treated nondiabetic rats had a significantly higher mean nuclei count in the inner nuclear layer and inner plexiform layer as compared with the other groups (p<0.05). The total mean retinal thickness of the cilostazol-treated nondiabetic rats was significantly higher than that of cilostazol-treated diabetic rats and untreated diabetic rats (p<0.05). Conclusion: By decreasing the loss of ganglion cells and reducing the sensorineural atrophy in the internal retinal layers, cilostazol had a protective effect against changes caused by diabetic retinopathy in diabetic rats.

RESUMO Objetivo: O objetivo deste estudo foi investigar o efeito protetor do cilostazol no desenvolvimento e na evolução da retinopatia diabética em ratos. Métodos: Sessenta ratos machos foram divididos em 4 grupos: ratos não-diabéticos não-tratados, ratos diabéticos não-tratados, ratos não-diabéticos tratados com cilostazol e ratos diabéticos tratados com cilostazol. A espessura da membrana limitante interna à membrana limitante externa, a camada plexiforme interna, a camada nuclear interna e a camada nuclear externa foram medidas. Para quantificar o grau de perda de células da retina, foi contado o número de núcleos de células por 50 μm de comprimento em secções retinianas. Resultados: O número de núcleos no GCL foi significativamente maior em Ratos não-diabéticos não-tratados com cilostazol (p<0,05). O número médio de núcleos em Ratos não-diabéticos tratados com cilostazol foi significativamente maior do que em Ratos diabéticos tratados com cilostazol (p<0,05). A contagem média de núcleos em camada nuclear interna e camada plexiforme interna de ratos não-diabéticos tratados com cilostazol foi significativamente maior do que nos outros grupos (p<0,05). A espessura retiniana média total de Ratos não-diabéticos tratados com cilostazol foi significativamente maior do que em Ratos diabéticos tratados com cilostazol e Ratos diabéticos não-tratados (p<0,05). Conclusão: Os resultados demonstraram que o cilostazol teve um efeito protetor contra as alterações causadas pela retinopatia diabética em ratos diabéticos, diminuindo a perda de células ganglionares e reduzindo a atrofia neurossensorial nas camadas retinianas internas.

Abstract Hydroxytyrosol (HT) is a natural phenolic compound found in leaves of olive trees. It is a photosensitive solid phytochemical constituent and an irritant substance to skin and mucosa as pure drug. HT is a lipophilic chemical constituent and shows low solubility in water, bitter taste, and instability to oxidizing atmosphere. Considering the limitations for its suitable therapeutic use, the aim of the present study was to obtain, to characterize, to quantify, and to evaluate the stability of HT-loaded poly(ԑ-caprolactone) nanocapsules for its further use in pharmaceutical and nutraceutical products. Nanocapsules were prepared by method of interfacial deposition of the preformed polymer. The formulations were characterized by morphological and spectroscopic methods. The UHPLC-PDA analytical method was developed and validated for quantifying the HT encapsulation. Physicochemical stability assay was performed for 120 days. Nanocapsules were successfully obtained by the proposed method. The morphological evaluation demonstrated the drug absence in the nanocapsule surface. Fourier-transform infrared spectroscopy demonstrated the drug nanoencapsulation. The analytical method was validated and confirmed a high HT encapsulation efficiency over 95%. Considering the physicochemical stability testing, the polydispersity index changed after 30 days for NC-2 (HT at 2 mg/mL) and after 90 days for NC-5 (HT at 5 mg/mL). Zeta potential changed after 60 days for both formulations (NC-2 and NC-5). pH values were modified after 60 days for NC-5 and after 90 days for NC-2. HT-loaded PCL nanocapsules demonstrate adequate physicochemical features and high drug encapsulation. The different formulations revealed stability changes from 30 to 90 days. Additional in vitro and in vivo evaluations may be performed in order to use these nanoformulations for treating several diseases based on oxidative-inflammatory environments.

RESUMO Objetivo: O objetivo deste estudo foi investigar o efeito protetor do cilostazol no desenvolvimento e na evolução da retinopatia diabética em ratos. Métodos: Sessenta ratos machos foram divididos em 4 grupos: ratos não-diabéticos não-tratados, ratos diabéticos não-tratados, ratos não-diabéticos tratados com cilostazol e ratos diabéticos tratados com cilostazol. A espessura da membrana limitante interna à membrana limitante externa, a camada plexiforme interna, a camada nuclear interna e a camada nuclear externa foram medidas. Para quantificar o grau de perda de células da retina, foi contado o número de núcleos de células por 50 μm de comprimento em secções retinianas. Resultados: O número de núcleos no GCL foi significativamente maior em Ratos não-diabéticos não-tratados com cilostazol (p<0,05). O número médio de núcleos em Ratos não-diabéticos tratados com cilostazol foi significativamente maior do que em Ratos diabéticos tratados com cilostazol (p<0,05). A contagem média de núcleos em camada nuclear interna e camada plexiforme interna de ratos não-diabéticos tratados com cilostazol foi significativamente maior do que nos outros grupos (p<0,05). A espessura retiniana média total de Ratos não-diabéticos tratados com cilostazol foi significativamente maior do que em Ratos diabéticos tratados com cilostazol e Ratos diabéticos não-tratados (p<0,05). Conclusão: Os resultados demonstraram que o cilostazol teve um efeito protetor contra as alterações causadas pela retinopatia diabética em ratos diabéticos, diminuindo a perda de células ganglionares e reduzindo a atrofia neurossensorial nas camadas retinianas internas.

ABSTRACT Purpose: This study investigates the protective effect of cilostazol on the development and evolution of diabetic retinopathy in rats. Methods: Sixty male rats were divided into four groups: untreated nondiabetic rats, untreated diabetic rats, cilostazol-treated nondiabetic rats, and cilostazol-treated diabetic rats. The thickness of the internal limiting membrane to the outer limiting membrane, inner plexiform layer, inner nuclear layer, and outer nuclear layer were measured. The number of cell nuclei per 50-μm length in retinal sections was counted to quantify the degree of retinal cell loss. Results: The number of nuclei in the ganglion cell layer was significantly higher in untreated nondiabetic rats (p<0.05). The mean number of nuclei in the cilostazol-treated nondiabetic rats was significantly higher than that in the cilostazol-treated diabetic rats (p<0.05). The cilostazol-treated nondiabetic rats had a significantly higher mean nuclei count in the inner nuclear layer and inner plexiform layer as compared with the other groups (p<0.05). The total mean retinal thickness of the cilostazol-treated nondiabetic rats was significantly higher than that of cilostazol-treated diabetic rats and untreated diabetic rats (p<0.05). Conclusion: By decreasing the loss of ganglion cells and reducing the sensorineural atrophy in the internal retinal layers, cilostazol had a protective effect against changes caused by diabetic retinopathy in diabetic rats.

OBJETIVO: Reconhecer a capacidade regenerativa influenciada pela administração de estradiol. MÉTODOS: Utilizaram-se 42 ratos Wistar, fêmeas, divididos em dois grupos controle e experimento. Realizou-se a ressecção de, aproximadamente, 70% do fígado destes animais. Ratos do grupo controle receberam injeção intramuscular de um mililitro de óleo de amendoim, enquanto que os do grupo experimento receberam hexaidrobenzoato de estradiol (50µg) diluídos em um mililitro de óleo de amendoim. Fizeram-se as aferições com 36 horas e 7 dias, com 3 métodos: Fórmula de Kwon et al.21 para reconhecer ganho de volume, contagem das figuras de mitose existentes em 5 campos e percentual dos núcleos PCNA positivos em 5 campos. RESULTADOS: O ganho de volume (massa) foi semelhante nos dois grupos com 36 horas (p=0,1873) e maior no grupo experimento com 7 dias (p=0,0447). À microscopia observou-se número de figuras de mitose em número semelhante com 36 horas (p=0,3528) e tendência a ser maior no grupo experimento com 7 dias (p=0,0883). A média de núcleos PCNA positivos foi maior no grupo experimento tanto com 36 horas (p=0,0009) quanto com 7 dias (p=0,0000). CONCLUSÃO: O hexaidrobenzoato de estradiol favoreceu a regeneração hepática em ratos submetidos à hepatectomia 70%.

PURPOSE: To recognize the regenerative capacity influenced by the administrating of estradiol. METHODS: 42 female Wistar rats were used, divided into two groups, the control and the experiment group. A resection of approximately 70% of the liver was made in the liver of these animals. The control group received an intramuscular injection of one ml of peanut oil. The experiment group were given estradiol hexahydrobenzoate (50µg) diluted in one ml of peanut oil. Calibrations were done after 36 hours and 7 days, using three methods: the formula of Kwon et al.21, to recognize gain in volume, counting of the mitosis figures in five fields and the percentage of positive PCNA nuclei. RESULTS: Gain in volume (mass) was similar in both groups after 36 hours (p=0.1873) and higher in the experiment groups after seven days (p=0.0447). Microscopy showed a similar number of mitosis figures after 36 hours (p=0.3528) and a tendency to be higher in the experiment group after 7 days (p=0.0883). The average of positive PCNA nuclei was higher in the experiment group both after 36 hours (p=0.0009) and 7 days (p=0.0000). CONCLUSION: The estradiol hexahydrobenzoate improved liver regeneration in rats submitted to a 70% hepatectomy.