This page has only limited features, please log in for full access.
During the development of polymeric wound coating materials, the in vivo reactions play a key role in the success of the material for tissue engineering. In this work, the chick chorioallantoic membrane (CAM) model is highlighted as an alternative test to evaluate the biocompatibility and changes through the vascular network topography of CAM after the incubation with polymeric membranes. The eggshells of fertilized chicken eggs are opened on the third day of embryo development and the CAM is exposed. Polymeric membranes are placed on the CAM surface and incubated for 96 h. After, the CAM is photographed, the vessels are quantified and histological sections are prepared. In the groups that received spunbonded and cellulose, it is possible to observe tissue growth of the CAM involving the membranes. Regarding the changes in the vascular metrics, a significant increase in the total vessel length and the number of junctions is detected for the spunbonded group. Also, there is an expressive decrease of the lacunarity for this group. Histological analysis revealed an increase of new vessels on the spunbonded, which corroborates to the results mentioned before. The results obtained point out that the CAM assay provides a new perspective on the biocompatibility tests.
Lays Fernanda Nunes Dourado; Carolina Nunes Da Silva; Raquel Gregorio Arribada; Armando Da Silva Cunha Júnior. Assessment of Biocompatibility and Vascular Effects of Polymeric Materials Using Chick Chorioallantoic Membrane, an Alternative Method. Macromolecular Symposia 2020, 394, 1 .
AMA StyleLays Fernanda Nunes Dourado, Carolina Nunes Da Silva, Raquel Gregorio Arribada, Armando Da Silva Cunha Júnior. Assessment of Biocompatibility and Vascular Effects of Polymeric Materials Using Chick Chorioallantoic Membrane, an Alternative Method. Macromolecular Symposia. 2020; 394 (1):1.
Chicago/Turabian StyleLays Fernanda Nunes Dourado; Carolina Nunes Da Silva; Raquel Gregorio Arribada; Armando Da Silva Cunha Júnior. 2020. "Assessment of Biocompatibility and Vascular Effects of Polymeric Materials Using Chick Chorioallantoic Membrane, an Alternative Method." Macromolecular Symposia 394, no. 1: 1.
Retinal ischemia, one of the most common cause of visual loss, is associated with blood flow inadequacy and subsequent tissue injury. In this setting, some treatments that can counteract glutamate increase, arouse interest in ischemic pathogenesis. Ketamine, a potent N-methyl-d-aspartate (NMDA) receptor antagonist, provides a neuroprotective pathway via decreasing the excitotoxicity triggered by excess glutamatergic. Thus, the goal of this study was to evaluate the safety of intravitreal use of ketamine and their potential protective effects on retinal cells in retinal ischemia/reperfusion model. Initially, ketamine toxicity was evaluated by cytotoxicity assay and Hen's egg chorioallantoic membrane (HET-CAM) method. Afterward, some ketamine concentrations were tested in rat's eyes to verify the safety of the intravitreal use. To investigate the neuroprotective effect on retinal, a single intravitreal injection of ketamine in concentrations of 0.059 mmol.L−1 and 0.118 mmol.L−1 was performed one day before the retinal injury by ischemia/reperfusion model. After 7 and 15 days, the retina activity was evaluated by electroretinogram (ERG) records and, lastly, by morphological analyzes. Cytotoxicity assay reveals that the maximum ketamine concentration that could reach retinal pigmented epithelium cells is 0.353 mmol.L−1. HET-CAM assay showed that concentrations above 0.237 mmol.L−1 are irritants to the eye. Thus, Ketamine in concentrations of 0.0237 mmol.L−1, 0.118 mmol.L-1, and 0.059 mmol.L-1 were selected for in vivo toxicity test. ERG records reveal a tendency of b-wave amplitude to decrease as the luminous intensity increased, in the group receiving ketamine at 0.237 mmol.L−1. Therefore, ketamine in concentrations at 0.059 mmol.L−1 and 0.118 mmol.L-1 were chosen for the following tests. In the ischemia retinal degeneration model, pretreatment with ketamine was capable to promote a recovery of retinal electrophysiological function minimizing the ischemic effects. In histological analysis, the groups that received intravitreal ketamine showed a number of retinal cells significantly higher than the vehicle group. In TUNEL assay a reduction on TUNEL-positive cells was observed in all the layers for both concentrations which allow to affirm that ketamine contributes to reducing cell death in the retina. Transmission electron microscopy (TEM) reaffirms this finding. Ketamine intravitreal pretreatment showed reduced ultrastructural changes. Our findings demonstrate that ketamine is safe for intravitreal use in doses up to 0.118 mmol.L−1. They seem to be particularly efficient to protect the retina from ischemic injury.
Lays Fernanda Nunes Dourado; Lucas Gomes Oliveira; Carolina Nunes da Silva; Cibele Rodrigues Toledo; Silvia Ligório Fialho; Rodrigo Jorge; Armando Silva-Cunha. Intravitreal ketamine promotes neuroprotection in rat eyes after experimental ischemia. Biomedicine & Pharmacotherapy 2020, 133, 110948 .
AMA StyleLays Fernanda Nunes Dourado, Lucas Gomes Oliveira, Carolina Nunes da Silva, Cibele Rodrigues Toledo, Silvia Ligório Fialho, Rodrigo Jorge, Armando Silva-Cunha. Intravitreal ketamine promotes neuroprotection in rat eyes after experimental ischemia. Biomedicine & Pharmacotherapy. 2020; 133 ():110948.
Chicago/Turabian StyleLays Fernanda Nunes Dourado; Lucas Gomes Oliveira; Carolina Nunes da Silva; Cibele Rodrigues Toledo; Silvia Ligório Fialho; Rodrigo Jorge; Armando Silva-Cunha. 2020. "Intravitreal ketamine promotes neuroprotection in rat eyes after experimental ischemia." Biomedicine & Pharmacotherapy 133, no. : 110948.
PnPa11 and PnPa13 are synthetic peptides derived from Phoneutria nigriventer spider venom, which display antinociceptive and neuroprotective properties. In this work, we evaluated the safety of intravitreal use and the neuroprotective effect of these peptides. The cytotoxicity and the antiangiogenic activity of these peptides were evaluated by the sulforhodamine-B method and chicken chorioallantoic membrane (CAM) assay, respectively. The in vivo safety was analyzed in Wistar rats that were intravitreally injected with different doses (0.50; 1.25; 2.50; 3.75 and 5.00 µg/mL) of these peptides (right eye, n = 6). The retinal function was assessed by electroretinography exams (ERG), intraocular pressure (IOP), and histological analyzes. In order to investigate the neuroprotective effect, Wistar rats received intravitreal injections (right eye, n = 6) of peptides at 1.25 µg/mL and then were exposed to blue LED light. In addition, the visual function and the retinal microstructure were verified. Cytotoxicity analyses demonstrated that the peptides did not present any toxicity over ARPE-19 (adult retinal pigmented epithelial) cell line and the antiangiogenic study highlighted that the peptides promoted the reduction of blood vessels. The intravitreal injection did not cause major changes, neither induced any irreversible damage. In the retinal degeneration assay, the ERG records demonstrated that the prior treatment with PnPa11 and PnPa13 protected the retina from damage. Morphological analyses confirmed the ERG findings. Immunoblotting analyses revealed that PnPa11 increased Erk1/2, NR2A, and NR2B retinal expression after the light stress model, but did not cause Akt1 activation, while PnPa13 prevented Erk1/2 and Akt1 dephosphorylation. The intraocular administration of these peptides was well tolerated and presented protective activity against retinal degeneration, suggesting the potential use of these peptides as neuroprotectors in the ophthalmological field.
Lays Fernanda Nunes Dourado; Flavia Rodrigues Da Silva; Cibele Rodrigues Toledo; Carolina Nunes Da Silva; Cleildo Pereira Santana; Bruna Lopes Da Costa; Maria Elena De Lima; Armando Da Silva Cunha Junior. Intravitreal injection of peptides PnPa11 and PnPa13, derivatives of Phoneutria nigriventer spider venom, prevents retinal damage. Journal of Venomous Animals and Toxins including Tropical Diseases 2020, 26, 1 .
AMA StyleLays Fernanda Nunes Dourado, Flavia Rodrigues Da Silva, Cibele Rodrigues Toledo, Carolina Nunes Da Silva, Cleildo Pereira Santana, Bruna Lopes Da Costa, Maria Elena De Lima, Armando Da Silva Cunha Junior. Intravitreal injection of peptides PnPa11 and PnPa13, derivatives of Phoneutria nigriventer spider venom, prevents retinal damage. Journal of Venomous Animals and Toxins including Tropical Diseases. 2020; 26 ():1.
Chicago/Turabian StyleLays Fernanda Nunes Dourado; Flavia Rodrigues Da Silva; Cibele Rodrigues Toledo; Carolina Nunes Da Silva; Cleildo Pereira Santana; Bruna Lopes Da Costa; Maria Elena De Lima; Armando Da Silva Cunha Junior. 2020. "Intravitreal injection of peptides PnPa11 and PnPa13, derivatives of Phoneutria nigriventer spider venom, prevents retinal damage." Journal of Venomous Animals and Toxins including Tropical Diseases 26, no. : 1.
Some recent studies have shown that pirfenidone (PFD) has favorable results in the healing process of the cornea. However, PFD in solution exhibits short half-life after topical application, and in this context, a liquid crystal nanoparticle system containing PFD (PFD-LCNPs) was developed. The nanoparticles were characterized by transmission electron microscopy, atomic force microscopy, small angle X-ray diffraction and polarized light microscopy. The PFD-LCNPs had particle size and zeta potential of 247.3 nm and -33.60 mV (stores at 4 °C), respectively, and 257.5 nm and -46.00 mV (stored at 25 °C), respectively. The pH of the formulation was 6.9 and the encapsulation efficiency was 35.9%. The in vitro release profiles indicated that PFD sustained release from PFD-LCNPs for up to 12 h. In vitro study of ocular irritation (HET-CAM test) concluded that components of the formulation are well tolerated for ocular administration. Corneal re-epithelialization time after chemical burning was significantly reduced in rabbits treated with PFD-loaded LCNPs when compared to the group treated with a vehicle. In addition, the anti-inflammatory action of pirfenidone was observed by reducing myeloperoxidase activity (MPO) and inflammatory cells in the histology of the tissues of animals treated with PFD-LCNPs. These findings indicated that the PFD-LCNPs might have the potential for effective ocular drug delivery.
Rummenigge Oliveira Silva; Bruna Lopes da Costa; Flavia Rodrigues da Silva; Carolina Nunes da Silva; Mayara Brandão de Paiva; Lays Fernanda Nunes Dourado; Ângelo Malachias; Adriano Antunes De Souza Araújo; Paula Santos Nunes; Armando Silva-Cunha. Treatment for chemical burning using liquid crystalline nanoparticles as an ophthalmic delivery system for pirfenidone. International Journal of Pharmaceutics 2019, 568, 118466 .
AMA StyleRummenigge Oliveira Silva, Bruna Lopes da Costa, Flavia Rodrigues da Silva, Carolina Nunes da Silva, Mayara Brandão de Paiva, Lays Fernanda Nunes Dourado, Ângelo Malachias, Adriano Antunes De Souza Araújo, Paula Santos Nunes, Armando Silva-Cunha. Treatment for chemical burning using liquid crystalline nanoparticles as an ophthalmic delivery system for pirfenidone. International Journal of Pharmaceutics. 2019; 568 ():118466.
Chicago/Turabian StyleRummenigge Oliveira Silva; Bruna Lopes da Costa; Flavia Rodrigues da Silva; Carolina Nunes da Silva; Mayara Brandão de Paiva; Lays Fernanda Nunes Dourado; Ângelo Malachias; Adriano Antunes De Souza Araújo; Paula Santos Nunes; Armando Silva-Cunha. 2019. "Treatment for chemical burning using liquid crystalline nanoparticles as an ophthalmic delivery system for pirfenidone." International Journal of Pharmaceutics 568, no. : 118466.
Bacterial keratitis is an ocular infection that can lead to severe visual disability. Staphylococcus aureus is a major pathogen of the eye. We recently demonstrated the strong antimicrobial activity of LyeTxI-b, a synthetic peptide derived from a Lycosa erithrognatha toxin. Herein, we evaluated a topical formulation (eye drops) containing LyeTxI-b to treat resistant bacterial keratitis. Keratitis was induced with intrastromal injection of 4 × 105 cells (4 µL) in New Zealand female white rabbits. Minimum inhibitory concentration (MIC) and biofilm viability were determined. LyeTxI-b ocular toxicity was evaluated through chorioallantoic membrane and Draize tests. One drop of the formulation (LyeTxI-b 28.9 µmol/L +0.5% CMC in 0.9% NaCl) was instilled into each eye four times a day, for a week. Slit-lamp biomicroscopy analysis, corneal histopathological studies and cellular infiltrate quantification through myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) detection were performed. LyeTxI-b was very effective in the treatment of keratitis, with no signs of ocular toxicity. Planktonic bacteria MIC was 3.6 µmol/L and LyeTxI-b treatment reduced biofilm viability in 90%. LyeTxI-b eliminated bacteria and reduced inflammatory cellular activity in the eyes. Healthy and treated animals showed similar NAG and MPO levels. LyeTxI-b is a potent new drug to treat resistant bacterial keratitis, showing effective antimicrobial and anti-inflammatory activity.
Carolina Nunes Da Silva; Flavia Rodrigues Da Silva; Lays Fernanda Nunes Dourado; Pablo Victor Mendes Dos Reis; Rummenigge Oliveira Silva; Bruna Lopes Da Costa; Paula Santos Nunes; Flávio Almeida Amaral; Vera Lúcia Dos Santos; Maria Elena De Lima; Armando Da Silva Cunha Júnior. A New Topical Eye Drop Containing LyeTxI-b, A Synthetic Peptide Designed from A Lycosa erithrognata Venom Toxin, Was Effective to Treat Resistant Bacterial Keratitis. Toxins 2019, 11, 203 .
AMA StyleCarolina Nunes Da Silva, Flavia Rodrigues Da Silva, Lays Fernanda Nunes Dourado, Pablo Victor Mendes Dos Reis, Rummenigge Oliveira Silva, Bruna Lopes Da Costa, Paula Santos Nunes, Flávio Almeida Amaral, Vera Lúcia Dos Santos, Maria Elena De Lima, Armando Da Silva Cunha Júnior. A New Topical Eye Drop Containing LyeTxI-b, A Synthetic Peptide Designed from A Lycosa erithrognata Venom Toxin, Was Effective to Treat Resistant Bacterial Keratitis. Toxins. 2019; 11 (4):203.
Chicago/Turabian StyleCarolina Nunes Da Silva; Flavia Rodrigues Da Silva; Lays Fernanda Nunes Dourado; Pablo Victor Mendes Dos Reis; Rummenigge Oliveira Silva; Bruna Lopes Da Costa; Paula Santos Nunes; Flávio Almeida Amaral; Vera Lúcia Dos Santos; Maria Elena De Lima; Armando Da Silva Cunha Júnior. 2019. "A New Topical Eye Drop Containing LyeTxI-b, A Synthetic Peptide Designed from A Lycosa erithrognata Venom Toxin, Was Effective to Treat Resistant Bacterial Keratitis." Toxins 11, no. 4: 203.
Investigate the potential application of corosolic acid (CA) in the treatment of diseases causing retinal neovascularization. CA cytotoxicity effect was evaluated in ARPE-19 cells by sulforhodamine B colorimetric method, and antiangiogenic activity was studied using chorioallantoic membrane (CAM) assay. An amount of 0.01 mL of CA formulations at 5, 10 and 25 µM was injected in the right eyes of Wistar rats, and the contralateral eyes received the vehicle to verify the safety of ophthalmic use. Electroretinography (ERG) was performed before, 7 and 15 days after CA administration. Animals were killed on the 15th day, and the histological analysis of retina was carried out under light microscopy. CA did not present cytotoxicity at concentrations below 35.5 μM after 48 h of treatment. The antiangiogenic activity was confirmed by CAM assay, since CA (range from 5 to 25 µM) induced a significant reduction in vascularity without any signs of toxicity. ERG recordings and histological evaluation did not show any signs of retinal toxicity. CA was effective in reducing vascularity in a CAM model and was found to be safe for potential ophthalmic use.
Cibele Rodrigues Toledo; Vinícius Viana Pereira; Lays Fernanda Nunes Dourado; Mayara Rodrigues Brandão Paiva; Armando Silva-Cunha. Corosolic acid: antiangiogenic activity and safety of intravitreal injection in rats eyes. Documenta Ophthalmologica 2019, 138, 181 -194.
AMA StyleCibele Rodrigues Toledo, Vinícius Viana Pereira, Lays Fernanda Nunes Dourado, Mayara Rodrigues Brandão Paiva, Armando Silva-Cunha. Corosolic acid: antiangiogenic activity and safety of intravitreal injection in rats eyes. Documenta Ophthalmologica. 2019; 138 (3):181-194.
Chicago/Turabian StyleCibele Rodrigues Toledo; Vinícius Viana Pereira; Lays Fernanda Nunes Dourado; Mayara Rodrigues Brandão Paiva; Armando Silva-Cunha. 2019. "Corosolic acid: antiangiogenic activity and safety of intravitreal injection in rats eyes." Documenta Ophthalmologica 138, no. 3: 181-194.